Graduate School of Agriculture, Kindai University, Nara, Nara, 631-8505, Japan.
Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata, 997-0017, Japan.
Theor Appl Genet. 2021 Sep;134(9):2947-2964. doi: 10.1007/s00122-021-03870-7. Epub 2021 Jun 3.
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 F 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.
通过图谱克隆和功能表征,从辣椒中鉴定出一个编码信使 RNA 监测因子 Pelota 的黄化曲叶病毒抗性基因 pepy-1。由黄化曲叶病毒引起的辣椒黄化曲叶病严重影响了全球许多地区的辣椒(Capsicum spp.)生产。目前已克隆的抗番茄黄化曲叶病毒的 Ty 基因是唯一的黄化曲叶病毒抗性基因。在这项研究中,我们专注于鉴定辣椒中的黄化曲叶病毒抗性基因。BaPep-5 被鉴定为一种对辣椒黄化曲叶印尼病毒(PepYLCIV)和辣椒黄化曲叶 Aceh 病毒(PepYLCAV)的新型抗性来源。在 BaPep-5 和感病 accession BaPep-4 杂交衍生的 F2 群体中,我们在第 5 号染色体上鉴定到一个单隐性基因座,负责 BaPep-5 对 PepYLCAV 的抗性,我们将其命名为 pepper yellow leaf curl disease virus resistance 1 (pepy-1)。在跨越 34 kb 的目标区域内,我们鉴定到一个单一的候选基因,即信使 RNA 监测因子 Pelota。对 BaPep-4 和 BaPep-5 的全基因组重测序,并比较它们的基因组 DNA 序列,发现 BaPep-5 中 CaPelota 的第 9 内含子剪接位点发生了单个核苷酸多态性(A 到 G),导致第 9 内含子插入到转录本中,使 CaPelota 蛋白增加了 28 个氨基酸,但没有导致移码。在感病辣椒品种 No.218 中,CaPelota 的病毒诱导基因沉默导致对 PepYLCIV 的抗性获得,这一表型与 BaPep-5 一致。本研究中开发的 DNA 标记将极大地促进辣椒中黄化曲叶病毒抗性的标记辅助选择。