---

作 者 ：李明亮 韩一凡 邱德有 李凝 田颖川

期 刊 ：林业科学 1999年 35卷 3期 页码：10-15

关键词：ACC(l-aminocyclopropane-l-carboxylate)合酶；基因；反义RNA；美洲黑杨；

Keywords:ACC(l-aminocyclopropane-l-carboxylate)synthase, Gene, Antisense RNA, Populus deltoides.,

摘 要 ：利用RT-PCR技术克隆了大豆ACC合酶基因GMCACCSI编码区1.5kb的cDNA片段,经酶切图谱分析和序列分析鉴定后,反向插入到2元载体pBin438中,构建了表达ACC合酶反义RNA的植物表达载体,转化农杆菌后用该农杆菌侵染美洲黑杨叶片,在含卡那霉素的MS培养基上选择转化子和植株再生,通过PCR检测从抗卡那植株中选到15株转基因植株,Southern杂交分析初步确证了外源基因是以单拷贝插入到杨树基因组DNA中;对杨树幼苗乙烯释放的测定结果表明转基因杨树幼苗的乙烯释放量为对照的22%。

Abstract:A 1.5 kb fragment of ACC synthase cDNA fragment prepared from total RNA of．soybean was amplified by polymerase chain reaction(PCR)and cloned into pGEM^R-T vector．The cloned ACC synthase gene was further inserted into a binary vector，pBin438，in an inverted orientation between the CaMV 35 S promoter and Nos 3′termination sequence(pBACS)．Transgenic poplar plants were obtained by regenera— tion of Agrobacterium-mediated transformation of leaves of Populus deltoides．PCR and Southern blotting analyses confirmed the integration of a single antisense ACC synthase gene in transformed poplar genome． The results form reverse transcription PCR(RT-PCR)of RNAs isolated from transgenic poplar leaves confimed that the antisense RNA of ACC synthase presented in these transgenic plants．The amount of ethylene released from transgenic poplar was reduced significantly to about 22％of that released from non-transformed control poplars.

---