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1.
Theor Appl Genet ; 134(9): 2947-2964, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34081151

RESUMO

KEY MESSAGE: A begomovirus resistance gene pepy-1, which encodes the messenger RNA surveillance factor Pelota, was identified in pepper (C. annuum) through map-based cloning and functional characterization. Pepper yellow leaf curl disease caused by begomoviruses seriously affects pepper (Capsicum spp.) production in a number of regions around the world. Ty genes of tomato, which confer resistance to the tomato yellow leaf curl virus, are the only begomovirus resistance genes cloned to date. In this study, we focused on the identification of begomovirus resistance genes in Capsicum annuum. BaPep-5 was identified as a novel source of resistance against pepper yellow leaf curl Indonesia virus (PepYLCIV) and pepper yellow leaf curl Aceh virus (PepYLCAV). A single recessive locus, which we named as pepper yellow leaf curl disease virus resistance 1 (pepy-1), responsible for PepYLCAV resistance in BaPep-5 was identified on chromosome 5 in an F2 population derived from a cross between BaPep-5 and the begomovirus susceptible accession BaPep-4. In the target region spanning 34 kb, a single candidate gene, the messenger RNA surveillance factor Pelota, was identified. Whole-genome resequencing of BaPep-4 and BaPep-5 and comparison of their genomic DNA sequences revealed a single nucleotide polymorphism (A to G) located at the splice site of the 9th intron of CaPelota in BaPep-5, which caused the insertion of the 9th intron into the transcript, resulting in the addition of 28 amino acids to CaPelota protein without causing a frameshift. Virus-induced gene silencing of CaPelota in the begomovirus susceptible pepper No.218 resulted in the gain of resistance against PepYLCIV, a phenotype consistent with BaPep-5. The DNA marker developed in this study will greatly facilitate marker-assisted breeding of begomovirus resistance in peppers.


Assuntos
Begomovirus/fisiologia , Capsicum/genética , Cromossomos de Plantas/genética , Resistência à Doença/imunologia , Genes Recessivos , Doenças das Plantas/imunologia , Proteínas de Plantas/metabolismo , Capsicum/crescimento & desenvolvimento , Capsicum/virologia , Mapeamento Cromossômico/métodos , Resistência à Doença/genética , Endonucleases/genética , Endonucleases/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/virologia , Proteínas de Plantas/genética
2.
Arch Virol ; 165(12): 3017-3021, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33037939

RESUMO

In 2017, a leaf sample from a single chili pepper (Capsicum annuum) plant exhibiting yellowing was collected from Aceh province, Indonesia. Total RNA was extracted from this sample, and RNA-Seq analysis was conducted. Putative infecting viruses were detected by mapping the obtained reads to the full-length viral genome sequences available in the GenBank database (7457 sequences) and the de novo-assembled contigs. RNA-Seq analysis detected polerovirus, begomovirus, and amalgavirus sequences, and the polerovirus-like sequences showed strong similarity to those of previously reported pepper vein yellows viruses (PeVYVs). The complete viral genome sequence obtained by RT-PCR had a length of 6023 nt, had the typical genome organization of a polerovirus and showed a high degree of sequence similarity to PeVYV-2 from Israel. Moreover, the predicted amino acid sequence of the P0 protein of the Indonesian isolate was 85.1% to 88.8% identical to those of other PeVYVs. In accordance with the polerovirus species demarcation criteria, this isolate should be assigned to a new polerovirus species, and we propose the name "pepper vein yellows virus 9" (PeVYV-9) for this virus.


Assuntos
Capsicum/virologia , Genoma Viral , Luteoviridae/classificação , Filogenia , Indonésia , Luteoviridae/isolamento & purificação , Doenças das Plantas/virologia , RNA Viral/genética , Recombinação Genética , Análise de Sequência de DNA
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