番茄ACO基因的克隆及其RNAi对乙烯产量的抑制.pdfVIP

农业生物技术学报 Journal ofAgriculturalBiotechnology 2007,15 (3): 464~468  ·研究论文 · Cloningof  GeneandInhibition ofEthylene  Evolution inTomatoeswith RNA Interfernce*  1,3,4  1  2  1,2  CHENYin­hua  ,OUYANGBo ,LIHan­xia ,YEZhi­biao  **  A fragment of 1018bp of  genecDNA sequencewascloned fromtomato (  ) leavesincu­  bated withpathogen usingRT­PCR withtwo PCR primersdesigned accordingto the sequence oftomato cDNA clone (  ).There­  sultthrough BLASTsearch showed the sequencepresenting averyhighmatchwith the  genesinthe otherplantsand itshomolo­  gywas from 83%to 99%.Usingthis sequence,a RNA interference (RNAi) transformation vector (pD311)was constructed through  thewayofBP cloning andtransformed intotomato. Twenty­sevenregeneratedplantswith kanamycinresistancewere obtained,show­  ing that the transgene integrated into tomato genome was provedwith PCR and Southern blot. The ethylenerespiration amount of the  RNAitransgenictomato plantswas measuredby gaschromatography,resulting inthat ethylene evolutionwas specifically inhibitedin  leavesand fruitsofthe transgenicplants.  tomato;ACCoxidasegene;RNA Interfernce;transformation 番茄 基因的克隆及其 RNAi 对乙烯释放的抑制 1,3,4 1 2 1,2 ** 陈银华  ，欧阳波  ，李汉霞  ，叶志彪  1. ， 武汉 430070 2. ）， 武汉 430070 （ 华中农业大学作物遗传改良 国家重点实验室 ； 华中农业大学国家蔬菜改良中心 （华中分中心 ； 3. ， 海口 570228