摘要
为了揭示辣椒胞质雄性不育(CMS)机制,以辣椒胞质雄性不育系9704A和其相应保持系9704B为材料,对其花蕾转录组进行了Solexa测序,经过生物信息学分析鉴别出1个花药特异性表达的查尔酮合酶基因(CHS),该基因在保持系9704B中的表达水平为其对应不育系9704A的16倍;克隆后的序列比对分析表明,该基因属于CHS基因家族,与菸草花药特异表达的同源基因CHSLK的同源性达90%;用实时荧光定量RT-PCR对9704A和9704B的根、茎、叶中CHS基因的表达进行了定量分析,结果表明该基因在2种材料的茎中几乎不表达,在9704B的根和叶中表达量也极低,证实了在正常情况下该基因只在辣椒花药中有较高水平的表达;对不育系的定量PCR结果表明,CHS基因在9704A的根和叶中有异常的较高水平表达,结合其在花药中的低表达,推测辣椒不育系中CHS的异常表达可能与其CMS性状相关。
To investigate the molecular mechanism of cytoplasmic male sterile (CMS) in Capsicum annuum L., transcriptome Solexa sequencing was conducted on pepper flower buds of the CMS line 9704A and its correspondence homonuclear maintainer line 9704B. A gene that expressed 16 times higher in the maintainer liner 9704A than it is in the CMS line 9704B was identified. The cloned cDNA of this gene showed similarity to Chalcone Synthase Family genes with an overall sequence identity of 90% to Nicotiana sylvestris anther-specific gene-CHSLK. The expression of the gene in root, stem and leaf of the two peppers are tested by real-time fluorescence quantitative (RTFQ) RT-PCR. The result showed that the expression level of the gene in the stems of the two peppers was barely detectable, and in the root and leaf of the maintainer liner was very low, which confirmed that the gene is highly expressed specifically only in anther under normal condition. But the expression of this gene in roots and leaves of the CMS line 9704A is abnormally high, thus the abnormal CHS expression pattern in the CMS line may correlate to the male sterility.
出处
《湖南农业大学学报(自然科学版)》
CAS
CSCD
北大核心
2013年第3期259-264,共6页
Journal of Hunan Agricultural University(Natural Sciences)
基金
国家自然科学基金项目(31101425)
关键词
辣椒
查尔酮合酶基因(CHS)
定量分析
雄性不育
Capsicum annuum L.
chalcone synthase gene (CHS)
quantitative analysis
male sterility