细菌几丁质酶基因转化烟草的研究

Study on Transformation of Tobacco with a Chitinase ChiA Gene from Serratia marcescens

为了提高烤烟品种K 32 6对真菌病害的抗性 ,以烟草再生植株的叶片为受体 ,采用农杆菌介导法将细菌几丁质酶基因导入烟草 ,获得了抗卡那霉素的转化植株 ,经PCR检测 ,初步证明了细菌几丁质酶基因已整合到烟草的基因组中 ;同时研究了再生烟草叶片的耐卡那霉素水平、受体预培养对转化的影响以及促进转化植株的生根等。结果表明 :培养基中卡那霉素浓度为 5 0mg/L时 ,能完全抑制烟草叶片的再生 ;烟草叶片预培养 2d的转化率提高 ;添加 0 .2mg/LIAA(吲哚乙酸 )能显著地提高转化植株芽梢的生根率。

In order to enhance the resistance to fungi diseases of K 326, a cultivar of flue\|cured tobacco, a chitinase chiA gene from Serratia marcescens was introduced into tobacco by an agro\|bacterium mediating method using tobacco lamina of regenerated plant as an accepter and transgenic plants with kanamycin\|resistance were obtained. The PCR examination confirmed that the chitinase chiA gene was incorporated in tobacco genome. Meanwhile, the anti\|Kan level of tobacco, the effect of pre\|culture time of accepter on transformation and anti\|Kan shoots rooting were studied. The results showed that kanamycin at the level of 50mg/L could restrain tobacco lamina from regeneration completely, the highest transformation rate was got when the lamina was pre\|cultured for 2 days and rooting rate of the transformed shoots cultured in MS containing 0.2mg/L IAA was increased remarkably.