摘要
以农杆菌介导法将黄瓜开花整合子CFL基因分别对仙客来幼嫩叶片、叶柄及愈伤组织进行遗传转化,比较了三种转化受体的遗传转化效率。应用超声波辅助的农杆菌介导法,对以叶片为转化受体的仙客来遗传转化体系进行了优化。结果表明:幼嫩叶片是最佳的转化受体,再生植株GUS阳性率达27.25%。叶片在农杆菌浸染时辅以60s的超声波处理,经超声波处理的转化效率与未经超声波处理的相比,叶片组织的GUS染色阳性率提高4倍。经GUS染色、PCR和RT-PCR检测,初步证明CFL基因已整合到仙客来再生植株的基因组中。本研究成功将CFL基因转入仙客来,并建立了高效的仙客来遗传转化体系,这不仅为培育早花仙客来新品种提供新的途径,也为进一步用分子手段对仙客来进行遗传改良奠定了基础。
Leave, petioles and callus in Cyclamen persicum were transformed with cucumber flowering integrator gene CFL by Agrobacterium tumefaciens. The transformation efficiency of three kinds of receptors have been compared. Furthermore, the leaf genetic transformation system was optimized by SAAT （Sonication-assisted Agrobacterium-mediate Transformation）. The results showed that GUS positive frequency of regenerated plants was highest, up to 27.25% when young leave were used as transformation receptors. By adding 60 seconds of Sonication-assisted treatment at Agrobacterium infecting sametime, GUS positive frequency of leave was four times higher than control. By histochemical assays of GUS and analysis of PCR, RT-PCR, it was confirmed preliminarily that CFL gene integrated into genome of the regenerated plants. CFL gene was transformed into Cyclamen persicum successfuliy in this study, and a high-efficiency genetic transformation system was established. It not only provide a new way for breeding early flowering variety, but also lay the foundation for the further genetic improvement by molecular means in Cyclamen persicum.
出处
《分子植物育种》
CAS
CSCD
2011年第5期629-634,共6页
Molecular Plant Breeding
基金
国家自然科学基金项目(31071818)资助
关键词
仙客来
花期
黄瓜CFL基因
农杆菌介导法
Cyclamen persicum
Flowering time
CFL gene
Agrobacterium-mediated transformation
