Callosobruchus Chinensis Research Articles

Screening of Comprehensive Panel of Cultivated and Wild Vigna Species for Resistance to Pulse Beetle, Callosobruchus chinensis L.

Pulses are a key source of dietary proteins in human nutrition. Despite several efforts to increase the production, various constraints, such as biotic and abiotic factors, threaten pulse production by various means. Bruchids (Callosobruchus spp.) are the serious issue of concern, particularly in storage conditions. Understanding host-plant resistance at morphological, biochemical and molecular levels is the best way to minimize yield losses. The 117 mungbean (Vigna radiata L. Wilczek) genotypes, including endemic wild relatives, were screened for resistance against Callosobruchus chinensis; among them, two genotypes, PRR 2008-2 and PRR 2008-2-sel, which belong to V. umbellata (Thumb.), were identified as highly resistant. The expression of antioxidants in susceptible and resistant genotypes revealed that the activity of phenylalanine ammonia lyase (PAL) was upregulated in the highly resistant wild Vigna species and lower in the cultivated susceptible genotypes, along with other biomarkers. Further, the SCoT-based genotyping revealed SCoT-30 (200 bp), SCoT-31 (1200 bp) and SCoT-32 (300 bp) as unique amplicons, which might be useful for developing the novel ricebean-based SCAR markers to accelerate the molecular breeding programme.

Evaluation of Plant Powder/ Oil Against Pulse Beetle (<i>Callosobruchus chinensis</i> L.) on Green Gram

Six plant powders viz. neem leaf powder, neem seed powder, neem cake powder, curry leaf powder, chili fruit powder and bael leaf powder each at 5 g/ kg and mustard oil at 5 ml/ kg of powder grains were evaluated for their effectiveness as grain protectants against Callosobruchus chinensis on green gram in storage. The results revealed that mustard oil and neem seed powder were more effective. The order of efficacy of remaining treatments was neem cake > neem leaf powder > curry leaf powder > chili fruit powder > bael leaf powder which were significantly superior over control. It was also found that treatments do not affect the germination of seeds.

Varietal susceptibility of certain broad bean seeds to infestation with Callosobruchus maculatus (F.) and Callosobruchus chinensis (L.) (Coleoptera: Bruchidae)

Seeds susceptibility of eight broad bean varieties to Callosobruchus maculatus (F.) and Callosobruchus chinensis (L.) infestation were studied for the first time in free- and no-choice methods in the laboratory of Plant Protection Department, Faculty of Agriculture, Zagazig University. The relation between certain seed physical characteristics and some biological and and infestation parameters of both insects in the two studied methods were evaluated. None of these varieties were resistant to both insects, showing various levels of susceptibility. Biological and infestation parameters were significantly different among varieties except the developmental period. In free- choice method, Giza 3 was the most susceptible variety to both insects, since produced the highest progeny of 246.67 and 75.67 adults and susceptibility index (SI) of 10.25 and 7.42, respectively, while the least susceptible variety was Giza 716. In no– choice method, Nubaria 5 and Sakha 1 were the most susceptible varieties to C. chinensis, while Nubaria 3 and Giza 3 to C. maculatus. Differences between physical characters of varieties were significant. Seed hardness were correlated negatively and seed coat thickness positively with laid eggs, progeny and (SI) of both insects in free-choice method. Also seed coat thickness correlated positively with weight loss and seed damage (%) of C. chinensis and negatively of C. maculatus. To reduce seed losses the cultivation of the least susceptible variety (Giza 716) is encouraged and considered for breeding purposes to avoid insecticide usage.

ELUCIDATING THE HOST PREFERENCE BY THE PULSE BEETLE <i>CALLOSOBRUCHUS CHINENSIS</i> (L)

Callosobruchus chinensis (Bruchidae: Coleoptera) known as pulse beetle is a serious pest of pulses causing economic losses. The present work reports the effect of pulses on life cycle parameters (oviposition, emergence, longevity, total development period) of Callosobruchus chinensis (L.), and also on the nutritional loss incurred by it. Grains of seven pulses viz. Vigna radiata (green gram), Vigna aconitifolia (moth bean), Cicer arietinum (desi chickpea), Vigna unguiculata (cowpea), Cajanus cajan (pigeon pea), Vigna mungo (black gram) and Pisum sativum (yellow pea) were used as host with three replications. The results exhibit alterations in the developmental period as it was recorded least in green gram (23± 0.58 days) and maximum in peas (42± 1 days). The number of eggs were maximum on pea (310± 2) and least in moth (180± 2). Maximum longevity was recorded on green grams (16 days) and least on pea (9± 1 days). A significant correlation between weight loss and adult emergence was observed. Loss of nutritional content like protein and carbohydrate was significant.

Association of traits promoting resistance to the adzuki bean beetle, Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) in pea genotypes grown at various soil fertility levels

Adzuki bean beetle, Callosobruchus chinensis (L.) (Coleoptera: Bruchidae), is one of the most important pests of pea (Pisum sativum L.) crops in Ethiopia. The study focused on the association of resistance potential in the no-choice test of pea genotypes managed at different fertility levels and trait contributions. Based on the significance of fertility levels, genotypes were grouped into four, six, and five clusters, viz. Under neither rhizobium and phosphorus, rhizobium alone and rhizobium and phosphorus, respectively. Regardless of fertility levels, the inter-cluster distance (D2) values of the two potential clusters were highly significant (P < 0.01). At all fertility levels, the average performance of genotypes in each cluster for individual traits to infestation varied significantly. Genotype distribution patterns tended to group together into a small number of clusters. Eighty genotypes of the pea (Pisum sativum L. subsp. sativum and Pisum sativum L. subsp. abyssinicum A. Braun) were systematically managed under three fertility levels, and the first four principal components accounted for 94%, 92.3%, and 94.2% of the total variation. The primary trait that determines the resistance potential of pea genotypes is the trait susceptibility index (SI), which exhibits highly significant and adverse associations with critical traits such as the date of adult emergency and the percentage of seed coat, while exhibiting highly significant and favorable associations with the remaining traits at all fertility levels. The remaining characteristics showed highly significant positive or negative correlations within and particularly with the characteristics that determine resistance. Therefore, the cultivar "Adi" from "Pisum sativum L. subsp. sativum" had higher susceptibility compared to other genotypes, while the small-seeded pea genotypes "Pisum sativum L. subsp. abyssinicum A. Braun"; fpcoll-1/07, fpcoll-2/07, fpcoll-21/07, and fpcoll-43/07 were moderately resistant.

Post-fumigation sub-lethal activities of phosphine and ethyl formate on survivorship, fertility and female sex pheromone production of Callosobruchus chinensis (L.)

Phosphine (PH3) and ethyl formate (EF), the two popular fumigant disinfectants of stored product insect pests, are primarily evaluated for their knock down effects without considering their post-fumigation sub-lethal activities. The sub-lethal activities (adult survivorship, fecundity, sterility and female sex pheromone production) of the fumigants were evaluated on a field-to-storage insect pest adzuki bean beetle, Callosobruchus chinensis (L.). The adults’ survivorship and female fecundity, both were dose-dependently affected by sub-lethal PH3 and EF fumigation exposures. Hatchability of the eggs laid by fumigated female adults were also significantly affected. Gas-chromatography mass-spectrometry analysis of solid-phase micro-extraction from virgin fumigated C. cinensis females revealed that the PH3 LC25 (the lethal concentration required to kill the 25% of the population) fumigated female C. chinensis released significantly less amount of the pheromone components. In contrast, EF LC25 exposure did not affect the pheromone release. This study unveils the facts that the EF and PH3 fumigation have detrimental bioactivities against C. chinensis. Notably, this suggests to consider the sub-lethal EF and PH3 fumigation rather than the dose required to instantly kill all the C. chinensis individuals for disinfestation of stored adzuki bean.

Bioefficacy of eight different plant powders applied as fumigants against the adzuki bean weevil, Callosobruchuschinensis

The adzuki bean weevil, Callosobruchus chinensis (L.), is a highly destructive pest of stored legumes. This study aims to evaluate the bioefficacy of eight plant powders applied as fumigants against C. chinensis. Among the evaluated plant powder fumigants, clove, holy basil, lemongrass, and turmeric powder fumigant elicited high mortality up to 100% of C. chinensis and simultaneously showed high oviposition deterrence and F1 progeny inhibition against the pest, which led to a lower percentage of bean damage and weight loss. The antifeedant activity of the plant powder fumigants is reflected by the high feeding deterrence value of up to 82.92%. Additionally, based on a two-choice test assay, it is revealed that clove, holy basil, lemongrass, and turmeric powder fumigant also have adult repellent activity. Analysis of the volatile organic compounds of those four plant powder fumigants indicated that the major constituents responsible for the anti-pest feature were eugenol and caryophyllene for clove; estragole for holy basil; α-Citral and β-Citral for lemongrass; and α-zingiberene and β-sesquiphellandrene for turmeric. However, clove powder was proved to be the most efficacious fumigant in all parameters observed since it showed high bioefficacy even when applied at the lowest dose. Our findings highlight the potential use of plant powder fumigant as an eco-friendly and cost-effective management method of stored product pests, which could be a valuable option for small-hold farmers in developing countries.

Comparative transcriptome analysis of Callosobruchus chinensis (L.) (Coleoptera: Chrysomelidae-Bruchinae) after heat and cold stress exposure

Callosobruchus chinensis is regarded as one of the cosmopolitan pests of legume crops and can cause tremendous losses to a variety of beans. This study focused on comparative transcriptome analyses of C. chinensis exposed to 45 °C (heat stress), 27 °C (ambient temperature) and −3 °C (cold stress) for 3 h to investigate the gene differences and underlying molecular mechanisms. There were 402 and 111 differentially expressed genes (DEGs) identified in the heat and cold stress treatments, respectively. “cell process”, “cell” and “binding” were the main enriched functions and biological processes revealed by gene ontology (GO) analysis. The clusters of orthologous genes (COG) showed that DEGs were assigned to the categories: “posttranslational modification, protein turnover, chaperones”, “lipid transport and metabolism”, and “general function prediction only”. With respect to the Kyoto Encyclopedia of Genes and Genomes (KEGG), the “longevity regulating pathway-multiple species”, “carbon metabolism”, “peroxisome”, “protein processing in endoplasmic”, “glyoxylate and dicarboxylate metabolism” pathways were significantly enriched. The annotation and enrichment analysis revealed that genes encoding heat shock proteins (Hsps) and cuticular proteins were significantly upregulated under high and low-temperature stresses, respectively. In addition, some DEGs encoding “Protein lethal essential for life”, “Reverse transcriptase”, “DnaJ domain”, “Cytochrome” and “Zinc finger protein” were also upregulated to varying degrees. Transcriptomic data were validated using qRT‒PCR, which confirmed that they were consistent. In this paper, the temperature tolerance of C. chinensis adults was evaluated and the results showed that female adults were more sensitive to heat and cold stress than males, and the upregulation of heat shock protein and epidermal protein was the largest in DEGs after heat and cold stress, respectively. These findings provide a reference for further understanding the biological characteristics of C. chinensis adults and the molecular mechanisms underlying the response to low and high temperatures.

Response of different mung bean varieties against pulse beetle (Callosobruchus chinensis L.)

Response of different mung bean varieties against pulse beetle (Callosobruchus chinensis L.)

Female-limited responses in remating rate and mating duration in the experimental evolution of a beetle Callosobruchus chinensis.

Mating rate optima often differ between the sexes: males may increase their fitness by multiple mating, but for females multiple mating confers little benefit and can often be costly (especially in taxa without nuptial gifts or mala parental care). Sexually antagonistic evolution is thus expected in traits related to mating rates under sexual selection. This prediction has been tested by multiple studies that applied experimental evolution technique, which is a powerful tool to directly examine the evolutionary consequences of selection. Yet, the results so far only partly support the prediction. Here, we provide another example of experimental evolution of sexual selection, by applying it for the first time to the mating behaviour of a seed beetle Callsorobruchus chinensis. We found a lower remating rate in polygamy-line females than in monogamy-line (i.e. no sexual selection) females after 21 generations of selection. Polygamy-line females also showed a longer duration of first mating than monogamy-line females. We found no effect of male evolutionary lines on the remating rate or first mating duration. Though not consistent with the original prediction, the current and previous studies collectively suggest that the observed female-limited responses may be a norm, which is also consistent with the conceptual advances in the last two decades of the advantages and limitations of experimental evolution technique.

Differential fumigant and contact biotoxicities of biorational essential oil of Indian sweet basil and its active constituent against pulse beetle, Callosobruchus chinensis

Pulses are important source of nutrition. Safe and effective storage of pulses till consumption is a challenging task. Spoilage of pulses by invertebrate pests occurs at both pre and post-harvest conditions. Plant based food protectants is an emerging area of grain protection. This study assessed the differential biotoxicities of Indian sweet basil (Ocimum basilicum essential oil) and its major chemical constituent estragole against the pulse beetle, Callosobruchus chinensis. GC-MS analysis of O. basilicum essential oil (EO) showed estragole (69.77%) and linalool (18.29%) as the main chemical constituent. O. basilicum EO and estragole exhibited a strong fumigant and contact toxicity giving 100% mortality against C. chinensis within an exposure period of 24 h in both without and with food conditions. LC50 of O. basilicum EO and estragole in presence of grain, were 83.33 μL/L air and 185.66 μL/L air and the LD50 values were 0.81 μL/cm2 and 1.67 μL/cm2 respectively. Furthermore, SEM analysis of EO and estragole treated C. chinensis exhibited significant changes in topology of scales on elytron. In-vitro acetylcholinesterase inhibition of C. chinensis, was moderately higher in the treatment with estragole (43.01%) than O. basilicum EO (29.03%). Besides, a significant percent of oviposition deterrence effect was noticed in EO and estragole treated grains. Thus, Indian Ocimum chemotype (O. basilicum) and its active biological compound estragole showed effective potentiality as biorational insecticide for the control of pulse beetle. The main aim of the work was to develop biorational alternative for chemical pesticides by exploring the potential insecticidal activity of O. basilicum essential oil as an ecofriendly approach.

Insecticidal activities of long pepper (Piper retrofractum Vahl) fruit extracts against seed beetles (Callosobruchus maculatus Fabricius, Callosobruchus chinensis Linnaeus, and Sitophilus zeamais Motschulsky) and their effects on seed germination

Bruchid beetles (Callosobruchus maculatus and Callosobruchus chinensis), and maize weevil (Sitophilus zeamais) are important insect pests during the postharvest period. Botanical insecticide is an alternative solution for controlling these insects, and long pepper (Piper retrofractum) has been reported as having insecticidal potential against general insect pests. Film seed coatings with various concentrations of hexane extracts were made for mung bean (Vigna radiata) and corn (Zea mays) seeds. Insecticidal activities of these treatments were assessed at before and after storage period of six months, and seed germination was also evaluated. The hexane extract was subjected to analysis of the bioactive components by using Gas Chromatography-Mass Spectrometry (GC-MS). Results revealed that the hexane extract presented extreme toxicity to both bruchid beetles higher compared to maize weevil at 24 h with LC50 values of 5.57–6.75 and 58.04 μg⋅cm−2, respectively. Bruchid beetles presented significant response to ethanol, acetone and hexane extracts, whereas maize weevil showed relatively low responsibility. Film seed coating with hexane extract at 1% and 3% concentrations with six-month storage presented high insecticidal activity against bruchid beetles by more than 88% mortality but had low kill rates against maize weevil. The coated mung bean seeds presented non-seed germination effect, whereas high effect was observed on coated corn. Isolation of bioactive components demonstrated that there were 74 compounds, where pentadecane was the main compound. Film seed coating technology for mung bean seed preservation by using 1% hexane extract from long pepper fruit presented to be an extremely effective method to control bruchid beetles without any seed germination effect. It could serve as one of the green insecticides of the future.

Response of field pea genotypes to the infestation of Adzuki bean beetle (Callosobruchus chinensis L.) under different soil fertility regimes and locations in Ethiopia

Response of field pea genotypes to the infestation of Adzuki bean beetle (Callosobruchus chinensis L.) under different soil fertility regimes and locations in Ethiopia

Effect of Solarization against Bruchid Infestation and Seed Quality in Greengram (Vigna radiata L. Wildzek) During Storage

Among the storage insect pests the Pulse beetle, Callosobruchus chinensis is predominant storage pest of green gram which infests during pre and post-harvest stages. The ovipositional potential, adult emergence, seed damage decreased at above 35ºC. After exposing to solar heat, it is better to store the seeds in moisture vapour proof containers like polythene bags having 700 gauge to maintain the quality of seeds during storage. To know the effect of solarization cum packaging in a clear polyethylene (700 gauge) packets on the storability of seeds and bruchid management in greengram. A laboratory study was conducted during June 2018 to April 2019 at the Seed Research and Technology Centre, PJTSAU. The experiment was conducted with eight treatments with three replications. The data on seed germination percentage, seed moisture content, adult emergence and insect infestation were recorded at every three months upto nine months of storage and statistically analyzed by using completely randomized block design (CRD). Solarization of fresh seeds in clear polyethylene (700 gauge) packet for 4 h for 6 days was able to maintain per cent insect infestation (0.49%) below permissible limit and maintained germination percentage (90.67) &gt;IMSCS upto six months of storage as compared with 4.22 per cent seed damage and 83.33 per cent germination in untreated control (Fresh seeds). Seeds packed in a clear polyethylene (700 gauge) packet serves as a moisture vapour proof containers and able to reduce moisture by 22.16 per cent over the untreated control.

Evaluation of Cowpea [Vigna unguiculata (L) Walp.] Germplasm against Pulse Beetle, Callosobruchus chinensis (L.) and Correlation with Morphological Seed Characters

Background: In India, cowpea (Vigna unguiculata (L) Walp) is a minor pulse and is cultivated mainly in arid and semi arid tracts. Cowpea grains are often completely destroyed by bruchids both quantitatively and qualitatively making them unfit for planting, marketing and human consumption. Therefore, development and use of tolerant/resistant cowpea cultivars offer a simple, cheap and attractive way for reducing bruchids damage. Hence, the present study was aimed to identify new sources of insect resistant traits against Callosobruchus chinensis in cowpea germplasm so as to incorporate them in cowpea crop improvement programmes to evolve resistant or tolerant varieties against bruchids. Methods: Twenty seven cowpea genotypes were evaluated for resistance or susceptibility to pulse beetle, C. chinensis (Coleoptera: Bruchidae) using no-choice artificial conditions. Various parameters viz., oviposition, adult emergence/20 seeds, percent adult emergence, mean developmental period, per cent seed weight loss and growth index were determined for each genotype. Result: Significant differences were observed among cowpea accessions in terms of growth index (GI) of bruchid. Based on GI, one accession (IC 257844) was found moderately resistant (MR). Although, cowpea accessions showed variation in physical seed characters viz., seed coat colour, shape, seed length, seed width, seed thickness and 100 seed weight, they could not exert significant influence on any of the insect biological characters. Based on the study, it was demonstrated that IC 257844 (categorized as moderately resistant) could be used as potential donor for the development of bruchid tolerant cultivars.

Control of Stored Agro-Commodity Pests Sitophilus granarius and Callosobruchus chinensis by Nitrogen Hypoxic Atmospheres: Laboratory and Field Validations

Given the complexity of the practical usage of controlled atmospheres for the protection of agro-commodities, several researchers have pointed out that there is not enough robust scientific documentation regarding the usage of inert gases for their widespread practical application. Therefore, this work evaluated various regimes of hypoxic and anoxic nitrogen atmospheres for the control of two key stored-product pests, in laboratory and under field conditions in silos. Sitophilus granarius and Callosobruchus chinensis were selected as the tested species since they are important pests of grain/rice or legumes in Europe and Asia. Under laboratory conditions, we tested nitrogen (N2) concentrations (from 95 to 100%) and exposure times (1–20 days) on the developmental stages of both pest species. In most developmental stages of S. granarius and C. chinensis, the shortest effective exposure was found for nitrogen concentration of 99%. Based on our laboratory tests, validation studies were subsequently carried out in semi-hermetic steel silos (25t) using continuous nitrogen saturation by on-site built swing pressure generators. It was found that a full control of all stages of S. granarius and C. chinensis was achieved in 11 days of nitrogen exposure, using concentrations ranging above 99% and below 100%. Our work shows that hypoxic nitrogen treatment can be effectively achieved in small steel silos under proper technological and environmental conditions.

Chemical characterisation, insecticidal and antioxidant activities of essential oils from four Citrus spp. fruit peel waste

Citrus peel is a major fruit waste generated during the processing of citrus fruits. We extracted and chemically characterised essential oil from four different species of citrus peel waste and evaluated its insecticidal, antioxidant and phytotoxic activities. GC-MS/MS characterisation revealed D-limonene as the major constituent of citrus peel waste essential oil. The essential oils from the four citrus peel wastes showed significant contact, fumigant and repellent activities against two key stored grain pests Tribolium castaneum (Herbst) and Callosobruchus chinensis (L.). DPPH, ABTS, and FRAP assays revealed the antioxidant potential of the citrus peel waste essential oil. The essential oil did not affect the germination of wheat seeds and revealed the safety of essential oil to germinating seeds. Hence, the essential oil extracted from citrus peel waste can be valorised and utilised as a botanical insecticide against T. castaneum and C. chinensis.

Assessment of Adzuki bean beetle (Callosobruchus chinensis) resistance in undamaged grains of bulk stored lentil

Assessment of Adzuki bean beetle (Callosobruchus chinensis) resistance in undamaged grains of bulk stored lentil

Efficacy of Some Botanical Powders as Grain Protectants Against Pulse Beetle (Callosobruchus chinensis L.) on Ricebean (Vigna umbellata Thunb.)

An experiment was conducted during January−June of 2019 and 2020 in the laboratory of the Department of Entomology, School of Agricultural Sciences and Rural Development, Nagaland University, Medziphema Campus, Dimpaur district, Nagaland, India to evaluate the efficacy of some botanical powders as grain protectants against pulse beetle, Callosobruchus chinensis (L.) on ricebean, Vigna umbellata (Thunb.). The experiment was conducted in a Completely Randomized Design (CRD) with 3 replications. A total of 7 plant materials viz., Azadirachta indica, Piper nigrum, Ocimum tenuiflorum, Eucalyptus globules, Allium sativum, Pongamia pinnata and Litsea citrata were evaluated for their effect on oviposition reduction, adult emergence, infestation and weight loss by C. chinensis. All the plant products were mixed with susceptible ricebean seeds @ 5% w/w, Malathion 5% dust @ 1% w/w was used as standard check and untreated seed was used as control. Among the botanical powder treatments, the highest reduction in oviposition was found in L. citrata followed by A. indica and P. nigrum. The % adult emergence was also lowest in L. citrata treatment (55.23%) compared to the other botanical treatments. L. citrata seed powder also provided effective protection up to 2 months of storage with an infestation of 16.12% and weight loss of 2.40%, respectively followed by P. nigrum seed powder with an infestation of 31.67% and weight loss of 6.93%, respectively. The least effective was found in P. pinnata leaf powder. The findings revealed the potential of L. citrata and P. nigrum as grain protectants against pulse beetle in storage.

Tandemly duplicated genes encoding polygalacturonase inhibitors are associated with bruchid (Callosobruchus chinensis) resistance in moth bean (Vigna aconitifolia)

Bruchids are stored-grain insect pests responsible for serious seed loss in legume crops. A previous study using an F2 population (F2OA) derived from a cross between wild moth-bean (Vigna aconitifolia [Jacq.] Maréchal) accession TN67 (resistant) and cultivated moth-bean accession ICPMO056 (susceptible) revealed that resistance to the azuki bean weevil (Callosobruchus chinensis L.) in TN67 was regulated by a single gene located in the major quantitative trait locus—qVacBrc2.1. In this study, qVacBrc2.1 was finely mapped and candidate genes in this locus were identified using F2OA and another large F2 population (F2NB) derived from the cross mentioned previously. In contrast to the previous study, segregation analysis in the F2NB population revealed that resistance against this pest was controlled by two genes. Furthermore, the addition of novel markers to qVacBrc2.1 and reanalysis of the QTL in the F2OA population demonstrated that qVacBrc2.1 constituted two linked QTLs—qVacBrc2.1-A and qVacBrc2.1-B. The presence of qVacBrc2.1-B was verified using the population F2NB. Comparative genomics using three Vigna spp. strongly suggested the presence of two tandemly duplicated genes, VacPGIP1 and VacPGIP2, which encoded polygalacturonase inhibitors (polygalacturonase-inhibiting proteins) as the candidates for conferring resistance, but only VacPGIP1 could be successfully cloned and sequenced. The alignment of VacPGIP1 coding sequences of TN67 and ICPMO056 revealed eight single nucleotide polymorphisms, three of which altered the amino-acid sequence of the predicted domains of polygalacturonase inhibitors in ICPMO056. Overall, these findings indicate that VacPGIP1 and VacPGIP2 regulated C. chinensis resistance in TN67.