Cucumber Mosaic Virus Resistance Research Articles

Transcriptional Comparison Reveals Differential Resistance Mechanisms between CMV-Resistant PBC688 and CMV-Susceptible G29.

The Cucumber mosaic virus (CMV) presents a significant threat to pepper cultivation worldwide, leading to substantial yield losses. We conducted a transcriptional comparative study between CMV-resistant (PBC688) and -susceptible (G29) pepper accessions to understand the mechanisms of CMV resistance. PBC688 effectively suppressed CMV proliferation and spread, while G29 exhibited higher viral accumulation. A transcriptome analysis revealed substantial differences in gene expressions between the two genotypes, particularly in pathways related to plant-pathogen interactions, MAP kinase, ribosomes, and photosynthesis. In G29, the resistance to CMV involved key genes associated with calcium-binding proteins, pathogenesis-related proteins, and disease resistance. However, in PBC688, the crucial genes contributing to CMV resistance were ribosomal and chlorophyll a-b binding proteins. Hormone signal transduction pathways, such as ethylene (ET) and abscisic acid (ABA), displayed distinct expression patterns, suggesting that CMV resistance in peppers is associated with ET and ABA. These findings deepen our understanding of CMV resistance in peppers, facilitating future research and variety improvement.

Integrated transmission electron microscopy and proteomic analyses reveal the cytoarchitectural response to cucumber mosaic virus infection in tobacco

Cucumber mosaic virus (CMV) causes huge economic losses to agriculture every year; thus, understanding the mechanism of plant resistance to CMV is imperative. In this study, an integrated analysis of transmission electron microscopy (TEM) observations and proteomic results was used to identify cytoarchitectural differences in Nicotiana tabacum cv. NC82 (susceptible) and cv. Taiyan 8 (T.T.8; resistant) following infection with CMV. The TEM observations showed that the structure of the chloroplasts and mitochondria was severely damaged at the late stage of infection in NC82. Moreover, the chloroplast stroma and mitochondrial cristae were reduced and disaggregated. However, in T.T.8, organelle structure remained largely intact Selective autophagy predominated in T.T.8, whereas non-selective autophagy dominated in NC82, resembling cellular disorder. Proteomic analysis of T.T.8 revealed differentially expressed proteins (DEPs) mostly associated with photosynthesis, respiration, reactive oxygen species (ROS) scavenging, and cellular autophagy. Biochemical analyses revealed that ROS-related catalase, autophagy-related disulfide isomerase, and jasmonic acid and antioxidant secondary metabolite synthesis-related 4-coumarate:CoA ligase (Nt4CL) exhibited different trends and significant differences in expression in the two cultivars after CMV inoculation. Furthermore, mutant phenotyping verified that reduced Nt4CL expression impaired resistance in T.T.8. The identified DEPs are crucial for maintaining intracellular homeostatic balance and likely contribute to the mechanism of CMV resistance in tobacco. These findings increase our understanding of plant cytological mechanisms conferring resistance to CMV infection.

Host Resistance to Virus Diseases Provides a Key Enabler towards Fast Tracking Gains in Grain Lupin Breeding.

Four lupin species, Lupinus angustifolius, L. albus, L. luteus, and L. mutabilis, are grown as cool-season grain legume crops. Fifteen viruses infect them. Two of these, bean yellow mosaic virus (BYMV) and cucumber mosaic virus (CMV), cause diseases that threaten grain lupin production. Phytosanitary and cultural control measures are mainly used to manage them. However, breeding virus-resistant lupin cultivars provides an additional management approach. The need to develop this approach stimulated a search for virus resistance sources amongst cultivated lupin species and their wild relatives. This review focuses on the progress made in optimizing virus resistance screening procedures, identifying host resistances to BYMV, CMV, and additional viral pathogen alfalfa mosaic virus (AMV), and the inclusion of BYMV and CMV resistance within lupin breeding programs. The resistance types found in different combinations of virus and grain lupin species include localized hypersensitivity, systemic hypersensitivity, extreme resistance, and partial resistance to aphid or seed transmission. These resistances provide a key enabler towards fast tracking gains in grain lupin breeding. Where studied, their inheritance depended upon single dominant genes or was polygenic. Although transgenic virus resistance was incorporated into L. angustifolius and L. luteus successfully, it proved unstable. Priorities for future research are discussed.

Novel microRNAs associated with the immune response to cucumber mosaic virus in hot pepper (Capsicum annuum L.)

MicroRNAs (miRNAs) are a category of short, highly conserved, noncoding RNAs that modulate various physiological activities correlated with plant growth, stress response, and virus-host interaction. However, the miRNAs associated with cucumber mosaic virus (CMV) resistance in hot pepper (Capsicum annuum L.) remain unexplored. The present work employed small RNA sequencing to compare CMV-infected pepper seedlings with the control seedlings at different time points. The analysis using miRDeep2 software identified 216 known and 34 novel miRNAs from the 12 pepper libraries generated in this study. Further research predicted 3914 targets for these novel miRNAs, of which 3502 were conserved miRNA target genes and 412 were novel miRNA target genes. Functional analysis revealed significant enrichment of these miRNA in ‘plant-pathogen interaction’, ‘ubiquitin-mediated proteolysis’, and ‘metabolic pathways’. Finally, quantitative RT-PCR validated the sRNA sequencing data. These findings set the groundwork for further studies on the CMV resistance mechanisms of pepper.

Identification of positive and negative regulators of antiviral RNA interference in Arabidopsis thaliana

Virus-host coevolution often drives virus immune escape. However, it remains unknown whether natural variations of plant virus resistance are enriched in genes of RNA interference (RNAi) pathway known to confer essential antiviral defense in plants. Here, we report two genome-wide association study screens to interrogate natural variation among wild-collected Arabidopsis thaliana accessions in quantitative resistance to the endemic cucumber mosaic virus (CMV). We demonstrate that the highest-ranked gene significantly associated with resistance from both screens acts to regulate antiviral RNAi in ecotype Columbia-0. One gene, corresponding to Reduced Dormancy 5 (RDO5), enhances resistance by promoting amplification of the virus-derived small interfering RNAs (vsiRNAs). Interestingly, the second gene, designated Antiviral RNAi Regulator 1 (VIR1), dampens antiviral RNAi so its genetic inactivation by CRISPR/Cas9 editing enhances both vsiRNA production and CMV resistance. Our findings identify positive and negative regulators of the antiviral RNAi defense that may play important roles in virus-host coevolution.

Unravelling cucumber resistance to several viruses via genome-wide association studies highlighted resistance hotspots and new QTLs.

The mapping and introduction of sustainable resistance to viruses in crops is a major challenge in modern breeding, especially regarding vegetables. We hence assembled a panel of cucumber elite lines and landraces from different horticultural groups for testing with six virus species. We mapped 18 quantitative trait loci (QTL) with a multiloci genome wide association studies (GWAS), some of which have already been described in the literature. We detected two resistance hotspots, one on chromosome 5 for resistance to the cucumber mosaic virus (CMV), cucumber vein yellowing virus (CVYV), cucumber green mottle mosaic virus (CGMMV) and watermelon mosaic virus (WMV), colocalizing with the RDR1 gene, and another on chromosome 6 for resistance to the zucchini yellowing mosaic virus (ZYMV) and papaya ringspot virus (PRSV) close to the putative VPS4 gene location. We observed clear structuring of resistance among horticultural groups due to plant virus coevolution and modern breeding which have impacted linkage disequilibrium (LD) in resistance QTLs. The inclusion of genetic structure in GWAS models enhanced the GWAS accuracy in this study. The dissection of resistance hotspots by local LD and haplotype construction helped gain insight into the panel's resistance introduction history. ZYMV and CMV resistance were both introduced from different donors in the panel, resulting in multiple resistant haplotypes at same locus for ZYMV, and in multiple resistant QTLs for CMV.

Genome scan for quantitative trait loci underlying cucumber mosaic virus resistance in Capsicum annuum ‘CA2106’

Genome scan for quantitative trait loci underlying cucumber mosaic virus resistance in Capsicum annuum ‘CA2106’

Pepper Crop Improvement Against Cucumber Mosaic Virus (CMV): A Review.

Cucumber mosaic virus (CMV) is a prevalent virus affecting the quality and yield of pepper, resulting in yield losses of greater than 80% during severe local epidemics. Cultural practices and the heavy use of agrochemicals are the most common control measures for CMV. Sources of resistance provide a practical reference and a basis for breeding for CMV resistance. Genetic factors underlying CMV resistance have been studied and advanced breeding lines and cultivars with improved resistance have been developed by traditional breeding methods. Additionally, QTLs or genes for CMV resistance have been identified and can be utilized for marker-assisted resistance breeding. This review focuses on status and prospect of CMV against different virus strains, host resistance, and its applied genetics. With the advent of novel technologies, more useful markers and precise approaches can facilitate the progress for improving CMV resistance in Capsicum.

Transcriptome analysis of Luffa cylindrica (L.) Roem response to infection with Cucumber mosaic virus (CMV)

Cucumber mosaic virus (CMV) can cause serious losses in Luffa cylindrica (L.) Roem. Chemical application to control CMV is ineffective and environmentally unfriendly. The development of resistant hybrids is the best way to control CMV disease. Elucidating the virus-host interaction of CMV and molecular basis underlying Luffa spp. resistance against CMV would undoubtedly facilitate breeding for resistance against CMV disease. Transcriptome sequencing was used to analyze differentially expressed genes (DEGs) caused by CMV infection. A total of 138,336 unigenes were assembled, and 74,525 unigenes were annotated. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that the three major enrichment pathways (according to the p-values) were flavonoid biosynthesis, sulfur metabolism, and photosynthesis. Genes involved in basal defenses, probably R genes, were determined to be related to CMV resistance. Using quantitative real-time PCR, we validated the differential expression of 8 genes. A number of genes associated with CMV resistance were found in this study. This study provides transcriptomic information regarding CMV-Luffa spp. interactions and will shed light on our understanding of host-virus interactions.

Single Amino Acid Substitutions in the Cucumber Mosaic Virus 1a Protein Induce Necrotic Cell Death in Virus-Inoculated Leaves without Affecting Virus Multiplication.

When Arabidopsis thaliana ecotype Col-0 was inoculated with a series of reassortant viruses created by exchanging viral genomic RNAs between two strains of cucumber mosaic virus (CMV), CMV(Y), and CMV(H), cell death developed in the leaves inoculated with reassortant CMV carrying CMV(H) RNA1 encoding 1a protein, but not in noninoculated upper leaves. In general, cell death in virus-infected plants is a critical event for virus survival because virus multiplication is completely dependent on host cell metabolism. However, interestingly, this observed cell death did not affect either virus multiplication in the inoculated leaves or systemic spread to noninoculated upper leaves. Furthermore, the global gene expression pattern of the reassortant CMV-inoculated leaves undergoing cell death was clearly different from that in hypersensitive response (HR) cell death, which is coupled with resistance to CMV. These results indicated that the observed cell death does not appear to be HR cell death but rather necrotic cell death unrelated to CMV resistance. Interestingly, induction of this necrotic cell death depended on single amino acid substitutions in the N-terminal region surrounding the methyltransferase domain of the 1a protein. Thus, development of necrotic cell death might not be induced by non-specific damage as a result of virus multiplication, but by a virus protein-associated mechanism. The finding of CMV 1a protein-mediated induction of necrotic cell death in A. thaliana, which is not associated with virus resistance and HR cell death, has the potential to provide a new pathosystem to study the role of cell death in virus–host plant interactions.

Cucumber mosaic virus resistance: Comparative proteomics of contrasting Cucumis sativus cultivars after long-term infection

Plant viruses are a significant threat to a wide range of host species, causing substantial losses in agriculture. Particularly, Cucumber mosaic virus (CMV) evokes severe symptoms, thus dramatically limiting yield. Activation of plant defense reactions is associated with changes in the cellular proteome to ensure virus resistance. Herein, we studied two cultivars of cucumber (Cucumis sativus) resistant host Heliana and susceptible host Vanda. Plant cotyledons were mechanically inoculated with CMV isolate PK1, and systemic leaves were harvested at 33 days post-inoculation. Proteome was profiled by ultrahigh-performance liquid chromatography and comprehensively quantified by ion mobility enhanced mass spectrometry. From 1516 reproducibly quantified proteins using a label-free approach, 133 were differentially abundant among cultivars or treatments by strict statistic and effect size criteria. Pigments and hydrogen peroxide measurements corroborated proteomic findings. Comparison of both cultivars in the uninfected state highlighted more abundant photosynthetic and development-related proteins in resistant cucumber cultivar. Long-term CMV infection caused worse preservation of energy processes and less robust translation in the susceptible cultivar. Contrary, compatible plants had numerous more abundant stress and defense-related proteins. We proposed promising targets for functional validation in transgenic lines: A step toward durable virus resistance in cucurbits and other crops. SignificanceSustainable production of crops requires an understanding of natural mechanisms of resistance/susceptibility to ubiquitous viral infections. We report original findings of comparative analysis of plant genotypes exposed to CMV. Deep discovery proteomics of resistant and susceptible cucumber cultivars, inoculated with widespread phytovirus, allowed to suggest several novel molecular targets for functional testing in plant protection strategies.

Transcription factor WRKY30 mediates resistance to Cucumber mosaic virus in Arabidopsis

WRKY transcription factors are key regulators in regulating abiotic or biotic stress response in Arabidopsis. Previous studies showed that WRKY30 expression was induced by oxidative stress treatment, fungal elicitor, SA and ABA. However, functions of WRKY30 on viral defense are not well studied. Here, we found that Arabidopsis WRKY DNA binding protein 30 (WRKY30) plays essential roles in regulating Cucumber mosaic virus (CMV) resistance. The expression of WRKY30 was induced by CMV infection and wrky30 mutant displayed more susceptibility (including higher oxidative damages, induced reactive oxygen species synthesis and more PSII photochemistry compromise), while WRKY30 overexpression plants (WRKY30OX) exhibited more resistance to CMV infection. Moreover BRs-induced CMV tolerance is partly dependent on WRKY30. And WRKY30 expression increased after BL treatment. All these demonstrated that WRKY30 works as a positive regulator in plant CMV resistance process.

Identification and characterisation of a glycine-rich RNA-binding protein as an endogenous suppressor of RNA silencing from Nicotiana glutinosa.

This study shows that NgRBP suppresses both local and systemic RNA silencing induced by sense- or double-stranded RNA, and the RNA binding activity is essential for its function. To counteract host defence, many plant viruses encode viral suppressors of RNA silencing targeting various stages of RNA silencing. There is increasing evidence that the plants also encode endogenous suppressors of RNA silencing (ESR) to regulate this pathway. In this study, using Agrobacterium infiltration assays, we characterized NgRBP, a glycine-rich RNA-binding protein from Nicotiana glutinosa, as an ESR. Our results indicated that NgRBP suppressed both local and systemic RNA silencing induced by sense- or double-stranded RNA. We also demonstrated that NgRBP could promote Potato Virus X (PVX) infection in N. benthamiana. NgRBP knockdown by virus-induced gene silencing enhanced PVX and Cucumber mosaic virus resistance in N. glutinosa. RNA immunoprecipitation and electrophoretic mobility shift assays showed that NgRBP bound to GFP mRNA, dsRNA rather than siRNA. These findings provide the evidence that NgRBP acts as an ESR and the RNA affinity of NgRBP plays the key role in its ESR activity. NgRBP responds to multiple signals such as ABA, MeJA, SA, and Tobacco mosaic virus infection. Therefore, it could participate in the regulation of gene expression under specific conditions.

Plant defense systems against cucumber mosaic virus: lessons learned from CMV–Arabidopsis interactions

To protect against infection, plants have evolved multi-layered defense systems such as RNA silencing, salicylic acid (SA)-mediated resistance, resistance (R) protein-conferred resistance and defense priming. Antiviral defense systems seem to act in a coordinated manner in response to viral infection, but the underlying mechanisms are unknown except in a limited number of specific virus–plant combinations. Cucumber mosaic virus (CMV) infection of Arabidopsis thaliana is a well-known sophisticated pathosystem often used to investigate antiviral defense reactions. On the basis of extensive studies on the interaction between CMV and A. thaliana, RNA silencing plays a key role in suppressing CMV infection in the host plant. However, several strains of CMV are able to overcome the RNA silencing system by producing an RNA silencing suppressor protein. If the virulent CMV strain is indeed able to overcome basal antiviral RNA silencing, host plants carrying the R gene exhibit a resistance response, generally accompanied by necrotic local lesions at the primary infection sites. RCY1, which encodes an NB-LRR class R protein, was isolated from A. thaliana and identified as the first R gene conferring resistance to CMV. SA-dependent and -independent signaling pathways also seem to be associated with CMV resistance, while in A. thaliana, resistance induced by a plant activator treatment is thought to activate defense priming against a broad range of pathogens. In this review, we summarize current knowledge on the multiple defense systems of A. thaliana against CMV infection.

Cucumber mosaic virus resistance and reproductive biology of brazilian melon accessions

Abstract In melon (Cucumis melo) the resistance to Cucumber Mosaic Virus (CMV) and the knowledge about reproductive characteristics are important for selection of genitors for breeding population. So, the objective was to investigate if different accessions of melon from a Brazilian germplasm bank have variability in resistance to CMV (FNY strain) and polymorphism in resistance loci markers, as well as whether there is genetic diversity on resistance and floral morphology descriptors and reproductive biology, aiming at genetic improvement. The symptoms induced by CMV were evaluated in 52 melon genotypes which were genotyped with three markers of single nucleotide polymorphisms related to CMV resistance. A sample of 24 genotypes was characterized for floral and reproductive descriptors. The significant variation in resistance levels allows classifying the accessions in five groups. The polymorphisms detected by the markers were independently distributed in these groups, consistent with the oligogenic quantitative expression of the melon’s resistance. There were significant differences between accessions in floral and reproductive descriptors. Through the Ward-MLM multivariate strategy, these accesses were distributed into seven groups with distinct reproductive characteristics, including variability in pollen viability. Therefore, there are Brazilian accessions of melon that have resistance to CMV, as well as variability in reproductive characteristics.

Identification of Cucumber mosaic resistance 2 (cmr2) That Confers Resistance to a New Cucumber mosaic virus Isolate P1 (CMV-P1) in Pepper (Capsicum spp.).

Cucumber mosaic virus (CMV) is one of the most devastating phytopathogens of Capsicum. The single dominant resistance gene, Cucumber mosaic resistant 1 (Cmr1), that confers resistance to the CMV isolate P0 has been overcome by a new isolate (CMV-P1) after being deployed in pepper (Capsicum annuum) breeding for over 20 years. A recently identified Indian C. annuum cultivar, “Lam32,” displays resistance to CMV-P1. In this study, we show that the resistance in “Lam32” is controlled by a single recessive gene, CMV resistance gene 2 (cmr2). We found that cmr2 conferred resistance to CMV strains including CMV-Korean, CMV-Fny, and CMV-P1, indicating that cmr2 provides a broad-spectrum type of resistance. We utilized two molecular mapping approaches to determine the chromosomal location of cmr2. Bulked segregant analysis (BSA) using amplified fragment-length polymorphism (AFLP) (BSA-AFLP) revealed one marker, cmvAFLP, located 16 cM from cmr2. BSA using the Affymetrix pepper array (BSA-Affy) identified a single-nucleotide polymorphism (SNP) marker (Affy4) located 2.3 cM from cmr2 on chromosome 8. We further screened a pepper germplasm collection of 4,197 accessions for additional CMV-P1 resistance sources and found that some accessions contained equivalent levels of resistance to that of “Lam32.” Inheritance and allelism tests demonstrated that all the resistance sources examined contained cmr2. Our result thus provide genetic and molecular evidence that cmr2 is a single recessive gene that confers to pepper an unprecedented resistance to the dangerous new isolate CMV-P1 that had overcome Cmr1.

Inheritance and QTL mapping of cucumber mosaic virus resistance in cucumber (Cucumis Sativus L.)

The commercial yield of cucurbit crops infected with Cucumber mosaic virus (CMV) severely decreases. Chemical treatments against CMV are not effective; therefore, genetic resistance is considered the primary line of defense. Here, we studied resistance to CMV in cucumber inbred line ‘02245’ using a recombinant inbred line (RIL) population generated from a cross between ‘65G’ and ‘02245’ as susceptible and resistant parents, respectively. Genetic analysis revealed that CMV resistance in cucumber is quantitatively inherited. Analysis of the RIL population revealed that a quantitative trait locus (QTL) was found on chromosome 6; named cmv6.1, this QTL was delimited by SSR9-56 and SSR11-177 and explained 31.7% of the phenotypic variation in 2016 and 28.2% in 2017. The marker SSR11-1, which is close to the locus, was tested on 78 different cucumber accessions and found to have an accuracy of 94% in resistant and moderately resistant lines but only 67% in susceptible lines. The mapped QTL was delimited within a region of 1,624.0 kb, and nine genes related to disease resistance were identified. Cloning and alignment of the genomic sequences of these nine genes between ‘65G’ and ‘02245’ revealed that Csa6M133680 had four single-base substitutions within the coding sequences (CDSs) and two single-base substitutions in its 3’-untranslated region, and the other eight genes showed 100% nucleotide sequence identity in their exons. Expression pattern analyses of Csa6M133680 in ‘65G’ and ‘02245’ revealed that the expression levels of Csa6M133680 significantly differed between ‘65G’ and ‘02245’ at 80 h after inoculation with CMV and that the expression in ‘02245’ was 4.4 times greater than that in ‘65G’. The above results provide insights into the fine mapping and marker-assisted selection in cucumber breeding for CMV resistance.

Construction of a high-density genetic map and identification of QTLs for cucumber mosaic virus resistance in pepper (Capsicum annuum L.) using specific length amplified fragment sequencing (SLAF-seq).

Pepper (Capsicum) is one of the most important vegetable and spice crops. Aphid-transmitted cucumber mosaic virus (CMV) causes significant damage to pepper crops across the world. The genetic basis of CMV resistance in pepper is complex, and the mechanisms underlying resistance remain largely unknown. Here, we employed a SLAF-seq approach to generate a high-density genetic map of pepper. The map spanned 1,785.46 cM, containing 12,727 markers on 12 chromosomes, with a mean marker distance of 0.16 cM between adjacent markers. We used this map and the interval mapping (IM) and multiple QTL mapping (MQM) procedures to detect genetic regions associated with quantitative trait for CMV resistance. Three QTLs, qcmv11.1, qcmv11.2 and qcmv12.1, conferred resistance to CMV and showed trait variation of 10.2%, 19.2% and 7.3% respectively. Our results will help to develop markers linked to CMV-resistant QTLs to improve pepper resistance to CMV.

Callus Induction and Differentiation on Melon From In Vitro Culture with The Addition of Indole Acetic Acid and Benzyl Amino Purine Growth Regulator

Melon plants (Cucumis melo L.) are susceptible to disease attacks primarily caused by viruses. One of the viruses often attacking melon plants is Cucumber mosaic virus (CMV). Previous studies have shown that melon callus produced from in vitro culture is detected to have CMV resistance gene (Creb-2). However, the trait of resistance to CMV in that melon (F2) is still segregated. To know the stability of the Creb-2 gene, further research is needed to detect the gene in the differentiated callus. This study aims to callus induce and callus differentiate melon. Firstly, melon seeds were grown on ¼ MS medium with addition of 2 mg / L 2.4-Dichlorophenoxyacetate and 1 mg / L BAP (Benzyl Amino Purine) to grow callus. Secondly, the callus was differentiated in MS medium with the addition of 2,4-D , BAP and IAA in different doses. 0 mg / L; 0.01 mg / L 2.4-D and 0.1 mg / L BAP, and MS medium with 0.01 mg / L; 0.1 mg / L; 1 mg / L IAA (Iodole Acetic Acid) and 0.1 mg / L BAP. The results showed that callus was successfully differentiated into root and it grew optimally in MS medium with 0.01 mg / L 2.4-D: 0.1 mg / L BAP and 1 mg / L IAA: 0.1 mg / L BAP . Therefore the roots can be used as isolation samples and DNA amplification to detect stability of the Creb-2 gene.

Tri-species bridge crosses (<i>C. annuum L.</i> x <i>C. chinense</i> Jacq.) x (<i>C. chinense</i> Jacq. x <i>C. frutescens</i> L.) as an alternative approach for introgression of Cucumber Mosaic Virus (CMV) and Chilli Veinal Mosaic Virus (CVMV) resistance

Cucumber mosaic virus (CMV) and Chilli Veinal Mosaic Virus (CVMV) are among the most destructive viruses affecting chilli crop in Sri Lanka. Identification of resistant sources and combining them in to cultivated forms is essential in resistance breeding. Capsicum frutescens L. has been reported as a source of variation for many different traits including disease resistance to improve chilli (Capsicum annum L.). However, strong inter-specific hybridization barriers exist between them. In the present study, wide hybridization approach for introgressing C. frutescens L. genes into C. annuum L. was performed through genetic bridging using C. Chinense Jacq. as a bridge species. Diverse collection of 115 accessions from three cultivated species of C. annuum L. (28), C. Chinense Jacq. (63) and C. frutescens L. (24) was screened for CMV and CVMV resistance. Two C. frutescens L. accessions were resistant to both viruses and six C. Chinense Jacq. Accessions were resistant to CVMV. In Genetic bridge approach three way hybrids and double crosses were produced among these three species. The double crosses [(C. annuum L. x C.chinense) x (C. Chinense Jacq.x C. frutescens L.)] and [(C. Chinense Jacq.x C. annuum L.) x (C. Chinense Jacq. x C. frutescens L.)] were more successful than the three way crosses when considering the combining of C. frutescens L. traits into C. annuum L. and development of resistance to CMV and CVMV.