农杆菌介导大豆子叶节系统的两个问题突破.pdfVIP

第27卷 第1期 大 豆 科 学 V01．27 No．1 2008矩 2月 SOYBEAN SCIENCE Feb． 2oo8 农杆菌介导大豆子叶节系统的两个问题突破 李文霞’，李文滨 ，吕文河’，宁海龙 ( 东北农业大学农学院，黑龙江哈尔滨150030； 东北农业大学大豆研究所，教育部大豆生物学重点试验室，黑龙江哈尔滨150030) 摘 要：为解决大豆子叶节系统存在的再生和检测问题，对影响丛生芽伸长和植株检测的因素进行研究。以大豆 品种黑农35的子叶节为外植体，利用k3 正交试验确定影响再生系统丛生芽伸长的因素及水平，将转基因小植株 的叶片培养愈伤组织以提取基因组DNA进行Southem检测。结果表明，丛生芽伸长的最佳组合为继代培养基与初 始芽诱导培养基的6-BA浓度比为1／5，芽伸长培养基中添加50 mg L～Glu、0．5 mg L—GA3、0．5 mg L—ZT；South— em杂交信号说明利用叶片培养的愈伤组织能够提取足够量的基因组DNA。 关键词：大豆；子叶节；丛生芽伸长；愈伤组织 Breakthrough of Two Questions on the Agrobacterium-mediated Soybean Coty- ledonary Node Systems LI Wen．xia ，LI Wen．bin ，L U Wen．he ，and NING Ha1．1ong2 ( College ofAgronomy，Northeast A cultural University，Harbin 150030，Heilongjiang； Soybcan Research Institute ofNortheast A cultural UniversityI Key Laboratory ofSoybean Biology in Chinese Ministry ofEducation，Harbin 150030，Heilongiiang，China) Abstract：The factors influencing soybean(G1)vine，懈 L．)cotyledonary node regeneration and detection from soybean culti- vats“Heinong 35”were studied in this research SO as to solve the two problems．k3 orthogonal design was used to discuss effect of four key factors and levels on multiple shoot elongation．DNA for Southern blot were extracted from calli tissue which were induced from putative transgenic plants leaves．The results indicated that the better conditions for elongation were got when concentration of 6-Benzylaminopurine in subculture medium for shoot induction Was dropped to 1／5 to the preceding shoot induction medium，and 50 mg L—Glutamic acid，0．5 mg L—Gibberellin A3and 0．5 mg L‘ Zeatin were added to shoot elongation medium；Southem blot signal indicated that calli tissue from leaves could extract enough genomic DNA． Key words：Soybean；Cotyledonary node；Multiple shoots elongation；Calli tissue 大豆(Clyc