【摘要】：為建立適用于大規(guī)模應(yīng)用功能性分子標(biāo)記輔助選擇(MAS)選育光周期敏感核雄性不育(PGMS)水稻的安全、高通量和廉價(jià)的光敏基因分型體系,本研究通過將基于競(jìng)爭(zhēng)性擴(kuò)增差異熔解擴(kuò)增子(CADMA)的高分辨熔解曲線(HRM)分析與水稻基因組DNA快速提取方法相結(jié)合,創(chuàng)建了一種安全、廉價(jià)和高通量的PGMS基因分型體系。利用該基因分型體系,從約8 000株花培苗中檢測(cè)出了2 027株P(guān)GMS水稻,并對(duì)117個(gè)DH_1光敏不育單株的DH_2材料進(jìn)行跟蹤檢測(cè),檢出了異交單株。在該體系中,96個(gè)水稻材料的取樣、DNA提取和PGMS基因分型可在2.5 h左右完成。從樣本DNA提取到基因分型均不用有毒化學(xué)物質(zhì),所有操作均在基于96孔PCR板的操作平臺(tái)進(jìn)行,閉管操作、無(wú)交叉污染。本研究建立的PGMS基因分型體系必將極大提高水稻光敏不育系選育的效率。
[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.
【作者單位】： 浙江大學(xué)農(nóng)業(yè)與生物技術(shù)學(xué)院作物研究所/國(guó)家水稻生物重點(diǎn)實(shí)驗(yàn)室;無(wú)錫哈勃生物種業(yè)技術(shù)研究院;浙江大學(xué)新農(nóng)村發(fā)展研究院;
【基金】：浙江省農(nóng)業(yè)(糧食)新品種選育重大科技專項(xiàng)(2016C02050-2) 杭州市重大科技創(chuàng)新項(xiàng)目(2015012A09)
【分類號(hào)】：S511
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本文編號(hào)：2457618