摘要
MADS-box基因在植物花发育的整个阶段起到了极为关键的作用。根据山核桃Carya cathayensis雌花芽454测序得到的基因片段,采用c DNA末端快速扩增(RACE)技术,获得了1条Cc MADS-like基因。序列分析结果表明:Cc MADS-like开放阅读框(ORF)长度为609 bp,编码202个氨基酸,具有典型的MADS-box结构域,包括高度保守的MADS盒和K区以及I区、C区。Cc MADS-like与拟南芥Arabidopsis thaliana,金鱼草Antirrhinum majus,美洲黑杨Populus deltoides,欧洲大叶杨Populus trichocarpa等的同源基因所编码的蛋白质同源性分别达到48%,65%,69%,71%。山核桃不同组织实时荧光定量聚合酶链式反应(q RT-PCR)结果表明:Cc MADS-like基因在花芽中表达量远远高于其他组织,暗示该基因极有可能参与山核桃成花发育。酵母单杂交实验表明,Cc MADS-like蛋白具有转录激活活性。
MADS-box family genes play key roles in all phases of floral development. Based on a partial fragment acquired from 454 sequencing of Carya cathayensis （hickory） floral buds, a full-length of a MADS- like gene was obtained by rapid amplification of cDNA ends （RACE）. Afterwards, a quantitative real-time （qRT） polymerase chain reaction （PCR） analysis, an amino acid sequence alignment, and a yeast one-hybrid assay were performed. Results of the sequence analysis revealed the open reading frame （ORF） of the Cc- MADS-like gene was 609 bp in length and encoded a protein of 202 aa. The protein presented typical characteristic of MADS-box family genes and contained MADS domain, I, K, and C, with the MADS domain was considerably conserved. The amino acid sequence alignment showed that the putative protein CcMADS-like gene had an identity to homologies of Arabidopsis thaliana （48%）, Antirrhinum majus （65%）, Populus deltoides （69%）, and Populus trichocarpa （71%）. The qRT-PCR analysis revealed that the expression level of CeMADS-like gene in flower buds was mueh higher than in other tissues. The yeast one-hybrid showed that the CcMADS-like gene had an activation activity as a transcription factor. Overall, the qRT-PCR analysis suggested that CeMADS-like gene possibly partieipated in floral development of hickory.
出处
《浙江农林大学学报》
CAS
CSCD
北大核心
2015年第1期33-39,共7页
Journal of Zhejiang A&F University
基金
国家高技术研究发展计划("863"计划)项目(2013AA102605)
国家重点基础研究发展计划("973"计划)项目(2011CB111500)
国家自然科学基金资助项目(31170637)
浙江农林大学科研发展基金资助项目(2010FR072)
