抗逆基因GmUBC2的大豆胚尖遗传转化以及抗性材料的鉴定  

Genetic transformation of soybean with resistance gene GmUBC2 and the identification of resistant materials

作　　者：张洁[1] 赵甜甜[1] 郑文永 商蕾 王月影[1] 邱丽娟[4] 王冬梅[1] 

机构地区：[1]河北农业大学生命科学学院/河北省植物生理与分子病理学重点实验室,河北保定071001 [2]河北省任丘市农业局,河北任丘062550 [3]河北省邢台市卫生局,河北邢台054000 [4]中国农科院作物所,北京100081

出　　处：《河北农业大学学报》2016年第2期105-111,116,共8页Journal of Hebei Agricultural University

基　　金：转基因生物新品种培育重大专项子课题(2014ZX0800402B-001)

摘　　要：本研究利用前期建立的大豆胚尖遗传转化体系,借助农杆菌转化技术将抗逆基因GmUBC2导入河北省推广大豆品种中,获得了抗逆能力强的优良大豆新材料。结果表明,6个供试河北省优质大豆品种中,‘冀豆16号’的胚尖再生率较高,可达到75.57%,‘五星2号’转化植株的PCR阳性率较高,可达到9.91%。不同农杆菌菌株中,农杆碱型的EHA105菌株侵染效率明显高于胭脂碱型的GV3101和C58C1。在侵染阶段和共培养阶段添加600mg/L L-Cys和2.0mmol/L DTT能够明显提高‘冀豆16号’胚尖的发芽率和平均重生芽数,提高转化效率。经PCR以及RT-PCR检测,获得了转抗逆基因GmUBC2的‘冀豆16号’和‘冀豆12号’转化植株及其T3代种子。经200mmol/L NaCl胁迫25d,T1代阳性转化材料仍然能够正常生长,而对照植株已经完全枯黄。经0～300mmol/L NaCl胁迫处理的T2代阳性植株,随着NaCl浓度的升高,植株叶片的脯氨酸积累量比对照明显增高,而丙二醛含量均明显降低。结果表明转GmUBC2基因的植株能够降低盐胁迫对膜质造成的伤害,具有一定的抗盐胁迫能力。In this study,Agrobacterium tumefaciens transformation technology was used to get the soybean varieties with resistance gene GmUBC2,which are popularized in Hebei provine.The main results were as follow：Jidou16had the highest regeneration rate of embryonic tipswith the average of 75.57% in the six tested varieties.The Wuxing2 had the highest PCR positive rate up to 9.91%.In different Agrobacteriumstrains,the infection efficiency of agropine type EHA105 was significantly higher than that of the nopaline type GV3101 and C58C1.In the infection and co-culture stage,adding a 600 mg/L L-cys and 2.0 mmol/L DTT could obviously improve the germination rate and average regeneration bud numbers,increasing the efficiency of jigou16 transformation.PCR and RT-PCR results showed that the GmUBC2 gene was integrated into the genome of soybean.The T1 positive plants after 200 mmol/L 25 d NaCl stress was still growing normally,and the negative plants had been completely dead.T2 seeldings of positive plants were taken into the 0-300 mmol/L NaCl salt stress.The results showed that the proline accumulation in the transgenic plants increased significantly than control,while MDA content was significantly lower.These proved that the GmUBC2 transgenic plants could reduce the damage of salt stress on membrane,and have certain resistance to salt stress.

关 键 词：大豆 胚尖 农杆菌 遗传转化 GmUBC2基因 抗盐 

分 类 号：S336[农业科学—作物遗传育种]