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雄性不育相关基因msLTP的反义RNA验证
引用本文:田爱梅.雄性不育相关基因msLTP的反义RNA验证[J].西北植物学报,2012,32(5):890-894.
作者姓名:田爱梅
作者单位:西安文理学院生物技术学院,西安,710065
基金项目:国家自然科学基金(30671426)
摘    要:根据普通白菜雄性不育相关的脂质转移蛋白基因(msLTP)的cDNA序列设计引物,从普通白菜花蕾的cDNA中扩增出312bp的片段,然后将该片段连接至双元载体pBI12l中,得到反义RNA植物表达载体并导入农杆菌LBA4404菌株中,通过农杆菌介导法转化菜心;利用PCR和Southern blot分析检测得到了25株转基因植株,转基因植株的花粉部分畸形或空瘪,花粉离体萌发率为38.56%,较未转化植株的萌发率(76.32%)降低了37.76个百分点.研究表明,反义RNA技术使msLTP基因沉默而导致了菜心转基因植株的部分花粉发育不良,说明msLTP基因在普通白菜和菜心等花粉发育中具有重要作用.

关 键 词:脂质转移蛋白  雄性不育  反义RNA  花粉

Validation of a msLTP Gene Related to Male Sterility by Antisense RNA
TIAN Ai mei.Validation of a msLTP Gene Related to Male Sterility by Antisense RNA[J].Acta Botanica Boreali-Occidentalia Sinica,2012,32(5):890-894.
Authors:TIAN Ai mei
Institution:TIAN Ai-mei(College of Biological Technology,Xian University of Arts and Science,Xi’an 710065,China)
Abstract:Based on the cDNA sequence of msLTP related to male sterility of Chinese cabbage-pak-choi(Brassica campestris ssp.chinensis var.communis,syn.B.rapa ssp.chinensis var.communis),a pair of specific primers were designed and a 312 bp fragment was amplified from the cDNA of flower buds of Chinese cabbage-pak-choi.The fragment was introduced to binary vector pBI121,and the generated antisene RNA vector was then mobilized into Agrobacterium tumefaciens strain LBA4404.The A.tumefaciens harboring the msLTP fragment was transformed to flowering Chinese cabbage(B.campestris ssp.chinensis var.parachinensis) via tissue culture and Agrobacterium-mediated transformation.The transgenic plants were confirmed by PCR and Southern blot,and 25 independent transgenic plants were obtained.The pollen grains from transgenic plants exhibited abnormal shapes,and only 38.56% of pollen grains from these plants germinated normally.The germination rate was reduced 37.76 percentage points than that of pollen grains from nontransgenic plants(76.32%).These results indicated that msLTP gene interrupted by antisense RNA resulted in partial pollen grains abortion in flowering Chinese cabbage,suggesting that msLTP gene possesses an important role in pollen development of Chinese cabbage crops such as Chinese cabbage-pak-choi and flowering Chinese cabbage.
Keywords:LTP  male sterility  antisense RNA  pollen
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