摘要
研究了西瓜品种ZKM的再生体系和农杆菌介导小西葫芦黄花叶病毒(Zucchini yellowmosaic virus,ZYMV)的外壳蛋白(cp)基因导入西瓜的遗传转化体系。结果表明,质量分数0.6%琼脂是适宜种子萌发的基本培养基,切取5d龄的西瓜无菌苗子叶为外植体,接种在MS+BA2.0mg/L诱导培养基上,诱导不定芽效率最高。选用75mg/L卡那霉素筛选西瓜抗性芽较为合适,外植体经预培养2～3d,菌液浓度以OD600nm值为0.4,共侵染10min有利于提高西瓜转化的效率。经PCR检测,ZYMVcp基因已经导入西瓜植株,转化频率为0.15%。
The regeneration and transformation of watermelon with Zucchini yellow mosaic virus coat protein gene by Agrobacterium tumefaciens were studied. The results showed that 0.6% agar was suitable to watermelon seed germination. The explants taken from 5 day-old seedlings were better for regeneration and transformation. MS medium containing 2.0 mg/L benzyladenine（BA） was the best induction medium. To induce resistant adventitious shoots, 75 mg/L kanamycin was suitable for selection. Pre-culturing explants for 2 to 3 days and infection with Agrobacterium tumefaciens LBA 4404 with OD60nm of 0.4 for 10 minutes could improve transformation rate. The transformation frequency was 0.15%. The integration of ZYMV cp gene into the regenerated progenies was confirmed by PCR.
出处
《果树学报》
CAS
CSCD
北大核心
2007年第4期496-501,共6页
Journal of Fruit Science
基金
国家重大基础研究973前期专项(2005CCA01700)
国家863项目(2004AA241171)
河南省自然科学基金(0511030800)
关键词
西瓜
外壳蛋白
遗传转化
Watermelon
Coat protein
Transformation
