空间诱变育成的茄子“HANGQIE-5”的RAPD体系优化及RAPD分析  

Optimization of RAPD Reaction System and Analysis of "HANGQIE-5" Eggplant Generated by Space Mutagenesis

作　　者：任春晓[1,2] 刘敏[3] 鹿金颖[3] 吴玉环[1] 

机构地区：[1]中国科学院沈阳应用生态研究所,辽宁沈阳110016 [2]中国科学院研究生院,北京110049 [3]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《航天医学与医学工程》2010年第1期52-56,共5页Space Medicine & Medical Engineering

基　　金：国家高技术研究发展计划(863计划)-果菜类蔬菜航天工程育种技术与品种创新(2007AA100104-3)

摘　　要：目的对空间诱变育成的"航茄"5号(HQ-5)进行RAPD分析,寻找特异扩增条带,试图为空间诱变育种提供分子生物学证据。方法"天水长茄"(TSCQ)和"龙果圆茄"(LGYQ)种子分别搭载"神舟"4号和"神舟"3号飞船进行空间诱变后,通过4代地面自交选育得到"天水长茄诱变第4代(TSCQ-MUT-4)"和"龙果圆茄诱变第4代(LGYQ-MUT-4)"2个突变系;利用后两者分别作为父母本进行杂交得到HQ-5。采用正交设计对其RAPD体系进行优化并进行RAPD分析。结果采用25μL扩增体系(DNA:40 ng,dNTPs:7.5 nmol,primer:10 ng和rTaq聚合酶:1.25 U)进行RAPD反应,发现"TSCQ-MUT-4"与野生型TSCQ相比产生4条差异带,LGYQ-MUT-4与野生型LGYQ相比产生1差异带;HQ-5与父母本相比均有6条差异带,有父母本共有带和特有带,并且保持航天诱变产生的变异带。结论空间诱变可能引起植物DNA水平的变异,有助于选育出新的农作物品种(系)。Objective To analyze the new breeding of eggplant ＂HANGQIE-5＂（HQ-5） generated by space mutagenesis with RAPD,identify the differential RAPD bands,and try to provide molecular proofs for space mutation breeding.Methods Dry seeds of two eggplants ＂Tianshuichangqie（TSCQ）＂ and Longguoyuanqie（LGYQ） were carried by ＂shenzhou-4＂ and ＂shenzhou-3＂spaceship.Two mutant lines ＂TSCQ-MUT-4＂ and ＂LGYQ-MUT-4＂ were selected after four generations of self-cross of TSCQ and LGYQ.A new variety of ＂HQ-5＂ was selected by crossing the two lines of ＂TSCQ-MUT-4＂ and ＂LGYQ-MUT-4＂,which were male and female,respectively.The RAPD reaction system was optimized with orthogonal design and further RAPD analysis was applied.Results The RAPD reaction system was determined as follows： DNA： 40 ng,dNTPs： 7.5 nmol,primer： 10 ng,and rTaq polymerase： 1.25 U.The RAPD results displayed that TSCQ-MUT-4 produced four differential bands compared with its wild type,while LGYQ-MUT-4 produced one,through space flight.Six differential fragments were amplified between HQ-5 and the paternal/maternal.HQ-5 not only shared the common fragments with its parents,but also held some particular ones,which were stably inherited.Conclusion The results indicate that space mutagenesis could cause mutations in DNA of plants,and is helpful to generate a new breeding.

关 键 词：空间诱变 RAPD 优化 茄子 

分 类 号：Q693[生物学—生物物理学]