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Molecular mapping of dominant gene responsible for leaf curl virus resistance in chilli pepper (Capsicum annuum L.).

ABSTRACT: A resistant source (S-343) having monogenic dominant resistance to chilli leaf curl virus disease (ChiLCVD) has been identified at Punjab Agricultural University (PAU), Ludhiana. The F2 mapping population of 204 plants was derived from the cross MS-341 (susceptible) × S-343 (resistant) to identify the linked marker with the disease-resistant gene. Out of the 685 single-sequence repeats (SSRs) used, only 160 primers showed parental polymorphism. These 160 polymorphic primers were used for bulk segregant analysis and only eight SSR primers were able to differentiate the resistant and susceptible bulks. The linkage analysis revealed that the two markers CA 516044 and PAU-LC-343-1 were found linked with the disease-resistant gene covering a total distance of 15.7 centimorgan (cM). The two primers CA 516044 and PAU-LC-343-1 were found located on chromosome 6 of the pepper genome at a genetic distance of 6.8 cM and 8.9 cM, respectively, from the resistant gene. The validation of linked markers was performed using 26 resistant and susceptible genotypes developed at PAU, Ludhiana by former researchers. The validation of the primers revealed that there was a correlation between phenotypic and genotypic data of the used genotypes, and these markers can be used for the marker-assisted breeding procedures for transferring ChiLCVD resistance until the gene-based markers will be developed. The markers described in this study are the first-ever molecular markers identified as linked to the ChiLCVD-resistant gene.

SUBMITTER: Thakur H

PROVIDER: S-EPMC7103024 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundIn Oman tobacco (Nicotiana tabacum; family Solanaceae) is a minor crop, which is produced only for local consumption. In 2015, tobacco plants exhibiting severe downward leaf curling, leaf thickening, vein swelling, yellowing and stunting were identified in fields of tobacco in Suhar Al-Batina region, Oman. These symptoms are suggestive of begomovirus (genus Begomovirus, family Geminiviridae) infection.MethodsCircular DNA molecules were amplified from total DNA extracted from tobacco plants by rolling circle amplification (RCA). Viral genomes were cloned from RCA products by restriction digestion and betasatellites were cloned by PCR amplification from RCA product, using universal primers. The sequences of full-length clones were obtained by Sanger sequencing and primer walking. Constructs for the infectivity of virus and betasatellite were produced and introduced into plants by Agrobacterium-mediated inoculation.ResultsThe full-length sequences of 3 begomovirus and 3 betasatellite clones, isolated from 3 plants, were obtained. Analysis of the full-length sequences determined showed the virus to be a variant of Chilli leaf curl virus (ChiLCV) and the betasatellite to be a variant of Tomato leaf curl betasatellite (ToLCB). Both the virus and the betasatellite isolated from tobacco show the greatest levels of sequence identity to isolates of ChiLCV and ToLCB identified in other hosts in Oman. Additionally clones of ChiLCV and ToLCB were shown, by Agrobacterium-mediated inoculation, to be infectious to 3 Nicotiana species, including N. tabacum. In N. benthamiana the betasatellite was shown to change the upward leaf rolling symptoms to a severe downward leaf curl, as is typical for many monopartite begomoviruses with betasatellites.ConclusionsThe leaf curl disease of tobacco in Oman was shown to be caused by ChiLCV and ToLCB. This is the first identification of ChiLCV with ToLCB infecting tobacco. The study shows that, despite the low diversity of begomoviruses and betasatellites in Oman, the extant viruses/betasatellites are able to fill the niches that present themselves.

Project description:Endornaviruses include viruses that infect fungi, oomycetes, and plants. The genome of plant endornaviruses consists of linear ssRNA ranging in size from approximately 13-18 kb and lacking capsid protein and cell-to-cell movement capability. Although, plant endornaviruses have not been shown to cause detectable changes in the plant phenotype, they have been associated with alterations of the host physiology. Except for the association of cytoplasmic vesicles with infections by Vicia faba endornavirus, effects on the plant cell ultrastructure caused by endornaviruses have not been reported. Bell pepper endornavirus (BPEV) has been identified in several pepper (Capsicum spp.) species. We conducted ultrastructural analyses of cells from two near-isogenic lines of the bell pepper (C. annuum) cv. Marengo, one infected with BPEV and the other BPEV-free, and found cellular alterations associated with BPEV-infections. Some cells of plants infected with BPEV exhibited alterations of organelles and other cell components. Affected cells were located mainly in the mesophyll and phloem tissues. Altered organelles included mitochondrion, chloroplast, and nucleus. The mitochondria from BPEV-infected plants exhibited low number of cristae and electron-lucent regions. Chloroplasts contained plastoglobules and small vesicles in the surrounding cytoplasm. Translucent regions in thylakoids were observed, as well as hypertrophy of the chloroplast structure. Many membranous vesicles were observed in the stroma along the envelope. The nuclei revealed a dilation of the nuclear envelope with vesicles and perinuclear areas. The organelle changes were accompanied by membranous structure rearrangements, such as paramural bodies and multivesicular bodies. These alterations were not observed in cells from plants of the BPEV-free line. Overall, the observed ultrastructural cell alterations associated with BPEV are similar to those caused by plant viruses and viroids and suggest some degree of parasitic interaction between BPEV and the plant host.

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Molecular mapping of dominant gene responsible for leaf curl virus resistance in chilli pepper (Capsicum annuum L.).

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