Unraveling the reasons for failure to control Amaranthus albus: insights into herbicide application at different growth stages, temperature effect, and herbicide resistance on a regional scale.

BACKGROUND: This study investigates factors contributing Amaranthus albus control failure in processing tomato fields in northern Israel. The study region is characterized by a significant climate gradient from east to west, providing the opportunity to investigate the effect of critical elements of the agricultural environment, e.g., temperature. Eight populations were collected from commercial fields in this region. Post-emergence herbicide efficacy of metribuzin, a photosystem II inhibitor, and rimsulfuron, an acetolactate synthase (ALS) inhibitor, was assessed through dose-response analyses at various growth stages. Temperature effects on control efficacy and resistance mechanisms were also explored. RESULTS: Standard metribuzin dose (X) was ineffective on A. albus plants with more than six true-leaves, whereas 2X dose proved effective. Rimsulfuron at 16X dose was ineffective on plants with more than four true-leaves. We report here the first case of target site resistance to ALS inhibitors in A. albus, due to point mutation in the ALS gene (Pro197 to Leu). Furthermore, our findings suggest potential involvement of CYT P450 enzymes in enhanced metabolizing of rimsulfuron. An overall decrease in dry weight was observed in response to both herbicides at 16/22 °C (P < 0.0001). Rimsulfuron was effective against only one population when applied at 28/34 °C. A possible fitness cost associated with target site-resistant biotypes was observed under low temperature conditions, leading to effective control. CONCLUSION: This regional-scale study highlights the challenges faced by growers, emphasizes the need for adapting management practices to the local climatic conditions and lays the groundwork for implementing location-specific weed management strategies in commercial fields. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The Invasive Weed Trianthema portulacastrum in Israel.

Trianthema portulacastrum L. (Aizoaceae), commonly known as desert horse purslane or black pigweed, is a C4 dicot succulent annual herb that is widespread in Southeast Asia, tropical America, Africa, and Australia. In Israel, it is an invasive weed of increasing importance in agricultural fields. The aim of this study was to investigate the biology of this invasive weed and its spread in the Hula Valley of Israel. Initial studies included the investigation of the T. portulacastrum specimens held at the Israel National Herbarium. On-site surveillance for the identification of weed infestation locations was conducted in the Hula Valley throughout 2019-2022, and an infestation map was assembled. In a study of the plant biology, greenhouse pot experiments revealed that T. portulacastrum seeds emerge best from the upper soil levels, and as seed depth increases, the emergence rate decreases, so that at 6 cm soil depth, there was no emergence. In controlled-environment growth chamber studies, there were no significant differences in germination with or without light. A maximum germination of 81% was observed for a 12 h night/day of 25/35 °C regime. Germination rates decreased with the decrease in temperature. A seed germination thermal time model that was developed for estimating the minimum temperature required for germination (Tbase) computed this temperature to be 10 °C. This study revealed the biology, in particular seed germination and emergence requirements, of the invasive weed T. portulacastrum that has spread in the Hula Valley in Israel and beyond. Future research will focus on an examination of control measures to combat this invasive weed.

Ambrosia grayi as a new alien causal species in Israel: plant biology and chemical management.

BACKGROUND: Ambrosia grayi is a perennial weed native to northern Mexico, which can also be found in the Great Plains of the US. Outside the Americas, A. grayi has only been documented in Israel, where it is currently categorized as a casual species at advanced eradication stages. Here, we studied the plant biology and chemical weed management options of A. grayi. RESULTS: Only large achenes of A. grayi (~5% of all achenes) contain seeds; moreover, the viability of seeds extracted from large achenes was ~25%. Examination of plant anatomy revealed that underground vegetative segments show an anatomical structure of stems (rhizomes) with anomalous secondary growth. The optimal (night/day) temperature for the emergence of A. grayi rhizomes was 20/30 °C, and the emergence rate increased under elevated temperatures. Emergence may occur at different soil moisture content (25-60%); rhizomes were able to emerge even after 1 month of drought conditions (20%, 25% and 30%). Herbicide combinations, such as fluroxypyr + glufosinate, fluroxypyr + glyphosate, and glyphosate + saflufenacil + surfactant, were tested under quarantine conditions and showed high efficacy for the control of A. grayi. However, the efficiency of these treatments was highly correlated with plant growth stage. CONCLUSION: In Israel, the spread of A. grayi occurs mainly via rhizomes that can emerge under a wide range of temperatures and soil moisture conditions. Data regarding herbicide efficacy will aid in improving the eradication efforts taken by Israel's Plant Protection and Inspection Services. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Variation in seed properties and germination capabilities among populations of the invasive weed Parthenium hysterophorus L. (Asteraceae).

Introduction: Parthenium hysterophorus (Asteraceae) is an invasive weed species that has invaded over 50 countries worldwide. It was first detected in 1980 at Tirat-Zvi, in eastern-northern Israel. In recent years, there has been an increasing concern over the spread of this weed in agricultural and non-agricultural habitats across the country. However, very little is known about the biology of P. hysterophorus and its variation among populations. Methods: Seeds collected from five locations across Israel were germinated and plants were grown in pollen-proof cages under uniform conditions to produce the progeny populations. Spatial parameters, weight and germination under different environmental conditions were recorded for field and progeny populations. Results: Seeds originating from field populations were significantly smaller and lighter than seeds of the progeny populations. Germination occurred in the range of 10°C to 30°C (To ranges from 19°C to 22.3°C, Tb ranged from 9°C to 15°C, Tc ranged from 24 ℃ to 30.5°C), depending on generation and population. A water potential-based model was developed to estimate germination under different soil water content using specific parameters (b - slope, d - upper limit, e - infliction point). The model suggests a correlation between germination and water potential. Indeed, reduced germination was recorded for the lower water potentials especially for the progeny populations. Spatial parameters, weight and germination under different environmental conditions were recorded for field and progeny populations. Discussion: We identified differences in seed size and weight, germination under different temperatures, and osmotic potential among P. hysterophorus Israeli populations. Differences across generations may arise due to the transgenerational effects. Our results, may shed light on the germination abilities of P. hysterophorus populations and provide vital insight into understanding the invasive capabilities of this highly noxious weed.

Comparative study of bacterial community dynamics in different soils following application of the herbicide atrazine.

Microbial communities in cultivated soils control the fate of pollutants associated with agricultural practice. The present study was designed to explore the response of bacterial communities to the application of the widely-used herbicide atrazine in three different crop fields that differ significantly in their physicochemical structure and nutritional content: the nutrient-rich (with relatively high carbon and nitrogen content) Newe Yaar (NY) and Ha-Ogen (HO) soils and the nutrient-poor, sandy Sde-Eliyahu (SE) soil. The 16 S rRNA gene amplicon sequencing revealed the nutrient poor HO soil differs in its response to atrazine in comparison to the two nutrient-rich soils both in the shortest persistence of atrazine and its effect on community structure and composition. Potential reported bacterial degraders of atrazine such as Pseudomonas, Clostridium and Bacillus were more abundant in contaminated sandy/poor soils (HO) whereas bacteria known for nitrogen cycling such as Azospirillum, Sinorhizobium, Nitrospira and Azohydromonas were significantly more abundant in the nutrient rich contaminated SE soils. No significant increase of potential indigenous degrader Arthrobacter was detected in SE and NY soils whereas a significant increase was recorded with HO soils. An overall shift in bacterial community composition following atrazine application was observed only in the nutrient poor soil. Understanding atrazine persistence and microbiome response to its application of in dependence with soil types serve the design of precision application strategies.

Modeling-Guided Amendments Lead to Enhanced Biodegradation in Soil.

Extensive use of agrochemicals is emerging as a serious environmental issue coming at the cost of the pollution of soil and water resources. Bioremediation techniques such as biostimulation are promising strategies used to remove pollutants from agricultural soils by supporting the indigenous microbial degraders. Though considered cost-effective and eco-friendly, the success rate of these strategies typically varies, and consequently, they are rarely integrated into commercial agricultural practices. In the current study, we applied metabolic-based community-modeling approaches for promoting realistic in terra solutions by simulation-based prioritization of alternative supplements as potential biostimulants, considering a collection of indigenous bacteria. Efficacy of biostimulants as enhancers of the indigenous degrader Paenarthrobacter was ranked through simulation and validated in pot experiments. A two-dimensional simulation matrix predicting the effect of different biostimulants on additional potential indigenous degraders (Pseudomonas, Clostridium, and Geobacter) was crossed with experimental observations. The overall ability of the models to predict the compounds that act as taxa-selective stimulants indicates that computational algorithms can guide the manipulation of the soil microbiome in situ and provides an additional step toward the educated design of biostimulation strategies. IMPORTANCE Providing the food requirements of a growing population comes at the cost of intensive use of agrochemicals, including pesticides. Native microbial soil communities are considered key players in the degradation of such exogenous substances. Manipulating microbial activity toward an optimized outcome in efficient biodegradation processes conveys a promise of maintaining intensive yet sustainable agriculture. Efficient strategies for harnessing the native microbiome require the development of approaches for processing big genomic data. Here, we pursued metabolic modeling for promoting realistic in terra solutions by simulation-based prioritization of alternative supplements as potential biostimulants, considering a collection of indigenous bacteria. Our genomic-based predictions point at strategies for optimizing biodegradation by the native community. Developing a systematic, data-guided understanding of metabolite-driven targeted enhancement of selected microorganisms lays the foundation for the design of ecologically sound methods for optimizing microbiome functioning.

Good News for Cabbageheads: Controlling Phelipanche aegyptiaca Infestation under Hydroponic and Field Conditions.

Phelipanche aegyptiaca (Orobanchaceae) is a parasitic weed that causes severe yield losses in field crops around the world. After establishing vascular connections to the host plant roots, P. aegyptiaca becomes a major sink that draws nutrients, minerals, and water from the host, resulting in extensive crop damage. One of the most effective ways to manage P. aegyptiaca infestations is through the use of herbicides. Our main objective was to optimize the dose and application protocol of herbicides that effectively control P. aegyptiaca but do not damage the cabbage crop. The interactions between the cabbage roots and the parasite were first examined in a hydroponic system to investigate the effect of herbicides on initial parasitism stages, e.g., germination, attachment, and tubercles production. Thereafter, the efficacy of glyphosate and ethametsulfuron-methyl in controlling P. aegyptiaca was examined in five cabbage fields naturally infested with P. aegyptiaca. The herbicides glyphosate and ethametsulfuron-methyl were applied on cabbage foliage and in the soil solution, both before and after the parasite had attached to the host roots. A hormesis effect was observed when glyphosate was applied at a dose of 36 g ae ha-1 in a non-infested P. aegyptiaca field. Three sequential herbicide applications (21, 35, and 49 days after planting) effectively controlled P. aegyptiaca without damaging the cabbages at a dose of 72 g ae ha-1 for glyphosate and at all the examined doses for ethametsulfuron-methyl. Parasite control with ethametsulfuron-methyl was also effective when overhead irrigation was applied after the herbicide application.

Wild sunflower goes viral: Citizen science and comparative genomics allow tracking the origin and establishment of invasive sunflower in the Levant.

Globalization and intensified volume of trade and transport around the world are accelerating the rate of biological invasions. It is therefore increasingly important to understand the processes through which invasive species colonize new habitats, often to the detriment of native flora. The initial steps of an invasion are particularly critical, as the introduced species relies on limited genetic diversity to adapt to a new environment. However, our understanding of this critical stage of the invasion is currently limited. We used a citizen science approach and social media to survey the distribution of invasive sunflower in Israel. We then sampled and sequenced a representative collection and compared it with available genomic data sets of North American wild sunflower, landraces and cultivars. We show that invasive wild sunflower is rapidly establishing throughout Israel, probably from a single, recent introduction from Texas, while maintaining high genetic diversity through ongoing gene flow. Since its introduction, invasive sunflower has spread quickly to most regions, and differentiation was detected despite extensive gene flow between clusters. Our findings suggest that rapid spread followed by continuous gene flow between diverging populations can serve as an efficient mechanism for maintaining sufficient genetic diversity at the early stages of invasion, promoting rapid adaptation and establishment in the new territory.

Biological and Transcriptomic Characterization of Pre-Haustorial Resistance to Sunflower Broomrape (Orobanche cumana W.) in Sunflowers (Helianthus annuus).

Infestations with sunflower broomrape (Orobanche cumana Wallr.), an obligatory root parasite, constitute a major limitation to sunflower production in many regions around the world. Breeding for resistance is the most effective approach to reduce sunflower broomrape infestation, yet resistance mechanisms are often broken by new races of the pathogen. Elucidating the mechanisms controlling resistance to broomrape at the molecular level is, thus, a desirable way to obtain long-lasting resistance. In this study, we investigated broomrape resistance in a confectionery sunflower cultivar with a robust and long-lasting resistance to sunflower broomrape. Visual screening and histological examination of sunflower roots revealed that penetration of the broomrape haustorium into the sunflower roots was blocked at the cortex, indicating a pre-haustorial mechanism of resistance. A comparative RNA sequencing between broomrape-resistant and -susceptible accessions allowed the identification of genes that were significantly differentially expressed upon broomrape infestation. Among these genes were ß-1,3-endoglucanase, ß-glucanase, and ethylene-responsive transcription factor 4 (ERF4). These genes were previously reported to be pathogenesis-related in other plant species. This transcriptomic investigation, together with the histological examinations, led us to conclude that the resistance mechanism involves the identification of the broomrape and the consequent formation of a physical barrier that prevents the establishment of the broomrape into the sunflower roots.

Strategies for Enhancing in vitro Degradation of Linuron by Variovorax sp. Strain SRS 16 Under the Guidance of Metabolic Modeling.

Phenyl urea herbicides are being extensively used for weed control in both agricultural and non-agricultural applications. Linuron is one of the key herbicides in this family and is in wide use. Like other phenyl urea herbicides, it is known to have toxic effects as a result of its persistence in the environment. The natural removal of linuron from the environment is mainly carried through microbial biodegradation. Some microorganisms have been reported to mineralize linuron completely and utilize it as a carbon and nitrogen source. Variovorax sp. strain SRS 16 is one of the known efficient degraders with a recently sequenced genome. The genomic data provide an opportunity to use a genome-scale model for improving biodegradation. The aim of our study is the construction of a genome-scale metabolic model following automatic and manual protocols and its application for improving its metabolic potential through iterative simulations. Applying flux balance analysis (FBA), growth and degradation performances of SRS 16 in different media considering the influence of selected supplements (potential carbon and nitrogen sources) were simulated. Outcomes are predictions for the suitable media modification, allowing faster degradation of linuron by SRS 16. Seven metabolites were selected for in vitro validation of the predictions through laboratory experiments confirming the degradation-promoting effect of specific amino acids (glutamine and asparagine) on linuron degradation and SRS 16 growth. Overall, simulations are shown to be efficient in predicting the degradation potential of SRS 16 in the presence of specific supplements. The generated information contributes to the understanding of the biochemistry of linuron degradation and can be further utilized for the development of new cleanup solutions without any genetic manipulation.

Simulating sulfosulfuron fate in soil under different weather scenarios to support weed management decisions.

BACKGROUND: Residual herbicides are an important component in many weed control strategies. Their herbicidal activity depends on their fate in soil, with respect to the required concentration for weed control in space and time. In this study, the effect of weather conditions on sulfosulfuron fate in soil, following pre-planting incorporation, and the predicted control efficacy of Egyptian broomrape in tomato, were analyzed for two sites using simulations in Hydrus-1D modeling software. Simulated concentration was compared to measured data from field experiments. RESULTS: Model evaluation against measured data from two fields, with weakly alkaline clay soils, showed high correlations between simulated and measured sulfosulfuron concentrations (r = 0.98 and 0.89). The ratio of measured to simulated concentration was relatively low (1.03) at the top 10-cm layer, in which the mean measured concentration was high (29.6 ng g-1 ). This ratio was higher (12.5) at the 30-60 cm depth, in which the mean measured concentration was lower (0.3 ng g-1 ). Simulations of sulfosulfuron fate in each site, using weather data from the years 2009 to 2019, revealed substantial variations in transport patterns. Thirty days after treatment, 16 out of the 22 years simulated for the two sites (11 at each site) resulted in concentrations lower than the critical value for Egyptian broomrape control throughout the soil profile. The data indicates that variation in sulfosulfuron fate is mainly due to differences in the cumulative precipitation. According to simulation results, cumulative precipitation above 20 or 10 mm during the first 10 or 20 days after treatment, respectively, is expected to reduce the efficiency of broomrape control. CONCLUSION: Considering weather effects when planning herbicide application could optimize herbicide use efficiency. A decision-support tool is presented, whose factors are the time gap and precipitation amount between sulfosulfuron application and tomato planting.

Genome-scale reconstruction of Paenarthrobacter aurescens TC1 metabolic model towards the study of atrazine bioremediation.

Atrazine is an herbicide and a pollutant of great environmental concern that is naturally biodegraded by microbial communities. Paenarthrobacter aurescens TC1 is one of the most studied degraders of this herbicide. Here, we developed a genome scale metabolic model for P. aurescens TC1, iRZ1179, to study the atrazine degradation process at organism level. Constraint based flux balance analysis and time dependent simulations were used to explore the organism's phenotypic landscape. Simulations aimed at designing media optimized for supporting growth and enhancing degradation, by passing the need in strain design via genetic modifications. Growth and degradation simulations were carried with more than 100 compounds consumed by P. aurescens TC1. In vitro validation confirmed the predicted classification of different compounds as efficient, moderate or poor stimulators of growth. Simulations successfully captured previous reports on the use of glucose and phosphate as bio-stimulators of atrazine degradation, supported by in vitro validation. Model predictions can go beyond supplementing the medium with a single compound and can predict the growth outcomes for higher complexity combinations. Hence, the analysis demonstrates that the exhaustive power of the genome scale metabolic reconstruction allows capturing complexities that are beyond common biochemical expertise and knowledge and further support the importance of computational platforms for the educated design of complex media. The model presented here can potentially serve as a predictive tool towards achieving optimal biodegradation efficiencies and for the development of ecologically friendly solutions for pollutant degradation.

Broomrape infestation in carrot (Daucus carota): Changes in carotenoid gene expression and carotenoid accumulation in the parasitic weed Phelipanche aegyptiaca and its host.

Carotenogenesis has been intensively studied in carrot roots, and transcriptional regulation is thought to be the major factor in carotenoid accumulation in these organs. However, little is known about the transcriptional regulation of carotenoid biosynthetic genes concerning carotenoid accumulation during infestation by the obligate parasite Phelipanche aegyptiaca. HPLC analysis revealed a decrease in carotenoid levels of the different carrot cultivars when parasitized by P. aegyptiaca. Besides, we isolated and analyzed P. aegyptiaca tubercles parasitizing the various carrot root cultivars and show that they accumulate different carotenoids compared to those in non-infested carrot roots. Expression analysis of PHYTOENE SYNTHASE (PSY1) and CAROTENOID ISOMERASE (CRTISO) as well as the strigolactone apocarotenoid biosynthetic genes DWARF27 (D27), CAROTENOID CLEAVAGE DIOXYGENASE 7 (CCD7) and CCD8 revealed that their transcript levels showed significant variation in P. aegyptiaca infested carrot roots. After parasite infestation, the expression of these genes was strongly reduced, as were the carotenoid levels and this was more pronounced in the uncommon non-orange varieties. We also analyzed the parasite genes encoding D27, CCD7 and CCD8 and show that they are expressed in tubercles. This raises important questions of whether the parasite produces its carotenoids and apocarotenoids including strigolactones and whether the latter might have a role in tubercle development.

3-D Image-Driven Morphological Crop Analysis: A Novel Method for Detection of Sunflower Broomrape Initial Subsoil Parasitism.

Effective control of the parasitic weed sunflower broomrape (Orobanche cumana Wallr.) can be achieved by herbicides application in early parasitism stages. However, the growing environmental concerns associated with herbicide treatments have motivated the adoption of precise chemical control approaches that detect and treat infested areas exclusively. The main challenge in developing such control practices for O. cumana lies in the fact that most of its life-cycle occurs in the soil sub-surface and by the time shoots emerge and become observable, the damage to the crop is irreversible. This paper approaches early O. cumana detection by hypothesizing that its parasitism already impacts the host plant morphology at the sub-soil surface developmental stage. To validate this hypothesis, O. cumana- infested sunflower and non-infested control plants were grown in pots and imaged weekly over 45-day period. Three-dimensional plant models were reconstructed using image-based multi-view stereo followed by derivation of their morphological parameters, down to the organ-level. Among the parameters estimated, height and first internode length were the earliest definitive indicators of infection. Furthermore, the detection timing of both parameters was early enough for herbicide post-emergence application. Considering the fact that 3-D morphological modeling is nondestructive, is based on commercially available RGB sensors and can be used under natural illumination; this approach holds potential contribution for site specific pre-emergence managements of parasitic weeds and as a phenotyping tool in O. cumana resistant sunflower breeding projects.

Use of a visible reporter marker- myb-related gene in crop plants to minimize herbicide usage against weeds.

Weeds, a main threat to agricultural productivity worldwide, are mostly controlled by herbicides. To minimize herbicide usage by targeting only weedy areas, we developed a new methodology for robust weed detection that relies on manipulating the crop plant's leaf hue, without affecting crop fitness. We generated transgenic tobacco (Nicotiana tabacum Xanthi) lines overexpressing the anthocyanin pigment as a traceable marker that differentiates transgenes from the surrounding weeds at an early stage. Transformation with the anthocyanin VlmybA1-2 gene produced purple-colored leaves. Subsequent gene silencing with vector pTRV2:VlmybA1-2 significantly reduced anthocyanin pigments in tobacco leaves 40 days after agroinfiltration, with a concomitant reduction in VlmybA1-2 transcript levels. Purple hue faded gradually, and there were no fitness costs in terms of plant height or leaf number in the silenced vs. non-silenced tobacco transgenes. These results could lead to a new sustainable weed-control method that will alleviate weed-related ecological, agricultural and economic issues.

Modeling microbial communities from atrazine contaminated soils promotes the development of biostimulation solutions.

Microbial communities play a vital role in biogeochemical cycles, allowing the biodegradation of a wide range of pollutants. The composition of the community and the interactions between its members affect degradation rate and determine the identity of the final products. Here, we demonstrate the application of sequencing technologies and metabolic modeling approaches towards enhancing biodegradation of atrazine-a herbicide causing environmental pollution. Treatment of agriculture soil with atrazine is shown to induce significant changes in community structure and functional performances. Genome-scale metabolic models were constructed for Arthrobacter, the atrazine degrader, and four other non-atrazine degrading species whose relative abundance in soil was changed following exposure to the herbicide. By modeling community function we show that consortia including the direct degrader and non-degrader differentially abundant species perform better than Arthrobacter alone. Simulations predict that growth/degradation enhancement is derived by metabolic exchanges between community members. Based on simulations we designed endogenous consortia optimized for enhanced degradation whose performances were validated in vitro and biostimulation strategies that were tested in pot experiments. Overall, our analysis demonstrates that understanding community function in its wider context, beyond the single direct degrader perspective, promotes the design of biostimulation strategies.

Phelipanche aegyptiaca parasitism impairs salinity tolerance in young leaves of tomato.

The parasite Phelipanche aegyptiaca infests tomato, a crop plant that is commonly cultivated in semi-arid environments, where tomato may be subject to salt stress. Since the relationship between the two stresses -salinity and parasitism - has been poorly investigated in tomato, the effects of P. aegyptiaca parasitism on tomato growing under moderate salinity were examined. Tomatoes were grown with regular or saline water irrigation (3 and 45 mM Cl- , respectively) in soils infested with P. aegyptiaca. The infested plants accumulated higher levels of sodium and chloride ions in the roots, shoots and leaves (old and young) under both salinity levels vs. non-infected plants. There was a positive linear correlation between P. aegyptiaca biomass and salt accumulation in young tomato leaves, and a negative linear correlation between parasite biomass and the osmotic potential of young tomato leaves. Concentrations of the osmoprotectants proline, myoinositol and sucrose were reduced in infected tomato plants, which impaired the host's osmotic adjustment ability. The sensitivity of P. aegyptiaca to salt stress was manifested as a decrease in biomass. In conclusion, P. aegyptiaca parasitism reduced the salt tolerance of tomato plants by promoting the accumulation of salts from the rhizosphere and impairing the host's osmotic adjustment ability.

Control of Egyptian Broomrape in Processing Tomato: A Summary of 20 Years of Research and Successful Implementation.

The obligate root parasitic weeds commonly known as broomrape (Orobanche and Phelipanche spp.) cause severe damage to vegetable and field crops worldwide. Efficient control of these parasites is difficult due to their development and attachment to the host plant (via a specialized organ, the haustorium) under the soil surface and to their unique biological traits of massive seed production, facile seed dispersal, germination only under specific conditions, and seed longevity. The major damage inflicted by the parasites takes place underground, making control extremely challenging. Egyptian broomrape (Phelipanche aegyptiaca) is a devastating pest in the Mediterranean basin, parasitizing a wide host crop range, including tomato, sunflower, legumes, and carrot, resulting in severe crop losses. Twenty years of research have led to the development of integrated smart management strategies for combating this parasite in processing tomato fields. In particular, an explicit decision support system (DSS) designated PICKIT has been developed; this DSS is based on predicting parasitism dynamics and employing a range of selective targeted chemical applications (preplanting incorporation, foliar application, and herbigation). In this feature article, we describe the evolution of this research from the laboratory, through greenhouse and experimental field trials, to large scale commercial fields and the successful assimilation of PICKIT into agricultural practice. The use of PICKIT in fields of processing tomatoes in northern Israel has led to effective control of Egyptian broomrape, even in fields with high infestation levels, resulting in a tomato yield increase of an average of 40 tons ha-1 compared with nontreated plots. In 2016, PICKIT was commercially implemented in 33 fields, totaling 400 ha, giving 95% Egyptian broomrape control and tomato yields of 115 to 145 tons ha-1. The outcome of this research is now enabling farmers to grow tomatoes in Egyptian broomrape-infested fields with assured increased yields and hence high profits.

Enhanced Host-Parasite Resistance Based on Down-Regulation of Phelipanche aegyptiaca Target Genes Is Likely by Mobile Small RNA.

RNA silencing refers to diverse mechanisms that control gene expression at transcriptional and post-transcriptional levels which can also be used in parasitic pathogens of plants that Broomrapes (Orobanche/Phelipanche spp.) are holoparasitic plants that subsist on the roots of a variety of agricultural crops and cause severe negative effects on the yield and yield quality of those crops. Effective methods for controlling parasitic weeds are scarce, with only a few known cases of genetic resistance. In the current study, we suggest an improved strategy for the control of parasitic weeds based on trans-specific gene-silencing of three parasite genes at once. We used two strategies to express dsRNA containing selected sequences of three Phelipanche aegyptiaca genes PaACS, PaM6PR, and PaPrx1 (pma): transient expression using Tobacco rattle virus (TRV:pma) as a virus-induced gene-silencing vector and stable expression in transgenic tomato Solanum lycopersicum (Mill.) plants harboring a hairpin construct (pBINPLUS35:pma). siRNA-mediated transgene-silencing (20-24 nt) was detected in the host plants. Our results demonstrate that the quantities of PaACS and PaM6PR transcripts from P. aegyptiaca tubercles grown on transgenic tomato or on TRV-infected Nicotiana benthamiana plants were significantly reduced. However, only partial reductions in the quantity of PaPrx1 transcripts were observed in the parasite tubercles grown on tomato and on N. benthamiana plants. Concomitant with the suppression of the target genes, there were significant decreases in the number and weight of the parasite tubercles that grew on the host plants, in both the transient and the stable experimental systems. The results of the work carried out using both strategies point to the movement of mobile exogenous siRNA from the host to the parasite, leading to the impaired expression of essential parasite target genes.