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.
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
关键词
四倍体刺槐
甜菜碱醛脱氢酶基因
农杆菌
转化
tetraploid clone of black locust( Robinia pseudoacacia L.)
betaine aldehyed dehydrogenase (BADH)
anrobacterium( Agrobacterium tumefaciens Conn)
transformation
