反义蜡质基因不同整合位点对降低稻米直链淀粉含量的影响及整合位点分析

The Effect of Different Integration Sites of anti-Waxy Gene on the Decrease of Rice Amylose Content and Analysis of the Integration Sites

本研究以含有反义蜡质基因双拷贝整合转化子Sh3为材料,通过后代分离获得由单拷贝及仍然呈双拷贝整合转基因植株后,采用TAIL-PCR对这些后代进行T-DNA整合位点分析。结果显示,两个外源基因分别整合在Sh3转化子基因组中第3号染色体9127056～91266603bp处和第4号染色体9977226～9976939bp处。检测由Sh3转化子后代分离获得外源基因单拷贝以及双拷贝整合植株糙米直链淀粉含量,结果显示,非转基因对照糙米直链淀粉平均含量为12.83%,外源基因整合在第3和第4染色体上的单拷贝植株糙米直链淀粉平均含量分别为5.31%和11.83%,双拷贝整合植株糙米直链淀粉含量平均为3.8%。通过本研究不仅明确了反义蜡质基因不同整合位点对于降低糙米直链淀粉含量程度有不同的影响,同时也获得了一个能够使转基因水稻糙米直链淀粉含量降低较明显的位点。

We enployed the rice transgenic line Sh3 with two copies of the anti-Waxy gene to be as material,and obtained transgenic palnts with a single anti-Waxy gene copy and still double anti-Waxy gene copies,which derived from the progeny separation.Then,we analyzed their integration sites of the T-DNAs by using TAIL-PCR in this study.The results revealed that two exogenous genes were integrated in a range from 9 127 056 bp to 91 266 603 bp of the chromosome 3 as well as a range from 9 977 226 bp to 9 976 939 bp of the chromosome 4 in the genome of the line Sh3,respectively.The results,which amylose contents in the brown rice of the transgenic plants with a single exogenous gene and two copies of exogenous genes were assayed,showed that the brown rice amylose content of non-transgenic control was averaged at 12.83%,and the brown rice amylose contents of the transgenic plants with a single copy of the anti-Waxy gene integrated in the chromosome 3 and in the chromosome 4 were averaged at 5.31% and 11.83%,respectively,and that of the transgenic plants with two anti-Waxy gene copies was averaged at 3.8%.In this study,that different integration sites of the anti-Waxy gene could affect the decrease extent on brown rice amylose content was confirmed,meanwhile,a integration site in which the amylose content of transgenic brown rice decreased more significantly was found.