[利用AFLP技术分析三个小麦细胞质雄性不育系线粒体DNA]

[Analysis of three wheat cytoplasmic male sterile lines mitochondrial DNA by AFLP].

作者信息

Zhu Qidi, Zhang Xinbo, Ejaz M, Zhang Gaisheng, Che Huixue, Wang Shuping, Song Qilu, Yang Shuling, Zhang Longyu

机构信息

College of Agronomy, Northwest A&F University, Yangling 712100, Shaanxi, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2013 May;29(5):646-56.

PMID:24010362

Abstract

Cytoplasmic male sterility is an important way to utilize wheat heterosis. The purpose of thisstudy was to identify cytoplasmic type of three wheat male sterile lines. Amplified fragment length polymorphism (AFLP) marker technique was used to analyze the wheat mitochondrial DNA. We isolated mitochondria by differential centrifugation and density gradient ultracentrifugation. The results show that the extracted mitochondrial DNA was pure. It was suitable for PCR and genetic analysis. We got 4 pairs of specific primers from 64 primers combinations. Primer E1/M7 amplified 3 specific fragments in ms(Kots)-90-110. Primer E4/M2 generated 2 specific fragments in ms(Ven)-90-110. Primer E7/M6 amplified 2 specific fragments in ms(S)-90-110. Primer E6/M4 produced 2 specific fragments in ms(Kots)-90-110. Four specific primers could be used to identify three cytoplasmic types of Aegilops kotschyi, Ae. ventricosa and Triticum spelta. It provided the molecular basis to further study the mechanism of wheat cytoplasmic male sterility.

摘要

细胞质雄性不育是利用小麦杂种优势的重要途径。本研究的目的是鉴定三个小麦雄性不育系的细胞质类型。采用扩增片段长度多态性（AFLP）标记技术分析小麦线粒体DNA。我们通过差速离心和密度梯度超速离心分离线粒体。结果表明，提取的线粒体DNA纯度高，适合进行PCR和遗传分析。从64对引物组合中获得了4对特异性引物。引物E1/M7在ms(Kots)-90-110中扩增出3个特异性片段。引物E4/M2在ms(Ven)-90-110中产生2个特异性片段。引物E7/M6在ms(S)-90-110中扩增出2个特异性片段。引物E6/M4在ms(Kots)-90-110中产生2个特异性片段。4对特异性引物可用于鉴定三种细胞质类型的粘果山羊草、偏凸山羊草和斯卑尔脱小麦。这为进一步研究小麦细胞质雄性不育的机制提供了分子基础。