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将玉米的转座子系统Ac/Ds导入水稻基因组中诱导产生突变体的方法已成为构建水稻插入突变体库的主要策略之一.本研究通过水稻Ds-T-DNA转化纯合体和Ac-T-DNA转化纯合体的杂交,构建水稻的Ac/Ds双因子转座系统,对F1～F5代的Ds转座行为进行了研究.通过转座检测,发现1例Ds转座引起约170bpT-DNA载体序列的缺失,另1例Ds转座插入基因组时带有一段272bp的T-DNA载体序列.对F1～F5代Ds转座的连续检测结果表明,配子体转座频率随世代的增加而提高,F3代的配子体转座频率达54.2%.采用TAIL-PCR的方法扩增Ds元件旁侧的基因组序列,对17个TAIL-PCR产物的序列进行了分析,发现有5例Ds插入到基因序列中,Ds既可发生邻近位置的转座,也可发生不连锁的转座,Ds在染色体间的转座频率为33.3%.
The method of introducing Ac / Ds into the rice genome to induce mutants has become one of the main strategies to construct a rice mutant library.In this study, Ds-T-DNA transformation of homozygotes and Ac-T -DNA transformation homozygotes to construct Ac / Ds two-factor transposon system in rice, and to investigate the Ds transposon behavior in F1-F5 generation.A transposon assay showed that one case of Ds transposition caused about 170 bp T-DNA And the other 1 case of Ds transposon was inserted into the genome with a sequence of 272bp T-DNA vector.The continuous test results of Ds transposition of F1 ~ F5 showed that the frequency of gametocyte transposition increased with the increase of generation, The transposition frequency of F3 generation was 54.2%. TAIL-PCR was used to amplify the genomic sequence flanking the Ds element, and the sequence of 17 TAIL-PCR products was analyzed. It was found that 5 Ds were inserted into the gene sequence , Ds can occur either adjacent transposon, also can occur unlinked transposition, Ds inter-chromosome transposition frequency was 33.3%.