| 摘要: |
| 【目的】对蕙兰CfMADS1基因cDNA全长进行克隆和时空表达特性研究，为研究成花相关基因的功能提供参考。【方法】以蕙兰萼片cDNA为模板，利用反转录RT PCR和RACE的方法，克隆获得CfMADS1全长cDNA序列，并对其进行生物信息学分析。以蕙兰各时期的器官组织为材料，利用荧光定量PCR进行该基因的时空表达分析。【结果】 成功克隆了CfMADS1基因（GenBank登录号为KC148540），其cDNA序列全长1 061 bp，开放阅读框长744 bp，编码247个氨基酸，分子式为 C1 244H2 040N370O377S9。其编码的氨基酸序列与AP1/FUL亚家族中金钗石斛MADS1具有较高的同源性 (85.43％)，与AP1/FUL转录因子亚家族中的蛋白聚为一类。该基因编码蛋白没有明显的信号肽和跨膜域，极可能位于细胞核内，具有MADS保守域(1－61aa)和相对保守的K区（87－178aa），而且在C末端具有AP1/FUL特征基序（LPPWML），二级结构中α-螺旋所占比例较高 (55.63％)，三级结构与月季、水稻和水仙MADS1蛋白结构非常相似。该基因在盛花期叶中表达最强烈，花蕾期叶、葶、蕾及营养期叶、盛花期葶中表达较为强烈，子房和营养期根中表达较弱，其余器官组织中表达痕量。【结论】从蕙兰萼片中克隆得到了CfMADS1 cDNA全长序列，CfMADS1与蕙兰的成花诱导、花发育以及果实形成有关。 |
| 关键词: 蕙兰 AP1/FUL-like基因 时空表达 |
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| 基金项目:河南省科技攻关项目（092102110128）；河南省教育厅科学技术研究重点项目（14A22005） |
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| Cloning and spatiotemporal expression of CfMADS1 gene in orchid (Cymbidium faberi) |
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TIAN Yun-fang,YUAN Xiu-yun,JIANG Su-hua,et al
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| Abstract: |
| 【Objective】This study cloned a full-length APETALA1/FRUITFULL like (AP1/FUL like) MADS box cDNA from Cymbidium faberi and analyzed its spatiotemporal expression to provide a reference for studying function of flower related genes.【Method】CfMADS1 gene was isolated from sepals of C.faberi by RT-PCR and RACE.Then spatiotemporal expression was detected by real time PCR.【Result】CfMADS1 gene was cloned (GenBank number:KC148540).The full length of cDNA sequence is 1 061 bp,with open reading frame of 744 bp,encoding 247 amino acids.The molecular formula is C1 244H2 040N370O377S9.The deduced amino acid sequence of CfMADS1 shared 85.43% homology with a member of the AP1/FUL like group of MADS box genes (Dendrobium nobile MADS1).CfMADS1 belonged to the AP1/FUL transcription factor subfamily.The deduced protein has no obvious signal peptide and transmembrane domain,and probably located in the nucleus.It had a MADS domain (1－61aa),a relatively conservative K region（87－178aa）and a C terminal conserved motif LPPWML.The secondary structure of CfMADS1 mainly consisted of alpha helices (55.63%),and the three-dimensional structure of the protein was similar to that of homologues in Roza hybrida,Oryza sativa and Narcissus tazetta.Real-time quantitative PCR (qRT-PCR) results revealed low levels of its mRNA in ovaries and roots during vegetative period,and in other tissues.Highest levels of CfMADS1 expression were observed in leaves at full-blossom stage followed by pedicels at reproductive period,leaves at vegetative period,and leaves,pedicels,and buds at bud stage.【Conclusion】A full-length AP1/FUL like MADS box cDNA was cloned from C.faberi sepals and CfMADS1 involved in floral induction,floral development and fruit formation. |
| Key words: Cymbidium faberi AP1/FUL-like gene spatiotemporal expression |
