胡钟东;乔玉山;王三红;姚泉洪;章镇;

• 1:南京农业大学园艺学院
• 2:南京农业大学园艺学院
• 3:南京农业大学园艺学院
• 4:上海市农业科学院生物技术研究中心

摘要(Abstract)：

以若光梨成熟果实为材料,提取总RNA,在ACC氧化酶基因(ACO)cDNA序列同源性较高区域设计一对特异引物,利用RT-PCR克隆了ACC氧化酶(ACO)基因片段。将ACO反义基因、与副球菌中类胡罗卜素合成有关的间隔基因YYT和ACO正义基因3个片段串联在一起,经鉴定后,插入到植物表达载体中,构建成能够表达ACC氧化酶基因的双链RNA的植物载体pYF028,为耐贮砂梨的遗传转化创造条件。

关键词(KeyWords)： 梨;ACC氧化酶(ACO);RT-PCR;克隆;RNAi载体构建

Abstract:

Keywords:

基金项目(Foundation): 江苏省高技术研究项目(BG2006313);; 南京农业大学青年科技创新基金项目(KJ05007)。

作者(Author): 胡钟东;乔玉山;王三红;姚泉洪;章镇;

Email:

参考文献(References)：

• [1]BOLITHO K M,LAY-YEE M.Antisense apple ACC-oxidase RNA reduces ethylene production in transgenic tomatofruit[J].Plant Sci-ence Limerick,1997,122(1):91-99.
• [2]SCORIA R.Progress in tree fruit improvement through molecular genetics[J].HortScience,2001,36:855-857.
• [3]XIONG A S,YAO Q H,LI X,et al(熊爱生,姚泉宏,李贤,等).Double antisense ACC oxidase and ACC synthasefusion gene intro-duced into tomato by agrobacterium-mediated transformation and analysis the ethylene production of transgenic plants[J].Acta Biolo-giae Experimentalis Sinica(实验生物学报),2003,6:428-434.(in Chinese)
• [4]FIRE A,XU S Q,MONTGOMERY MK,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhaditis ele-gans[J].Nature,1998,391:806-811.
• [5]ELBASHIR S,HARBORTH J,LENDECKEL W,et al.Duplexed of21-nucleotide RNA mediate RNA interference in cultured mam-malian cells[J].Nature,2001,411:494-498.
• [6]SHINJIROO,HIROTAKAU,YUBE Y,et al.Producing decaffeinat-ed coffee plants[J].Nature,2003,423:823.
• [7]XIONG A S,YAO Q H,PENG R H,et al.Different effects on ACC oxidase gene silencing triggered by RNA interference in transgenic tomato[J].Plant Cell Report,2005,23(9):639-646.
• [8]JAAKOLA L,PIRTTILA A,HALONEN M,et al.Isolation of high quality RNA from bilberry(Vaccirrium myrtillus L.)fruit[J].Molecu-lar biotechnology,2001,19:201-203.
• [9]LOEB K R,JEROME K R,GODDARD J,et al.High-throughput quantitativeanalysisofhepatitisBvirusDNAinserumusingthe Taq-Manfluorogenicdetectionsystem[J].Hepatology,2000,32:626-629.
• [10]CAOYH,ZHANGZ,YAOQH,et al(曹艳红,章镇,姚泉洪,等).Research on apple polyphenol oxidase double-stranded RNA(DSR-NAi)[J].ActaBiologiaeExperimentalisSinica(实验生物学报),2004,37(6):487-493.(in Chinese)
• [11]DONGJ G,FERNANDEZM,YANGS F.Purification and character-isation of1-aminocyclopropane-l-carboxylic acid oxidase from ripe apple fruit[J].Proc Natl Acad Sci USA,1992,89:9789-9793.
• [12]ROTTMANN W H,PETER G F,OELLER P W,et al.1-aminocy-clopropane-l-carboxylate synthase in tomato is encoded by a multi-gene family whose transcription is induced during fruit and floral senescence[J].Mol Biol,1991,222:937-961.
• [13]VANDER GEEST A H,HALL T C.Theβ-phaseolin5'matrix at-tachment region acts as an enhancer facilitator[J].Plant Mol Biol,1997,3(3):553-557.