西藏拟南芥InDel分子标记开发与株高调控基因初步定位

Development of InDel Molecular Marker and Initial Location of Plant Height Regulatory Genes in Arabidopsis thaliana(Tibet)

西藏拟南芥是近年于青藏高原地区发现的一个新的拟南芥生态型,现已成为研究植物高原适应性的模式材料。图位克隆是分离目标基因的常用手段之一,然而有效分子标记的缺少严重阻碍了西藏拟南芥对高海拔适应机制的研究。本研究对西藏拟南芥基因组进行组装,获得了相对完整的西藏拟南芥基因组,并通过BLAST序列比对筛选和重新设计了一套均匀分布于5条染色体的InDel分子标记引物。结果表明,实验验证的119对InDel分子标记引物中,54对引物在西藏型和哥伦比亚型之间具有多态性;其中37对引物扩增效果最好,在2%琼脂糖凝胶中即可产生清晰的多态性条带,并用于后续研究。初定位连锁图表明,西藏拟南芥株高矮化基因定位于4号染色体9.3~16.2 Mb区间。本研究开发的分子标记能够有效对西藏拟南芥进行基因分型,为西藏拟南芥的正向遗传研究奠定了基础。

Arabidopsis thaliana(Tibet), the n ew-discovered Arabidopsis ecotype in Qinghai-Tibet Plateau, is a model plant for high-altitude acclimatization research. Map-based cloning is one of the commonly used methods to isolate target genes. However, the lack of effective molecular markers severely hinders the study of the high-altitude adaptation mechanism of A. thaliana(Tibet). In this study, the A. thaliana(Tibet) genome was assembled, and a set of evenly-distributed InDel markers were developed by a BLAST-based method. Results showed that among 119pairs of InDel marker primers verified by our experiments, 54 pairs of primers are polymorphic between Tibet ecotype and Col ecotype;among them, 37 pairs of primers had the best amplification effect and could generate clear polymorphic bands in 2% agarose gel, which were used for subsequent studies. Preliminary mapping linkage map showed that the dwarfing gene of A. thaliana(Tibet) was located in 9.3~16.2 Mb of chromosome 4. The molecular markers developed in this research were effective for A. thaliana(Tibet) genotyping, and laid a foundation for the positive genetic research of A. thaliana(Tibet).