香蕉ACC合成酶反义基因转化香蕉的研究

Genetic Transformation of ACC Antisense Gene into Banana

香蕉是重要的热带、亚热带地区作物,典型的呼吸跃变型果实,不耐贮藏。传统的保鲜方法存在诸多弊端,利用现代生物技术培育耐贮藏新品种,已经成为我们解决香蕉保鲜问题的重要课题。本研究利用果实特异性ACC合成酶基因反义植物表达载体,采用基因枪和农杆菌介导法转化海南主栽的巴西香蕉及红香蕉,并比较了不同转化方法及不同栽培品种的转化效果。结果表明农杆菌侵染法好于基因枪法,在农杆菌侵染方法中,对红香蕉的转化效果好于巴西香蕉。经PCR检测获得了9株转化植株,进一步采用Southern blot分析的方法,其中4株杂交信号较强,确认ACC合成酶反义基因已经整合进香蕉基因组中。本研究为香蕉的转基因研究和耐储藏新品种的培育奠定了一定的基础。

Banana （Musa AAA group） is one of important tropical and subtropical fruits, with the feature of typical climacteric fruit, The climacteric fruit cannot be stored for a long time because of the aspiratory climacteric. Most cultivars of banana are triplets and hard to be genetically improved by using conventional breeding methods. As a result, genetic engineering is a potential powerful tool to improve banana and develop new banana cultivars, In this report, a construct with a fruit-specific expressed ACC synthase gene inserting in reverse direction, which could deliver and express antisense RNA of ACC synthase gene, named as pBBB-aACC, was constructed. The construct was transformed to two cultivars of banana, each by using both particle bombardment and Agribacteriurn-mediated transformation. The results showed that transformation ofpBBB-aACC into banana mediated by A - gribacterium was more effective than that by particle bombardment. Genotype of red banana （a cultivar with red peel） was transformed easier than that of Brazilian Banana genotype （a cultivar from Brazil）, By PCR detection, we obtained 9 transformed plants, 4 plants of which had the strong hybridization＇ s signal using Southern blot analysis, then we would affirm that antisense RNA of ACC synthase gene had been integrated into banana genome. These studies would provide the basis for the transgenic research of banana and developing of new storage-tolerance cultivars.