Reduce Pesticide Application Research Articles

Economics of herbicide‐free crop production

AbstractAmbitious pesticide policies aiming to reduce pesticide applications and risks have been introduced across Europe. Herbicides represent a major part of pesticide use, but the uptake of mechanical or agronomic alternatives remains low. We here explore underlying reasons and provide policy insights for supporting a transformation to herbicide‐free production, accounting for both economic and agronomic drivers. We develop detailed extensions to an existing bio‐economic modeling approach and use stochastic dominance analysis to assess the performance of non‐chemical alternatives to herbicides under different production and market conditions exante. We apply our approach to Swiss wheat production and find that herbicide‐free production is not viable without financial support, and thus requires additional per‐hectare agri‐environmental payments and price markups to be economically viable. We find that currently available support payments in Switzerland are sufficient in achieving economic viability of herbicide‐free production. Moreover, we confirm the relevance of risk and risk preferences and identify a risk‐reducing character of herbicide‐free production systems with support payments. Our analysis provides insights on potential drivers, trade‐offs, decision‐making factors, and policies for a transition to non‐chemical weed control.

Effects of pesticides on soil bacterial, fungal and protist communities, soil functions and grape quality in vineyards

Abstract Pesticides can have unintentional effects on non‐target organisms and change biotic communities. Such changes might be particularly important in soil microbial communities, which drive many ecosystem functions and may affect crop quality. Here, we investigated, in a three‐year study, how vegetation control (by herbicide application) and soil copper content (from long‐term copper‐based fungicide application) affect biodiversity and the community structure of soil bacteria, fungi and protists, and associated soil functions (respiration, decomposition) in Swiss vineyards. Furthermore, we determined the effects of these two management practices on grape quality as the most direct ecosystem service to farmers. Across all study years, the community composition of microorganisms was affected by herbicide application, however, a significant loss of operational taxonomic units (OTUs) was only observed in fungi and protists. Soil copper content reduced OTU richness of bacteria and protists in some years but had no significant effect on fungal richness. Copper changed the community composition in all three groups of soil microorganisms. While we found no effect of copper on soil functions, herbicide application reduced microbial respiration and biomass by about 39% and 45%, respectively. However, decomposition rates remained virtually unchanged by any pesticide. Yeast assimilable nitrogen (YAN) levels in grape were below the critical threshold of 140 mg/L in 40% of the vineyards without herbicide application and the variety Chasselas, whereas in vineyards with herbicide application, it was only 20%. Application of pesticides led to changes in richness and composition of soil microbial communities and directly reduced some soil functions (microbial biomass and respiration) but not all (decomposition). Some grape quality parameters can be indirectly enhanced by pesticide application, highlighting the trade‐off between the interests of nature conservation and the interests of the farmer. Balancing these two diverging interests requires the establishment of alternative vineyard management allowing reduced pesticide application.

Perfect cooperative pest control via nano-pesticide and natural predator: High predation selectivity and negligible toxicity toward predatory stinkbug

The improper use of synthetic pesticides has caused adverse effects on global ecosystems and human health. As a part of sustainable pest management strategy, natural predators, along with nano-pesticides, have made significant contributions to ecological agriculture. The cooperative application of both approaches may overcome their limitations, substantially reducing pesticide application while controlling insect pests efficiently. Herein, the current study introduced a cationic star polymer (SPc) to prepare two types of nano-pesticides, which were co-applied with predatory stinkbugs Picromerus lewisi to achieve perfect cooperative pest control. The SPc exhibited nearly no toxicity against predatory stinkbugs at the working concentration, but it led to the death of predatory stinkbugs at extremely high concentration with the lethal concentration 50 (LC50) value of 13.57 mg/mL through oral feeding method. RNA-seq analysis revealed that the oral feeding of SPc could induce obvious stress responses, leading to stronger phagocytosis, exocytosis, and energy synthesis to ultimately result in the death of predatory stinkbugs. Then, the broflanilide and chlorobenzuron were employed to prepare the self-assembled nano-pesticides via hydrogen bond and Van der Waals force, and the complexation with SPc broke the self-aggregated structures of pesticides and reduced their particle sizes down to nanoscale. The bioactivities of prepared nano-pesticides were significantly improved toward common cutworm Spodoptera litura with the corrected mortality increase by approximately 30%. Importantly, predatory stinkbugs exhibited a strong predation selectivity for alive common cutworms to reduce the exposure risk of nano-pesticides, and the nano-pesticides showed negligible toxicity against predators. Thus, the nano-pesticides and predatory stinkbugs could be applied simultaneously for efficient and sustainable pest management. The current study provides an excellent precedent for perfect cooperative pest control via nano-pesticide and natural predator.

The efficacy of spatio‐temporal predictors in forecasting the risk of Cydia nigricana infestation

AbstractThe ability to estimate the risk of pest infestation can help farmers to reduce pesticide application and provide guidance that would result in better management decisions. This study tested whether different combinations of spatial and temporal risk factors may be used to predict the damage potential of pea moth, Cydia nigricana Fabricius (Lepidoptera: Tortricidae), a major pest in field pea (Pisum sativum L., Fabaceae). Over four consecutive years, the abundance of pea moth was monitored by placing pheromone traps at various field pea‐cultivation sites. We also assessed the phenological development stages and the percentage of damaged seeds per 100 pods collected from each growing pea field in a region of approximately 30 km in diameter. The study found the significant infestation risk indicators to be the time of flowering, the date on which male pea moths are first detected in the monitoring traps and the minimum distance to pea fields that were planted and harvested in the previous growing season. The combination of all three factors using a general additive model approach yielded the best results. The model proposed by this study accurately discriminated between low‐infestation and high‐infestation fields in 95% of cases.

Who is willing to pay a premium for leafy greens produced by indoor agriculture? A comparative choice experiment with US consumers

AbstractIndoor agriculture (IA) presents a pathway to producing leafy greens more sustainably by strictly controlling the growing environment, increasing the efficiency of water and land resources use, reducing pesticide application, and enhancing quality characteristics. However, the commercial success of adopting this production method depends on the technology's widespread consumer acceptance paired with a price premium for the improved quality it provides. This paper characterizes American consumer preferences for leafy greens, reports a positive WTP for leafy green attributes, and estimates how consumer attitudes toward IA affect their leafy green choices. The findings are drawn from an online survey of over 2000 US leafy green consumers conducted in 2021. The respondents faced a hypothetical discrete choice experiment (DCE) designed to simulate purchasing leafy green produce. The alternatives presented in the DCE varied over production systems (IA, field farming, or greenhouses) and quality levels (taste, freshness, nutrient level, and food safety). Estimates identify significant preference heterogeneity for leafy green attributes and facilitate classifying consumers into three preference‐based groups: “Quality Seekers,” “Price Conscious,” and “Focused Practicals.” Preference variation is significantly greater for production methods than quality characteristics, indicating nascent production method preferences. Results suggest most US consumers view IA positively; however, WTP estimates vary significantly between consumer segments [EconLit Citations: D12, D13].

Unveiling the importance of VOCs from pesticides applicated in main crops for elevating ozone concentrations in China

Volatile organic compounds (VOCs) are considered as important precursors of ozone in the air, while the contribution of VOCs from pesticide application (PVOCs) to ozone production is unknown. Utilizing data from the Ministry of Agriculture and Rural Affairs of the People's Republic of China and ChinaCropPhen1km, this paper developed PVOC emission inventories with a resolution of 1 km for the main crops (rice, maize, and wheat) from 2012 to 2019 in China. The results revealed that pesticide application is an important VOC emission source in China. Specially, the PVOC emissions from the major grain-producing regions in June accounted for approximately 30% of the annual total PVOC emissions in the local regions. The simulation with the Weather Research and Forecasting Community Multiscale Air Quality model (WRF-CMAQ) indicated that the PVOC emissions increased the mean maximum daily 8-hour average (MDA8) ozone concentration across China by 2.5 ppb in June 2019. During the same period, PVOCs in the parts of North China Plain contributed 10% of the ozone formation. Under the comprehensive emission reduction scenario, it is anticipated that by 2025, the joint implementation of measures including reducing pesticide application, improving pesticide utilization efficiency and promoting solvent substitution will decrease PVOC emissions by 60% compared with 2019, thereby mitigating ozone pollution.

Assessment of cultivation intensity can improve the correlative strength between agriculture and the ecological status in rivers across Germany

Agriculture has been identified as a main cause for more than 90% of Germany´s rivers still not meeting good ecological status in 2021. While many large-scale studies observed a negative effect of catchment agricultural land use on river biota, they rarely considered differences in cultivation intensities, although small-scale studies highlight clear differences between the effects of agricultural crops. Here we used Germany-wide and spatially explicit information on crop types to calculate agricultural intensity indices for nutrients and pesticides, weighting different crop types based on average pesticide treatment and nutrient application rates. These indices were then used as explanatory variables for the ecological status of n = 7677 biological sampling sites. Pesticides were more important than nutrient pollution for macroinvertebrates and macrophytes, while diatoms were more sensitive to nutrients. Considering the most relevant intensity index (pesticide or nutrient) slightly increased the correlative strength with ecological status, as compared to the correlation with agricultural land or cropland cover by up to R² = 0.14 for diatoms. Correlative strength of agricultural intensity indices was substantially larger in small mountain and (pre)-alpine streams compared to lowland streams, with an R² up to 0.43 for macroinvertebrates. These results not only confirm previous large-scale studies by demonstrating the detrimental effects of present-day agriculture on river biota, but also shed light on the main pathways involved, particularly highlighting the adverse impacts of agrochemicals. Consequently, to protect river biota, a shift to more sustainable agricultural practices, like reducing pesticide application, is urgently required.

Anti-fungal bioactive terpenoids in the bioenergy crop switchgrass (Panicum virgatum) may contribute to ecotype-specific microbiome composition

Plant derived bioactive small molecules have attracted attention of scientists across fundamental and applied scientific disciplines. We seek to understand the influence of these phytochemicals on rhizosphere and root-associated fungi. We hypothesize that – consistent with accumulating evidence that switchgrass genotype impacts microbiome assembly – differential terpenoid accumulation contributes to switchgrass ecotype-specific microbiome composition. An initial in vitro Petri plate-based disc diffusion screen of 18 switchgrass root derived fungal isolates revealed differential responses to upland- and lowland-isolated metabolites. To identify specific fungal growth-modulating metabolites, we tested fractions from root extracts on three ecologically important fungal isolates – Linnemania elongata, Trichoderma sp. and Fusarium sp. Saponins and diterpenoids were identified as the most prominent antifungal metabolites. Finally, analysis of liquid chromatography-purified terpenoids revealed fungal inhibition structure – activity relationships (SAR). Saponin antifungal activity was primarily determined by the number of sugar moieties – saponins glycosylated at a single core position were inhibitory whereas saponins glycosylated at two core positions were inactive. Saponin core hydroxylation and acetylation were also associated with reduced activity. Diterpenoid activity required the presence of an intact furan ring for strong fungal growth inhibition. These results inform future breeding and biotechnology strategies for crop protection with reduced pesticide application.

Design and Spray Performance Evaluation of an Air–Ground Cooperation Stereoscopic Plant Protection System for Mango Orchards

With the aim of solving the problems of high labor intensity, low operational efficiency, and poor deposition distribution uniformity in the mango canopy associated with traditional plant protection devices, an air-ground co-operation stereoscopic plant protection system consisting of an orchard caterpillar mist sprayer and a six-rotor plant protection UAV was designed to jointly undertake plant protection operations in mango orchards. We tested the spraying performance of the system on mango trees, compared with the single-machine operation, the air–ground co-operation system could significantly increase the droplet coverage on the upperside of mango leaves in each part of the canopy (a 14.7% increase for the mist sprayer and 12.9% for the UAV). This increased the active component deposition distribution uniformity in the mango canopy but could not significantly improve the deposition and coverage of droplets on the underside of leaves compared with the mist sprayer and plant protection UAV. Due to the characteristics of the mango canopy such as large leaf length and thickness and complex leaf inclination distribution, this led to poor deposition distribution uniformity of the two spray units, and the overall CV was over 150%. The pesticide active ingredients were almost uniformly distributed in the vertical direction when the application ratios (ground implements/plant protection drones) were 8/2 and 7/3, offering a promising protocol for reduced pesticide application in mango orchards. This study presents promising data that support the innovative integration of drones into crop protection programs for large canopy crops (e.g., mango) and provides guidance for the ACSPPS system in reduction and precision application research.

Differential responses of Orthoptera in vineyards to organic farming, pesticide reduction, and landscape heterogeneity

Conservation measures such as those under the European Green Deal aim to counteract the biodiversity loss by increasing the share of organic farming and reducing pesticide use, as well as increasing the proportion of semi-natural habitats (SNH) in agricultural landscapes. Given the large environmental impacts of agriculture, it is important to thoroughly understand effects of such measures on organisms to provide evidence-based and effective implications for conservation. In this study, we analysed how vineyard management, pesticide reduction, and landscape composition affect Orthoptera densities and species composition. Therefore, we sampled herb- and vine-dwelling orthopterans in a paired design of classic and fungus-resistant grape (FRG) varieties in conventionally and organically managed vineyards along a landscape heterogeneity gradient. Here, FRG varieties allowed us to study the effect of 44% reduced pesticide applications under real-world conditions. Total densities of herb-dwelling Orthoptera did not differ between grape varieties in conventional vineyards, but were 2.9 times higher in FRG varieties under organic management. In contrast, total densities of vine-dwelling Orthoptera, mainly driven by the dominant species Phaneroptera falcata, were similar between grape varieties in organic vineyards, but tended to be 1.4 times higher in classic grapes under conventional management. Furthermore, the management system and SNH in a radius of 500 m in the surrounding landscape influenced species composition.

Fruit Bats: Their Importance, Threats and Conservation

Bats are nocturnal mammals which lives in large aggregations as colonies, hang their feet to twig of tree upward side, provide widespread ecological and monetary services via pollination, seed dispersal, rejuvenation of forests, guano as nutrient rich fertilizer and agricultural pest control. Different factors like exposure to heavy metals, pesticides, hunting, diminishing food supply, habitat destruction, forest fires and diseases are responsible for decreasing bat population, but urbanization greatly affects the composition and structure of inhabiting animal communities by transforming the natural habitats into environments dominated by human constructions. Micro bats feed on insect pests and help to reduce pesticide application in agricultural crops, whereas fruit bats feed on horticultural orchards like guava, ber and litchi thus act as pest. Bats are reservoirs of many pathogens like hendra, nipah, tioman, European lyssa and ebola viruses which cause several epidemic diseases among humans and domestic animals. Among 60 countries around the world, more than 200 bat species are considered to be threatened (critically endangered, endangered or vulnerable) by the International Union for the Conservation of Nature (IUCN). Using different eco-friendly methods like artificial lights and netting we can save our orchards and conserve fruit bats which will be helpful in the maintenance of ecological balance of nature.

Measuring the Impact of Relative Deprivation on Tea Farmers’ Pesticide Application Behavior: The Case of Shaanxi, Sichuan, Zhejiang, and Anhui Province, China

Reducing chemical interaction within core farming tactics has gained much attention worldwide due to ever-increasing water, soil, and air pollution trends caused by various agricultural activities. Since, in the developing countries, tea is primarily produced conventionally, clarifying the impact of relative deprivation on the pesticide application rate of tea farmers is conducive to promoting the reduction of pesticides and the green development of the tea industry. Thus, based on extensive literature reviews, the study constructs a theoretical framework of relative deprivation and pesticide application rate by tea farmers. Moreover, the moderating effect of external intervention and behavioral factors has also been assessed. A data set of 786 tea farmers’ responses from Shaanxi, Sichuan, Zhejiang, and Anhui provinces has been utilized to test, outline and validate the proposed framework. We utilized the Ordered Probit model to measure the psychological fluctuation of tea farmers. The results are as follows. (i) The more substantial their perception of relative deprivation, the more tea farmers tend to increase the application rate. (ii) In external interventions, the degree of government regulation can not only directly promote the reduction of pesticide application but also play a negative regulatory role between the relative deprivation and the amount of pesticide applied by tea farmers. Although the degree of community control can directly promote the reduction of pesticide application by tea farmers, its regulating effect is insignificant. (iii) Regarding behavioral ability, the management scale can directly promote the reduction of pesticide application by tea farmers, but its regulating impact is not significant. Thus, government should highlight the importance of organic and environmentally friendly tea cultivation and encourage tea farmers to reduce pesticide application. Along with the market regulation, point-of-sale testing and traceability of pesticide residues should continue to be consolidated, strengthened and improved.

Exploring Floridians' perceptions of pollinator-friendly gardening to identify critical adoption barriers and strategies

Developed landscapes in urban and peri-urban environments significantly influence the diversity, abundance, and overall health of pollinator populations such as bees, butterflies, moths, and birds. Considering growing evidence demonstrating that pollinator health can be more robust in urban habitats than rural ones, the landscape development and management practices adopted in urban and suburban areas have a significant potential impact on pollinator health. Given their influence with respect to urban and suburban landscape management in Florida, it is critical to better understand the knowledge, attitudes, and perceptions of Florida residents regarding pollinator-friendly gardening practices such as the flowering plant installation or reduced pesticide application. To explore the most salient factors influencing residents’ adoption of pollinator gardening practices, we facilitated three elicitation focus groups with 20 residents representing three distinct regions in Florida. Data analysis generated a series of salient sub-themes across two thematic categories: ‘barriers’ and ‘solutions and strategies’. Major ‘barrier’ themes included ‘limited knowledge or information access’ about pollinator gardening, and ‘social norms/community expectations’. Major solution and strategy’ themes included ‘developing neighborhood/homeowner association buy-in,’ and ‘pre-installation research and planning’. Overall, findings demonstrate that Florida residents have strong views regarding both the most influential barriers to adopting or engaging in pollinator-friendly gardening and the most promising solutions or strategies to mitigate those barriers. These findings have tangible implications for educational practitioners interested in expanding distribution of targeted educational outreach to further promote the adoption of pollinator-friendly landscape management practices in Florida. Findings also have implications for researchers interested in further examining the socio-economic and socio-cultural factors most influential to residents’ pollinator gardening practices.

Evaluating, Screening and Selecting Yardlong Bean [Vigna unguiculata subsp. sesquipedalis (L.) Verdc.] for Resistance to Common Cutworm (Spodoptera litura Fabricius)

The yardlong bean [Vigna unguiculata subsp. sesquipedalis (L.) Verdc.] is an important vegetable crop, but it is prone to pest infestation. Therefore, breeding insect-resistant varieties is essential to reduce pesticide applications and to increase bean quality and yield. In the present study, 64 yardlong bean varieties were screened for their resistance to the common cutworm (Spodoptera litura Fabricius). In the greenhouse, leaves, pods, and seeds of yardlong beans were harmed by naturally occurring common cutworms. Seventeen insect-resistant and four insect-susceptible yardlong bean varieties were identified based on the weight of the nine-day-old larvae and 72 h weight increases of 4th instar larvae through feeding newly hatched and 4th instar larvae, respectively. Subsequent verification feeding experiments with newly hatched larvae showed that Zhuzaidou and Pingtangjiangdou’s insect resistance are the weakest and Jiangdou No.5, j-1, Zhijiangtezao No. 30, and Changcaidou have the strongest insect resistance. In 21 yardlong bean varieties, starch content and larval weight showed negative correlation and there was positive correlation between crude protein and larval weight, but almost neither of them reached significant levels. Through organ antibiotic and antixenotic experiments, it was concluded that common cutworms preferred feeding on yardlong bean leaves, and the weight increase of common cutworms feeding on leaves was higher than that of pods and seeds. These insect-resistant yardlong bean varieties warrant further investigation in basic antibiosis mechanism research in yardlong beans and can serve as germplasm resources for breeding programs engaged in reducing pesticide usage.

Management of take-all disease caused by Gaeumannomyces graminis var. tritici in wheat through Bacillus subtilis strains.

Wheat (Triticum aestivum) is the second largest grain crop worldwide, and one of the three major grain crops produced in China. Take-all disease, caused by Gaeumannomyces graminis var. tritici (Ggt) infection, is a widespread and devastating soil-borne disease that harms wheat production. At present, the prevention and control of wheat take-all depend largely on the application of chemical pesticides. Chemical pesticides, however, not only lead to increased drug resistance of pathogens but also leave significant residues in the soil, causing serious environmental pollution. In this study, we investigated the application of Bacillus subtilis to achieve take-all disease control in wheat while reducing pesticide application. Antagonistic bacteria were screened by plate test, species identification of strains was performed by Gram staining and sequencing of 16s rDNA, secondary metabolite activity of strains was detected by clear circle method, strain compatibility and effect of compounding on Ggt were detected by plate, and the application prospects of specific strains were analyzed by greenhouse and field experiments. We found that five B. subtilis strains, JY122, JY214, ZY133, NW03, Z-14, had significant antagonistic effects against Ggt, and could secrete antimicrobial proteins including amylase, protease, and cellulase. Furthermore, Z-14 and JY214 cultures have also been shown to change the morphology of Ggt mycelium. These results also showed that Z-14, JY214, and their combination can control take-all disease in wheat at a reduced level of pesticide use. In summary, we screened two Bacillus spp. strains, Z-14 and JY214, that could act as antagonists that contribute to the biological control of wheat take-all disease. These findings provide resources and ideas for controlling crop diseases in an environmentally friendly manner.

Intelligent spray technology to control pests in nursery and orchard systems with reduced pesticide application rates

Intelligent spray technology to control pests in nursery and orchard systems with reduced pesticide application rates

Genetically Modified (GM) Crop Use 1996–2020: Environmental Impacts Associated with Pesticide Use Change

ABSTRACT This paper assesses the environmental impacts associated with changes in pesticide use with GM crops at a global level. The main technologies impacting on pesticide use have been crops modified to be tolerant to specific herbicides so as to facilitate improved weed control and crops resistant to a range of crop insect pests that otherwise damage crops or typically require the application of insecticides to control them. Over the 24 year period examined to 2020, the widespread use of GM insect resistant and herbicide tolerant seed technology has reduced pesticide application by 748.6 million kg (−7.2%) of active ingredient and, as a result, decreased the environmental impact associated with insecticide and herbicide use on these crops (as measured by the indicator, the Environmental Impact Quotient (EIQ)) by a larger 17.3% between 1996 and 2020. The technology that has delivered the largest change in pesticide use has been insect resistant cotton, where a 339 million kg of active ingredient saving has occurred and the associated environmental impact (as measured by the EIQ indicator) has fallen by about a third.

Prospects of fungal endophytes for stress mitigation in rice (Oryza sativa L.)

Fungal endophytes are highly diverse in their functions and impart functional advantage to the host plants. Application of fungal endophytes is gaining relevance in field of agriculture as an ecofriendly approach and means of reducing pesticide application in crop plants. Rice production is known to be affected by both biotic and abiotic stresses and application of microbes could be one of the means of reducing the stresses. Fungal endophytes are isolated and studied from various parts of rice plants and they are reported to be involved in improving the growth and reducing various stresses during plant growth. This review discusses about relevance and potential of fungal endophytes in rice on mitigating various stresses like drought, salinity, heat, heavy metals and biotic elements such as diseases and pests.

Physiological and Molecular Response Modifications by Ultraviolet-C Radiation in Plutella xylostella and Its Compatibility with Cordyceps fumosorosea

Ultraviolet-C (UV-C) radiation significantly impacts living organisms. UV-C radiation can also be used as a pest management tool. Therefore, this study was designed to investigate the effect of UV-C radiation on the physiology and gene expression level of Plutella xylostella, a destructive vegetable pest. Results showed that, after exposure to UV-C radiation for 3, 6, 12, and 24 h, the activity of SOD (superoxide dismutase) and CAT (catalase) of P. xylostella increased, while the activity of PPO (polyphenol oxidase), POD (peroxidase), AChE (acetylcholinesterase), CarE (carboxylesterase), and ACP (acid phosphatase) decreased with increased exposure time. Correlation coefficient analyses indicated that the activity of CAT correlated positively, while PPO and CarE correlated negatively, with exposure time. Gene regulation analysis via qRT-PCR confirmed a significant increase in regulation in CAT, CarE, and PPO-related genes. We also investigated the effect of UV-C exposure on the virulence of Cordyceps fumosorosea against P. xylostella. Here, results indicated that when the fungal treatment was applied to larvae before UV-C radiation, the virulence of C. fumosorosea was significantly reduced. However, this decline in virulence of C. fumosorosea due to UV-C exposure remained only for one generation, and no effect was observed on secondary infection. On the other hand, when larvae were exposed to UV-C radiation before fungal application, the mortality rate significantly increased as the exposure time to UV-C radiation increased. From the current study, it could be concluded that UV-C exposure suppressed the immunity to P. xylostella, which later enhanced the virulence of entomopathogenic fungi. Moreover, the study also suggested that UV irradiation is an effective pest management tool that could be incorporated into pest management strategies, which could help reduce pesticide application, be economically beneficial for the farmer, and be environmentally safe.

Efficient fluorescence-based localization technique for real-time tracking endophytes route in host-plants colonization.

Bacterial isolates that enhance plant growth and suppress plant pathogens growth are essential tools for reducing pesticide applications in plant production systems. The objectives of this study were to develop a reliable fluorescence‐based technique for labeling bacterial isolates selected as biological control agents (BCAs) to allow their direct tracking in the host‐plant interactions, understand the BCA localization within their host plants, and the route of plant colonization. Objectives were achieved by developing competent BCAs transformed with two plasmids, pBSU101 and pANIC‐10A, containing reporter genes eGFP and pporRFP, respectively. Our results revealed that the plasmid‐mediated transformation efficiencies of antibiotic‐resistant competent BCAs identified as PSL, IMC8, and PS were up 84%. Fluorescent BCA‐tagged reporter genes were associated with roots and hypocotyls but not with leaves or stems and were confirmed by fluoresence microscopy and PCR analyses in colonized Arabidopsis and sorghum. This fluorescence‐based technique's high resolution and reproducibility make it a platform‐independent system that allows tracking of BCAs spatially within plant tissues, enabling assessment of the movement and niches of BCAs within colonized plants. Steps for producing and transforming competent fluorescent BCAs, as well as the inoculation of plants with transformed BCAs, localization, and confirmation of fluorescent BCAs through fluorescence imaging and PCR, are provided in this manuscript. This study features host‐plant interactions and subsequently biological and physiological mechanisms implicated in these interactions. The maximum time to complete all the steps of this protocol is approximately 3 months.