A Basic Guide to the Propagation and Manipulation of the Clubroot Pathogen, Plasmodiophora brassicae.

Clubroot caused by the obligate parasite Plasmodiophora brassicae is a devastating disease affecting the canola industry worldwide. The socio-economic impact of clubroot can be significant, particularly in regions where Brassica crops are a major agricultural commodity. The disease can cause significant crop losses, leading to reduced yield and income for farmers. Extensive studies have been conducted to understand the biology and genetics of the pathogens and develop more effective management strategies. However, the basic procedures used for pathogen storage and virulence analysis have not been assembled or discussed in detail. As a result, there are discrepancies among the different protocols used today. The aim of this article is to provide a comprehensive and easily accessible resource for researchers who are interested in replicating or building upon the methods used in the study of the clubroot pathogen. Here, we discuss in detail the methods used for P. brassicae spore isolation, inoculation, quantification, propagation, and molecular techniques such as DNA extraction and PCR. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Extraction of Plasmodiophora brassicae resting spores and propagation Support Protocol 1: Evans blue staining to identify resting spore viability Support Protocol 2: Storage of Plasmodiophora brassicae Basic Protocol 2: Generation of single spore isolates from P. brassicae field isolates Basic Protocol 3: Phenotyping of Plasmodiophora brassicae isolates Basic Protocol 4: Genomic DNA extraction from Plasmodiophora brassicae resting spores Basic Protocol 5: Molecular detection of Plasmodiophora brassicae.

The Cellular and Subcellular Organization of the Glucosinolate-Myrosinase System against Herbivores and Pathogens.

Glucosinolates are an important class of secondary metabolites in Brassicales plants with a critical role in chemical defense. Glucosinolates are chemically inactive but can be hydrolyzed by myrosinases to produce a range of chemically active compounds toxic to herbivores and pathogens, thereby constituting the glucosinolate-myrosinase defense system or the mustard oil bomb. During the evolution, Brassicales plants have developed not only complex biosynthetic pathways for production of a large number of glucosinolate structures but also different classes of myrosinases that differ in catalytic mechanisms and substrate specificity. Studies over the past several decades have made important progress in the understanding of the cellular and subcellular organization of the glucosinolate-myrosinase system for rapid and timely detonation of the mustard oil bomb upon tissue damage after herbivore feeding and pathogen infection. Progress has also been made in understanding the mechanisms that herbivores and pathogens have evolved to counter the mustard oil bomb. In this review, we summarize our current understanding of the function and organization of the glucosinolate-myrosinase system in Brassicales plants and discuss both the progresses and future challenges in addressing this complex defense system as an excellent model for analyzing plant chemical defense.

Detection of enteroparasites in foliar vegetables commercialized in street- and supermarkets in Aparecida de Goiânia, Goiás, Brazil / Detecção de enteroparasitas em hortaliças comercializadas em feiras e supermercados em Aparecida de Goiânia, Goiás, Brasil

Foliar vegetables contaminated with fecal residues are an important route of transmission of intestinal parasites to humans. The aim of this study was to evaluate the presence of parasitic forms of protozoa and helminths on lettuces (Lactuca sativa) and collard greens (Brassica oleracea) sold in street- and supermarkets in the city of Aparecida de Goiânia, Goiás, Brazil. A total of 30 samples of each vegetable (15 samples from each supermarkets and street markets) was analyzed. All samples were processed by spontaneous sedimentation method and centrifugal flotation. In 45% of the samples, immature forms of intestinal parasites were identified with 66.7% helminths eggs and 33.3% protozoan cysts or oocysts. Significantly more lettuce samples were contaminated with eggs, cysts or oocyst of at least one parasite than collard green samples (U=216; Z=-3.45; P <0.001). The parasitic forms were identified morphologically up to the family level with eggs of Ancylostomatidae, Strongyloididae, Ascarididae and Taeniidae, or oocysts of Eimeriidae, to the genus with Cystoisospora sp. and Toxocara sp., and to the species level with Cystoisospora canis, Dipylidium caninum and Hymenolepis nana. The presence of these infective agents in lettuce and collard green from both street- and supermarkets highlights the high risk of spreading parasites by eating raw vegetables sold in Aparecida de Goiânia.

Hortaliças contaminadas com resíduos fecais são importantes vias de transmissão de parasitas intestinais ao homem. O objetivo deste estudo foi avaliar a presença de formas parasitárias de protozoários e helmintos em alface (Lactuca sativa) e couve (Brassica oleracea) vendidas em feiras e supermercados na cidade de Aparecida de Goiânia, Goiás, Brasil. Um total de 30 amostras de cada vegetal (15 amostras de supermercados e 15 de feiras livres) foi analisado. Todas as amostras foram processadas pelos métodos de sedimentação espontânea e centrífugo-flutuação. Em 45% das amostras foram identificadas formas imaturas de parasitas intestinais sendo 66,7% ovos de helmintos e 33,3% de cistos ou oocistos de protozoários. Significativamente, mais amostras de alface estavam contaminadas com ovos, cistos ou oocistos de pelo menos um parasita do que as amostras de couve (U = 216; Z = -3,45; P <0,001). As formas parasitárias foram identificadas morfologicamente ao nível de família com ovos de Ancylostomatidae, Strongyloididae, Ascarididae e Taeniidae ou oocistos de Eimeriidae, dos gêneros Cystoisospora sp. e Toxocara sp., e ao nível de espécies com Cystoisospora canis, Dipylidium caninum e Hymenolepis nana. A presença desses agentes infecciosos em alface e couve, provenientes tanto de feiras quanto de supermercados, ressalta o alto risco de veiculação de parasitas pela ingestão de hortaliças cruas comercializadas em Aparecida de Goiânia.

Application of exogenous salicylic acid reduces disease severity of Plasmodiophora brassicae in pakchoi (Brassica campestris ssp. chinensis Makino).

Clubroot is one of the most serious diseases affecting Brassicaceae plants worldwide. However, there is no effective control method for clubroot. Salicylic acid (SA) is a plant hormone that plays a critical role in plant defense. In our study, we found the disease severity of a clubroot-sensitive cultivar of pakchoi, Xinxiaqing, was reduced with 0.6mM exogenous SA after the infection of P. brassicae. To investigate the mechanism of SA-reduced disease severity against clubroot, then we analyzed the plant growth, alteration of antioxidant enzyme system, and related gene expression of Xinxiaqing. Results showed that the clubroot incidence rate and disease index were decreased after being treated with 0.6 mM exogenous SA. Furthermore, plant growth, reactive oxygen species (ROS) contents, and membrane lipid peroxidation were changed. The activities of antioxidant enzymes, including superoxide dismutase (SOD), ascorbic acid-peroxidase (APX), catalase (CAT), and glutathione reductase (GR), were increased. Additionally, the production rates of malondialdehyde (MDA), hydrogen peroxide (H2O2), and superoxide anion (O2·-) were also inhibited. The expression levels of genes, encoding SOD, APX, CAT, and GR, were increased. By summering all results, we conclude that 0.6 mM SA contributes to the reduction of disease severity to clubroot by increasing the activities of antioxidant enzymes, abilities of osmotic regulation, and ROS scavenging to reduce the clubroot-induced damage in pakchoi.

Assessing the Single and Combined Toxicity of Chlorantraniliprole and Bacillus thuringiensis (GO33A) against Four Selected Strains of Plutella xylostella (Lepidoptera: Plutellidae), and a Gene Expression Analysis.

Concerns about resistance development to conventional insecticides in diamondback moth (DBM) Plutella xylostella (L.), the most destructive pest of Brassica vegetables, have stimulated interest in alternative pest management strategies. The toxicity of Bacillus thuringiensis subsp. aizawai (Bt GO33A) combined with chlorantraniliprole (Chl) has not been documented. Here, we examined single and combined toxicity of chlorantraniliprole and Bt to assess the levels of resistance in four DBM strains. Additionally, enzyme activities were tested in field-original highly resistant (FOH-DBM), Bt-resistant (Bt-DBM), chlorantraniliprole-resistant (CL-DBM), and Bt + chlorantraniliprole-resistant (BtC-DBM) strains. The Bt product had the highest toxicity to all four DBM strains followed by the mixture of insecticides (Bt + Chl) and chlorantraniliprole. Synergism between Bt and chlorantraniliprole was observed; the combination of Bt + (Bt + Chl) (1:1, LC50:LC50) was the most toxic, showing a synergistic effect against all four DBM strains with a poison ratio of 1.35, 1.29, 1.27, and 1.25. Glutathione S-transferase (GST) and carboxyl-esterase (CarE) activities showed positive correlations with chlorantraniliprole resistance, but no correlation was observed with resistance to Bt and Bt + Chl insecticides. Expression of genes coding for PxGST, CarE, AChE, and MFO using qRT-PCR showed that the PxGST and MFO were significantly overexpressed in Bt-DBM. However, AChE and CarE showed no difference in the four DBM strains. Mixtures of Bt with chlorantraniliprole exhibited synergistic effects and may aid the design of new combinations of pesticides to delay resistance in DBM strains substantially.

Clubroot in Brassica: recent advances in genomics, breeding, and disease management.

Clubroot disease, caused by Plasmodiophora brassicae, affects Brassica oilseed and vegetable production worldwide. This review is focused on various aspects of clubroot disease and its management, including understanding the pathogen and resistance in the host plants. Advances in genetics, molecular biology techniques, and omics research have helped to identify several major loci, QTL, and genes from the Brassica genomes involved in the control of clubroot resistance. Transcriptomic studies have helped to extend our understanding of the mechanism of infection by the pathogen and the molecular basis of resistance/susceptibility in the host plants. A comprehensive understanding of the clubroot disease and host resistance would allow developing a better strategy by integrating the genetic resistance with cultural practices to manage this disease from a long-term perspective.

The botanical eurycomanone is a potent growth regulator of the diamondback moth.

Eurycomanone is a quassinoid compound that is derived from Eurycoma longifolia, and it is often used as an indicator to evaluate the active ingredients of Eurycoma longifolia. However, Eurycomanone has rarely been reported to have biological activity toward pests. In this study, we evaluated the antifeedant activity of eurycomanone against the diamondback moth(Plutella xylostella), with a non-selective AFC50(the concentration that corresponds to 50% antifeedant action) value and selective AFC50 of 17.5 mg/L and 14.2 mg/L, respectively, which were 2.1-fold (36.9 mg/L) and 2-fold (28.5 mg/L) lower than that of azadirachtin, respectively. In addition, eurycomanone was used to treat the roots of Brassica chinensis L. at a concentration of 100 µg/g for 72 h. The antifeedant index was found to reach 93% by tracking the leaves. After feeding with 20 µg/g eurycomanone, no pupae or eclosion were observed. To explore this mechanism, we used scanning electron microscopy to discover that eurycomanone could prevent the development of taste receptors on the maxillary palp of diamondback moth larvae. Additional electrophysiological measurements showed that eurycomanone exhibited excitatory action to the central taste neurons of diamondback moth and significantly inhibited the GABAA receptor current. Eurycomanone exhibited significant activity as an antifeedant, inhibited growth and excelled at systemic absorption.

Exogenous inoculation of endophytic bacterium Bacillus cereus suppresses clubroot (Plasmodiophora brassicae) occurrence in pak choi (Brassica campestris sp. chinensis L.).

MAIN CONCLUSION: The presence of Bacillus cereus plays a key role in clubroot suppression and improves plant biomass in pak choi. B. cereus is reported for the first time as a novel biocontrol agent against clubroot. Plasmodiophora brassicae Woronin causes a devastating infectious disease known as clubroot that is damaging to cruciferous vegetables. This study aimed to isolate beneficial bacteria from the rhizosphere soil of pak choi (Brassica campestris sp. chinensis) and to evaluate the ability of the isolate to reduce the severity of clubroot. Strains obtained from the rhizosphere of symptomless pak choi were first selected on the basis of their germination inhibition rate and effects on the viability of P. brassicae resting spores. Eight bacterial isolates had inhibitory effects against the resting spores of clubroot causing pathogen. However, MZ-12 showed the highest inhibitory effect at 73.4%. Inoculation with MZ-12 enhanced the plant biomass relative to plants grown without MZ-12 as well as P. brassicae infected plants. Furthermore, enhanced antioxidant enzymatic activities were observed in clubroot-infected plants during bacterial association. Co-inoculation of the plant with both P. brassicae and MZ-12 resulted in a 64% reduction of gall formation in comparison to plants inoculated with P. brassicae only. Three applications of MZ-12 to plants infected with P. brassicae at 7, 14 and 21 days after seeding (DAS) were more effective than one application and repressed root hair infection. According to 16S rDNA sequence analysis, strain MZ-12 was identified as had a 100% sequence similarity with type strain Bacillus cereus. The findings of the present study will facilitate further investigation into biological mechanisms of cruciferous clubroot control.

Root-associated entomopathogenic fungi manipulate host plants to attract herbivorous insects.

Root-associated entomopathogenic fungi (R-AEF) indirectly influence herbivorous insect performance. However, host plant-R-AEF interactions and R-AEF as biological control agents have been studied independently and without much attention to the potential synergy between these functional traits. In this study, we evaluated behavioral responses of cabbage root flies [Delia radicum L. (Diptera: Anthomyiidae)] to a host plant (white cabbage cabbage Brassica oleracea var. capitata f. alba cv. Castello L.) with and without the R-AEF Metarhizium brunneum (Petch). We performed experiments on leaf reflectance, phytohormonal composition and host plant location behavior (behavioral processes that contribute to locating and selecting an adequate host plant in the environment). Compared to control host plants, R-AEF inoculation caused, on one hand, a decrease in reflectance of host plant leaves in the near-infrared portion of the radiometric spectrum and, on the other, an increase in the production of jasmonic, (+)-7-iso-jasmonoyl-L-isoleucine and salicylic acid in certain parts of the host plant. Under both greenhouse and field settings, landing and oviposition by cabbage root fly females were positively affected by R-AEF inoculation of host plants. The fungal-induced change in leaf reflectance may have altered visual cues used by the cabbage root flies in their host plant selection. This is the first study providing evidence for the hypothesis that R-AEF manipulate the suitability of their host plant to attract herbivorous insects.

Genome-Wide Identification and Expression Analysis of Udp-Glucuronosyltransferases in the Whitefly Bemisia Tabaci (Gennadius) (HemipterA: Aleyrodidae).

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is an important agricultural pest worldwide. Uridine diphosphate (UDP)-glucuronosyltransferases (UGTs) are one of the largest and most ubiquitous groups of proteins. Because of their role in detoxification, insect UGTs are attracting increasing attention. In this study, we identified and analyzed UGT genes in B. tabaci MEAM1 to investigate their potential roles in host adaptation and reproductive capacity. Based on phylogenetic and structural analyses, we identified 76 UGT genes in the B. tabaci MEAM1 genome. RNA-seq and real-time quantitative PCR (RT-qPCR) revealed differential expression patterns of these genes at different developmental stages and in association with four host plants (cabbage, cucumber, cotton and tomato). RNA interference results of selected UGTs showed that, when UGT352A1, UGT352B1, and UGT354A1 were respectively silenced by feeding on dsRNA, the fecundity of B. tabaci MEAM1 was reduced, suggesting that the expressions of these three UGT genes in this species may be associated with host-related fecundity. Together, our results provide detailed UGTs data in B.tabaci and help guide future studies on the mechanisms of host adaptation by B.tabaci.

Population viability in a host-parasitoid system is mediated by interactions between population stage structure and life stage differential susceptibility to toxicants.

The effects of toxicants, such as pesticides, may be more severe for some life stages of an organism than others. However, in most toxicity studies, data is developed for only one life stage, which may lead to misleading interpretations. Furthermore, population stage-structure may interact with differential susceptibility, especially when populations consist of higher proportions of individuals in more susceptible stages at the time of toxicant exposure. We explore the interaction of differential stage susceptibility and stage distribution using a stage-structured Lefkovitch matrix model. We incorporate lab-derived toxicity data for a common parasitoid, the braconid Diaeretiella rapae (M'Intosh), a common natural enemy of the cabbage aphid (Brevicoryne brassicae L.), exposed to the pesticide imidacloprid. We compare population outcomes of simulations in which we vary both the population stage structure along with the susceptibility of each stage to toxicants. Our results illustrate an interaction between differential susceptibility and initial stage distribution, highlighting the fact that both of these demographic features should be considered in interpreting toxicity data and the development of ecological risk assessments.

Demystifying biotrophs: FISHing for mRNAs to decipher plant and algal pathogen-host interaction at the single cell level.

Plant-pathogen interactions follow spatial and temporal developmental dynamics where gene expression in pathogen and host undergo crucial changes. Therefore, it is of great interest to detect, quantify and localise where and when key genes are active to understand these processes. Many pathosystems are not accessible for genetic amendments or other spatially-resolved gene expression monitoring methods. Here, we adapt single molecule FISH techniques to demonstrate the presence and activity of mRNAs at the single-cell level using phytomyxids in their plant and algal host in lab and field material. This allowed us to monitor and quantify the expression of genes from the clubroot pathogen Plasmodiophora brassicae, several species of its Brassica hosts, and of several brown algae, including the genome model Ectocarpus siliculosus, infected with the phytomyxid Maullinia ectocarpii. We show that mRNAs are localised along a spatiotemporal gradient, thus providing a proof-of-concept of the usefulness of single-molecule FISH to increase knowledge about the interactions between plants, algae and phytomyxids. The methods used are easily applicable to any interaction between microbes and their algal or plant host, and have therefore the potential to rapidly increase our understanding of key, spatially- and temporally-resolved processes underpinning complex plant-microbe interactions.

Genome-wide analysis of diamondback moth, Plutella xylostella L., from Brassica crops and wild host plants reveals no genetic structure in Australia.

Molecular studies of population structure can reveal insight into the movement patterns of mobile insect pests in agricultural landscapes. The diamondback moth, Plutella xylostella L., a destructive pest of Brassica vegetable and oilseed crops worldwide, seasonally colonizes winter canola crops in southern Australia from alternative host plant sources. To investigate movement, we collected 59 P. xylostella populations from canola crops, Brassica vegetable and forage crops and brassicaceous wild host plants throughout southern Australia in 2014 and 2015 and genotyped 833 individuals using RAD-seq for genome-wide analysis. Despite a geographic sampling scale > 3,000 km and a statistically powerful set of 1,032 SNP markers, there was no genetic differentiation among P. xylostella populations irrespective of geographic location, host plant or sampling year, and no evidence for isolation-by-distance. Hierarchical STRUCTURE analysis at K = 2-5 showed nearly uniform ancestry in both years. Cluster analysis showed divergence of a small number of individuals at several locations, possibly reflecting an artefact of sampling related individuals. It is likely that genetic homogeneity within Australian P. xylostella largely reflects the recent colonization history of this species but is maintained through some level of present gene flow. Use of genome-wide neutral markers was uninformative for revealing the seasonal movements of P. xylostella within Australia, but may provide more insight in other global regions where the species has higher genetic diversity.

Induced tolerance to abiotic and biotic stresses of broccoli and Arabidopsis after treatment with elicitor molecules.

The plant hormones salicylic acid (SA) and jasmonic acid (JA) regulate defense mechanisms capable of overcoming different plant stress conditions and constitute distinct but interconnected signaling pathways. Interestingly, several other molecules are reported to trigger stress-specific defense responses to biotic and abiotic stresses. In this study, we investigated the effect of 14 elicitors against diverse but pivotal types of abiotic (drought) and biotic (the chewing insect Ascia monuste, the hemibiotrophic bacterium Pseudomonas syringae DC 3000 and the necrotrophic fungus Alternaria alternata) stresses on broccoli and Arabidopsis. Among the main findings, broccoli pre-treated with SA and chitosan showed the highest drought stress recovery in a dose-dependent manner. Several molecules led to increased drought tolerance over a period of three weeks. The enhanced drought tolerance after triggering the SA pathway was associated with stomata control. Moreover, methyl jasmonate (MeJA) reduced A. monuste insect development and plant damage, but unexpectedly, other elicitors increased both parameters. GUS reporter assays indicated expression of the SA-dependent PR1 gene in plants treated with nine elicitors, whereas the JA-dependent LOX2 gene was only expressed upon MeJA treatment. Overall, elicitors capable of tackling drought and biotrophic pathogens mainly triggered the SA pathway, but adversely also induced systemic susceptibility to chewing insects. These findings provide directions for potential future in-depth characterization and utilization of elicitors and induced resistance in plant protection.

Alyssum (Lobularia maritima) selectively attracts and enhances the performance of Cotesia vestalis, a parasitoid of Plutella xylostella.

The use of nectar-providing plants to nourish natural enemies of pest species has become a widely-used approach in conservation biological control to reduce pest damage without the indiscriminate use of insecticides. Choice of plant species is crucial to maximize benefits, but suitable species are yet to be identified for many important crop-pest systems. Here we explored the suitability of three candidate nectar plants for use in brassica vegetables to suppress the globally significant pest, Plutella xylostella L. (Lepidoptera: Plutellidae), using the widely-distributed parasitoid, Cotesia vestalis (Haliday) (Hymenoptera: Braconidae). Volatiles of alyssum (Lobularia maritima (L.) Desv) (Brassicaceae) were attractive to the parasitoid and access to flowering shoots increased adult longevity and realized fecundity of C. vestalis. Moreover, adult diamondback moth derived no benefit from this flower. In contrast, buckwheat (Fagopyrum esculentum Moench) (Polygonaceae), a species widely used in conservation biological control in other systems, increased the longevity and fecundity of both pest and parasitoid, rendering it less suitable. A third plant, heronsbill (Portulaca grandiflora Hook.) (Portulacaceae) denied benefit to the pest and promoted longevity of the parasitoid under no-choice conditions but did not improve fecundity and was repellent to female parasitoids under choice conditions. The contrasting effects of this set of plants illustrate the need to test multiple response variables and effects on both pest and natural enemy when seeking optimal nectar plants for use in a novel conservation biological control system.

Host plant nutrient contents influence nutrient contents in Bradysia cellarum and Bradysia impatiens.

The chive maggot Bradysia cellarum and the fungus gnat B. impatiens are two primary root pests of plants, which can coexist on the same host plants and are the devastating pests on liliaceous crops and edible fungi. Their growth and development are affected by the nutrient contents of their host plants. In this study, we assessed the effects of different host plant nutrients on the nutrient contents of these two Bradysia species. The nutrients of the chive (Allium tuberosum Rottl. ex Spreng.), board bean (Vicia faba L.), lettuce (Lactuca sativa L. var. ramosa Hort.), cabbage (Brassica oleracea L.), wild cabbage (Brassica oleracea var. capitata rubra) and pepper (Capsicum annuum L.) roots were determined, and their effect on nutrient content of the two Bradysia species after feeding on the host plant for three continuous generations were evaluated. The results show that chive and B-bean contained higher levels of protein, free amino acid, soluble sugar and starch than others. As a result, the soluble sugar, fat and protein contents were significantly higher in both Bradysia species reared on chive and B-bean than on cabbage, lettuce, W-cabbage and pepper, suggesting nutritional preference of these insects. Based on our results, we concluded that the two Bradysia species displayed nutrient preference toward chive and B-bean, which provides a reference for understanding their host plant range and for control of the insect species via field crop rotations.

Copitarsia decolora Guenée (Lepidoptera: Noctuidae) females avoid larvae competition by detecting larvae damaged plants.

Herbivory insects can discriminate the quality of a host plant for food or oviposition, by detecting the volatile organic compounds (VOC's) released by the plant, however, damaged plants may release a different VOC's profile modifying the insects' response. We tested if the VOC's profile from damaged plants affected the response of Copitarsia decolora as these moths oviposit preferably around undamaged host plants. We assessed the response in wind tunnel conditions of C. decolora mated females to volatiles collected by dynamic headspace from 30-40 d old cabbage undamaged plants and mechanical and larval damaged plants. Headspace volatile compounds from undamaged cabbage plants were more attractive to mated females than those from larval and mechanical damaged cabbage plants. Moths stimulated with headspace volatiles from undamaged plants performed more complete flight and ovipositor displays than those moths stimulated with headspace volatiles from damaged cabbage plants. A mixture of synthetic compounds identified from undamaged cabbages elicited similar antennal and wind tunnel responses in mated females as headspace volatiles from undamaged cabbage plants. C. decolora females may discriminate between damaged and undamaged host plants by detecting their VOC's profiles as a strategy to avoid unsuitable plants for their offspring increasing their fitness.

Myzus persicae (Hemiptera: Aphididae) infestation increases the risk of bacterial contamination and alters nutritional content in storage Chinese cabbage.

BACKGROUND: Aphids are common insect pests that feed on and excrete feces/honeydew on storage vegetables, especially in the temperate region of the northern hemisphere. The honeydew of aphids is an excellent growth medium for microorganisms. To explore the effects of aphid infestation on the risk of microbial contamination and food safety: (i) the bacterial diversity and community in aphid honeydew were investigated; (ii) the nutritional components of the cabbage were analyzed; and (iii) safety was evaluated. RESULTS: The results showed that the dominant bacteria in storage Chinese cabbage under different exposure times belonged to the phylum Proteobacteria, family Enterobacteriaceae. The richness of Enterobacteriaceae increased from 36.35% (1 day) to 39.70% (5 days) and to 50.74% (10 days) as the exposure time increased. Serratia was the genus with the highest abundance (23.38% for 1 day, 30.56% for 5 days and 37.85% for 10 days). The abundance of pathways associated with Staphylococcus aureus infection and Shigellosis increased significantly after prolonged storage. In addition, when the aphid density increased from 0 to 100 per 250 g of Chinese cabbage leaves, the protein content in Chinese cabbage decreased significantly, whereas the reducing sugar content increased significantly. CONCLUSION: These results demonstrate that the honeydew excreted by the green peach aphid Myzus persicae (Sulzer) on storage Chinese cabbage can serve as a medium for some foodborne disease pathogens. The present study may provide both a theoretical and practical basis for vegetable storage to reduce the risk of foodborne pathogen infection and to maintain the balance of nutrients. © 2020 Society of Chemical Industry.

Insecticidal activity and biochemical composition of Citrullus colocynthis, Cannabis indica and Artemisia argyi extracts against cabbage aphid (Brevicoryne brassicae L.).

Plant extracts contain many active compounds, which are tremendously fruitful for plant defence against several insect pests. The prime objectives of the present study were to calculate the extraction yield and to evaluate the leaf extracts of Citrullus colocynthis (L.), Cannabis indica (L.) and Artemisia argyi (L.) against Brevicoryne brassicae and to conduct biochemical analysis by gas chromatography-mass spectrometry (GC-MS). The results suggested that when using ethanol, C. colocynthis produced a high dry yield (12.45%), followed by that of C. indica and A. argyi, which were 12.37% and 10.95%, respectively. The toxicity results showed that A. argyi was toxic to B. brassicae with an LC50 of 3.91 mg mL-1, followed by the toxicity of C. colocynthis and C. indica, exhibiting LC50 values of 6.26 and 10.04 mg mL-1, respectively, which were obtained via a residual assay; with a contact assay, the LC50 values of C. colocynthis, C. indica and A. argyi were 0.22 mg mL-1, 1.96 and 2.87 mg mL-1, respectively. The interaction of plant extracts, concentration and time revealed that the maximum mortality based on a concentration of 20 mg L-1 was 55.50%, the time-based mortality was 55% at 72 h of exposure, and the treatment-based mortality was 44.13% for A. argyi via the residual assay. On the other hand, the maximum concentration-based mortality was 74.44% at 20 mg mL-1, the time-based mortality was 66.38% after 72 h of exposure, and 57.30% treatment-based mortality was afforded by A. argyi via the contact assay. The biochemical analysis presented ten constituents in both the A. argyi and C. colocynthis extracts and twenty in that of C. indica, corresponding to 99.80%, 99.99% and 97% of the total extracts, respectively. Moreover, the detected caryophylleneonides (sesquiterpenes), α-bisabolol and dronabinol (Δ9-THC) from C. indica and erucylamide and octasiloxane hexamethyl from C. colocynthis exhibited insecticidal properties, which might be responsible for aphid mortality. However, A. argyi was evaluated for the first time against B. brassicae. It was concluded that all the plant extracts possessed significant insecticidal properties and could be introduced as botanical insecticides after field evaluations.

The effect of combined application of Streptomyces rubrogriseus HDZ-9-47 with soil biofumigation on soil microbial and nematode communities.

Meloidogyne incognita causes significant damage to many different crops. Previous studies showed that Streptomyces rubrogriseus HDZ-9-47 is a promising biocontrol agent. Combining it with biofumigation improved its efficacy against M. incognita. In the present study, the reason for the improved efficacy of the combination was investigated by analyzing its impact on both the soil microbial and the nematode communities in the field. The results showed that the combined application reduced root galls by 41% and its control efficacy was greater than each treatment alone. Cultivation-based analyses showed that the combination treatment affected the soil microbial community. Actinomycetes and bacterial densities were negatively correlated with the root knot score. In contrast, the fungal densities were positively correlated with the root knot score. Denaturing gradient gel electrophoresis (DGGE) results showed that the combination of S. rubrogriseus HDZ-9-47 and biofumigation enriched beneficial microbes and reduced certain soil-borne fungal phytopathogens, thereby enhancing the efficacies of both S. rubrogriseus HDZ-9-47 and biofumigation against M. incognita. And HDZ-9-47 could colonize in soil. The total abundance of nematode and plant parasites, the ratio of soil fungivore nematode to fungivore plus bacterivore nematode, and the nematode diversity indices all decreased with the combination treatment. Overall, the results of this study demonstrate that combined application of HDZ-9-47 with biofumigation was a useful and effective approach to suppress M. incognita by manipulating soil microbial communities in field.