马铃薯薯形突变体T-DNA插入侧翼序列分析

Flanking Sequence Analysis of a Potato T-DNA Inserted Mutant of Tuber Shape

以‘鄂马铃薯3号’为材料,通过试管薯快速转基因技术获得一个编号为pCL2b-2薯形突变系,本研究通过表型鉴定、侧翼序列扩增和生物信息学等方法对其进行了研究,目的是分析该突变株表型变化控制机制。结果表明,该突变体的试管薯和温室收获块茎长宽比在p<0.05水平均显著大于野生型受体薯,其中试管薯长宽比为1.78,野生型试管薯长宽比为1.33;温室收获块茎长宽比为1.50,野生型块茎为1.29;利用hiTAIL-PCR技术对T-DNA插入位点两侧的侧翼序列分析表明,T-DNA插入编码异戊烯基转移酶(IPT)基因内部,该基因为细胞分裂素合成酶基因(StIPT),暗示StIPT基因可能参与马铃薯薯形发育。

A tuber shape mutant, pCL2b-2, was obtained by T-DNA inserted transformation on microtuber of potato （So/anum tuberosum L.） variety ＇E-potato 3＇ （E3）. In this study, we identified the tuber shape, amplified the T-DNA flanking sequences and did some bioinformatics work to analyze the tuber shape control mechanism. The length-width ratio of in vitro grown microtubers of the mutant （1.78） was significantly greater than that of the wild type （1.33） （p 〈 0.05）. The shape of tubers grown in pots was also significantly different between pCL2b-2 mutant and the wild type, 1.50 vs. 1.29. Flanking sequences of inserted location of the T-DNA were amplified by hiTAIL-PCR. Bioinformatics analysis showed that the T-DNA was inserted into the isopentenyltransferase （IPT） gene which encodes a cytokinins synthetase StIPT. It is proposed that St/PTmay participate in morphogenesis of potato tubers.