转果聚糖蔗糖转移酶基因(Sac B)美丽胡枝子的获得  

Regeneration of Transgenic Lespedeza thunbergii Expressing Levansucrase Gene (Sac B) from Bacillus subtilis

作　　者：杜金友[1] 陈晓阳[2] 张桂荣[1] 李伟[2] 胡冬南[2] 胡赞民[3] 

机构地区：[1]河北科技师范学院,秦皇岛066600 [2]北京林业大学,北京100083 [3]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《生物工程学报》2006年第6期940-944,共5页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划(863计划)项目(No.2002AA241111);河北科技师范学院博士基金资助。~~

摘　　要：采用农杆菌介导的遗传转化方法,将来自枯草杆菌的果聚糖蔗糖转移酶基因(SacB)导入美丽胡枝子,以提高胡枝子抵御干旱胁迫和盐胁迫的能力。以美丽胡枝子子叶节为外植体,通过与含有植物双元表达载体pKP的农杆菌LBA4404共培养,将SacB基因导入美丽胡枝子基因组。经卡那霉素筛选后,共获得62株卡那霉素抗性植株。经PCR特异性扩增和PCR-Southern杂交,证明有5株再生植株基因组DNA中整合了SacB基因。通过RT-PCR分析,结果表明SacB基因均获得表达。经过200mmol/LNaCl和5%PEG模拟胁迫,发现转基因植株美丽胡枝子中,可溶性糖含量在任何时候均高于未转化植株,并比对照拥有更高的抗干旱胁迫和盐胁迫能力。The Bacillus subtilis Sac B gene with the vacuolar targeting signal sequence driven by 35S promotor was transferred into Lespedeza thunbergii by Agrobacterium mediated method. Total 62 Kan-resistant plants were obtained, of which 5 plants were proved to be transgenic plants. The transgenic plants were characterized by PCR amplification, PCR-Southern hybridization and RT-PCR. The physiological assay results showed that the transgenic plants were more tolerant to stress than the controls under the condition of 200mmol/L NaC1 and 5 % PEG, respectively, and that the content of soluble sugar in trnsgenic plants was significantly higher than that of controls in the period of tests （5 - 15 days） under salt and PEG stress.

关 键 词：美丽胡枝子 果聚糖蔗糖转移酶基因 农杆菌介导转化 干旱胁迫 盐胁迫 

分 类 号：S541.5[农业科学—作物学] Q943.2[生物学—植物学]