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2025年4月5日 星期六
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粘类小麦雄性不育系线粒体DNA的AFLP标记及SCAR转化
朱启迪1,翁群珠1,张改生1,张姣2,巨岚3,于永昂3,牛娜3,王军卫3,马守才3
1. 西北农林科技大学
2.
3. 西北农林科技大学,国家杨凌农业生物技术育种中心
Mitochondrial DNA AFLP and SCAR Marker Linked to the Male Sterile Lines with Aegilops kotschyi Cytoplasm in Wheat (Triticum aestivum L.)
2, 2, 2, 2, 2, 2, 2, 2
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摘要 为了能在分子水平上有效鉴定具有粘果山羊草(Aegilops kotschyi)、偏凸山羊草(A. ventricosa)、普通小麦变种斯卑尔脱(Triticum spelta)细胞质雄性不育系及其保持系90-110和8222,提高其在杂交小麦(Triticum aestivum L.)研究与应用中的定向遗传改良,本研究对其线粒体DNA进行了扩增片段长度多态性(amplified fragment length polymorphism, AFLP)标记和序列特征性片段扩增区域(sequence characterized amplified region, SCAR)标记。通过AFLP标记方法,应用64对引物组合EcoRⅠ-NNN/MseⅠ-NNN对小麦同核异质雄性不育系和保持系进行扩增,共扩增出682条带,其中113条为多态性条带。引物E-AGG/M-CTA组合在粘果山羊草细胞质雄性不育系中扩增出一条大小约300 bp的特异性条带,对该特异条带进行回收、测序,利用 Primer Premier 5.0软件重新设计SCAR引物,并对这3种类型同核异质小麦细胞质雄性不育系和保持系进行扩增,其中引物YW1在3种细胞质雄性不育系和保持系中都扩增出条带,而引物YW2仅在粘果山羊草细胞质雄性不育系扩增出一条198 bp的特异性片段,结果表明,已成功地将AFLP标记转化为操作简便、表现稳定的SCAR标记。此片段与小麦线粒体基因组有很高的同源性(同源性为99%),为烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶基因(GenBank登录号: EU534409.1)上的序列,该酶是线粒体中氧化磷酸化的入口酶,与小麦细胞质雄性不育密切相关。本研究可以用于粘类小麦细胞质雄性不育系分子标记辅助育种,也为小麦细胞质种性鉴定提供了技术支撑和理论依据。
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朱启迪
翁群珠
张改生
张姣
巨岚
于永昂
牛娜
王军卫
马守才
关键词 小麦粘类细胞质雄性不育同核异质线粒体DNASCAR标记    
Abstract:The purpose of this study is to identify cytoplasmic type of isonucleur allocytoplasm cytoplasmic male sterile lines with Aegilops kotschyi, Ae. Ventricosa cytoplasm, Triticum spelta cytoplasm and their maintainer lines (90-110 and 8222) and to improve its directional genetic improvement in the research and application of hybrid wheat(Triticum aestivum L.). Sequence characterized amplified region(SCAR) molecular marker technique and amplified fragment length polymorphism(AFLP) marker molecular technique were used to analyze the wheat mitochondrial DNA. Three isonucleur allocytoplasm cytoplasmic male sterile lines with Aegilops kotschyi, Ae. Ventricosa cytoplasm, Triticum spelta cytoplasm and their maintainer lines were selectively amplified using 64 primer combinations EcoRⅠ-NNN/MseⅠ-NNN, and the results showed that 113 out of 682 bands were polymorphic. Primer E-AGG/M-CTA was found to amplify a special 300 bp fragment in the cytoplasmic male sterile lines with Aegilops Kotschyi cytoplasm. Specific fragment was collected and sequenced, SCAR molecular markers were redesigned with Primer Premier 5.0, and SCAR marker(YW1) amplified a fragment in 3 isonuclear alloplasmic cytoplasmic male sterile lines and their maintainer lines. SCAR marker(YW2) was successfully designed and validated, and it did not amplify fragment in isonuclear alloplasmic cytoplasmic male sterile lines with Ae. Ventricosa cytoplasm, Triticum spelta cytoplasm and their maintainer lines (90-110 and 8222), only one specific 198 bp fragment was amplified with YW2 in ms(Kots)-90-110 and ms(Kots)-8222 which showed sequence similarity was as high as 99%, and it was sequence of nicotinamide adenine dinucleotide(NADH) dehydrogenase gene (GenBank: EU534409.1). Oxidoreductase (complexⅠ) is the entry enzyme of mitochondrial oxidative phosphorylation, it was closely associated with the cytoplasmic male sterility in wheat. This SCAR marker can be used in molecular marker-assisted breeding for identifying wheat male sterile lines and provide theoretical and technical support for identifying cytoplasmic types of wheat.
Key wordsWheat    Cytoplasmic male sterility of NIAN type    Isonucleur and Allocytoplasm    Mitochondrial DNA    SCAR
收稿日期: 2014-03-03     
通讯作者: 张改生   
引用本文:   
朱启迪1,翁群珠1,张改生1,张姣2,巨岚3,于永昂3,牛娜3,王军卫3,马守才3. 粘类小麦雄性不育系线粒体DNA的AFLP标记及SCAR转化[J]. , 2014, 22(8): 958-965.
2, 2, 2, 2, 2, 2, 2, 2. Mitochondrial DNA AFLP and SCAR Marker Linked to the Male Sterile Lines with Aegilops kotschyi Cytoplasm in Wheat (Triticum aestivum L.). , 2014, 22(8): 958-965.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I8/958
 
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