四倍体刺槐转甜菜碱醛脱氢酶基因的研究  

Research for Betaine Aldehyde Dehydrogenase(BADH)Gene Transformation of Tetraploid Clone of Black Locust

作　　者：夏阳[1] 梁慧敏[1] 孙仲序[2] 陈受宜[3] 王太明[1] 李小玉[4] 付玲玲[4] 黄剑[4] 孙道英[1] 

机构地区：[1]山东省林业科学研究院,济南250014 [2]山东农业大学园艺学院 [3]中国科学院遗传研究所 [4]甘肃农业大学草业工程学院

出　　处：《山东林业科技》2003年第3期1-3,共3页Journal of Shandong Forestry Science and Technology

摘　　要：试验以所建立的四倍体刺槐高频再生体系为基础,通过农杆菌介导法转化甜菜碱醛脱氢酶(BADH)基因,以GUS染色组织分析法为依据探讨了影响转化效率的各种因素,建立了优化转化体系,结果如下:20mg·L^(-1)乙酞丁香酮可显著提高转化效率;菌液浓度OD_(600)值0.3～0.7、预培养2d为宜;转化后暗培养对转化效率没有影响。并在以上研究基础上,成功地建立了高效、可重复的遗传转化体系,选择培养基上头孢霉素400mg·L^(-1)可有效地抑制农杆菌;卡那霉素50mg·L^(-1)时愈伤组织的白化死亡率达96.4％。经PCR检测,外源基因已成功的整合到植株的基因组DNA中,获得了15个转基因株系。Gene transformation system was optimized by GUS coloation analysis and betaine aldehyde dehydrogenase (BADH)gene was transferrde into tetraploid clone of black locust ( Robinia pseudoacacia L. ) by agrobacterium (Agrobacterium trefaciens Conn) mediated transformation. The results was following:20mg·L-1 As could increased the transformation effciencg significantly;0.3 ~ 0.7 OD600 for agrobacteriun concentration,2d for precul-ture time was suitable for the transformation;dark cultivation after the bacterium infection had no effect on transformation efficiency ;400mg·L-1 cef for selected medium could control the agrobacterium effectively; the albino rate for buds on callus reached 96.4% under50 mg·L-1 Kan stress.The PCR analysis showed that the BADHgene was integrated into genome of many plants.

关 键 词：四倍体刺槐 甜菜碱醛脱氢酶基因 农杆菌 转化 

分 类 号：S792.27[农业科学—林木遗传育种]