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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 rice‘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.
水稻 / 籼粳交杂株 / 大青棵 / InDel标记 / 籼粳属性鉴定 {{custom_keyword}} /
rice / indica-japonica untypical plants / abnormal heading / InDel marker / indica-japonica identification {{custom_keyword}} /
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【目的】随着籼粳杂交稻育种技术的发展与完善,直接利用籼粳亚种间杂种优势培育高产的水稻品种已成为可能。利用InDel分子标记及其成熟的技术,准确地分析品种的籼粳基因型频率、籼粳稻属性、品种类型与竞争优势水平,对籼粳杂交稻的育种与品种类别的甄别具有重要的意义。【方法】征集浙江省多家育种单位培育的24个籼粳杂交组合和粳粳杂交组合,选用18对InDel分子标记引物,采用王林友等报道的方法,检测水稻样品在InDel位点上的粳型基因频率,判别参试品种的籼粳属性。采用Nei的方法求算参试品种的InDel遗传距离,用UPGMA法进行遗传相似性聚类,用SPSS 17.0统计分析软件对InDel条带赋值进行主成分分析,依据第一和第二主成分特征向量的平均分量值作平面散点图。调查或考察全生育期、株高、每穗总粒数、千粒重等10项主要农艺性状,测定小区籽粒产量及产量的竞争优势。测定InDel遗传距离与籼粳杂交组合和粳粳杂交组合竞争优势的相关性。【结果】18对InDel引物可扩增获得每个材料的籼稻特异带、粳稻特异带和籼粳杂合带型,经计算粳型基因频率,所有参试材料准确地判别出籼粳属性,并归为6类,其中12个为籼粳杂交组合,另外13个为粳粳杂交组合。经InDel标记的聚类分析,验证了上述判别的正确性。PCA分析把所有品种归入3个不重叠的区域,即籼稻区、典型粳稻与粳稻区、偏粳型与中间型区。籼粳杂交稻品种的2年小区籽粒产量分别比粳粳杂交稻品种增产12.47%和14.89%,表现十分明显的产量竞争优势。对比2类组合的10项农艺性状,籼粳杂交稻组的每穗总粒数2年分别比粳粳杂交稻品种增加32.0%和37.1,每穗实粒2年分别增加26.8%与34.4%,说明大穗是籼粳杂交稻增产的主要贡献因子。分析参试组合与嘉优2号的遗传距离(GD)及其各性状的竞争优势的相关性,结果粳粳杂交稻组合未发现有相关性,籼粳杂交稻表现为穗长和千粒重极显著呈正相关,着粒密度呈现为显著负相关。【结论】InDel分子标记技术可以有效地测定各种类型品种的粳型基因频率,并判别参试品种的籼粳属性,甄别品种的类型。籼粳杂交稻籽粒产量的竞争优势十分明显,其主要贡献因子是较大的每穗粒数。
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