发根农杆菌介导不同基因型大豆转化效率的筛选

摘要

基因型是影响发根农杆菌转化大豆的主要因素之一,为筛选转化效率高的大豆基因型,利用不同基因型大豆的下胚轴接种发根农杆菌K599(舍GUS基因),待长出毛状根后,根据不同基因型大豆发根诱导率和发根数的差异,选择适宜大豆基因型进行GUS基因的组织化学染色和RT-PCR分析.结果表明,34个大豆基因型发根诱导率和发根数有很大差异,选择发根诱导率大于60％以及发根数大于1.6条的20个大豆基因型毛状根进行GUS基因组织化学染色,其中Y091、Z184、P108、L010等4个大豆基因型的GUS基因转化效率较高,分别为96.2％、91.7％、87.5％、86.4％；进一步对以上4个大豆基因型毛状根进行RT-PCR分析,结果表明,GUS基因均得到了正确转录.表明大豆基因型Y091、Z184、P108、L010是发根农杆菌转化大豆较为理想的受体材料.%Soybean genotype might be one of the most important factors that influenced Agrobac-terium rhizogenes transformation. In order to screen soybean genotypes with high transformation efficiency,K599 Agrobacterium rhizogenes harboring a binary vector pBI121 with GUS gene was used to inoculate different hypocotyls of different soybean genotypes. Suitable soybean genotypes were chosen to conduct histochernical staining for GUS gene and RT - PCR analysis according to the differences of hairy roots inducing rate and the number of roots when hairy roots grown. The results showed that there were great differences among 34 soybean genotypes in hairy roots inducing rate and the number of roots. Among them 20 soybean genotypes whose hairy roots inducing rate over 60% and number of roots per plant more than 1. 6,were chosen to detect GUS histo-chemical staining,indicating Y091, Z184?P108 and L010 showed higher transformation frequencies of 96. 2% ,91. 7% ,87. 5% and 86. 4%. Further RT - PCR analysis of those 4 soybean genotypes revealed GUS gene were transcribed correctly. It was indicated that soybean genotypes Y091 ,Z184?P108 and L010 were ideal receptor materials for Agrobacterium rhizogenes transformation.