Hemp Growing

Awan et al. did observe a decrease in plant height from pendimethalin treated plots in a dry-seeded system with no recovery by the final evaluation.Pendimethalin is currently not available for water-seeded rice; however, these results support the introduction of pendimethalin in California water-seeded rice. These results indicate rice injury is reduced with a post-emergence application after the 3- to 4-leaf stage rice in a water-seeded system compared to an application at 1- to 2-leaf stage. Pendimethalin is not a stand-alone herbicide and will need to be accompanied with other available herbicides to achieve season long weed control. In general, most rice cultivars tested were relatively tolerant to pendimethalin when treated after the 3-leaf stage rice; furthermore, cultivars with lower seedling vigor scores may become more injured from a pendimethalin post-emergence application. The results provide supporting data for registration of pendimethalin in water-seeded rice and provide a base knowledge from which further work should be conducted to enhance its use in this system.The authors would like to acknowledge the California Rice Research Board and BASF for providing funding for this project, the California Rice Experiment Station for their support in field management, and the various past and present lab members who assisted with this project, in particular Saul Estrada and Dr. Alex R. Ceseski. The authors also acknowledge the D. Marlin Brandon Rice Research Fellowship by the California Rice Research Trust, the Horticulture and Agronomy Graduate Group scholarships including the Bert and Nell Krantz Fellowship and the Jack Pickett Agricultural Scholarship, and the Department of Plant Sciences, UC Davis for the award of a GSR scholarship funded by endowments, indoor weed growing accessories particularly the James Monroe McDonald Endowment, administered by UCANR, which supported the student.

Rice is a staple crop produced worldwide of cultural and economic value . The export exchange of rice has become a prominent market for many countries worldwide . In the US, the export value rice production was nearly 1.7 billion USD in 2022 . Therefore, worldwide rice production must be upheld to current or superior standards to continuously fulfill the global rice demands. There are various common rice production systems used worldwide like transplanted paddies, dry-seeded seasonally flooded and continuously flooded systems . Water-seeded rice is not common worldwide but is the primary method in some geographical areas such as the Sacramento Valley of California . Waterseeded rice is the practice of seeding pregerminated seeds onto fields with a 7- to 15-cm flood, then, typically continuously flooded for the remaining of the season. The water-seeded rice production is popular in areas with ample water for irrigation or where early flood occurrence and poor drainage lead to continuously flooded fields . The flood in water seeded rice helps to control weedy rice, weedy grasses and non-aquatic weed species . However, flood-adapted and herbicide-resistant weeds have further intensified the weed management challenges in many rice fields . Historically, there has been a limited number of herbicide modes of action available for water-seeded rice . Continuous rice cultivation is common in many growing regions because of soil types and economic limitations . Overuse of the same herbicides and continuous rice cultivation have selected for herbicide-resistant weeds which reduce weed control with the currently available herbicides. To support herbicide resistance management, additional herbicides would be beneficial for growers to practice herbicide mode of action rotations . Pendimethalin is a mitotic inhibiting pre-emergence herbicide from the dinitroaniline chemistry that halts seedling growth shortly after germination . In previous surveys and preliminary greenhouse work, pendimethalin has been successful in controlling herbicide-resistant grass populations . Therefore, pendimethalin was evaluated for rice response in water-seeded rice to understand its applicability in this system .

Results from Becerra-Alvarez and Al-Khatib, in chapter 2 of this dissertation, demonstrated rice injury from pendimethalin was reduced in a post-emergence application at the 4-leaf stage rice and in a capsule suspension formulation within 1.1 to 3.4 kg ai ha-1 . However, at the suggested rice stage timing, many grasses have already emerged and control with pendimethalin is reduced. Therefore, if applied post-emergence in herbicide mixtures to control the emerged grasses, then, greater season-long weed control can be achieved. Additionally, herbicide mode of action mixtures are important strategies for herbicide resistance management which help delay resistance development and can control herbicide-resistant populations . It is hypothesized that the residual pendimethalin soil activity when applied postemergence at 4-leaf stage water-seeded rice could assist in control of late-emerging grasses. Economically important late-emerging grasses in California rice include bearded sprangletop [Leptochloa fusca Kunth ssp. fascicularis N. Snow] and watergrass populations. Bearded sprangletop is characterized as a late-emerging grass weed when compared to barnyardgrass [E. crus-galli Beauv] . While the majority of watergrass will emerge early in the season, there are subpopulations that can emerge later and are characterized as prolonged emergence throughout the season . Populations of multiple-resistant late watergrass [E. phyllopogon Koss] have demonstrated evidence of biphasic emergence with the majority emerging early in the season followed by late-emerging cohorts within the population . There is potential benefit from a pendimethalin post-emergence application for control of late-emerging grasses in water-seeded rice. Preliminary field studies evaluating water-seeded rice response were conducted on a continuous 10-cm flood with application onto the water and demonstrated timing after the 3- to 4-leaf stage reduced injury . However, some growers lower the flood depth to encourage rice seedling establishment, or when irrigation water is limited that year.

Decreasing the flood depths can influence pre-emergence herbicide rice injury in water-seeded rice as observed with available herbicides . Therefore, knowledge of rice response as affected by pendimethalin applications at different flood depths in water-seeded rice is important to develop appropriate application methods and recommendations. The objective of the field study was to evaluate the weed control and rice response of a post-emergence application of pendimethalin alone and in mixtures with currently available herbicides. The objective of the greenhouse study aimed to characterize rice response from pendimethalin applications at two flood depths.The study was conducted at the Rice Experiment Station in Biggs, CA in 2022 and 2023. The field soil is characterized as an Esquon-Neerdobe , silty clay, made up of 27% sand, 39% silt, and 34% clay, with a pH of 5.1, and 2.8% organic matter. During the off-season months, the field stubble was burned in spring 2022 prior to a pass with a single offset stubble disc. Field preparation for both years consisted of one pass with a chisel plow to dry the upper soil surface and then two passes with a single offset disc, followed by a land plane to smooth the soil surface. A granule fertilizer starter mixture application of ammonium sulfate and potassium sulfate was applied at 336 kg ha-1 . Then, a corrugated roller was used to pack the soil and eliminate large clods on the soil surface. Individual 3-m wide by 6-m long plots surrounded by 2.2-m wide shared levees were made after fertilizing and prior to flooding to prevent contamination from adjacent treatments in a replication. Seeds of the rice cultivar ‘M-209’ were pregerminated in water. For disease control, a 5% sodium hypochlorite solution was used for the first hour, then drained and refilled with only water for the remaining 24 hours. The seed was then drained until dry up to 12 hours, and seeded at 170 kg ha-1 both years onto the field with a 10-cm standing flood. The flood was maintained the whole season with the exception of a temporary lowering for the post-emergence herbicide treatments but was reflooded back to 10 cm 48 hours after the application. Copper sulfate crystals were applied by plane at 17 kg ha-1 three days after seeding for control of algae. Standard agronomic and pest management practices were followed based on the University of California rice production guidelines . Seeding dates were May 23, 2022 and May 31, 2023. The herbicides and adjuvants used in the field study are outlined in Table 1. Pendimethalin with 0.4 kg L-1 of active ingredient, vertical grow rack system was applied alone and in mixture with foliar active herbicides at the four-leaf stage rice. The pendimethalin application rates were 1.1, 2.3 and 4.6 kg ai ha-1 . The selection of these rates was based on preliminary studies on pendimethalin rates and timings, where 1.1 and 2.3 kg ha-1 were most appropriate rates for water-seeded rice as a post-emergence application . The 4.4 kg ha-1 rate was included in this study to provide rice response data at 2X of the proposed rate for waterseeded rice. The treatment herbicide mixtures with each pendimethalin rate were propanil, cyhalofop-butyl , and bispyribac-sodium . The applications were carried out with a CO2 backpack sprayer calibrated to deliver 187 L ha-1 at 206 kPa traveling at 4.8 km h-1 . The sprayer boom was 3-m wide equipped with six flat-fan 8003VS tips . At time of herbicide applications, the flood water was lowered 24 hours before treatment and reflooded back to 10 cm 48 hours after the treatment. A non-treated control and a grower standard treatment of clomazone applied at day of rice seeding were included for comparison . The treatments were arranged in a randomized complete block design with four replications both years. A follow-up herbicide application of propanil plus triclopyr was applied for sedge and broadleaf control at the midway of full tiller formation rice stage on all treatments except the non-treated . The treatments with pendimethalin alone had a follow-up treatment of cyhalofop plus florpyrauxifen-benzyl at the mid-tiller stage to control all remaining weeds after the initial assessment date . The metabolites have not been labeled of environmental concern and for the most part the pendimethalin parent molecule remains intact when bound to organic matter . In plants, metabolites are also not common and the majority remain as pendimethalin parent molecule when absorbed . The metabolites are also not documented as of concern to the environment by the US EPA ; however, quantifying metabolites helps in understanding the partitioning behavior of an herbicide in an agricultural or environmental system.Visual weed control was recorded for Echinochloa spp., bearded sprangletop, ricefield bulrush [Schoenoplectus. mucronatus Palla], smallflower umbrella sedge , ducksalad , water hyssop and redstem on 14, 24 and 56 days after pendimethalin treatment , on a scale of 0 to 100, where 0=no control and 100=complete control. Weed density counts for Echinochloa spp., sedges and broadleaves were conducted 30 DAT by sampling twice in each plot with a 30-cm by 30-cm quadrat and data scaled to a meter squared area for presentation. Bearded sprangletop counts were conducted for the whole plot after heading of the grass due to a relatively low population density in the field. Visual rice injury assessments were conducted at 20 DAT and 40 DAT by observing present symptomology, which included chlorosis and stunting on a scale of 0 to 100, where 0=no injury and 100=plant death. Rice tiller counts were conducted at 75 days after seeding by sampling twice in each plot with a 30-cm by 30-cm quadrat and data scaled to a meter squared area for presentation. Plant height was recorded at 100 DAS. Rice grain yield was collected both years and adjusted to 14% moisture. The rice grain was harvested from a 2-m by 6-m area in the plots with a small-plot combine on November 2, 2022 and October 30, 2023 .A greenhouse study was conducted at the Rice Experiment Station in Biggs, CA to characterize rice growth as affected by two flood depths after a pendimethalin application. The greenhouse study allowed more accurate management of flood depths than feasible in the field study and direct side by side treatment comparison. Plastic containers with 34-cm by 20-cm by 12-cm dimensions, with openings for drainage were filled with soil from the field study and placed inside larger 58-cm by 41-cm by 31-cm plastic containers, with no drain holes. ‘M-206’ rice seeds were pregerminated by placing the seeds inside cloth bags, and submerging in five gallon buckets for 24 hr. Then, the seeds were air-dried and ten seeds were placed on the soil surface of each smaller container, which would later be thinned to five evenly spaced plants per plot.

The growth benefits in height, canopy formation and leaf numbers per plant is a reflection of the nutrient use efficiency from the cover cropping treatments. In contrast, some previous researchers have observed higher mean number of leaves and heavier stem dry weight from a bare soil than when a rye cover crop used suggesting that rye cover cropping treatments resulted in broccoli marketable yield losses. The negative consequences from cover cropping may have been from cover crop vegetable intercropping, hence live competition for available resources. In this experiment I did not observe any negative consequences of cover cropping on any of the three year growth or yield components of the subsequent vegetable crop. Broccoli shoot biomass determination from destructive crop sampling at harvest time showed that there was no significant broccoli shoot biomass gain from cover cropping for the first year rotation. The observation once again suggests that a single year cover cropping rotation is not sufficient enough to benefit dry mass accumulation by a subsequent vegetable crop. Shoot biomass gain from cover cropping of the latter years was consistent with the observation of increased soil and crop nutrition. The increase in crop biomass with increasing years of cover cropping reveals that repeated cover cropping results in the buildup of cover crop effects. As for crop shoot dry biomass, vertical cannabis broccoli marketable yields were not significantly different between the cover cropping and fallow treatments for the first year cropping. Such responses were seen in almost all broccoli growth parameters and suggest that a one-year cover cropping rotation is of no net and ultimate benefit to broccoli. Increase in marketable yield from cover cropping was significant in the subsequent study years.

Similar to the higher marketable yield observed from cover crop residue supplemented tomatoes , I observed vigorous growth, higher shoot biomass accumulation and higher marketable yield of cover crop residue supplemented broccoli.In contrast, Hoyt observed a reduction in yield of broccoli planted into desiccated barley cover crop and attributed it to lower soil temperatures in the cover crop treatment. The reduction of soil temperature with the use of cover crop mulches and residues has been discussed as a possible limitation delaying crop harvest for several days . However, I observed that broccoli crops grown following summer cover cropping were heavier and had vigorous crop appearances compared these on a fallow field. These benefits however were more eminent after the second year of cover cropping rotations, indicating a buildup effect of the cover crops. Broccoli seems to have benefited from previous summer cover cropping during its second and third year trials. Higher fresh broccoli marketable heads were obtained during the second year from both harvest times and the total marketable heads. An increase in marketable yields starting the second year indicates the necessity of repeated cover cropping rotations to be beneficial. The increase in both marketable head numbers and fresh weights of the marketable heads occurred during the third year, further revealing the importance of longer and repetitive cover cropping rotation. The generally lower yield for the 2009 trial, relative the previous year, however was due to crop damage by other herbivorous pests and unexpected flowering, reducing number and fresh weights of marketable broccoli heads. Yet relative yield comparisons were valid.The more inclusion of the cover crops in the cropping rotations, the higher was the crop yield benefit both in number and weights of marketable broccoli yield. These findings confirm the recommendations that vegetable farmers can grow cover crops during the off-season and benefit from the harvest of the subsequent crop.

Ngouajio et al. suggests that cover crops can be used in diverse cropping conditions as they are compatible with both organic and conventional farming practices by either incorporating or using them as surface mulches. Improvements in soil physical, chemical, and biological environment from the use of cover crops are the reasons for the improved yields of subsequent crops, although crop yields may vary from crop to crop and agroecological regions. The positive response of the subsequently grown crop is also attributed to the transfer of nutrients from cover cropping and less immobilization nutrients . Similar to our findings, Hively and Cox ; Fageria et al. 2005 observed a higher corn yield following white clover and red clover cover crops. Marketable yield of sweet corn was approximately doubled by hairy vetch in 2 of 3 years compared to an unfertilized, no cover crop control . Burket et al. observed a 58% higher average broccoli yield when grown with no fertilizer N, but following a legume cover crop. In general, the response of broccoli as a vegetable crop to cover cropping rotations was positive associated with nutrient, growth and yield output of the crop. If properly managed, then it is most likely that the cover cropping system can sponsor its own soil fertility, crop protection and productivity. Such low input farming systems with improved crop productivity and profitability can be easily adopted by farmers and becomes very useful in organic farming systems where the use of synthetic fertilizers in not acceptable. Cover crops in farming systems improve soil health, reduce environmental pollution, and improve crop yields and maintain sustainability of crop production . Such sustainable production of agricultural products achievable through cover cropping must be based on holistic agricultural management that encourages interdependent and diverse properties. For higher cover crop use efficiency farmers should also deal with selection of appropriate cover crop species with desirable socioeconomic considerations and ultimate vegetable crop yield improvement. It must also involve lower production costs with no adverse effect on crop health and the environment.Management of weeds is a challenge in crop production.

Weeds can interfere with the cultivated crop by competing for light, water and nutrients, which can lead to reduced yields and reduced economic return on investment to the grower . The approach for integrated management of weeds consists of combined inputs from cultural, mechanical, biological and chemical control methods. Cultural practices like clean seed, clean equipment, and proper field preparation are commonly integrated with mechanical practices like tillage, mowing or cultivation for control of weeds. Biological practices are less common in weed management. Therefore, chemical control is the following option to integrate for weed management . Chemical practices are the use of herbicides to prevent weed emergence or to cease growth of weeds until plant death, in most cases. Herbicides continue to be important tools to integrate in weed management programs because of their cost effectiveness, rapid action and flexibility with management, when used appropriately, which have allowed for increased crop yields to be achieved . A successful weed management program can be accomplished when cultural, mechanical and chemical management are integrated. In the California rice production system, herbicide resistance has been a continuing challenge due to continuous rice cultivation year after year, a historically limited number of herbicides available and the overuse of the available herbicides for weed control . From 2015 to 2021, there were 661 suspected herbicide-resistant weed reports and nearly 53% of watergrass populations recorded multiple-resistance to up to four modes of action . The presence of herbicide-resistant weeds leads to a reduction in weed control with the available herbicides and reduced yield. The most recent herbicides registered in California rice include pyraclonil in 2024, florpyrauxifen-benzyl in 2023, benzobicylcon in 2017, carfentrazone in 2006 and clomazone in 2004 . However, grow racks these herbicides have varying degrees of control over different weed species and producers are limited in control options . There is a need for new herbicide tools to maintain the viability of the current herbicides for future years by practicing herbicide rotations and mixtures . However, the registration of new modes of action in a crop or region is influenced by many factors like the crop injury potential, weed control efficacy, environmental concerns or lack of economic incentive by the manufacturing companies . Because not many new herbicide modes of action have been developed recently and herbicide resistance is increasing, new potential rice herbicides can be evaluated from other cropping systems or by revaluating or reformulating older chemistries. There has been success in introducing herbicides from larger agronomic crops to high value specialty crops through the Interregional Project Number 4, a US federal program . Similarly, evaluating older chemistries for new crops can be successful; however, the environmental effects are of greater concern because old chemistries tend to be less environmentally safe . Various characteristics are important to consider when evaluating a potential herbicide for a new crop like crop safety, weed control spectrum and persistence in the environment.

To ensure a greater potential for success when evaluating new herbicides, a hypothesis-driven research approach should be taken. Pendimethalin [N–2,6-dinitro-3,4-xylidine] is a mitotic inhibiting herbicide from the dinitroaniline chemistry that inhibits seedling growth shortly after germination . Pendimethalin controlled herbicide-resistant grass populations in the greenhouse and has relatively few reports of resistant weed populations . Preliminary greenhouse work indicated pendimethalin was effective in controlling several recently collected herbicide-resistant grasses from California rice fields . Therefore, pendimethalin could be a valuable addition for management of herbicide-resistant weedy grasses. Pendimethalin is registered for use in dryseeded rice and commonly applied to the soil surface after drill-seeding rice relatively deep in the soil . In dry-seeded systems; however, rice injury from pendimethalin is influenced by soil moisture, where higher soil moisture leads to greater injury levels . Characterization of pendimethalin in water-seeded rice, where moisture is always present, has not been evaluated because of the perceived risk of rice injury . There is no previous research that has evaluated pendimethalin formulations at different rates and timings in water-seeded rice. Therefore, the objective of these studies was to evaluate and characterize pendimethalin in water-seeded rice. The objective of this chapter is to review the literature on pendimethalin, pendimethalin use in rice production systems and background related to characterizing pendimethalin for water seeded rice. This review will provide greater background to the research studies outlined in the following dissertation chapters. The review will begin with a history and background of the dinitroaniline chemical family, pendimethalin use in rice, environmental fate of pendimethalin in rice production systems, and future directions for characterization in water-seeded rice.The dinitroaniline chemistries have been available herbicides since the 1960’s. The first dinitroanilines were synthesized by the Eli Lilly Research Laboratories and included trifluralin, benefin, nitralin, isopropalin, oryzalin, profluralin, butralin, ethalfluralin, fluchloralin andprosulfalin . Trifluralin was the first commercialized compound in the US and was used in soybean and cotton as a pre-plant incorporation for grass weed control at rates of 1,000 to 2,000 g ai ha-1 . Currently, trifluralin and pendimethalin are the most commonly used dinitroanilines in the US and worldwide for weed control in cereals, cotton, soybeans, vegetables, ornamentals and fruit and nut trees . Pendimethalin was developed by American Cyanamid in the 1970’s, previously named penoxalin . BASF would later purchase the American Cyanamid’s agrichemical business and take possession of pendimethalin in the 2000’s. Pendimethalin was moderately less volatile than trifluralin, which lead to relatively greater soil persistence and longer weed control activity .Characteristics of Dinitroanilines. The physical, chemical and biological properties of herbicides can help broadly predict their behavior in the environment, weed control efficacy and handler safety. The 2,6-dinitroaniline chemical structure is the base structure that defines the compoundsin the dinitroaniline chemical group . The additional chemical structures on the base structures will affect the specific characteristics of each compound. The common characteristics in all dinitroaniline compounds is a low water solubility, typically <1 parts per million , and most are soluble in organic solvents. The nitro groups decrease the water solubility by creating hydrogen bonds with alkyl groups of other compounds like soil or organic sediments, which creates lipophilic aggregates . The lipophilic nature appears to make the compounds susceptible to bioaccumulation in the environment; however, they have a high affinity for organic sediments or organic matter in the soil . Most compounds are non-ionizable, except for oryzalin . These characteristics tend to lead to the classification of dinitroanilines including pendimethalin as low risk for contaminating surface or ground water and low risk to environment contamination and human health .

The sugar beet cyst nematodes were not even as much sensitive as the RKN to the cover crop treatments was not variable among the cropping treatments, although were slightly higher in cowpea and marigold cover crops at the ABH sampling of the second year and the ACCP and ABH samplings of the third year. The increase in SCN mainly at the ABH samplings than at other sampling periods, may indicate that broccoli is a host to the SCN. Potter and Olth of actually show that broccoli is a potential host to the cyst nematodes. Infection of broccoli roots and broccoli root gall formation was very minimum and unaffected by the cropping treatments. Based on my current over all findings therefore, the usefulness of cowpea and marigold as offseason cover crops does not confirm their nematode suppression potentials in the subsequent winter broccoli crop.There are various reasons documented for variation in nematode suppressing efficiency of cover crops. Ploeg and Maris state that the life cycle of Meloidogyne incognita complete between average soil temperatures of 16°C and 30°C on tomato, but only at 30°C on marigold . Furthermore, motility of M. incognita J2 and its subsequent root penetration may decrease with decreased soil temperatures below 18°C . These findings suggest that the effectiveness of cover crops to suppress nematodes depends on the condition under which they are utilized. Ploeg and Maris further suggested the need for information on thermal-time relationships of plant parasitic nematodes to predict geographical distributions, nematode population dynamics and effects of cover crops on the subsequent crops. Effectiveness of a cover crop for the purpose of nematode suppression may also depend on the type of target nematode itself.

Wang and McSoreley pointed out that Iron Clay‘, cowpea failed to suppress root-knot nematodes where there were mixed species of Meloidogyne. Ploeg and Maris also identified nematode suppression of marigold being influenced by crop plant variety, nematode species, hydroponic trays and soil temperature. Marigold while suppressive to root-knot nematode, it enhanced the population densities of other nematodes such as stubby-root, spiral and sting nematodes on the other hand. Therefore, the evidence suggests that the type of nematode can determine the effectiveness of a cover crop.Others observed that nematode suppression of cover crops may depend on how the cover crops were utilized. Wang and McSorley observed that cover crop mulch was more effective than live crops. On the other hand, Ploeg and Maris state that live marigold suppress nematodes, because of the release of alpha-terthienyl, a toxic chemical compounds from its live roots that have nematicidal characteristics . These nematicidal compound released by active, living marigold roots may not be available if marigold is used as an organic mulch . Since my research was based on the off season cover cropping system and employed their residues as surface mulch and soil incorporation, the observation of poor or no nematode suppression can be justified. Similarly, Ploeg did not observe any significant suppression from preceding vegetable crops or amending a planting site with marigold plant parts. Furthermore, while cowpea incorporation as a green manure has been observed to suppress Meloidogyne incognita , the suppression was short-lived, and the numbers of M. incognita were not different from a fallow treatment . Another factor determining cover crop effectiveness was the type of the subsequent vegetable crop that may determine the potential incidence of plant parasitic nematodes.

If the subsequent indicator crop is a nematode susceptible plant, it may be possible to detect nematode suppression of cover crops, otherwise, the effects of the cover crops can be masked if the indicator crop is nematode resistant. However, if the vegetable crop isresistant to nematodes by itself, nematode suppression potential of a cover crop could be masked. Accordingly, I may not have observed any significant nematode suppression by the cover crops, because the broccoli used in this research was resistant or a poor host to most nematodes. Most broccoli cultivars contain sulphur compounds such as methanethiol, dimethyl sulphide, methyl thiocyanate, dimethyl disulphide, dimethyl trisulphide, dimethyl tetrasulphide that may be toxic to nematodes . The presence of nematode antagonizing organisms such as bacteria and fungi in a soil may also contribute to the reduction of nematode population densities , regardless of nematode suppressive treatments. Kerry observed that the second-stage juveniles of root-knot nematodes encumbered with spores of the bacterium Pasteuria penetrans are less able to invade the roots of host than the unencumbered nematodes. The most significant outcome of the cover cropping treatment was the enhancement of saprophytic nematodes. Saprophytic nematode populations were significantly enhanced at ACCP sampling of the second year and the ACCI sampling of the third year in the cover cropped plots, relative to the fallow plots. Since these nematode populations became higher at after cover crop incorporation, the increase in saprophytes may have come from the accumulation and decomposition of cover crop residues. The relatively lower saprophytic nematode populations in the fallow plots may have been associated to the lower input of organic matter from such cropping system.Therefore, the results confirm that preceding vegetable crops with cover crop could enhance beneficial saprophytic nematode populations. Saprophytic nematode population density for the first year was not significantly different for the cropping treatments, indicating that a one year cover cropping rotation is not sufficient to enhance populations of free-living nematodes. On the other hand, the increase in saprophyte population with repeated years of cover cropping suggests that there is accumulative effect of the cover cropping treatments.

The results clearly demonstrate that cover-cropping rotations must be repeated for several years in order to provide significant contributions to enhance saprophytic populations. The sharp decline in saprophyte populations at the ABH sampling of the third year might have been due to a complete decomposition and degradation of the organic matter to a level that no longer sustained high saprophytic populations at this stage. An increase in bacterial-feeding nematode population densities following soil treatment with sunn hemp as organic mulch was also observed by Wang and McSorley . Free living bacterial feeding taxa of nematodes constitute more than 60% of the nematode community . The presence of high population densities of saprophytic nematodes may provide an added advantage in soil biology . Saprophytic nematodes are useful in mineralization of plant nutrients and nutrient cycling and can be used as sensitive indicators of ecosystem change . Langat et.al. suggested that bacterivorous nematodes respond quickly to increased food supply. Therefore, cover crops play an indirect role of increasing population levels of the beneficial free living nematodes. The structure of the nematode assemblage offers an interesting instrument to assess changes in soil conditions . Enhancement of saprophytic nematodes and the mineralization and nutrient cycling benefit that such nematodes can provide to the subsequent crop indicates the profitability of cover cropping rotations. In general, the use of cowpea or marigold cover crops as an off-season cropping rotation may not provide a viable alternative as a nematode suppression strategy. Hence, the use of cover crops for nematode suppression must be considered carefully, accounting for the target nematode, how the cover crops are to be used, and the environmental conditions of the field. However, pipp mobile systems these cover crops can be used as off-seasoning cropping rotations to effectively enhance beneficial saprophytic nematode population densities in the subsequent vegetable crop. They do so as their residues decompose supporting nematode food webs. The increase in population levels of saprophytes and feeding on nutrientimmobilizing bacteria and fungi promotes nutrient mineralization and nutrient cycling. It is believed that with more knowledge about the mechanisms stimulating a beneficial nematode community, we may develop cover crop management plans to maximize the desirable effects associated with free living nematodes . Traditional insect pest management approaches utilize pesticides, but these are known tobe environmental pollutants , and in some cases carcinogenic . Insect pests may also develop resistance to insecticides making them inefficient . Those shortcomings of broad spectrum insecticides, encouraged attempts to replace them by ―soft‖ microbial based insecticides such as Bacillus thuringiensis . Yet, insect pests developed resistance to the soft insecticides as well and some ―soft‖ insecticides can be injurious to parasitoids . Therefore, there is an increasing demand for environmentally friendly and economical alternative pest management strategies . Many researchers have suggested crop diversification and cover crops as alternative insect pest management tactics . Cover cropping systems may adversely affect insect pests and, if effective could be used as an alternative insect pest management strategy as they are ecologically benign, minimize reliance on pesticides, reduce chemical exposure, and increase consumer confidence in food production . Although cover crops could potentially interfere with vegetable insect pests, not all cover crops are equally efficient in suppressing vegetable insect pests.

For example, sweet clover cover crop suppressed broccoli pest populations, but not pests of tomato or pepper . There are also concerns that cover crops used as simultaneous inter planting may compete with the main crop for growth resources and reduce vegetable crop yield . Consequently, this research was designed to evaluate the effectiveness of off-season summer cover cropping as an ecologically desirable pest management strategy for the subsequent winter vegetable crop of broccoli. It specifically evaluated the effect of two summer cover crops on population densities of broccoli insect pests and beneficial arthropods.A three-year field study was conducted from 2007-2009 at the University of California South Coast Research and Extension Center in Irvine, CA on a loamy-sandy soil. Three summer cropping treatments were employed: 1) French marigold , 2) cowpea , seeded at 56 kg/ha, and 3) a summer dry fallow as the untreated control. Each treatment plot was 12 m long x 10.7 m wide and laid out into 14 planting rows. The cover crops were direct-seeded in the last week of June in the center of the planting rows of each plot, watered through drip-tubing and grown for three months. The fallow control plots did not receive water during the summer. Each cover crop treatment plot was planted with the same cover crop in each of the three years of study. Plots were separated from each other with a 3 m wide buffer bare ground. The three treatments were replicated four times in a completely randomized design. At the end of the summer cropping period , the cover crops were mowed at the soil line, chopped, and the residues left on the ground. Concurrently, alternate rows of each of the cover crop treatments were incorporated into the soil at about 0.4 m intervals using a hand-pushed rotary tiller in preparation for broccoli transplanting. The fallow plots were not tilled. Plots for cover crop and broccoli planting are shown in Figure 1a. At the beginning of the subsequent cropping season , broccoli seedlings were transplanted in double rows into the tilled strips of the summer cover crop and fallow plots at an inter and intra-row spacing of 13 and 35 cm, respectively . Broccoli transplants were drip irrigated and fertilized with emulsified fish meal at 5 gallons/acre rate. Broccoli was chosen because it is a high-value vegetable crop that is sensitive to weeds, insect pests, nematodes , and requires high soil nutrients . All plot treatments were maintained in the same location for all three years of study in order to assess a cumulative effect of cover crops over time.Plants were non-destructively sampled for major broccoli insect pests beginning 15 days after broccoli transplanting and continuing every two weeks until broccoli harvest. On each sample date, 20 randomly-selected plants per plot were assessed visually for thepresence of insect pests following methods used by Costello and Altieri and Hooks and Johnson . The larvae of the insects actively feeding on the vegetable crop were identified to species level and recorded as the number of individuals per plant. Population density of each species was computed as average number of larvae per broccoli plant.Arthropod parasitoids were assessed by rearing field collected broccoli insect pests in the laboratory, beginning at 21 DAT and continuing once every two weeks until broccoli harvest. Five plants from the interior three rows of each plot were randomly selected and searched for insect larvae. Larvae were collected and placed in clear plastic cups with greenhouse grown broccoli leaves as source of food. Individual larvae were reared in the lab and the fate of each larva recorded.

Agronomic crops have distinct and uniform morphology, but for roadside invasive species, the high variation in plant morphology and the non-uniform backgrounds will lead to more significant detection errors. Previous studies and discussions supported that computer vision can replace human observers in species detection. However, we still need human observers to create a training dataset. Training, testing, and validation datasets are the essential components of a deep learning model, with the most time-consuming task being image labeling. A larger training dataset can increase model performance, but the size would determine the amount of labor for a single project. Abdulsalam & Aouf suggested that 1,000 images of a particular species are required to achieve high prediction accuracy. Yan & Ryu proposed that the training sizes would differ depending on the mapping species since they only used 400 training samples for corn, but the model could still perform with high accuracy. Compared to the traditional car survey, the AIbased survey method can be conducted by non-experts once the image detection model is trained. Johnsongrass is a common weed along the roads in the United States and is a good model plant for the AI-based survey method. It is native to the Mediterranean and North Africa and was introduced to the United States in the early 1800s . Johnsongrass is a perennial grass and can colonize and spread nearby landscapes through the underground rhizome system . The mature plant can grow up to 2.5 meters in height, and the height of the mature plant can vary based on the local condition . McWhorter reported that the reproductive stage of johnsongrass started around a month after seed emergence, cannabis drying system and the maximum rhizome growth was about 60 meters in 5 months. The flowering part of johnsongrass is a diffuse panicle, which is the primary feature in the image identification process.

The flowering head is orange and purple at the mature stage. Johnsongrass is a weedy relative of the cultivated sorghum , which compete for the same limiting resources, and the presence of johnsongrass will cause yield loss in sorghum or other crop fields . Kansas and Texas are the top two states in cultivated sorghum production regarding planting acreage . As a weedy relative, johnsongrass is widespread in sorghum fields and along the roads around Nebraska, Kansas, and Texas to conduct an automated road survey of johnsongrass, examined the cost-effectiveness, and discussed the potential application of the johnsongrass population map.The performance of the YOLOv2 model in detecting johnsongrass in GSV images was tested using a total of 2,040 test images. Based on the threshold value of 0.6 for presence , the confusion matrix shown in Table 1 was created. The YOLOv2 model achieved a recall value of 85% in the GSV testing dataset. Dang et al. reported similar recall values in their study which the average recall value of the YOLOv3 detection model on 12 different weed species was 87.93%. However, there is still about a 15% chance that johnsongrass could be undetected by our model when johnsongrass is present in the image. In the testing dataset, there were 153 images were classified as FN in the confusion matrix. FN in this project is the image that contained johnsongrass and was identified and labeled by human observers, but the model was not able to output the same result. Individuals that were at pre-flowering stage were not considered as FN since the training dataset only contained mature johnsongrass. The precision for the YOLOv2 model was 0.74, which is lower than the recall . Both the precision and FPR include false positive detection in the calculation. FPR implies that the model could wrongly detect other plant species as johnsongrass, with a 30% chance. Among the group of incorrect detection , FP had twice the number of images compared to FN in the test dataset . Sincemost studies in weed detection were conducted in the crop field, and their models were applied to distinguish weeds from the crop, the precision values were high and were about 85% to 95% . Yan & Ryu applied a CNN model on GSV images to detect roadside crop type, and the results denoted that most crops had detection precision above 90%, but only rice had a 76% precision. The study also reported that the misclassification of rice was more frequent in low-resolution images or if the object was far away from the camera . In our research, the quality of images and the distance between the target object and the camera might contribute to the high FP value. Future work will focus on decreasing the rate of FP in the image where johnsongrass is absent. More CNN models will be tested on the johnsongrass training database to compare the precision and accuracy of different models. More roads with high-resolution images from Google can help improve this survey method’s accuracy. The overall accuracy of the YOLOv2 model was 77.5% for detecting johnsongrass in the GSV images. This index provided an overall evaluation based on total correct detection and the total number of test images. Ringland et al. and Yan & Ryu both conducted image detection models on GSV images to survey different types of crop production along the roads but with different CNN networks from our model. The accuracy of detecting general crops like alfalfa , almond , corn , and rice in the GSV images could reach 92% . An explanation of high accuracy on crops is that major crops always have unique morphology or patterns because of domestication, row spacing, and field layout that might help to increase performance in computer vision. For roadside weedy species like johnsongrass, morphological variations under different environmental conditions were reported in many studies, and the variation could lead to low precision and overall accuracy . There were several challenges in the labeling process, and they can explain most of the incorrect detections. In some annotated training images, the target species were partially occluded by other objects, including other invasive species, traffic signs, and fences. In this case, we could only label either the flowering part or the basal part of johnsongrass. In this project, and for johnsongrass specifically, the panicle part of the plant would be labeled in most cases since we could not differentiate the basal part of johnsongrass from other grass species. The growing stages of the target species were another challenge in the labeling process. The juvenile stage of johnsongrass has no panicles and looks similar to many other grass species. Only the mature johnsongrasses were included in the training dataset, so the model was unlikely to detect individuals at their early vegetative stage.The trained model was applied to 269,489 images collected from Google Street View. In Figure 6, the red points denote the potential location of johnsongrass predicted by the model. The model identified a total of 2,031 images as having johnsongrass. The predicted distribution of johnsongrass suggested that johnsongrass is less widespread in Nevada than in the other states in this study. The location shown on this map is only the prediction. The johnsongrass individual might not be found in that location depending on the growing season since most images were taken 2 to 3 years ago. Deus et al. conducted a Google Street View study that surveyed E. globulus , and their results mentioned that environmental stresses could lead to variability in species abundance in a short period, from one to two years. Recent studies and our results suggested that integrating GSV and a deep learning image detection model can map species on a much larger scale. Yan & Ryu integrated GSV and other deep learning algorithms and produced cropping system maps of Central Valley in California and the state of Illinois. Another roadside crop survey in Thailand covered 572 km of road and examined about 57,000 panoramas . Our study covered more areas , longer roads , growing tray and more panoramas than studies used a similar road survey method . Future research will focus on the survey in other states in the US, and our goal is to survey all the roads in the US.YOLO has been tested in many studies to detect multiple plant species in a single image . Johnsongrass was the only detection target in this study, but other invasive species can be mapped by using our methods. A larger-scale species distribution map can be combined with environmental factors or land use to determine the conditions suitable for spreading the species. An example would be the habitat suitability model, which predicts how well species thrive and spread in a location given environmental conditions . According to Crall et al. , even though the habitat suitability model is a key tool for invasive species risk management, the model requires location data on a large spatial scale. Our method can provide a more prominent presence/absence dataset than the traditional local dataset. Habitat suitability models based on a larger scale can yield a more robust conclusion. As noted, the sampling created by our method is a biased sampling of the environment under which the species can thrive as we only search for species along the roadside habitats. AI-based surveys can provide accurate location data to build and test invasive species dispersal models. The AI-based mapping approach can only detect roadside weedy species. A dispersal model can be applied based on the johnsongrass location map. A typical dispersal model requires two primary parameters, reproduction rate and spread distance, and then for parameterization and calibration, a multiple-time-step map is required . The johnsongrass map was created based on images from a different date. Even though most of the images were taken in recent years, like 2020, a small portion was taken 8 or 10 years ago. Expenses and estimated time for the car survey, the human-based GSV survey, and the AI-based GSV survey were calculated . Expenses for the car survey were calculated based on the same scale as the AI-based survey. The cost breakdown of the car survey was calculated based on regular domestic travel daily expenses. Vehicle rentals and gas estimation are US$ 9,408 and US$ 7,350, respectively, and the accommodation accounts for a more significant portion of the costs, i.e., US$ 25,200 . The total travel time of a car survey per person requires 180 days, estimated based on a daily 750 km drive. Dues et al. conducted a 38-day car survey of 15,000 km of roads in Portugal, and a standard car survey would drive much less than 750 km per day. For the human- and AI-based GSV survey, US$1,890 is required for image purchase from Google, which is US$7 per 1000 images . Labor costs for all three types of surveys were calculated based on the minimum hourly wage in California and the total time spent on each method. In terms of cost, the AI-based GSV survey is 50% less than the human-based GSV survey, and the AI-based GSV survey is only 5.6% of the total cost of the car survey.Compared to car surveys, GSV-based surveys did not require outdoor work and driving, so GSV-based detection can minimize potential worker risks . Studies by Deus et al. and Kotowska et al. reported that the results produced by GSV resemble that produced by the field survey. Compared to the GSV survey by a human observer, the AI-based GSV survey had spent shorter time. Once the detection model is trained, the machine can work 24 hours per day, but on average, a human can process about 6000 ~ 8000 images per day based on our labeling experiences. A more detailed GSV survey might take more time. For example, a human-based GSV survey took 35 hours to examine 2,350 panoramas in Sicily, Italy, to assess invasive species abundance along the roads . As noted, once the training dataset is created, the labor cost of the AI-based GSV surveys is fixed and will not increase as the number of images increases. However, the relation between labor cost and image number in the human-based surveys is linear. As the sampling scales increase, AI-based surveys will outcompete human-based surveys, and the comparison between these two surveys in Table 3 is underestimated. Then in terms of errors, the detection errors by the AI-based model could be consistent, quantified, and improved, while errors in car surveys and humanbased GSV surveys are unable to quantify and inconsistent.

A key finding is that overnight charging currently has the lowest absolute level of CO2 emissions and a relatively low variance compared to other times of charging. This may not imply that an EV solution is categorically preferred on the grounds of net CO2 emissions compared to a baseline HEV, although the research does permit such quantitative comparisons. And it does convey actionable information given the vast temporal optionality currently allowed for charging. Going forward, we can envision a means of aggregating the impacts of individual analyses, meaning that these events will be weighted based on their likelihood and current behaviors. We can furthermore consider behaviors will evolve as time goes by and as larger shares of EVs are realized. However, within a transition period where EV growth and grid dynamics adapt iteratively, this study conducted novel simulations of the primary grid-vehicle scenarios which are reflective of current EV behavior and grid characteristics in recent years. The study’s consultations with experts, literature review, and data analysis revealed that about ¾ of events occur at home, with 50% on a level two charger and 25% on a level one charger. This would suggest, for the near and intermediate term, that EVs will act as a kind of aggregated demand in the evening/overnight hours and that as a block , EVs are more likely to require marginal resources because they act together to force demand projections out of the expected regime. The good news is that, for the foreseeable future, these are, on balance, curing cannabis hours that are moderately lower in terms of marginal resource carbon intensity, since they can be met by intermediate resources . Whereas workplace charging is substantially less common, perhaps the marginal mix or the hourly estimation is reasonable for the effective CO2 signatures.

As one thinks about the scale, and a situation where of EV reach double-digit shares of the fleet, it’s very likely that all the modes and locations of charging will eventually be subject to conditions where marginal assumptions prevail. How the transition period is defined and how the system boundaries between EV growth and grid resources are balanced are important, but open questions. Another essential unknown that will impact effective CO2 emissions from EVs are how predictable EV charging events become, with an emphasis on the high power, coincident peak events. More research into this question can help inform more accurate methods and models for simulating the environmental impacts of EVs. The present framework sets up an approach that will be valuable in estimating future impacts under such conditions. While additional focus and scope lie beyond this study, it is clear that a more complete understanding of popular EV charging profiles and EV driving behaviors will be essential inputs to better decision-making and resource planning. As EV use and charging habits become more predictable and well-known, the relevant data and insights can be critically valuable to utilities. For instance, foreknowledge of EV charging events will be needed at an aggregate level and could be beneficial to grid operators. The reason is that they can better plan and iterate their learning for those types of loads and events for which currently they lack visibility.In the last 40 years, 30 percent of the world’s arable land has become unproductive and 10 million hectares are lost each year due to erosion.1 Additionally, accelerated erosion diminishes soil quality, thereby reducing the productivity of natural, agricultural and forest ecosystems. Given that it takes about 500 years to form an inch of topsoil, this alarming rate of erosion in modern times is cause for concern for the future of agriculture.

This supplement explores the major causes of soil erosion and the social impacts it has on communities, underscoring the importance of agricultural practices that prevent or minimize erosion. Anthropogenic causes of accelerated soil erosion are numerous and vary globally. Industrial agriculture, along with overgrazing, has been the most significant contributor, with deforestation and urban development not far behind.2, 3, 4 Heavy tillage, fallow rotations, monocultures, and marginal-land production are all hallmarks of conventional agriculture as it is variably practiced around the world and significantly encourage accelerated soil erosion. Repeated tillage with heavy machinery destroys soil structure, pulverizing soil particles into dust that iseasily swept up by wind or water runoff. Fallow rotations, common with cash crops around the world and subsidized in bio-fuel production in the U.S., leave land vulnerable to the full force of wind gusts and raindrops. Monocultures tend to be planted in rows, exposing the soil between to erosion, and are commonly associated with fallow rotations. More and more marginal land, land that is steep and particularly susceptible to water erosion, is being planted by farmers either attracted by higher crop prices or forced by loss of productivity on flatter, but already eroded lands. In an increasingly complex global food web, seemingly separate causes of erosion begin to influence each other, magnifying their effects. For example, deforestation of tropical forests in Brazil clears the way for industrial soybean production and animal grazing to feed sprawling urban populations in the U.S. All the while, fertile topsoil is carried away by wind and water at alarming rates. Environmental harms resulting from accelerated erosion are well documented. Decreased soil fertility and quality, chemical-laden runoff and groundwater pollution, and increased flooding are just a few of these detrimental effects. There are, in addition, disproportionate social harms resulting from high rates of erosion that are less obvious, but no less directly linked. Hunger, debt, and disease are serious problems in mostly poor, rural communities around the world that are exacerbated by accelerated erosion.

As global agricultural development and trade have accelerated in the last half-century, mainly via the “green revolution” and the formation of the World Trade Organization , increasing trade pressures have raised export crop production in less developed countries. As a result, farmers mainly in Asia, Latin America, and sub-Saharan Africa are increasingly abandoning traditional farming techniques and locally significant crops in favor of adopting the industrial practices mentioned above that lead to high rates of erosion.5 While development institutions and governments proclaim concerns for the rural environment, agricultural policy supporting high commodity prices and limited credit access continually pushes farmers to intensify land use. Coupled with the fact that the total area of arable land in cultivation in these parts of the world is already very high , land degradation by soil erosion threatens food security by removing from cultivation land sorely needed for domestic food production. The majority of the world’s 868 million undernourished people live in Eastern and Southern Asia and sub-Saharan Africa. One of the international responses to soil degradation in the developing world has been to promote soil conserving tillage practices known as minimumor no-till agriculture. No-till agriculture protects soil by leaving crop residue on the field to decompose instead of plowing it into the ground before planting the next crop. Weed management is addressed with heavy herbicide use to make up for the loss of weed control from tillage. The practice, extensively adopted in the U.S., weed dryer has been popular in Brazil and Argentina, and much effort is being expended to expand no-till to Asia and Africa. There are, however, costs associated with no-till agriculture, both economic and social. First, no-till agriculture is expensive to adopt. Herbicides, seed drills, fertilizers, and other equipment require a high initial investment not possible for poor farmers without incurring significant debt. Second, heavier herbicide use increases human exposure to chemicals and contributes to water and air pollution. Third, weed pressures can change in unexpected ways as reliance on a handful of herbicides breeds resistance. Weed resistance to the popular herbicide, glyphosate, is an increasing concern in conventional agriculture and is leading to development of more harmful herbicides to compensate for glyphosate’s reduced effectiveness. Lastly, no-till agriculture also promotes monoculture cropping systems that, as described above, have a deleterious effect on soil quality. The techniques illustrated in this manual emphasize long-term soil stewardship using an integrated approach to soil health and management. For example, cover crops hold soil aggregates together in the wet season, protecting soil from the erosive effects of rain. Properly timed tillage limits its destructive effects on soil particles and soil structure. Compost promotes a healthy soil ecosystem, improving soil’s structure and its ability to more successfully withstand wind and water erosion. In addition to environmental benefits, agroecological systems are often based on traditional farming practices that promote soil-conserving techniques and varietal choices adapted to the particular region, stemming the tide of land consolidation and commodity crop production. Food security is enhanced and debt risk reduced by way of diverse cropping systems and labor-intensive, rather than input intensive, production methods. And there are public health benefits from eliminating exposure to harmful pesticides and herbicides. In sum, the serious challenge presented by accelerated soil erosion coupled with the uncertainty about whether no-till agriculture’s benefits outweigh its harms underscores the importance of employing an agroecological approach to farming that prevents soil erosion on farms.On vegetable farms in the Salinas Valley, a shrinking farm labor pool and rising minimum wages are driving innovation and adoption of machinery that can automate manual labor tasks — thinning, weeding and, for some crops, harvest. The technology is evolving quickly, led mainly by small engineering firms collaborating with large growers. Automation promises a number of benefits. Foremost, of course, is a reduced dependence on manual labor. But it could help in other ways too — for instance, automated weeding could remedy the declining effectiveness of some herbicides. UC researchers and advisors are helping to advance the basic technologies involved, and also serving as key evaluators of the technology . But the drive to automate also raises decades-old concerns about UC contributions to new technologies that are likely to primarily benefit only large-scale growers, at least in the short term.The automation of thinning and weeding involves two main steps: identifying each plant to be removed and then directing the killing of the undesired plant with a blade or a small dose of herbicide. It replaces work that would otherwise be done by hand with hoes. Figures on the acreage being thinned by machine aren’t available, but the use of automated thinners in some crops, notably lettuce, has been expanding in the Salinas Valley since its introduction in 2012 . The two in widest use in the Salinas Valley, according to several researchers and equipment suppliers, are made by two small northern European firms, Denmark-based F. Poulsen Engineering and Netherlands-based Steketee. Long-running concerns about farm labor cost and availability in Europe have driven automation innovation, and the technology has been more widely adopted there than in the United States, said Richard Smith, a UC Cooperative Extension farm advisor in Monterey County. While the weeding machines are costly — roughly $150,000 to $200,000 — their use appears to be limited more by availability than by price, according to equipment suppliers and UCCE staff. Poulsen and Steketee are small operations with limited production capacity. Britton Wilson of Pacific Ag Rentals, an equipment supplier to Salinas Valley farms, estimated that there are 15 to 20 Poulsen weeders in the United States, a figure Poulsen corroborated. “I’d love to get my hands on more” to meet local demand, he said.A crop like lettuce or broccoli represents a comparatively small market for major farm equipment makers like John Deere and Case IH. About 300,000 acres of lettuce are grown in the United States, for instance, compared with 12 million acres of cotton or 90 million acres of soybeans. As a result, vegetable crop automation is being led by small engineering and fabrication firms as well as growers themselves, often in close collaboration, said Mark Siemens, an associate specialist and associate professor of agricultural and biosystems engineering at the University of Arizona. Because the technology is somewhat modular, it’s possible to address the needs of a particular crop or grower by combining or modifying existing technologies and equipment. An example: Harvest Moon Automation, a four employee engineering firm with several clients in the Salinas Valley, recently received a patent on a modified version of a leafy greens harvester developed in partnership with two Salinas Valley growers. Steve Jens, Harvest Moon’s president, said the new machine uses a camera and pattern-recognition technology to spot foreign objects and diseased or damaged plants as the harvester moves across a field.

The intensification experiment was implemented in an established walnut orchard at the Plant Sciences Field Facility in Davis, CA, USA . The orchard was planted in the spring of 2015 with ‘Chandler’ walnuts. The entire orchard was 0.7 ha in area consisting primarily of Yolo silt loam soils . Orchard management included microsprinkler irrigation and weed-free tree strips maintained with preemergent herbicides. Experimental plots included the orchard alley between seven pairs of trees, approximately 6 m by 40 m. Cover crop programs were based on cereal rye, since it is known to be a competitive, weed-suppressing species that has desirable termination characteristics . Furthermore, this species thrives under various cultural management conditions and has cultivars that are well-adapted to grow as a winter cover crop in Central California. We used ‘Merced’ rye, which is a relatively tall cultivar. The whole experiment was conducted in one orchard over two growing seasons. Cover crops were established in the fall of each year, on November 11, 2019 and November 9, 2020, and terminated in the spring of each study year, on April 24, 2020 and April 9, 2021. Each plot received the same cover crop management program in both years of the experiment. Except for the forage treatment described below, rye was direct-planted with a seed drill at 22.5 kg planted ha-1 , drying rack and cover crop termination was performed with a flail mower. Planting and termination operations were planned to minimize equipment traffic in the orchard, and only one tractor pass was made across each orchard alley at each planting and termination date.

Flail mowers are practical for cover crop termination in California, since these implements are more common than other cover crop termination tools and they minimize crop residue ahead of nut harvest. We had five treatments which represented a range of different cover crop management intensities. The ‘sprayed’ treatment was used as our non-treated control, and the rye planted in these plots was terminated with a glyphosate application when rye plants reached 5 to 10 cm in height. These burn down applications occurred on January 13, 2020 and January 12, 2021, and included a broadcast application of Roundup Weather MAX at 1.607 L ha-1 with a carbon dioxide-propelled backpack sprayer. This treatment mimics a relatively intense commercial management system where orchard alleys are kept weed free. The ‘standard’ treatment included rye with no other cover crop management until termination. The ‘multi-species’ treatment included the base planting of rye and several additional cover crop species. The other cover crop species in the mix were common vetch 4.5 kg planted ha-1 , ‘PK’ berseem clover at 4.5 kg planted ha-1 , daikon radish at 2.25 kg planted ha-1 , and ‘Braco’ white mustard at 2.25 kg planted ha-1 . These seeds were broadcast spread immediately before rye was planted. We used these methods to establish the sprayed and multi-species treatments to minimize logistical challenges and orchard traffic, while also relying on the tractor and seed drill to enhance seed-to-soil contact of our additional cover crop species in the multi-species treatment. The multi-species treatment in this experiment has the same species and approximate planting rates as the multi-species mix in the planting date experiment described below.The ‘boosted’ treatment included a 45 kg ha-1 N top dress with granular urea after rye tillering which were made on February 25, 2020 and February 26, 2021.

The ‘forage’ treatment was managed as a rye hay intercrop. This treatment was planted at a rate of 45 kg planted ha-1 . At planting, we fertilized with 40 kg ha-1 N and 28 kg ha -1 P as granular urea and monoammonium phosphate at planting. We also top dressed with 45 kg ha-1 N after rye tillering. On the same day as topdressing, we broadcast-applied carfentrazone at 73 mL ha-1 with a backpack sprayer as a post emergent herbicide application for broadleaf weed control. The top dress and herbicide applications were applied on February 25, 2020 and February 26, 2021. The forage treatment was terminated with a swather, and the crop material was subsequently baled and removed. Immediately before cover crop termination, we destructively sampled cover crop and weed biomass. We collected biomass samples from two 0.25 m2 quadrat subsamples in each plot. Cover crops and weeds were separated before being dried in forced air drying ovens. Finally, we weighed dry plant biomass. Summer weed emergence was assessed after cover crop termination using point intercept transects. One transect was placed diagonally across the alley in each plot. Transects were 25 m long with 25 points spaced evenly along the transect. Plants were identified visually at each point. These summer weed transects were performed on June 17, 2020 and May 21, 2021, when summer weed emergence and potential cover crop regrowth might be scouted by a grower planning summer weed management. Planting date experiment. The planting date experiment was implemented in a non-bearing almond orchard at the Wolfskill Experimental Orchard near Winters, CA, USA . The orchard was established in the fall of 2017 with alternating rows of‘Nonpareil’ and ‘Aldrich’ almonds.

The entire site was about 1.1 ha in area with primarily Yolo loam soils . Orchard management included microsprinkler irrigation and weed-free tree strips treated with preemergent herbicides. The experiment was laid out as a randomized complete block design with five repetitions. Experimental plots were roughly 25 m long and 12 m wide, comprising five trees in length and two orchard alleys in width. We had five treatments, including a non-treated control and two multi-species cover crop mixes each planted at two different planting dates. The non-treated control had commercial standard vegetation management practices, which included several glyphosate applications throughout the winter months. We used cover crop mixes in this experiment because of their existing use by California orchard growers . Orchard growers frequently choose among cover crop mixes that support a variety of ecosystem services aside from vegetation management, such as pollinator health or improved soil structure, and multi-species cover crops can support some of these multifunctionality goals. Additionally, using different cover crop mixes allowed us to evaluate cover crops with different germination timings and a range of emergence phenologies. The two cover crop mixes used in this study were a ‘multi-species’ mix and a ‘brassica’ mix. The multi-species mix used the same species as the multi-species treatment in the intensification study, and it included a common combination of cover crop functional groups including a small grain, legumes, and mustards . The mix consisted of 10% ‘Braco’ white mustard, 10% daikon radish, 30% ‘Merced’ rye, 20% ‘PK’ berseem clover, and 30% common vetch planted at 56 kg planted ha-1 . Each of the cover crop mixes was planted at a relatively early planting date and a late planting date. These dates were chosen to represent a timely cover crop planting soon after nut harvest and coincidental with the onset of winter rains as well as a later cover crop planting coincidental with nut pruning, sanitation, and other winter management activities. This experiment was conducted in one orchard over three growing seasons. The early planting date occurred on October 15, 2018, October 24, 2019, and November 9, 2020. The late planting date occurred on January 31, 2019, February 10, 2020, and January 21, 2021. Cover crops were direct-seeded with a conventional grain drill. Ground preparation occurred before each planting date. Before the early planting date, the whole orchard received light tillage immediately before a glyphosate burndown. Before the late planting date, cannabis curing late planted plots and the non-treated control received an additional glyphosate burndown but no additional soil disturbance. Cover crops were terminated with a flail mower on April 19, 2019, April 27, 2020, and April 22, 2021. Weed emergence was monitored throughout the cover crop growing season using permanent point intercept transects. Each plot had one transect placed diagonally across one orchard alley. Each transect was 10 m long with 10 points along the transect. Plants were identified at each point along the transect, and monitoring took place weekly while cover crops were growing. This experiment did not have different residue management treatments, so summer weeds were not evaluated. Immediately before cover crop termination, we sampled cover crop and weed biomass using the methodology described above, including two 0.25 m2 quadrat subsamples in each plot.Data analysis. Analyses were performed in R 3.0.3 .

For biomass data from both experiments, we used ANOVA and performed multiple comparisons with Fisher’s LSD. ANOVA was performed by specifying a model with lm and entering it into Anova from the car package . The models we used had treatment, replicate, and their interaction as predictors and either weed biomass or cover crop biomass as a response variable. We inspected ANOVA assumptions visually using plot. Subsequently, weed biomass from the intensification experiment was analyzed with one outlier removed and a square root transformed response variable due to leptokurtosis. However, unabridged and non-transformed data are displayed in the figures. Finally, we performed Fischer’s LSD with LSD.test from agricolae using a significance level of P<0.05 . Summer weed emergence data were analyzed in the same manner but using cover crop regrowth and summer weed emergence as response variables. Weekly transect surveys were analyzed with multiple linear regression. We compared the slope of each regression line in to evaluate the relative rates of weed and cover crop emergence after each plant date. Cover crop emergence was represented as the change in ground cover as observed in weekly observations throughout the first ten weeks following the respective planting date of each treatment. There was only one non-treated plot in each repetition, and we evaluated ground cover following both the early and late planting dates in the same non-treated plots. Weed and cover crop emergence were modeled as functions of treatment, weeks after respective planting, and their interaction. These linear models were created using lm. We created additional linear models using other possible combinations of predictor variables and compared these various models using anova. However, we determined the model described above to be the most parsimonious. Parameter estimates for the slope of each line were compared with Tukey’s HSD using lstrends from the emmeans package . All figures were made with ggplot2 .In the intensification experiment, cover crop biomass varied with management treatment . While year was not a significant predictor of cover crop biomass , we detected an interaction between year and treatment . Furthermore, multiple comparison testing led to different conclusions from each year of the intensification experiment. With data pooled across years, the forage and boosted treatments resulted in higher cover crop biomass than multi-species or standard treatments. Within each year, the boosted treatment alone resulted in the highest cover crop biomass in 2020, while the forage treatment did so in 2021. Cover crop treatment and year both predicted weed biomass. The interaction term was also important . In general, the four cover crop programs resulted in less weed biomass compared to the sprayed treatment and similar weed biomass compared to each other. This conclusion was supported in both years of the study, but we observed less weed biomass overall in 2021. Intensified cover crop programs can increase cover crop biomass, but all of the cover crop programs we tested were similarly effective at reducing weed biomass. Less rainfall in 2021 could have contributed to differences between study years, and we attribute some decrease in boosted cover crop biomass to dry conditions after top dress fertilizer application which likely caused a reduction in plant-available nutrients from the applied fertilizer. Some cumulative effect of two years of cover cropping could have also contributed to these results.In the planting date experiment, cover crop biomass varied with cover crop treatment . Year was not significant , but the interaction between treatment and year was . In 2019 and 2020, the early planting treatments resulted in higher cover crop biomass than the late planting treatments. Differences between cover crop treatments were greatest in 2020, and the multi-species mix also resulted in greater cover crop biomass compared to the brassica mix in this year. There were no differences in cover crop biomass between treatments in 2021. Year , treatment , and their interaction all contributed to weed biomass. While we observed a lot of year-to-year variation, the late-planted multi-species treatment was consistently in the lowest statistical group for weed biomass, and the early-planted brassica treatment was consistently in the highest group.

In California, field bindweed survives in both irrigated and dryland environments, and it grows through much of the year in annual and perennial cropping systems as well as non-crop areas like roadsides . This species causes yield loss, reduces water use efficiency, disrupts irrigation infrastructure, impedes crop harvest, and creates multiple flushes of growth each growing season . These factors contribute to ongoing problems in California orchard systems. Field bindweed has a reproductive biology that includes sexual reproduction with large flowers and hard-coated seeds as well as asexual reproduction with an extensive root system, and this biology has helped it thwart many common weed management programs across California orchard systems . Because it is difficult to control, orchard weed managers often use several weed management operations against field bindweed each year, especially repeated application of systemic herbicides like glyphosate in mature orchards . Repeated applications of contact herbicides like glufosinate, paraquat, or PPO inhibitors are also common in young orchards where crop safety is a larger concern when using systemic herbicides. In fact, repeated herbicide applications are common in a variety of cropping systems . Systemic herbicides offer the apparent benefit of translocation to the extensive root system of field bindweed . However, translocation of some herbicides to field bindweed roots can be limited, and repeated herbicide applications have the potential to select for herbicide resistance . Furthermore, vertical farm equipment field bindweed frequently demonstrates capacity for regrowth following systemic herbicide applications, even when factors such as application timing, herbicide mixtures, or spray adjuvants are optimized .

Mechanical management practices that disturb the soil and underground tissues could likewise be more efficacious against field bindweed relative to mechanical practices that only affect above ground tissues . These practices, however, frequently allow regrowth from root cuttings, adventitious roots, or perennial buds . The light environment in young orchards compared to shady, mature orchards is more conducive to field bindweed . However, many young orchard trees are susceptible to injury from the systemic herbicides and intensive mechanical management practices that are commonly used against field bindweed . The compounded challenges of managing field bindweed in young orchards necessitate deeper understanding of how this species persists and can be managed in unique orchard environments. This knowledge could then inform more sustainable integrated pest management strategies with greater efficacy while reducing reliance on glyphosate and other systemic herbicides and increasing the number of management practices that are known to be safe for young trees . Integrated pest management relies, in part, on information about pest life cycles and phenology and how they relate to cropping system context. The development of integrated management programs for field bindweed necessitates greater understanding of this species’ population ecology, particularly within the context of current cropping system practices and limitations . Current management programs react to the presence of field bindweed vegetation, but integrated management programs could better account for the specific ways that this species reproduces and create site-specific management that targets the most susceptible life cycle stages . Recent advances in soil seed bank management highlight the importance of understanding all kinds of plant propagules and how these propagules differentially contribute to weed populations .

Given its relatively complex life history, field bindweed could be a useful study system for understanding perennial weed reproduction and its importance to integrated pest management. This species reproduces with both sexual and asexual propagules, creating a persistent seed bank as well as perennial roots and buds . Perfect, self-incompatible flowers bear hard, dormant seeds that maintain genetic diversity within the soil seed bank . Significant resources are also allocated towards prolific root systems, and individual plants can create underground systems that are several meters in diameter . The morphology and biomass of these root systems are influenced by light, cultivation, and other cropping system cultural factors . However, field bindweed exhibits varying degrees of phenotypic plasticity despite its morphological diversity . Integrated management programs could be strengthened by improved knowledge of how this diversity contributes to field bindweed survival in the face of varied management. Despite the importance of reproductive allocation and diversity, direct quantification of field bindweed reproduction remains challenging due to the inaccessibility of root structures and dehiscence of mature seeds .There is a need for research that addresses these challenges to help us answer questions about field bindweed reproduction in ways that contribute to commercially-relevant management. We are particularly interested in how weed management practices in orchards differentially affect above ground and below ground tissues in order to better understand how each contributes to field bindweed persistence in orchard cropping systems . Additionally, we focus on time to first flowering as a practically important reproductive trait, because it can help determine when to schedule sequential weed management treatments and how species adapt to changing environmental conditions .

California orchards are unique cropping systems that require dedicated development of integrated management programs, especially given the distinctive biology of field bindweed and the specific challenges of weed management in young orchards. Our overall aim is to use information about the reproductive biology of field bindweed to support improved ecological management of this species with practices that are feasible in California orchards. The experiments described in this study evaluated field bindweed flowering and biomass production using field and potted plant experiments. Our approach was to determine how a variety of common and prospective weed management practices in young orchards affect field bindweed reproductive resource allocation. In the field experiment, we tested various chemical and mechanical weed management practices to identify effects on field bindweed flowering timing and aboveground biomass production at timings relevant for commercial orchard production. In the potted plant experiment, we evaluated different mechanical disturbance treatments on field bindweed collected from different source populations to describe responses in flowering timing or root:shoot biomass ratios. Together, these experiments were designed to provide insight into how the distinctive reproductive morphology of field bindweed behaves and responds to orchard weed management programs.A field plot experiment was designed to evaluate how in situ field bindweed alters its reproductive response to various commercial management practices. A potted plant experiment was designed to create a common environment for testing field bindweed from a variety of source environments, as well as to provide opportunity for detailed assessment of root biomass. Together, these complementary experiments allowed us to use a broader array of methods for observing the reproductive response of field bindweed to management. Field experiment. The field experiment was a small plot study arranged in a randomized complete block design with four repetitions in each replicate. Various management programs were applied to 4.6 by 6.1 m rectangular plots in fallow fields with endemic field bindweed infestations. The whole experiment was replicated three times in time, each with a different experimental timing that coincides with different management periods for California orchard growers. Each of the replicates took place in separate fallow fields at the Plant Sciences Field Facility at UC Davis in Davis, CA . These fields consist of Yolo silt loam and had a history of orchards, agronomic crops, and field bindweed infestation before being fallowed. Field bindweed grows nearly year-round in central California, commercial indoor growing systems with a period of senescence in the winter months only; these replicate timings were chosen to mimic some of the primary periods where agricultural weed management practices already occur within that window. The first replicate was performed in the fall of 2020, to coincide with post harvest weed management timing in nut orchards . The second replicate was performed in early summer 2021, coincidental with weed management that targets summer weed emergence in orchards, especially in young orchards where there may be a greater need to manage small weeds and prevent weed establishment given a relative lack of registered herbicide options. Finally, the third replicate was performed in the mid-summer of 2021, at the timing of preharvest weed management in orchards. Management programs tested in this experiment involved sequential management steps, as is often necessary for growers dealing with field bindweed. Each of the treatments received discing and culti-packing as the first management step in order to eliminate emerged bindweed vegetation, as well as to create a uniform soil surface for treatment application. This tillage step occurred on August 19, 2020, March 25, 2021, and May 4, 2021 at each of the three replicates, respectively. The fields were subsequently monitored for bindweed reemergence, and mechanical and chemical treatments were applied to the replicates when stem regrowth reached approximately 10-15 cm in length, approximately when sequential weed management would be applied commercially.

Treatments were applied on September 15, 2020, April 20, 2021, and June 3, 2021. There were seven treatments, including one non-treated control, three herbicide treatments, and three mechanical treatments. The three herbicide treatments were broadcast glyphosate, strip-applied glyphosate, and glufosinate. Both herbicides are widely used for field bindweed management in California, but they have contrasting systemic or contact actions. The broadcast glyphosate was applied at a rate of 2.8 L ha-1 across the entire plot. The strips were applied at a rate of 5.6 L treated ha-1 in two 1.15 m-wide strips in the plot, leaving two 1.15 m-wide non-treated strips in the same plot. The glyphosate strips treatment usedthe same total amount of herbicide as the broadcast treatment, and this treatment was designed to evaluate potential impacts of glyphosate translocation when applied in strips as is common in orchards. The glufosinate treatment used 3.9 L ha-1 of Rely 280 . Each of the herbicide treatments was applied using a CO2 propelled backpack sprayer equipped with a three-nozzle boom and 80015XRVS nozzles and calibrated to apply 187 L ha-1 spray volume, based on 3.2 km hr-1 ground speed and 50.8 cm nozzle spacing. The three mechanical treatments were rototilling, flail mowing, and string trimming. These three treatments were chosen because they affect field bindweed roots and shoots differently, with rototilling representing deeper disturbance compared to string trimming and flail mowing. Each plot was monitored weekly for 10 weeks following treatment application. The first five plants to emerge in each plot were marked with stakes. In the glyphosate strip plots only, these first-to-emerge plants were in the non-treated strips within each plot. Individual plant subsamples were evaluated throughout the experiment. We used flower counts to determine average time to first flowering in each plot. No flowering was observed in the first replicate, likely since it was relatively late in the 2020 growing season, and we did not include that replicate in flower timing analysis. At the end of the 10-week observation period, we collected the above ground portion of the marked plants, dried them in a forced air oven, and weighed the dry biomass. Pot experiment. The potted plant experiment involved propagating field bindweed plants from several source populations into pots and subjecting them to different mechanical disturbance treatments. The plants were propagated vegetatively from annual crop, perennial crop, and non-agricultural home environments. The experiment used a factorial design with three disturbance treatments, including non-treated, clipping, and simulated tillage, and four field bindweed populations. Plants were collected in late 2020 from an almond orchard near Corning, California , an almond orchard in the Wolfskill Experimental Orchard near Winters, California , an annual crop field in Davis, California , and a vacant lot in Davis, California . Several dozen plants cuttings, each including both root and shoot tissue, were collected at each site and transplanted into greenhouse pots. Plant populations were maintained in the greenhouse and re-transplanted twice into new pots over a six-month period. Re-transplanting included shoot trimming to minimize powdery mildew pressure, ensure uniform plant size, and control for any legacy effects from respective environmental conditions at the time of collection. After growing in the greenhouse, we transplanted plants into 12 L pots on outdoor benches. Each pot was filled with greenhouse soil . Plants were uniformly trimmed to have 10 cm of root and 10 cm of shoot length, and only one plant was transplanted per pot. Pots were watered daily with drip irrigation. We replicated the experiment twice, and there were six repetitions of each population-treatment combination in each replicate. Transplanting occurred on April 29, 2021 for the first replicate and on May 26, 2021 for the second replicate.

However, competitive suppression of native seedlings by exotics eclipsed the feedback effect, highlighting the need for weed control in the restoration process.Understanding long-term plant community dynamics has been a key challenge in ecology , made even more challenging by the increased firequency of plant invasion and novel environmental conditions. A number of studies have demonstrated that new climatic extremes, such as severe and prolonged drought, may have long term legacies on plant communities by affecting resource availability and altering dominant species . In systems dominated by long-lived perennial plants, novel disturbances can disrupt generally predictable successional change . Understanding the impacts of novel disturbances on vegetation dynamics is even more challenging in annual-dominated plant communities, where composition can reset each year . Annual systems have high turnover in species composition due to multiple biotic and abiotic drivers with high inter-annual variability . Drivers such as the timing and amount of precipitation and temperature , fungal pathogens , and herbivores act on seed production, seed survival and germination, seedlings, and mature plants . In addition, vegetation can be influenced by the quantity of litter from the previous season . Thus, species composition in annual systems is difficult to predict and manage . These controlling variables are often strongly impacted by previous species composition, and thus predicting community changes in annual systems may benefit from considering priority effects. Priority effects are when the timing and order of species arrival during assembly determines which species can later establish in the community, leading to alternative stable states, vertical grow rack alternative transient states, or compositional cycles . Priority effects have been observed to affect composition and diversity in multiple systems .

In perennial systems, a long-lived species can physically hold space against competitors over time, occupying this niche to the exclusion of later arrivals . In fact, planting of perennial grasses is firequently used as a way to suppress annual weeds . Among annual plants, priority effects may occur through mechanisms such as litter build up, seed production and faster germination, and plant-soil feedbacks related to changes in pathogen and symbiont communities . Priority effects are stronger among species with high overlap in resource use, resulting in greater niche co-option and exclusion of later arrivals of similar function and phenology . Assessing vegetation community dynamics in California’s annual grasslands may thus benefit from defining functional groups in terms of phenology , which dictates when California grasses compete for the limited resource of soil moisture. California’s grasslands are dominated by early-season exotic annual species that are now so entrenched in the landscape they are considered naturalized; they generally outcompete native grass and forb seedlings, as they germinate and grow faster, depleting shallow soil moisture and creating light limitation . Several priority experiments have shown that native perennial grasses in this system benefit from a two-week head start over naturalized exotic species . A newer set of invading exotic annual species, including the grasses Elymus caputmedusae and Aegilops triuncialis, are classified as noxious weeds and are of particular concern because they produce thick layers of thatch that is slow to decompose and prevents germination of other species . These noxious weeds germinate at the same time as the early-season exotics, but their above-ground growth primarily occurs after rains have ended and the earlier season grasses have senesced .

In drier years, early-season naturalized exotic growth can limit noxious weeds by utilizing all soil moisture, but cannot prevent noxious weed growth if late-season moisture is present . Native perennial grasses, which are active during the late-season, may compete with noxious weeds for the dwindling late-season soil moisture. However, how long priority effects last on all three functional groups is unknown. We established field plots consisting of early-phenology naturalized exotic annual species, late-phenology annual noxious weeds, and native species commonly used in restoration mixes . Each group was seeded alone or simultaneously in mixtures with other groups to compare to assess the importance of being seeded without initial competition. All plots were then allowed to be naturally colonized by non-seeded species in the experiment, but with a subset of plots receiving a one-year weeding treatment to provide additional priority. We assessed composition over a twelve-year period that included an extreme multi-year drought followed by a historic wet year, and then another severe drought year. Such extreme weather events in other systems have been shown to disrupt plant community dynamics, reducing cover of resident species and increasing invasion . This experiment is particularly valuable because the majority of priority effect studies take place in the greenhouse and for less than a year in duration, and it is widely recognized that more field and long-term experiments are needed to determine the strength of priority effects in varying conditions . Of the few long-term studies, the current literature has shown both that priority effects either persist or disappear depending on the system and the species involved .

We hypothesized that all three functional groups of species, when seeded alone, will: have greater cover than when grown in competition, and limit recruitment of another naturally colonizing functional group. We also predict that the multi-year drought will weaken priority effects during and beyond the drought in favor of the early-season naturalized exotics, as they have will have first access to soil moisture.Plots were located in UC Davis Campbell Tract Experimental Site in Davis, CA . Prior to plot establishment, the land was used for agricultural purposes and then lay fallow for twenty-two years. The site was primarily on Reiff series soil, with a sandier lens of Brentwood soil series on 25% of the site . Under a Mediterranean climate, the site experienced hot, dry summers and wet, cold winters that correspond with the growing season of cool season grasses.Rainfall has high inter- and intra-annual variability, with mean annual precipitation of 445 mm . During the experiment, California experienced a historic drought between the 2012-2014 water years , followed by 2 years with precipitation levels that were substantially closer to the 30-year precipitation average, and then followed by one of the wettest years on record in 2017, then a dry year, followed by another historic wet year . Prior to initial seeding at the start of the experiment in fall 2007, the seed bank and resident vegetation were minimized by disking the soil, irrigating to germinate the seedbank, and spraying germinating seedlings with herbicide . Irrigation and herbicide treatments were performed twice before planting. Plots were seeded with one of the following vegetation treatments in a randomized block design : native perennial grasses and annual forbs, exotic annual noxious grasses, naturalized exotic annual grasses and forbs, and all possible combinations of these three groups , at a rate of 139 g seed/plot. Plots were 1.5 x 1.5 m with a 1m buffer between them . For each single functional group mix, equal proportion of seeds of each species were added. For functional group mixes, an equal proportion of each functional group seed was added, commercial vertical hydroponic systems with equal proportion of individual species within each functional group. Given that the common design of seeding equivalent weights across all treatments design results in the multi-group treatments having less seed per species group , short-term conclusions may be influenced by seed limitation as well as priority effects. However, the effects of seed limitation should be short-lived given the annual species in this experiment are highly prolific and able to rebound from extremely low to high cover from one year to the next, as well as naturally self-thinning . While natives are seed limited, low density can still result in high cover , and so changes in native cover can be attributed to changes in individual size as well as population response. Percent cover of each species present was assessed visually with a modified Daubenmire bin method of the inner 1m x 1m core.

Composition was measured twice each season annually from 2008 to 2019 to capture peak flowering species with different phenologies . The highest cover value across the two sample points each year for each species was used for analysis.There were multiple levels of priority in this experimental design. For a given functional group, the seeding treatment in which they were seeded alone in monotype is the highest level, as they accessed the space first with no competition from other functional groups. The seeding treatments in which that functional group were seeded simultaneously with another group are a secondary level of priority, as they shared first access to the space. The remaining seeding treatments are those in which the functional group in question was not seeded, thus they had no priority at all. If present later in the experiment, they recruited into the space after initial colonization by the planted functional group, and thus faced high competition for resources from established residents or high propagule pressure . Given the spatial distribution of the experimental plots, every plot was close to a seed source of another functional group from neighboring plots and the unplanted walkways between plots and blocks became quickly colonized, indicating that the species were dispersing throughout the site quickly. Superimposed on the above listed priority levels was a weeding treatment to determine the difference between one vs two years of priority. Natural colonization was either allowe dimmediately or postponed an extra year due to hand-weeding after the first year to maintain the initial species compositions of the seeding treatments . Since then, all plots were occasionally weeded for agricultural weeds that were not part of the study but species that self-recruited and are typical species in California grasslands were not weeded.Priority effects can be the outcome of the seeded species having both higher propagule pressure and biotically resisting the recruiting species and we are interested in the long-term trajectory of the initially planted communities and the extent to which they are naturally colonized by functional groups that have been planted nearby. To test our hypothesis, we performed two comparisons. First, for each year, we compared cover of the recruiting functional group in its monotypic seeding treatment, which presumably represents the best conditions for performance as it has the highest level of priority, to the three seeding treatments in which the functional group was not planted . If the recruiting functional group cover is higher in its own monotypic treatment, then it would suggest that the functional groups originally planted are providing some level of resistance. Second, we compared the recruiting functional group’s cover as it changed over time within each seeding treatment it was not planted into, such as whether recruiting species cover stays consistently low or increases with time. This took into consideration potential annual population fluctuations due to environmental conditions and provides detail on resistance not captured by the first comparison. Both comparisons are necessary to determine whether low presence of a recruiting functional group is due to bioticresistance or poor environmental conditions. We also compared cover between the one- and twoyear priority weeding treatments. Both comparisons were assessed with the same linear mixed effect model but differed in how we performed the post-hoc analysis. First, we fit linear mixed effect models with percent cover of each recruiting functional group as the dependent variable, specifying seeding treatment, priority length , year , and their 3-way interaction as fixed effects and plot and block as random effects. Significance was tested with analysis of variance . Both comparisons were assessed with post-hoc multiple comparison tests using estimated marginal means on the interaction of seeding treatment and year when significant in the ANOVA but differed in the order of terms used. The first compared cover across the different seeding treatments within each year while the second compared over across the 12 years within each seeding treatment. All statistical analyses were conducted in R 4.0.3 . Linear mixed effect model fitting was performed using “lme4” and “lmerTest” . Multiple comparison tests were conducted using “emmeans” .Our study assessed the role of priority effects in determining long-term community composition in a system that experienced high annual variability in weather, including a multiyear drought event followed by one of the wettest years on record, and then another historic dry year, and another historic wet year .

Wild fungus gardens contained similar peptaibols, indicating their ecological relevance, consistent with peptaibol-producing Trichoderma being an opportunistic pathogen of T. septentrionalis fungus gardens . In our laboratory experiments, two of the Trichoderma fractions with the highest abundance of peptaibols, fractions D and E, induced some of the strongest T. septentrionalis weeding responses , and purified peptaibols also induced a weeding response similar to that of theTrichoderma extract . Given that the Trichoderma fractions and purified peptaibols were derived from different strains and represent a diversity in peptaibol composition, the similarities in the observed ant behavioral activity are thus likely to be unrelated to specific peptaibols but rather generally attributable to the peptaibol class of metabolites. Furthermore, the presence of peptaibols in environmental fungus gardens lends credence to their ecological relevance and together with the experimental data parallels the logic of Koch’s postulates, suggesting that peptaibols are likely produced during Trichoderma infections of ant fungus gardens and induce ant defensive behaviors in response. Peptaibols are produced by fungi in the order Hypocreales, especially by members of the mycoparasitic family Hypocreaceae, which contains both Trichoderma and Escovopsis, a specialized ant fungus garden pathogen . Peptaibols have been hypothesized to be important for mycoparasitism , which we show here can include infections of ant fungus gardens. Interestingly, the genes needed to synthesize peptaibols are encoded within the genome of Escovopsis , vertical farming systems for sale and Escovopsis has been shown to induce a strong ant weeding response in tropical leafcutting fungus-growing ants , similar to the T. septentrionalis weeding behaviors that we observed in response to Trichoderma and its peptaibols.

We therefore hypothesize that peptaibolinduced weeding behaviors are conserved among diverse fungus-growing ants and may reflect an ancient means of pathogen detection and defense. Future work should test if ant weeding intensity is directly correlated with pathogen load, virulence, peptaibol production, or other contributing environmental factors, and also compare the behavioral responses of diverse fungus-growing ant species to diverse fungal pathogens with varying levels of virulence and specialization toward ant fungus gardens, e.g., as in ref. 58. This study demonstrates how T. septentrionalis ants protect their cultivar mutualist from opportunistic Trichoderma pathogens by sensing and responding to peptaibols as specific molecular cues that induce an ant weeding response. These cues included two previously undescribed bioactive peptaibol metabolites that we identified in this study. Future research will investigate whether ants directly sense peptaibols or indirectly respond to an intermediate signal produced by the cultivar in response to peptaibols, in addition to characterizing other potential signaling molecules that are unlikely to be present in our Trichoderma extracts . In contrast to the canonical logic of host immune responses, in which hosts directly respond to infections, T. septentrionalis responses to peptaibol signaling molecules comprise an extended defense response whereby T. septentrionalis ants respond to infections of their cultivar mutualist. Such extended defense responses may be a widespread but poorly recognized mechanism that increases host health indirectly by preventing harm to their beneficial symbionts.Vineyard-fl oor management strategies, such as weed control and cover-cropping, have wide-ranging impacts both inside the vineyard, in terms of crop management and productivity, and outside the vineyard, in terms of runoff and sediment movement into streams and rivers.

The increasing importance of water-quality issues statewide, including in Monterey County where the Salinas River drains into the Monterey Bay National Marine Sanctuary, highlights the need for management strategies that limit environmental impacts. Growers are interested in alternative weed-control practices and cover crops, but they need information in order to balance benefits with the economic realities of wine-grape production. We established a 5-year experiment in a commercial vineyard in Monterey County with the intent of identifying effective practices that can be integrated into the cropping system without negatively affecting winegrape production. The vineyard floor consists of two zones: the rows, a 2- to 4-foot-wide swath underneath the vines, which are managed primarily to control weeds by herbicide applications or cultural practices ; and the middles, interspersed between the rows, which are vegetated by cover crops or resident vegetation in the dormant season, and are tilled or left untilled in spring. Growers manage weeds in rows to reduce competition for water, nutrients and light , and to prevent tall-statured weeds such as horse weed from growing or climbing into the canopy, where they interfere with harvest. Growers transitioning to more sustainable production systems need information on how management practices affect the physical properties, health, organic matter and water retention of soil. We monitored soil microbial activity for arbuscular mycorrhizal fungi and soil microbial biomass, since weed control and cover-cropping can affect populations of beneficial soil microbes in annual crops . Dormant-season cover crops in the middles minimize runoff from winter rains . Many California growers are also willing to plant cover crops because they protect soil from nutrient and sediment loss in winter storms , suppress weeds , harbor beneficial arthropods , enhance vine mineral nutrition and increase soil organic matter .

Competition between vines and cover crops for soil moisture in spring, when both are actively growing, can lead to severe water stress and reduce grape production . However, wine-grape production is distinct from other cropping systems because water stress may be imposed to enhance wine composition ; this practice has been studied mostly in high-rainfall regions of California. The vineyard production region of Monterey County, in contrast, has low rainfall , and growers must weigh the benefits of cover crops with the possible need to replace their water use with irrigation. In addition, growers must decide on the type of vegetation to utilize in the middles. Resident vegetation is cheap and generally easy to manage. Cover crops can provide specific benefits such as nitrogen fixation or high biomass production and vigorous roots . There are many choices for cover crops in vineyard systems, ranging from perennial and annual grasses, to legumes . Each species has strengths and weaknesses, as well as associated seed and management costs.Row weed control treatments were: cultivation, post-emergence weed control only and pre-emergence herbicide , followed by post-emergence herbicide applications . Cultivations and herbicide applications were timed according to grower practices and label rates. Cultivations were carried out every 4 to 6 weeks during the growing season using a Radius Weeder cultivator . The cultivator used a metal knife that ran 2 to 6 inches below the soil surface cutting weeds off in the vine row; it had a sensor that caused it to swing around vines. Pre-emergence herbicides were applied in winter with a standard weed sprayer, and postemergence herbicides were applied in spring through fall as needed with a Patchen Weedseeker light-activated sprayer . An early and late-maturing cereal were chosen for the cover-crop treatments; legumes were not considered due to aggravated gopher and weed problems. Cover-crop treatments in the middles were: no cover crop , earlier maturing ‘Merced’ rye and later maturing ‘Trios 102’ triticale . Cover crops were planted with a vineyard seed drill in a 32-inch-wide strip in the middle of 8-foot-wide rows just before the start of the rainy season in November 2000 to 2004 . They were mowed in spring to protect vines from frost, and both cover-crop species senesced by summer. Prior to planting cover crops each November, vertical farming equipment row middles were disked to incorporate the previous year’s cover crop and stubble and prepare a seedbed. Weed control and cover-crop treatments were arranged in a 3 x 3 splitblock design with three replicate blocks covering a total of 23 vineyard rows . Each block contained six vine rows and six adjacent middles. Weed control treatments were applied along the entire length of each vine row ; cover-crop treatments were established along one-third of each middle and were continuous across the main plot treatments in each block.

Each replicate main plot-by-subplot treatment combination included 100 vines.Soil compaction. Soil compaction was measured in the vine row in November or December 2003, 2004 and 2005 with a Field Scout Soil SC-900 compaction meter . Ten sites in each plot were sampled to a depth of 15 inches. Soil moisture. Soil water storage was evaluated from volumetric soil moisture measurements taken in-row and adjacent middles to a depth of 3.5 feet at 1-foot intervals using a neutron probe. The neutron probe readings were calibrated with volumetric moisture measured from undisturbed soil cores collected at the site. Rainfall and runoff. A tipping bucket rain gauge with an 8-inch-diameter collector was used to monitor daily and cumulative rainfall at the field site. Runoff was collected at the lower end of the plots into sumps measuring 16 inches in diameter by 5 feet deep. Each sump was equipped with a device constructed from a marine bilge pump, a float switch and flow meter, to automatically record the runoff volume from the plots during storm events. During the second and third years the sampling devices were modified to collect water samples for sediment and nutrient analysis. Vine mineral nutrition. One-hundred whole leaves opposite a fruit cluster were collected from each plot at flowering in May 2003, 2004 and 2005. Petioles were separated from leaf blades, and tissue was immediately dried at 140°F for 48 hours and then sent to the ANR Analytical Laboratory for nutrient analyses. Petiole and leaf-blade tissue samples were analyzed for nitrate , ammonium , nitrogen , phosphorus , potassium , sulfur , calcium , magnesium , boron , zinc , manganese , iron and copper . Soil mineral nutrition. Composited samples from 10 soil cores taken to a depth of 1 foot were collected from the vine rows and middles at flowering as described above. Samples were air dried and sent to the ANR Analytical Laboratory for analyses. Soil samples were analyzed for pH, organic matter, cation exchange capacity , nitrate, Olsen-phosphorus, potassium, calcium, magnesium, sodium , chloride , boron and zinc. Soil microbial biomass. Due to the limited capacity of the laboratory, microbial biomass assays were conducted on selected treatments. Ten soil cores were collected to a depth of 1 foot and then composite samples were made from each replicate of the pre-emergence and cultivation weed-control treatments and the adjacent middles of the ‘Merced’ rye and bare treatments. Samples were collected about four times each year from November 2001 to November 2005 for a total of 14 sets of samples. Soil samples were immediately placed on ice and taken to the laboratory for soil microbial biomass carbon analysis according Vance et al. . Mycorrhizae. Roots were collected, stained and examined as previously reported on April 16, 2003, May 3, 2004, and June 2, 2005. Grape yield, fruit quality and vine growth. Fruit weight and cluster number were determined by individually harvesting 20 vines per subplot. Prior to harvest a 200-berry sample was collected from each subplot for berry weight and fruit composition. Berries were macerated in a blender and the filtered juice analyzed for soluble solids as Brix using a hand-held, temperature compensating refractometer. Juice pH was measured by pH meter and titratable acidity by titration with a 0.133 normal sodium hydroxide to an 8.20 pH endpoint. At dormancy, shoot number and pruning weights were measured from the same 20 vines. Statistical analysis. Analyses of variance were used to test the effects of cover crop, weed control and year on the vine, soil and microbial parameters, according to a split-block ANOVA model in SAS . Cover crop, weed control, year and their interactions were treated as fixed effects. The main and interactive effects of block were treated as random effects. Year was treated as a repeated measure. When necessary, data were log-transformed to meet the assumption of normality for ANOVA, although untransformed or reversetransformed means are presented. Changes in soil moisture among treatments during the winter and the irrigation seasons were determined from significant treatment-date interactions.We conducted evaluations with a penetrometer each fall to determine the impact of weed-control treatments on soil compaction. Soil compaction was not significantly different at any depth in 2003 . However, in 2004 and 2005 soil compaction began to increase in the cultivation treatment compared to the other two weed-control treatments. In 2004, soil compaction at the 4- to 7-inch depth was significantly greater in the cultivation treatment compared to the standard treatment , but not more so than in the post-emergence treatment . In 2005, the cultivation treatment had significantly greater soil compaction at the 4- to 7-inch depth than both the post emergence and standard weed-control treatments . At the 8- to 11-inch depth, soil compaction was significantly greater than the standard treatment , but not greater than in the post-emergence treatment .

There are several methods to make pollen grains functionally deficient and thereby reduce seed set . The most commonly used is ionizing irradiation with X-rays or gamma rays due to their ease of use, effective penetration, consistent results, and minimal disposal issues . Irradiated pollen grains can be physiologically alive, depending on the irradiation dosage, but are infertile. Irradiated grains can germinate on the stigma and even produce pollen tubes but cannot fertilize egg cells to produce embryos . Further, when sterile pollen grains are deposited on a stigma through artificial pollination, they can interfere with fertile pollen in the process of fertilization and disrupt seed production, as has been shown in apple , pear , citrus , cacao , and melon . The use of sterile pollen to reduce weed seed production is similar to the insect sterile technique , an environmentally-friendly and biologically-based method for controlling insect pest. This technique involves sterilizing male insects by irradiation and subsequently releasing the sterile males to mate with wild females , resulting in infertile eggs and reduced insect pest population sizes. Pollinating the female plants of Palmer amaranth with sterile pollen resulted in 40% reduction in the number of newly formed seeds . However, the sterile pollen technique has been rarely used as a weed control technique but could potentially be effective on dioecious weedy species because female and male flowers are in separate plants and pollen grains can be collected from male plants, sterilized and then released on female plants. The summer annual dioecious weed A. palmeri is one of the most devastating weeds in the US. It was ranked as the worst weed in US corn fields in a survey by the Weed Science Society of America . Furthermore, industrial drying rack it has evolved resistance to nine herbicide classes used and is able to produce up to one million seeds per plant .

This weed is a particularly suitable candidate for exploration of the sterilepollen technique for weed control. Being a dioecious species with separate male and female plants, it relies on cross-pollination for successful seed production. This makes it feasible to collect pollen grains from male plants, sterilize them, and subsequently release them onto female plants. The primary goal of this research was to examine the effectiveness of sterile pollen technique, SPT, as a means of disrupting seed production in A. palmeri. To this end, it was necessary to determine the optimal irradiation dose for pollen sterilization as excessively high doses may kill the pollen entirely, thereby eliminating their preventative effects on fertile pollen, while low doses may allow the treated pollen to maintain its fertility. Accordingly, a broad range of irradiance doses was tested in combination with an extensive array of artificial pollination treatments to fully explore the potential effects of the SPT on seed production in A. palmeri. Our hypothesis is that pollinating with sterile pollen, irradiated at an optimal dose, could reduce seed production in this weed. Furthermore, we speculated that the maximum reduction in seed output could be achieved when pollination with sterile pollen precedes open pollination.Seeds of A. palmeri collected from Kansas were planted in May 2020 into 3-L pots filled with UC Davis potting medium containing in a greenhouse set at a 24/32 ℃ night/day temperature regime and extended photoperiod . Fertilizers were applied as 80 ml of a general-purpose fertilizer solution weekly at 350 ppm N starting from the 2-true leaf plant stage with drip irrigation applied at 65 mL/min for two minutes twice per day. Once plants reached the flowering stage, 50 male and 50 female plants were isolated and grown in separate greenhouses .

Pollen collections were made from male plants by gentle tapping or shaking of the inflorescence. Pollen grains from all male plants were pooled and released onto aluminum foil held beneath the inflorescence. The collected pollen was then sieved through 250-mm mesh to remove large floral materials. Pollen was placed in Petri dishes covered with parafilm and then irradiated immediately with gamma rays from Cesium-137 at six dosages of 0 , 100, 200, 300, 400 and 500 Gray  at the UC Davis Center for Health & the Environment . Irradiated and untreated pollen were immediately used for pollen viability tests and hand -pollination experiments as described below.Pollen viability was assessed immediately after irradiation by using a test solution consisting of a 1% concentration of the substrate 2,5-diphenyl tetrazolium bromide in 5% sucrose. The MTT assay measures cellular metabolic activity as an indicator of cell viability and cytotoxicity . In this assay, viable pollen appears dark violet and non-viable pollen did not stain at all . Viability of 100 pollen grains for each dose at each irradiation dose was assessed by analyzing the brightness of the resulting tetrazolium stain using a digital camera and ImageJ software Version 1.46r . Grey values were used to indicate the brightness of a pixel. Because the range for grey values is 0- 255, grey value percentages were calculated by dividing the recorded grey values by 255 and multiplying by 100. Higher grey value percentages indicated lower pollen viability. Theeffects of irradiation dosage on grey value percentages were analyzed using ANOVA with Dunnett’s test.In the 2020 experiment, six lateral inflorescences of similar size from each female plant were selected to receive the following treatments: hand pollination with 1) non-irradiated pollen only, 2) irradiated pollen only, 3) non-irradiated pollen followed by irradiated pollen, 4) irradiated pollen followed by non-irradiated pollen, 5) no pollination, or 6) open pollination .

Each inflorescence was meticulously dusted with 1 ml of pollen, ensuring even and gentle distribution using a paintbrush. Thereafter, the inflorescence was immediately enclosed in a paper bag with the exception of inflorescences receiving the open pollination treatment. Hand pollination was conducted through a one-time application. About 6 weeks after pollination, inflorescences were harvested. Flower and seed numbers were measured on the above mentioned six inflorescences for each of five plants at each irradiation dosage. For each replicate, six 1-cm sections of plant branches were dissected and measured for flower and seed numbers. Two categories of seeds were identified and recorded: abnormal seeds and normal full seeds. Seed set was calculated by using the number of viable and full seeds divided by the number of flowers and expressed as percentage. To more closely simulate field conditions, this experiment was repeated in 2021 with three additional treatments: 1) hand pollination with irradiated pollen followed by open pollination , 2) open pollination for two weeks followed by hand pollination with irradiated pollen , and 3) open pollination for two weeks followed by hand pollination with irradiated pollen with no bagging, i.e., open pollination. These treatments began simultaneously when nine lateral inflorescences of similar size from five female plants reached full anthesis. As with 2020 experiment, the hand pollination was performed as a single, one-time application. Data from each year of study was analyzed separately because hand -pollination treatments differed slightly across the two years of experiment. Prior to ANOVA, in order to reduce heteroscedasticity of the residuals, seed set values were transformed using a square root transformation. Two factors, irradiation dose and pollination treatment, were firstly combined into a single factor and a one-way ANOVA was performed on seed set measurements by using aov functions in R . To better explore the interaction between the two factors, drying rack for cannabis the non-crossed treatments were removed to obtain a full-factorial design for a two-way ANOVA. The two-way factorial ANOVA was conducted using lm function followed by slicing each level of irradiation doses, with SLICE function in sasLM package , to perform the F-test for the effect of hand-pollination on seed set at each level of irradiation dose. Lastly, seed set data was back-transformed using the re_grid function in the emmeans package and confidence intervals were constructed using confint function in R .Pollen irradiated at the lowest dose exhibited the lowest grey value percentage while pollen irradiated at 500 Gy had the highest grey value .

The mean grey value of pollen irradiated at 500 Gy was significantly different from the other doses, which indicates this highest irradiation dose reduced pollen viability to a greater degree than the other doses . Under this high irradiation dose, pollen will likely be unable to produce a pollen tube and disrupt the process of double fertilization since it has lost its viability as determined by MTT staining.The viability of pollen is affected by factors such as genotype, pollen maturity, growth media composition , and environmental variables such as air temperature and humidity . Gamma ray irradiation can decrease water content in pollen, reducing the ability to transfer carbohydrate reserves, leading to changes in the cytoplasmic water, abnormal meiosis, irregular gamete formation, and ultimately decreased viability, which has been supported in studies on apples , pumpkins and winter squash , and citrus . The effect of radiation dose on pollen viability is species dependent. In some species irradiation effect is limited. For example, melon pollen can tolerate gammairradiation doses up to 3,600 Gy whereas in winter squash a 300 Gy dose reduced pollen viability by almost 80% . We found significantly reduced viability of A. palmeri pollen irradiated at 500 Gy compared with non-irradiated pollen, which indicates that seed production in this weed is sensitive to ionizing irradiation. However, our goal in the practice of irradiation is not the complete loss of viability. For effective implementation of SPT, it is essential to have semi-functional pollen that can outcompete and displace wild pollen while remaining incapable of fertilizing the ovule. Understanding the sexual reproduction process is important to gain insight about how to increase the competitiveness of irradiated pollen. When the pollen tube enters the female reproductive tissue, intensive communication occurs between the pollen tube and one synergid cell. After the contact of pollen tube and synergid cell, the receptive synergid degenerates . Following release of the two sperm cells from the pollen tube, they interact and fuse with the egg cell nucleus and the central cell nuclei, forming the major seed components embryo and endosperm, respectively. Any of the steps involved in double fertilization or a subsequent event could trigger the block of attraction of multiple pollen tubes to a single ovule . If fusion fails, one synergid can persist and continue to attract multiple pollen tubes until fertile sperm are delivered or the synergid senesces. The recovery of fertilization is limited to the second pollen tube, indicating that there is no third chance for fertilization in two synergid celled plants. The optimal irradiation dosage to sterilize pollen should maintain the function of the vegetative cell but induce failure in cell fusion. If the irradiated pollen can disrupt fertilization twice, there is no third chance for this ovule to produce a seed , thereby reducing overall seed production.Both in 2020 and 2021, the combined effect of irradiation dose and application treatment had a significantly different effect on seed set . Additionally, the effect of different irradiation doses, application treatments, and their interaction on seed set was significant in both years . Female plants that received no pollination did not produce seed in either year so this treatment will not be discussed further in the results. However, in contrast to this observation, a study has proposed apomixis as a potential mechanism for seed production in isolated female plants . In both years, regardless of the irradiation dose, all pollination treatments involving irradiated pollen consistently resulted in lower seed sets compared to open pollination . The mean seed set obtained from pollination treatments involving irradiated pollen never exceeded 35% and decreased to nearly 0% when using only irradiated pollen at doses of 300, 400, and 500 Gy . Seed set decreased with increasing irradiation dose up to 300 Gy in all pollination treatments with irradiated pollen. However, there was an increase in seed set beyond the 300 Gy dose when pollination with irradiated pollen followed by hand pollination with non-irradiated pollen. This suggests that pollen irradiated at 100 Gy and 200 Gy maintained some ability to fertilize egg cells and produce seeds, while pollen irradiated at the higher doses of 300 Gy to 500 Gy were functionally deficient and unable to complete sexual reproduction. The 300 Gy dose seems to be the optimal dose for disrupting seed production inA. palmeri as it produced the lowest seed set when interfering with non-irradiated pollen . Irradiation of pollen has also decreased seed production and seed set in other species.