大白菜高通量SSR标记鉴定体系的建立和应用

大白菜是重要的蔬菜作物,准确鉴定大白菜品种对于大白菜种质资源管理、新品种测试、种子质量检测等具有重要意义。本研究从已定位到大白菜10个连锁群的205个SSR标记中,筛选出30个在连锁群上分布均匀、PCR扩增稳定、带型简单的标记,用于大白菜品种DNA指纹鉴定。对入选的引物用4种荧光染料进行了标记,利用基于毛细管电泳荧光检测的DNA分析仪对SSR扩增产物进行检测。通过比较分析2种不同型号的3台DNA分析仪的扩增片段检测数据,明确了不同DNA分析仪的检测数据间一般存在明显的系统误差。系统误差的大小取决于引物,一般在1～4 bp之间。使用扩增产物的片段长度对各SSR位点的不同等位变异进行命名。通过使用...

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Bibliographic Details
Published in植物遗传资源学报 Vol. 15; no. 4; pp. 815 - 823
Main Author 张晗 王东建 孙加梅 郑永胜 姚凤霞 许金芳 宋国安 段丽丽 李华 王雪梅 李汝玉
Format Journal Article
LanguageChinese
Published 2014
Subjects
DNA指纹
SSR
品种鉴定
大白菜
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Summary:大白菜是重要的蔬菜作物,准确鉴定大白菜品种对于大白菜种质资源管理、新品种测试、种子质量检测等具有重要意义。本研究从已定位到大白菜10个连锁群的205个SSR标记中,筛选出30个在连锁群上分布均匀、PCR扩增稳定、带型简单的标记,用于大白菜品种DNA指纹鉴定。对入选的引物用4种荧光染料进行了标记,利用基于毛细管电泳荧光检测的DNA分析仪对SSR扩增产物进行检测。通过比较分析2种不同型号的3台DNA分析仪的扩增片段检测数据,明确了不同DNA分析仪的检测数据间一般存在明显的系统误差。系统误差的大小取决于引物,一般在1～4 bp之间。使用扩增产物的片段长度对各SSR位点的不同等位变异进行命名。通过使用一组参照品种,消除了不同批次、不同DNA分析仪型号间的系统误差,保证了检测数据的可重复性和可再现性。在此基础上,对184份大白菜品种进行了DNA分子数据采集。
Bibliography:Chinese cabbage is one of the most important vegetables in the world. Identifying and distinguishing Chinese cabbage varieties is critical for management of genetic resources,new variety testing,and seed quality supervision. In this study,we screened from 205 mapped SSR markers for those that amplified stably in PCR reaction and were easily scored and relatively evenly distributed on the ten linkage groups of Chinese cabbage. 30 makers were selected for Chinese cabbage identification and were labeled with four fluorescent dyes and allele sizing were conducted using DNA analyzers based on capillary electrophoresis and fluorescent detection. Allele sizes of same groups of samples determined using three DNA analyzers of two models were compared and analyzed. And it was found that systematic errors often occured among DNA analyzers. Values of the systematic errors depended on the markers and varied between 1 and 4 bp. Alleles of the SSR loci were named according to the length of the amplified DNA fragment determi
ISSN:1672-1810