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簇毛麦5VS染色体臂特异分子标记开发与遗传效应分析
付必胜1, 孟祥如2, 刘润然2, 卢春甜2, 闫丽娟1, 张巧凤1, 郭 炜1, 蔡 瑾1, 刘彩云1, 张瑞奇2, 吴纪中1
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(1.江苏省农业科学院种质资源与生物技术研究所/江苏省农业生物学重点实验室;2.南京农业大学农学院/作物生产省部共建协同创新中心)
摘要:
簇毛麦(Dasypyrum villosum 2n=14,VV)是小麦改良重要的三级基因源。簇毛麦5VS染色体臂上携带抗白粉病基因Pm55、抗条锈病基因Yr5V和籽粒硬度基因Dina/Dinb等优异基因。已创制的小麦-簇毛麦T5VS.5AL和T5VS.5DL易位系为小麦抗病和品质改良提供了优异新种质。为加快T5VS.5AL和T5VS.5DL易位系在育种改良中的高效利用并深入分析5VS染色体臂在小麦背景的遗传效应,本研究通过流式细胞染色体分离技术和二代测序结合的方法,获得了簇毛麦5VS的基因组序列信息。利用该基因组序列信息与中国春小麦染色体5AS、5BS、5DS上的基因进行比对,共开发了69个保守直系同源基因序列(conserved ortholog sequence,COS)标记,经过PCR扩增鉴定,发现59对引物扩增产物具有簇毛麦5VS特异条带,多态率为85.5%。其中,共显性标记40个,显性标记19个。结合生物信息学分析,明确了这些特异标记与普通小麦第五部分同源群短臂的共线性区间,为物理定位5VS上的优异基因及筛选鉴定5VS小片段易位系提供了标记信息。通过回交6次,将T5VS.5AL和T5VS.5DL易位染色体导入‘南农0686’遗传背景,利用BC6F2近等系群体结合白粉病抗性鉴定对扩增条带共显性标记的可靠性进行了验证,为分子标记辅助选择T5VS.5DL易位系提供了易于利用的特异新标记。在BC6F2:3近等系群体中,进一步明确了5VS染色体臂在‘南农0686’背景中的遗传效应,即降低籽粒硬度、株高和千粒重以及提高主穗粒数。本研究新开发的5VS特异分子标记及选育的新种质为高效利用T5VS.5AL和T5VS.5DL易位系培育优质、抗病弱筋小麦新品种奠定基础。
关键词:  小麦  T5VS.5AL和T5VS.5DL易位系  分子标记
DOI:10.13430/j.cnki.jpgr.20210605001
投稿时间:2021-06-05修订日期:2021-09-10
基金项目:国家自然科学基金面上项目(31871619, 31971938);江苏省政策引导类计划(引进外国人才专项)(BX2020009);江苏省农业科技自主创新资金项目(CX(18)1001)
Development of Molecular Markers Specific to 5VS Chromosome Arm of Dasypyrum villosum and Their Genetic Effects in Common Wheat
FU Bi-sheng1, MENG Xiang-ru2, LIU Run-ran2, LU Chun-tian2, YAN Li-juan1, ZHANG Qiao-feng1, GUO Wei1, CAI Jin1, LIU Cai-yun1, ZHANG Rui-qi2, WU Ji-zhong1
(1.Institute of Germplasm Resources and Biotechnology / Jiangsu Provincial Key Laboratory of Agrobiology, Jiangsu Academy of Agricultural Sciences;2.College of Agronomy / JCIC‑MCP, Nanjing Agricultural University)
Abstract:
Dasypyrum villosum (2n = 14, VV) is an important tertiary gene pool of bread wheat. Excellent genes including powdery mildew resistance gene Pm55, stripe rust resistance gene Yr5v, and grain hardness gene Dina/Dinb were located on chromosome arm 5VS of D. villosum. The newly developed Triticum aestivum - Dasypyrum villosum T5VS.5AL and T5VS.5DL translocation lines provided germplasms for bread wheat improvement in disease resistance and end-use quality. In order to make efficient use of the translocation lines in wheat breeding programmes and analyze the genetic effects of the 5VS chromosome arm in wheat background, sequence information of T5VS.5DL translocated chromosome was isolated by the combination of chromosome sorting and next-generation sequencing. A total of 69 COS markers were developed by comparing the 5VS sequence information with genes on wheat ‘Chinese Spring’ homologous chromosome arms 5AS, 5BS and 5DS. After PCR amplification, 59 COS markers were identified with D. villosum 5VS specific bands, and the polymorphism rate was 85.5%, of which 40 markers were co-dominant between 5VS and homoeologous chromosome arms 5DS/5AS of wheat, and 19 markers were dominant. These markers provided information for physical mapping of beneficial genes on 5VS, and would contribute to development of small segment of 5VS in wheat. The T5VS.5AL and T5VS.5DL translocated chromosomes were introgressed into the genetic background of ‘NAU0686’ through backcross for 6 times. Comparing of powdery mildew reaction and molecular marker identification in BC6F2 generation, the reliabilities between co-dominant markers with simple amplified bands and 5VS chromosome arm were verified. In BC6F2:3 near-isoline population, the genetic effects of 5VS chromosome arm in ‘NAU0686’ background were further clarified. The results indicated that T5VS.5AL and T5VS.5DL translocated chromosomes could reduce grain hardness, plant height and 1000-grain weight, and increase grain number per main spike. The newly developed 5VS-specific molecular markers and the new germplasms laid a foundation for the efficient use of the two-translocation lines to breed new weak gluten wheat varieties with high quality and disease resistance.
Key words:  Triticum aestivum  T5VS.5AL and T5VS.5DL translocation lines  molecular markers