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当前位置: 首页 > 文章 > 玉米胚乳DNA的快速提取方法及其在wx基因分子标记辅助选择中的应用 分子植物育种 2014,12 (5) 158-165
Position: Home > Articles > A Rapid DNA Extraction Protocol from Endosperm and Its Applications in Marker-assisted Selection for wx Gene in Maize Molecular Plant Breeding 2014,12 (5) 158-165

玉米胚乳DNA的快速提取方法及其在wx基因分子标记辅助选择中的应用

作  者：
彭勃;赵晓雷;付从贵;张巧云;王奕
单  位：
天津市农作物研究所
关键词：
玉米胚乳;DNA提取;分子标记辅助选择
摘  要：
模板DNA的制备是制约分子育种效率的瓶颈步骤之一。本研究介绍一种高通量快速的玉米基因组DNA提取方法,即蔗糖提取液法。该方法切取20~30 mg玉米种子胚乳,通过批量化研磨、加热和瞬时离心3个步骤,即可获得稳健PCR扩增的模板DNA,并且在光温培养箱条件下切取部分胚乳的剩余种子与对照完整种子发芽率无显著差异。通过蔗糖提取液法提取192粒F2种子胚乳DNA,鉴定WxWx、Wxwx和wxwx 3种基因型与表型鉴定结果相一致。蔗糖提取液法提取胚乳DNA简单、快速、成本低廉,可有效用于高通量分子标记辅助选择。
译  名：
A Rapid DNA Extraction Protocol from Endosperm and Its Applications in Marker-assisted Selection for wx Gene in Maize
作  者：
Peng Bo;Zhao Xiaolei;Fu Conggui;Zhang Qiaoyun;Wang Yi;Tianjin Crops Research Institute;
关键词：
Maize endosperm,DNA extraction,Marker-assisted selection
摘  要：
Extracting genomic DNA is one of the bottleneck steps for molecular breeding. In this study, a high throughput and rapid genomic DNA extraction protocol for maize, named as sucrose prep method, were developed. According to this method, 20~30 mg of endosperm cut from seed were suffered by 3 steps, i.e.grinding, heating and instantaneously centrifuging in batch, and then the genomic DNA template with robust PCR products were obtained. In addition, there are no significant difference for germination rates between endosperm sampled seeds and non-sampled control. Three genotypes with WxWx, Wxwx and wxwx identified by genotyping endosperm DNA were in completely accordance with the corresponding phenotypes in 190 F2 individuals.Therefore, endosperm DNA extracted through the Sucrose prep method was a simple, rapid and low cost method,and especially suitable for the application in high throughput marker-assisted selection.

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