利用InDel分子标记鉴定杂交籼稻中籼粳交杂株

Indica-japonica Untypical Plants Among Hybrid Indica Rice:Identification by InDel Markers

在杂交籼稻制种过程中,若混入粳稻花粉就会产生籼粳交杂株,这种类型的杂株常表现为大青棵而使得杂交稻纯度降低,最终影响产量。为避免因配组亲本选择不当或在粳稻区制种因隔离不严格而产生大青棵,找到能准确区分籼稻与粳稻的分子标记可为育种和制种生产提供参考。以三系杂交籼稻粤泰A/R2020和田间表现为大青棵的杂株为试验材料,利用两个亲本间的多态性SSR标记对疑似杂株进行基因型分析,并通过18对特异性的InDel标记对其进行籼粳属性鉴定。结果表明:在行业标准《水稻品种鉴定技术规程SSR标记法》(NY/T1433—2014)推荐的48对引物中,不育系‘粤泰A’与恢复系‘R2020’之间有22个位点差异;在6对标记下(RM72、RM85、RM224、RM253、RM424、RM481),8个疑似杂株全为串粉带型;13对籼粳间特异性InDel标记检验结果证明了8个串粉杂株均来源于籼粳亚种间杂交。本研究初步筛选出13对InDel标记可用于籼粳属性鉴定,为避免大青棵的产生提供了分子依据。

Indica-japonica untypical plants will be produced in the process of indica hybrid seeds production when japonica pollens are mixed,and this type of untypical plants often displays the character of abnormal heading(commonly known as Daqingke in China),and results in purity decrease of hybrid rice,which ultimately affects grains yield.In order to avoid the occurrence of abnormal heading due to improper choosing of parents in mating groups or poor isolation in seed production in japonica regions,finding out proper molecular markers that could accurately distinguish indica from japonica may provide a reference for breeding and hybrid seed production.In this study,the three-line indica hybrid ric‘e Yuetai A’‘/R2020’and untypical plants with character of abnormal heading in the field were used as experimental materials.Polymorphic SSR markers between two parents were used to analyze genotype of the untypical plants,and eighteen pairs of specific InDel markers were used to identify the indica-japonica attributes.The results showed that there were twenty-two loci differences between the male sterile line‘Yuetai A’and the restorer line‘R2020’among the forty-eight pairs of SSR markers recommended by the industry standard"Protocol for identification of rice varieties-SSR Marker Method"(NY/T 1433-2014);eight untypical plants all showed out-crossing type bands under six pairs of SSR markers(RM72,RM85,RM224,RM253,RM424 and RM481);the results of thirteen pairs of indica japonica specific InDel markers further proved that these eight out-crossing plants were all derived from hybridization of indica-japonica subspecies.Thirteen pairs of InDel markers for identification of indica-japonica attributes were preliminarily selected in this study,which provided a molecular basis for avoiding the generation of untypical plants of abnormal heading.