转甜菜碱醛脱氢酶基因豆瓣菜的耐盐性

Genetic Transformation of Watercress with a Gene Encoding for Betaine-aldehyde Dehydrogenase (BADH)

将山菠菜甜菜碱醛脱氢酶 (BADH)基因经根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)AGL1介导转入豆瓣菜 (NasturtiumofficinaleR .Br.) ,PCR和Southernblotting检测呈阳性的再生植株有 46株 ;对 6株再生植株的BADH活性和Northernblotting检测发现 ,有 5株BADH酶活性明显高于对照 ;膜的相对电导率测定结果说明 ,在盐胁迫下转基因豆瓣菜的膜结构所受损伤小于对照。转基因植株能够在 0 .5 %NaCl培养基上正常生长 ,而对照在相同的培养基上生根困难且生长缓慢。在 0 .8%NaCl的培养基上 ,部分转基因植株虽然生长减慢 ,但生长状况明显优于对照 ,且对照植株在相同培养基上培养 2 0d到 1个月左右后 ,叶片变黄并最终死亡。

Betaine_aldehyde dehydrogenase (BADH) gene was transferred into watercress (Nasturtium officinale R. Br.) by Agrobacterium_mediated transformation. PCR and Southern blotting analysis showed that the BADH gene was integrated into genome of 46 watercress plants. Among the 6 transgenic plants being examined, 5 showed higher BADH activity than the control. Northern blotting analysis of the 5 plants showed that 4 with higher BADH activity had BADH transcript expression. Relative electronic conductivity demonstrated that less membrane damage of transgenic plants. Some transgenic plants grew normally on the medium with 0.5% NaCl while the control plants had very poor roots and slow grew in the same medium. The growth of some transgenic plants were superior to control obviously on the medium with 0.8% NaCl, although they grew slower than on the medium with 0.5% NaCl. And the leaves of control turned yellow after about 20 days to 1 month of cultured and eventually died.