摘要
根据百脉根花对称性基因LjCYC3构建反义表达载体,经农杆菌介导正、反义LjCYC3基因转化烟草,以获得花型改变的转基因烟草植株,并根据表型选取正、反义各3株转基因植株和1株野生型植株进行Q-PCR分析,观察外源基因在远源植物中的作用和影响.PCR分析结果表明:转基因植株中正义基因和反义基因的转化率分别为62.5%和71%.表型观察显示:30%的正义转基因植株主要表现为花瓣和雄蕊数目减少;而40%的反义转基因植株表现为2朵花有一定程度的融合,花冠萼片有褪色现象,雄蕊端头有花瓣状结构,雌蕊2枚且与雄蕊有合生现象.Q-PCR分析结果表明:在正义转基因再生植株中,外源基因均高效表达,随着表达量的增加,花型变化更加明显;而在反义转基因再生植株中,外源基因也有表达,随着表达量的降低,花型变化更加明显;与正义基因表达量相比,反义基因表达量要小约150倍.本研究结果为下阶段用该基因转化花卉植物以改变花型、改良花卉品种打下了实验基础.
According to LjCYC3 gene controlling flower symmetry in Lotus japonicus,the antisense gene expression vector was constructed,and by using Agrobacterium-mediated transformation of tabacoo with justice-antisense LjCYC3 gene,flower patterns changed in transgenic tobacco plants were obtained,and then three transgenic plants and a wild-type plant were selected for Q-PCR analysis according to phenotype.PCR analysis showed that transformation rates of justice gene and antisense gene were 62.5% and 71% in transgenic plants,respectively.Phenotypic observations showed that 30% of justice transgenic plants mainly appeared to a decrease number in petals and stamens,however,40% of the antisense transgenic plants appeared to have two flowers fusion to some degree,and color fading for petals and sepals,the head of stamens having petal-shaped structures,and two pistils being connate with stamens.The Q-PCR analysis results showed that foreign genes were highly expressed in the justice transgenic plants,and the more expression increased,the more pronounced pattern changed.The foreign gene was also expressed in the antisense transgenic plants,and as the reduction of the expression,pattern change was even more evident.Compared with the justice gene expression quantity,the antisense gene expression quantity was 150 times lower.The results above lay a good foundation for transformation of flowering plants with the LjCYC3 gene to change flower pattern and improve flower varieties for the next stage.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2011年第1期13-21,共9页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
国家自然科学基金资助项目(30671059)
