摘要
采用cDNA-AFLP差异显示技术对大豆细胞质雄性不育系NJCMS2A与其保持系NJCMS2B间基因差异表达进行研究,结果从NJCMS2A花蕾中分离到一个差异表达片段,对该差异片段进行克隆、测序和序列比对分析,Blast检索结果显示它与大豆基因组中Gm13上g29510.1 cDNA片段的同源性达98.7%,与大豆中一个MADS-box基因的同源性达98%,氨基酸序列比对结果表明它与大豆中一个MADS-box蛋白有96%的同源性,与豌豆中MADS-box M7蛋白有83%的同源性,与苦瓜中MADS-box2蛋白有88%的同源性,与海岛棉典型的MADS-box基因编码的AGAMOUS蛋白保守区有83%的同源性,进一步对其氨基酸序列进行结构和功能预测显示该差异片段具有MADS-box转录因子的典型结构域K-box,证明其编码蛋白为一MADS-box转录因子,半定量RT-PCR分析结果显示其在NJCMS2A花蕾中表达量很高,而在NJCMS2B花蕾中表达量很低,推测该差异片段可能与大豆细胞质雄性不育有关。
CMS（cytoplasmic male sterility） plays an important role in the utilization of crop heterosis.It is of important significance on theory and practice to study the genetic base and mechanism of CMS.To reveal the molecular mechanism of soybean cytoplasmic male sterility,we analyzed the gene differential expression between the soybean cytoplasmic male sterile line NJCMS2A and its maintainer line NJCMS2B by the cDNA-AFLP differential display method.A differentially expressed fragment from the flower buds of NJCMS2A was cloned and sequenced.The Blast results showed that the differentially expressed fragment shared 98.7% homology with g29510.1 cDNA fragment on Gm13 of soybean genome,and 98% homology with a MADS-box gene of soybean.The Blast results of the amino acid sequences indicated that the differentially expressed fragment shared 96% homology with a MADS-box protein of Glycine max,83% homology with the MADS-box M7 protein of Pisum sativum,88% homology with the MADS-box 2 protein of Momordica charantia,and 83% homology with AGAMOUS protein of Gossypium barbadense.The structure and function prediction of the amino acid sequence showed that the protein encoded by the differentially expressed fragment was the transcription factor of the MADS-box gene which contained a typical K-box domain.The results of semi-quantitative RT-PCR showed that the expression quantity of the differentially expressed fragment in the flower buds of NJCMS2A was much higher than that of NJCMS2B.According to the above results,it was inferred that the differentially expressed fragment was probably related to the soybean cytoplasmic male sterility.
出处
《作物学报》
CAS
CSCD
北大核心
2010年第6期905-910,共6页
Acta Agronomica Sinica
基金
国家高技术研究发展计划(863计划)项目(2009AA101106)
作物遗传与种质创新国家重点实验室开放基金(ZW2007004)资助
