文章摘要
条斑紫菜单孢子形成相关基因(PyMFG)的特性及其表达差异分析
Characteristics of formation related genes (PyMFG) of monospores and analysis of differential expression in Pyropia yezoensis
投稿时间:2019-03-26  修订日期:2019-05-28
DOI:
中文关键词: 条斑紫菜,RNA-Seq,单孢子,PyMFG,TEA-ATTS超家族,qRT-PCR
英文关键词: Pyropia yezoensis  RNA-Seq  monospores  PyMFG  TEA-ATTS superfamily  qRT-PCR
基金项目:国家重点研发计划“蓝色粮仓科技创新”重点专项;国家自然科学基金;浙江省农业(水产)新品种选育重大科技专项;江苏省科技计划(现代农业)重点项目;福建省海洋生物增养殖与高值化利用重点实验室开放课题基金
作者单位E-mail
宋珊珊 水产种质资源发掘与利用教育部重点实验室上海海洋大学 xiaomusym@163.com 
丁洪昌 水产种质资源发掘与利用教育部重点实验室上海海洋大学  
严兴洪 水产种质资源发掘与利用教育部重点实验室上海海洋大学
水产科学国家级实验教学示范中心上海海洋大学
上海水产养殖工程技术研究中心上海海洋大学 上海 
xhyan@shou.edu.cn 
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中文摘要:
      本实验以条斑紫菜转录组测序筛选得到的差异性表达基因(Contig-21827)序列为基础,采用RT-PCR技术克隆得到了条斑紫菜单孢子形成相关基因PyMFG的完整开放阅读框。对其进行序列分析发现,PyMFG基因的ORF全长为1224 bp,编码一个包含407个氨基酸的多肽片段,其分子量为46.24 kDa,等电点为9.08。结构域分析结果显示,该蛋白含有保守的TEA结构域和YAP结构域,属于TEA-ATTS超家族结构域。通过多序列比对和系统进化分析发现,该蛋白与真菌分生孢子形成蛋白聚为一个大的分支,亲缘关系较近。此外,不同品系间的单孢子放散能力和qRT-PCR分析结果显示,PyMFG基因在PY26W和PY26W’ 品系中的表达趋势与宏观统计结果高度一致,在PY26R和PY26R’ 品系中的表达则与统计结果存在差异。由此推断,父本(PY26W)/偏父本品系(PY26W’)中调控单孢子形成、放散的相关基因种类和具体分子机制与母本(PY26R)/偏母本品系(PY26R’)存在差异,因而导致单孢子的宏观统计结果与荧光定量结果存在差异。
英文摘要:
      In our study, a complete open reading frame of PyMFG was obtained by RT-PCR based on the sequence of differentially expressed genes (Contig-21827) screened by transcriptome sequencing of Pyropia yezoensis. Sequence analysis showed that the ORF of the PyMFG was 1224 bp in length and encoded a polypeptide fragment of 407 amino acids with a molecular weight of 46.24 kDa, and theoretical pI of 9.08. Domain analysis revealed that the protein contains a conserved TEA domain and a YAP domain, which belongs to the TEA-ATTS superfamily domain. We found that the protein and fungal conidia forming proteins clustered into one large branch with close genetic relationship through multiple sequence alignment and phylogenetic analysis. In addition, the comparison of the ability of releasing monospores and qRT-PCR analysis between different strains indicated that the expression trend of PyMFG in PY26W and PY26W' was highly consistent with macroscopic statistical results, while the trend in PY26R and PY26R' were exactly existed difference. By inference, the gene type and molecular mechanism regulating the formation and release of monospores in the parental (PY26W) and partial parental strain (PY26W') and the female parental (PY26R) as well as partial female strain (PY26R') existed certain difference, leading to the divergence between the macroscopic statistical results of monospores and the qRT-PCR results.
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