粳糯谷子Waxy基因序列变异及其与禾谷类作物的相似性

Sequence Variation of Waxy Gene of Waxy and No-waxy Foxtail Millet and the Similarity on Cereal Crops

为了探讨糯谷子Waxy基因序列多态性及其与禾谷类作物的相似性,采用特异PCR扩增产物测序和DNAStar软件分析糯谷子"十里香"Waxy基因序列变异以及与小麦、玉米、水稻、糯谷子Waxy基因序列的相似性,以非糯类型"赤谷16"的Waxy基因作为参考序列,Gen Bank提取的糯谷子"十里香"Waxy基因为测试序列进行比对,结果表明,"十里香"Waxy基因(登录号为KF372879)序列总长度9 467 bp,直链淀粉含量1.22%。KF372879共检测到碱基颠换36个、转换21个、插入5个、缺失5个、插入序列3个。Waxy基因编码区第3外显子存在6 852(G-T)、6 854(G-T)、6 856(A-C)、6 857(T-A)、6 858(A-T)等5个颠换、6 814~6 815(A)的缺失;exon8的7 867(A-C)颠换。Intron1、exon8和exon13分别有484~512(28 bp)、7 811(54 bp)和9 274(130 bp)的插入片段。内含子Intron1检测到6 492(T-C)和6 493(C-T)的转换,Intron3和Intron7分别检测到6 931(G-T)和7 793(C-A)的颠换,intron9在8 384位点插入T。绘制了小麦、玉米、水稻、糯谷子4种禾谷类作物Waxy基因序列系统进化树;糯谷子"十里香"与小麦、玉米、水稻Waxy基因序列相似性分别为26.8%、25.8%、25.5%。

In order to explore Waxy gene polymorphism of waxy foxtail millet, and the similarity among waxy millet and the other cereal crops, specific PCR products sequencing technology and DNAStar software were used to analyze the Waxy gene variation between waxy Shilixiang and no-waxy millet sequence, associated with similarity among wheat, corn, rice and millet. Compared with the no-waxy Chigu 16, which was used for references sequences, KF372879（9 467 bp） from waxy Shilixiang was used for tested sequences, its amylase content was 1.22%.The result was showed that KF372879 was detected 36 single base transversions, 21 single base transitions, 5 single base indels, 5 single base deletions, 3 insertion sequences. In waxy gene encoding area, five single base transversions, associated 6 852 th, 6 854 th, 6 856 th, 6 857 th, 6 858 th with G-T, G-T, A-C, T-A, A-T and 6 814~6 815 th（A） deletion from exon3 were detected. One single base transversion of 7 867 th（A-C） was in exon8. Otherwise,484~512 th（28 bp）, 7 811 th（54 bp） and 9 274 th（130 bp） insertion fragments were detected in intron1, exon8 and exon13, respectively. Transitions of 6 492 th（T-C） and 6 493 th（C-T） were found in intron1, transversions of 6 931 th（G-T） and 7 793 th（C-A） were found in intron3 and intron7, respectively. 8 384 th（T） indel was in intron9. System evolutionary tree of Waxy gene sequences was drawn using Meg Align software in the four cereal crops, the results showed that sequence similarity of waxy millet over wheat, corn, rice were 26.8%, 25.8%, 25.5%, respectively.