一种基于HRM技术的安全及高通量水稻光敏不育基因分型体系的创建与应用
发布时间：2019-04-13 13:40
【摘要】：为建立适用于大规模应用功能性分子标记辅助选择(MAS)选育光周期敏感核雄性不育(PGMS)水稻的安全、高通量和廉价的光敏基因分型体系,本研究通过将基于竞争性扩增差异熔解扩增子(CADMA)的高分辨熔解曲线(HRM)分析与水稻基因组DNA快速提取方法相结合,创建了一种安全、廉价和高通量的PGMS基因分型体系。利用该基因分型体系,从约8 000株花培苗中检测出了2 027株PGMS水稻,并对117个DH_1光敏不育单株的DH_2材料进行跟踪检测,检出了异交单株。在该体系中,96个水稻材料的取样、DNA提取和PGMS基因分型可在2.5 h左右完成。从样本DNA提取到基因分型均不用有毒化学物质,所有操作均在基于96孔PCR板的操作平台进行,闭管操作、无交叉污染。本研究建立的PGMS基因分型体系必将极大提高水稻光敏不育系选育的效率。
[Abstract]:In order to establish a safe, high-throughput and cheap Guang Min genotyping system for the selection of photoperiod-sensitive genic male sterile (PGMS) rice by functional molecular marker-assisted selection (MAS) on a large scale. In this study, a safe, cheap and high-throughput PGMS genotyping system was established by combining the high resolution fusion curve (HRM) analysis based on competitive amplification differential fusion amplification (CADMA) with the rapid extraction method of rice genomic DNA. By using this genotyping system, 2 027 PGMS rice plants were detected from about 8 000 anther culture seedlings, and the DH_2 materials of 117 DH_1 Guang Min sterile plants were tracked and detected, and outbred single plants were detected. In this system, the sampling, DNA extraction and PGMS genotyping of 96 rice materials could be completed at about 2.5h. No toxic chemicals were used from DNA extraction to genotyping. All the operations were carried out on the operating platform based on 96-well PCR plate closed-tube operation without cross-contamination. The PGMS genotyping system established in this study will greatly improve the breeding efficiency of rice Guang Min sterile line.
【作者单位】： 浙江大学农业与生物技术学院作物研究所/国家水稻生物重点实验室;无锡哈勃生物种业技术研究院;浙江大学新农村发展研究院;
【基金】：浙江省农业(粮食)新品种选育重大科技专项(2016C02050-2) 杭州市重大科技创新项目(2015012A09)
【分类号】：S511
，
本文编号：2457618
[Abstract]:In order to establish a safe, high-throughput and cheap Guang Min genotyping system for the selection of photoperiod-sensitive genic male sterile (PGMS) rice by functional molecular marker-assisted selection (MAS) on a large scale. In this study, a safe, cheap and high-throughput PGMS genotyping system was established by combining the high resolution fusion curve (HRM) analysis based on competitive amplification differential fusion amplification (CADMA) with the rapid extraction method of rice genomic DNA. By using this genotyping system, 2 027 PGMS rice plants were detected from about 8 000 anther culture seedlings, and the DH_2 materials of 117 DH_1 Guang Min sterile plants were tracked and detected, and outbred single plants were detected. In this system, the sampling, DNA extraction and PGMS genotyping of 96 rice materials could be completed at about 2.5h. No toxic chemicals were used from DNA extraction to genotyping. All the operations were carried out on the operating platform based on 96-well PCR plate closed-tube operation without cross-contamination. The PGMS genotyping system established in this study will greatly improve the breeding efficiency of rice Guang Min sterile line.
【作者单位】： 浙江大学农业与生物技术学院作物研究所/国家水稻生物重点实验室;无锡哈勃生物种业技术研究院;浙江大学新农村发展研究院;
【基金】：浙江省农业(粮食)新品种选育重大科技专项(2016C02050-2) 杭州市重大科技创新项目(2015012A09)
【分类号】：S511
，
本文编号：2457618
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