新疆伽师瓜高效再生系统建立及抗真菌病基因转化

成熟子叶切块作为外殖体,经过组织培养获得再生植株。以MS作为基本培养基,取3日龄子叶块外殖体,选用MS 6-BA1.5 mg/L IAA0.3 mg/L诱导丛生芽,最高诱导率可达78%。而且分化再生过程,6-BA的浓度逐渐降低。在生根培养时,用MS 0.3 mg/L NAA诱导生根率较高,根系粗壮。在建立甜瓜高效再生体系的基础上,用根癌农杆菌介导法将几丁质酶基因和β-1,3-葡聚糖酶基因导入新疆甜瓜中,经卡那霉素的抗性筛选,获得转化的再生植株。用PCR反应及Southern-blot检测,进一步研究了再生植株的基因整合与表达情况。经PCR扩增反应和Southern-blot检测均得到了与阳性对照一致的特异性带,可初步证明外源基因已整合到了甜瓜基因组中。

Research on Construction of Regeneration System and Tranformation of Xinjiang 'Jiashi' Muskmelon with Anti-Fangal

In order to establish the transgenic plant system,the cotyledons of muskmelon as explants,regenerated plantlets were used in culture.Shoots regeneration were induced on MS medium as basic medium supplemented with 1.5 mg/L 6-BA 0.3 IAA mg/L.The highest frequency shoot regeneration observed is 78%.To achieve the high rate of rooting,the concentration of hormones 6BA was gradually decreased during the whole growth period.The 1/2 MS medium with 0.3 mg/L NAA was used for this propose.On the basis of regeneration protocol of Xinjiang muskmelon,two genes were transferred into muskmelon plant and selected of kanamycin in the medium.After kanamycin resistant plants were obtained,Integration of these genes into the genome of muskmelon plant was confirmed by PCR,Southern hybridization analyses.