四倍体刺槐转甜菜碱醛脱氢酶基因的研究  

Betaine Aldehyde Dehydrogenase (BADH) Gene Transformation of Tetraploid Clone of Black Locust Mediated by Agrobacterium

作　　者：夏阳[1] 梁慧敏[1] 陈受宜[2] 孙仲序[3] 王太明[1] 李小玉[1] 付玲玲[1] 黄剑[1] 燕丽萍[1] 刘德玺[1] 

机构地区：[1]山东省林业科学研究院,济南250014 [2]中国科学院遗传与发育生物学研究所,北京100101 [3]山东农业大学园艺学院,泰安271018

出　　处：《中国农业科学》2004年第8期1208-1211,F003,共5页Scientia Agricultura Sinica

基　　金：国家转基因植物研究与产业化开发专项资助项目(JY03-B-24-03);山东省科技发展计划资助项目(011100104)

摘　　要：利用农杆菌(Agrobacterium tumefaciens Conn)介导法,将山菠菜甜菜碱醛脱氢酶(BADH)基因转入四倍体刺槐(Robinia pseudoacacia L.)。结果表明,以愈伤组织作为外植体、液体分化培养基(MS+6-BA 2.0 mg·L-1+IBA 0.2 mg·L-1)活化农杆菌、转化培养基中2 0 mg·L-1乙酰丁香酮、菌液浓度OD600值0.3-0.7、预培养1～2d、浸染4～8 min、共培养4～8 d、延迟1 5 d选择的条件下,4 00 mg·L-1头孢霉素可有效抑制农杆菌,卡那霉素为5 0 mg·L-1时愈伤组织上芽的白化率达97.3％。经PCR和PCR-Southecn检测,外源基因已成功整合到植株基因组DNA中;转化植株丛生芽生长的NaCl相对抗性提高了2‰～3‰。Betaine aldehyde dehydrogenase (BADH) cDNA cloned from Atriplex hortensis L. was transferred into tetraploid clone of black locust (Robinia pseudoacacia L.) by agrobacterium (Agrobacterium tumefaciens Conn) mediated transformation. It was found that under the condition when callus was used as the explants for transformation, the liquid differentiation medium (MS+6-BA 2.0 mg·L-1+IBA 0.2 mg · L-1) was used for agrobacterium activation, 20 mg· L-1 AS in transformation medium, 0.3-0.7 OD600 for agrobacterium concentration, 1-2 days for preculture time, 4-8 min for infection , 4-8 days for co-cultivation , 15 days delay selection, 400 mg · L-1 Cef could control the agrobacterium effectively, the albino rate for buds on callus reached 97.3% under 50 mg · L-1 Kan. The PCR and PCR-Southern analysis showed that the BADH gene was integrated into genome of many plants and the NaCl resistance of the growth of transformation plants increased by 2‰-3‰.

关 键 词：四倍体刺槐 甜菜碱醛脱氢酶 转基因 菌液浓度 

分 类 号：S792[农业科学—林木遗传育种]