基于水稻内含子长度多态性开发禾本科扩增共有序列遗传标记

Development of Amplified Consensus Genetic Markers in Gramineae Based on Rice Intron Length Polymorphisms

通过在多态位点两侧的保守编码序列上设计引物,可以开发出在不同物种间通用的基于PCR的分子标记,称为扩增共有序列遗传标记(ACGM)。【目的】探讨利用已公布的水稻籼、粳两亚种的基因组序列开发禾本科ACGM的可行性。【方法】根据水稻两亚种间的内含子长度多态性,开发了38对ACGM引物。用这些引物对玉米、粟、大麦、小麦、竹子、旱稗草和大米草6个属共12个不同材料进行PCR实验。【结果】几乎所有引物都可在至少1种材料中扩增出特异条带,而1/3以上的引物可以在全部供试材料中获得特异扩增产物。在每一种供试材料中,平均大约有2/3的引物可以成功扩增出目的条带。这些引物在各属内不同种、亚种或品种(系)之间的多态性比例在24.1%～90.3%之间,平均为44.6%。【结论】利用水稻基因组序列信息开发禾本科通用型ACGM是可行的。

By designing primers in the conservative coding sequences flanking a polymorphic site, it is possible to exploit a PCR-based marker called amplified consensus genetic marker （ACGM）, which is transferable among different species. [Objective] The purpose of this study was to investigate the feasibility of developing ACGMs in Gramineae using the published genomic sequences of indica and japonica rice. [ Method] Based on the intron length polymorphisms between the two rice subspecies, 38 pairs of ACGM primers were developed. Twelve accessions representing 6 grass genus including Oryza, Zea, Setada, Triticum, Phyllostachys, Echinochloa crusgalli Beauv and Sparlina anglica were used to verify the candidate ACGM markers.[ Result ] Almost all primers could acquire specifically amplified products in at least one accession. More than 1/3 primers could obtain specifically amplified products in all accessions. For each accession, about 2/3 primers on average could obtain specifically amplified products. The proportion of polymorphisms between species or subspecies within genus was 24.1%-90.3%, with an average of 44.6%. [Conclusion] These results showed that developing ACGM in Gramineae using rice genome data is feasible.