药用野生稻TAC克隆转化籼稻的体系初探

采用已构建的药用野生稻TAC文库，比较研究4个籼稻品种在不同培养基的诱导率及愈伤组织对潮霉素抗性。通过2种不同诱导方法产生的愈伤组织，将携带药用野生稻大片段DNA的TAC克隆转化到籼稻品种中的结果表明，除了‘粤香占’以外，其它3个品种在心诱导培养基中添加微量B，后，其愈伤诱导率最高。不同品种的潮霉素筛选适合浓度存在差异，其中‘华粳籼74’的适合筛选浓度为50mg．L-1，‘粤香占’为40mg．L-1。用预培养5d的愈伤组织进行遗传转化，在潮霉素筛选之前先以头孢拉定抑菌处理的，容易获得转基因植株。4个品种中以‘粤香占’的抗性愈伤组织筛选率和分化率最高，分别为22．58％和14．86％。分子检测11株转基因水稻的结果表明，其中8株含有抗性标记基因。据此认为，药用野生稻TAC文库在水稻创新育种中可能有一定的利用前景。

Preliminary Study of Transformation System of indica Rice with TAC Clones Containing Large Genomic Sequences of Oryza officinalis Wall.

By using TAC library of Oryza officinalis, induction rates of four indica rice varieties in different media and resistance to hygromycin of calli were compared. TAC clones with large genomic sequences of Oryza officinalis were transferred into indica rice varieties by using calli generated by two different induction methods. The results showed that except ＇Yuexianzhan＇, the callus induction rates of three rice varieties were highest in N6 induction media plus Bs. The appropriate concentrations of hygromycin for four indica rice varieties were detected, 50 mg.L-1 hygromycin was appropriate for ＇Huajinxian 74＇ and 40 mg-L-1 for ＇Yuexianzhan＇. In the genetic transformation of 5-day precultured calli, it was more easily to get transgenic plants with anti-bacterium treatment of Cefotaxime before the screening by hygromycin. The transformation frequency and the regeneration frequency of ＇Yuexiangzhan＇ was 22.58% and 14.86% respectively, highest among four indica rice varieties. Molecular analysis of 11 transgenic plants showed that, 8 plants carried the transformed selective marker gene. The conclusion suggested that TAC library of Oryza officinalis might have the prospects of utilization in the innovation of rice breeding.