Department of Agronomy, Faculty of Agriculture at Kampaheng Saen, Kasetsart University, Nakhon Pathom, 73140, Thailand.
Institute of Vegetable Crops, Jiangsu Academy of Agricultural Science, 50 Zhongling Street, Nanjing, 210014, China.
Theor Appl Genet. 2016 Sep;129(9):1673-83. doi: 10.1007/s00122-016-2731-1. Epub 2016 May 24.
The Br locus confers bruchid resistance in mungbean; VrPGIP2 (encoding a polygalacturonase inhibitor) is a strong candidate gene for this resistance. The VrPGIP2 sequence differs between resistant and susceptible lines. Azuki bean weevil (Callosobruchus chinensis) and cowpea weevil (Callosobruchus maculatus) are serious insect pests of mungbean during storage. Bruchid resistance in mungbean is controlled by a single dominant locus, Br. Although the Br locus has been located on a genetic map, molecular basis and function of the gene remain unknown. In this study, high-resolution mapping using a BC11F2 population of 418 plants derived from a cross between 'Kamphaeng Saen 1' (KPS1; susceptible) and 'V2802' (resistant) using simple sequence repeat (SSR) markers delimited the Br locus to a genomic region of 38 Kb of chromosome 5 containing two annotated genes. EST-SSR marker DMB-SSR158 co-segregated perfectly with the Br locus. Bioinformatics analyses revealed that DMB-SSR158 corresponds to a gene encoding a polygalacturonase inhibitor (polygalacturonase-inhibiting protein PGIP) and was designated as VrPGIP2. Comparison of VrPGIP2 coding sequences between four bruchid-resistant (V2802, V1128, V2817 and TC1966) and four bruchid-susceptible (KPS1, Sulu-1, CM and an unknown accession) mungbean lines revealed six single nucleotide polymorphisms (SNPs) between the resistant and susceptible groups. Three of the six SNPs resulted in amino acid changes; namely, alanine (A) to serine (S) at position 320, leucine (L) to proline (P) at position 332, and threonine (T) to P at position 335 of the VrPGIP2 sequence in resistant lines, compared with that in susceptible lines. The A to S change at position 320 may affect the interaction between PGIP and polygalacuronase. These results indicate that VrPGIP2 is very likely the gene at the Br locus responsible for bruchid resistance in mungbean.
Br 基因赋予绿豆抗豆象侵害的能力;VrPGIP2(编码多聚半乳糖醛酸酶抑制剂)是该抗性的一个强候选基因。VrPGIP2 序列在抗性和敏感系之间存在差异。豇豆象(Callosobruchus chinensis)和绿豆象(Callosobruchus maculatus)是绿豆在储存期间的严重害虫。绿豆对豆象的抗性由一个单一的显性基因 Br 控制。虽然 Br 基因座已定位在遗传图谱上,但该基因的分子基础和功能仍然未知。在这项研究中,利用来自‘Kamphaeng Saen 1’(KPS1;敏感)和‘V2802’(抗性)杂交的 418 个 BC11F2 群体的高分辨率作图,使用简单序列重复(SSR)标记将 Br 基因座限定在包含两个注释基因的 5 号染色体的 38 Kb 基因组区域内。EST-SSR 标记 DMB-SSR158 与 Br 基因座完全共分离。生物信息学分析表明,DMB-SSR158 对应于一个编码多聚半乳糖醛酸酶抑制剂(多聚半乳糖醛酸酶抑制蛋白 PGIP)的基因,并被命名为 VrPGIP2。在四个抗豆象(V2802、V1128、V2817 和 TC1966)和四个抗豆象(KPS1、Sulu-1、CM 和一个未知品系)绿豆品系之间比较 VrPGIP2 编码序列,发现抗性和敏感组之间存在六个单核苷酸多态性(SNP)。六个 SNP 中有三个导致氨基酸变化;即在抗性系中,第 320 位的丙氨酸(A)突变为丝氨酸(S),第 332 位的亮氨酸(L)突变为脯氨酸(P),第 335 位的苏氨酸(T)突变为 P,而在敏感系中则为缬氨酸(V)。第 320 位的 A 到 S 变化可能影响 PGIP 和多聚半乳糖醛酸酶之间的相互作用。这些结果表明,VrPGIP2 很可能是绿豆 Br 基因座中负责抗豆象侵害的基因。