香蕉茎尖遗传转化法研究

通过对多茎尖繁殖方法和转基因研究，建立了香蕉茎尖外源基因的遗传转化方法。明确了卡那霉素或Ｇ－４１８在转化植株中所使用的合适浓度，即当使用卡那霉素时，其产梢的浓度为１００ｍｇ／Ｌ，导根的浓度为１５０ｍｇ／Ｌ；但当使用Ｇ－４１８时，其产梢浓度则为２５ｍｇ／Ｌ其导根浓度则为３０ｍｇ／Ｌ或３５ｍｇ／Ｌ。通过基因枪和农杆菌共培养法尝试将黄瓜花叶病毒的衣壳蛋白基因转化香蕉组织，其结果表明：经过卡那霉素或Ｇ－４１８的筛选，仅有５％～７％的芽能够存活下来。通过选择性继代培养，最后获得了７３株株转化植株；对其中的２４株进行ＰＣＲ检测，结果表明阳性率达８３．３％；进一步的Ｓｏｕｔｈｅｒｎ ｂｌｏｔ检测表明：ＰＣＲ检测阳性的８１．８％植株含有外源衣壳蛋白基因。

Genetic Transformation System of Banana Meristems

A genetic transformation system with banana meristems. was established by studying banana multi--meristem propagation and foreign gene transformation. The optimal concentrations of media, i.e. 100 mg/L Kanamycin or 25 mg/L G--418 for shooting, 150 mg/L Kanamycin or 30-35 mg/L G--418 for rooting of transformed banana plants, were determined. Coat Protein (CP) of cucumber mosaic virus was transformed into banana tissues by gene gun bombardment and agrobacterium - mediated culture. 5 % ~ 7 % of transformed banana buds survived in the selected media, and 73 putative transgenic plants were finally Obtained. 83.3 % of 24 transformed plants showed positive in PCR analysis, and further Southern blot analysis indicated that 81.8 % of PCR positive plants contained foreign CP gene.