转GAFP-NPI基因香石竹  

Transfering the GAFP-NPI Gene into Dianthus caryophyllus

作　　者：何秋伶[1] 陈英[1] 戴咏梅[1,2] 杨春霞[1] 黄敏仁[1] 

机构地区：[1]南京林业大学林木遗传和基因工程重点实验室 [2]上海绿化管理局,上海200001

出　　处：《分子植物育种》2007年第3期314-318,共5页Molecular Plant Breeding

基　　金：上海科技兴农项目(沪农科攻字2004第4-1号)资助。

摘　　要：将双价抗病基因转入同一植物,是增强植物抗病性、拓宽抗病谱的有效措施。在建立香石竹叶片离体培养再生体系的基础上,将兔防御素基因(NP-I)和天麻毒蛋白基因(gastrodia antifungal protein,GAFP)构建在同一个植物表达载体pBin35SGAFP-NPI上,通过农杆菌介导法,利用香石竹叶片作外植体,将GAFP-NPI双价抗病基因导入香石竹品种Asso、Tuareg、ciao Bianco,经卡那霉素筛选共获得300余株转基因植株,并进行多重PCR检测,证明GAFP-NPI双价抗病基因已整合进香石竹的基因组中。琼脂糖孔隙扩散实验证明转基因香石竹对绿色木霉具有抑菌活性,说明了GAFP和NPI双价抗病基因在转基因植株中得到了表达。Transformation of multi-genes of disease resistance into plants has been an efficient measure both for improving disease resistance and extending disease resistant spectnma. The high frequency regeneration system of carnation from leaf to callus and then to plantlets has been established. GAFP and NP-I genes were constructed on one vector （pBin35SGAFP-NPI）, performed via agrobacterium-mediated method and there are over 300 Kanamycin-resistant plants that were regenerated through Kanamycin selection at last. The results of the PCR analysis showed that the GAFP-NPI gene has integrated into genomes, in vitro antibiosis assay to Trichoderma of the extract of the transgenic plants suggested that the transgenic plants have the ability of disease-resistance and the two genes can be expressed efficiently in transgenic carnation.

关 键 词：转基因香石竹 抗病 GAFP-NPI基因 离体抑菌活性 

分 类 号：S682.19[农业科学—观赏园艺]