野生稻Wx基因(CT)n卫星序列和第一内含子SNP特征标签初探

Preliminary Study on the Characteristics SNP of Microsatellite Sequence (CT)n and Intron 1 in Wx Gene in Wild Rices

本研究利用特异引物对来源于云南的22份栽培稻及分别来源于海南、云南元江、江西东乡的3份普通野生稻(O.rufipogon Griff.)和1份长雄野生稻(O.longistaminata)Wx基因中第1内含子和第1内含子上游微卫星序列(CT)n的序列进行PCR扩增并测序。结果显示所有材料均能扩增出产物并获得碱基测序(包括长雄野生稻),4份野生稻的(CT)重复次数分别是(CT)8、(CT)10、(CT)10、(CT)11,较栽培稻少;第1内含子中+1位碱基均为G。相较于栽培稻,野生稻在第1内含子与(CT)n重复中有明显区别于栽培稻的SNP位点,包含基因突变的In Del、转换和颠换3种类型。说明野生稻和栽培稻在进化的过程中出现了差异。Wx基因中微卫星序列(CT)n和第1内含子的序列可作为检测野生稻遗传组分的一个遗传标签。

The sequence of （CT）n and intron 1 in Wx gene were obtained by specific primers from 22 Yunnan indigenous rice varieties, 30. rufipogon Griff. from Hainan, Yuanjiang, Dongxiang, and 10. longistaminata were analyzed. Results showed that all the rice varieties could amplify single PCR product （including O. longistaminata）. Sequence analysis revealed that 4 wild rice varieties had fewer CT repeats with repeat number ranged from 8 to 11, the first base pair of splice donor site of intron 1 in Wx gene were GWx. Compared with cultivated rice, wild rice has significant different SNPs in intron 1 and （CT）n repeats. All different SNPs include 3 types ofgene mutations like InDel, transform and transversion. The types of gene mutations showed evolutionary differences between cultiv- ated rice and wild rice. These nucleotides polymorphism differentiate wild rice from cultivated rice, suggesting that they can be used as SNP genetic marker for identifying wild rice varieties from indigenous rice varieties.