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真核生物翻译起始因子4E(eukaryotic translatoin initiation factor 4E,eIF4E)及其异构体(eIF(iso)4E)在植物和病毒相互作用过程中发挥重要的作用。大白菜(Brassica rapa ssp.pekinensis)广谱抗病毒性状由eIF(iso)4E.a和eIF(iso)4E.c两个基因同时发生功能缺失突变所致,目前仅在RLR22和BP8407两份材料中鉴别出了双位点功能缺失突变,且二者在eIF(iso)4E.c位点持有相同的突变类型。为了筛选遗传背景更加丰富的双突变体材料,本研究选择抗/感芜菁花叶病毒(Turnip mosaic virus,TuMV)差异较大的12份大白菜自交系组成微核心种质,采用候选基因重测序的方法从9份来自中国不同地区的高抗TuMV材料中筛选到一份eIF(iso)4E.a和eIF(iso)4E.c双突变体自交系材料He102,其eIF(iso)4E.a位点是外显子4和外显子5发生缺失的假基因突变、eIF(iso)4E.c位点则是编码区碱基缺失导致的移码突变。拟南芥lsp突变体功能互补实验表明:大白菜eIF(iso)4E.a和eIF(iso)4E.c野生型能够互补lsp突变体的功能、而对应的突变体基因则不能互补其病毒敏感的功能。本研究还开发了检测两个突变位点的共显性功能标记。本研究筛选的eIF(iso)4E.a和eIF(iso)4E.c双位点突变体材料是遗传背景更加丰富的抗源材料,所开发的共显性标记为利用分子标记辅助选择培育高抗TuMV大白菜新种质和新品种提供了的辅助选择工具。同时本研究也为利用微核心种质开展有效的候选基因多样性筛查提供了典型范例。
The eukaryotic translatoin initiation factor 4E (eIF4E) and its isoforms (eIF (iso) 4E) play an important role in plant-virus interaction. The broad-spectrum antiviral trait of Brassica rapa ssp.pekinensis is caused by the simultaneous deletion of eIF (iso) 4E.a and eIF (iso) 4E.c loss-of-function mutations. At present, only the two materials RLR22 and BP8407 Identified a double-site loss of function mutation, and both held the same mutation at the eIF (iso) 4E.c locus. In order to screen double mutant materials with more abundant genetic backgrounds, we selected 12 Chinese cabbage inbred lines with large difference between Turnip mosaic virus (TuMV) Re-sequencing method, a eIF (iso) 4E.a and eIF (iso) 4E.c double mutant inbred material He102 was screened from 9 high resistant TuMV materials from different regions in China. The eIF (iso) The 4E.a site is a pseudogenetic mutation with exon 4 and exon 5 deletions. The eIF (iso) 4E.c site is a frameshift mutation caused by a base deletion in the coding region. Functional complementation of Arabidopsis thaliana lsp mutants showed that wild type eIF (iso) 4E.a and eIF (iso) 4E.c of Chinese cabbage were able to complement the function of the lsp mutant while the corresponding mutant genes did not complement their virus sensitivity Function The study also developed co-dominant functional markers to detect two mutations. This study screened eIF (iso) 4E.a and eIF (iso) 4E.c double-site mutant material is more genetic background of the anti-source material, the development of codominant markers for the use of molecular marker assisted selection to cultivate high TuMV Chinese cabbage new germplasm and new varieties provide a complementary selection tool. At the same time, this study also provides a typical example for the effective screening of candidate gene diversity using micro-core germplasm.