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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2016, Vol. 42 Issue (3): 313-320    DOI: 10.3785/j.issn.1008-9209.2015.10.091
生物科学与技术     
金针菇疏水蛋白Fv-Hyd1及其潜在的转录因子Fv-Rtg3的共表达分析
陈仁良, 李肖, 龙莹, 张磊, 严俊杰, 黄千慧, 熊宏民, 谢宝贵
1.福建农林大学生命科学学院菌物研究中心,福州350002;2.福建农林大学园艺学院,福州 350002
Analysis on the co-expression of hydrophobin Fv-Hyd1 and its potential transcription factor Fv-Rtg3 in Flammulina velutipes
CHEN Renliang, LI Xiao, LONG Ying, ZHANG Lei, YAN Junjie, HUANG Qianhui, XIONG Hongmin, XIE Baogui
1. Mycological Research Center of Life Science College, Fujian Agricultural and Forestry University, Fuzhou 350002, China; 2. College of Horticulture, Fujian Agricultural and Forestry University, Fuzhou 350002, China
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摘要： 疏水蛋白(hydrophobin,Hyd)普遍存在于真菌的气生菌丝、侵染结构或子实体表面,且存在时空性高丰度表达。不同真菌疏水蛋白在其各个发育阶段发挥不同的功能,如气生菌丝的生长、子实体原基的形成及其形态分化等。虽然已有众多报道指出Hyd与生物或非生物环境刺激所产生的抗逆性有关,但是对它与细胞分裂分化等生物学功能及其信号转导调控的关系仍了解甚少。通过培养金针菇(Flammulina velutipes)菌丝以及金针菇出菇实验,结合基因组及转录组数据、蛋白结构预测、系统发育进化树聚类分析等生物信息学分析鉴定金针菇中一个编码疏水蛋白基因Fv-Hyd1,并利用其上游序列预测该疏水蛋白基因潜在的调控转录因子,辅以实时荧光定量PCR(real time fluorescence quantitative polymerase chain reaction,qRT-PCR)数据和皮尔森相关系数鉴定它们的表达关系。结果表明,Fv-Hyd1与金针菇菌丝扭结和子实体原基的形成具有一定的相关性,与其潜在的转录因子Fv-Rtg3之间存在较强的共表达规律,并以此结合蛋白磷酸化位点预测,推测基因Fv-Hyd1的表达调控机制。
Abstract: Flammulina velutipes, with high edible and medicinal value, was widely cultivated in China and Southeast Asia. Being worthy to be noted, the fruiting body formation and development of F. velutipes were influenced by many factors, such as the internal factors including transcription factor, protein kinase, sparse water protein and the cell pigment, and environmental factors including light, and temperature and carbon dioxide. It is very important to understand the response of genes to the influencing factors in transcription level, translation level and environmental stimuli for studying the regulation of F. velutipes fruiting body growth and development. 　　Among them, the hydrophobins were commonly found in fungal hyphae, infection structure or surface of the fruiting body, and there is a temporal and spatial expression with high abundance. Different fungal hydrophobins perform different functions in various stages, such as aerial hyphae growth, fruiting body formation and morphological differentiation of the primordium. Although there are numerous reports that the hydrophobins result from stress-related biological or non-biological environment, but the biological functions such as cell division, differentiation and its regulation of signal transduction are still poorly understood. For example, the analysis on Schizophyllum commune genome and expression profile showed that in S.commune fruiting body development process, the expression of energy and hydrophobins related genes were higher in knot than the other stages. Besides, we found that the expression of hydrophobins Fv-Hyd1 were upregulated in knot and mycelium. We hypothesized that hydrophobin Fv-Hyd1 played an important role in aggregation in mycelium and primordia formation. 　　Through the F. velutipes genome and transcriptome data, as well as the bioinformatics analysis, a gene coding hydrophobin Fv-Hyd1 encoded 131 amino acids, with 6 exons and 5 introns. Through physical and chemical properties, composition arrangement of cysteine C-X, protein tertiary structure and phylogenetic tree clustering analysis, it was showed that Fv-Hyd1 belongs to the type Ⅰ hydrophobins and is basidiomycetes-specific, which is different from the type Ⅱ hydrophobins (C-X9-C-C-X11-C-X16-C-X9 -C-C-X10-C-X6). 　　By acquiring the upstream sequences of genes, combined with yeast genome database SDG, YEASTRACT database and fungal transcription factors database FTFD, the predicted genes potentially regulated transcription factor. bHLH transcription factor associated with changes in the morphology of the fungus (Rtg3) was selected as the candidate transcription factor. Combined with the real-time fluorescent quantitative PCR and the Pearson correlation coefficient, the results showed that there was a strong co-expression rule between Fv-Hyd1 and its potential transcription factor Fv-Rtg3 with a high expression in F. velutipes mycelium knot and fruiting body primordium formation. In addition, according to NetPhos 2.0 SERVIER phosphorylation sites prediction for Fv-Hyd1 and Fv-Rtg3 and the above expression of co-expression analysis data, we speculated the gene expression regulation mechanism of Fv-Hyd1.
出版日期: 2016-05-20
CLC:  Q 785  
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黄千慧
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引用本文:

陈仁良, 李肖, 龙莹, 张磊, 严俊杰, 黄千慧, 熊宏民, 谢宝贵. 金针菇疏水蛋白Fv-Hyd1及其潜在的转录因子Fv-Rtg3的共表达分析[J]. 浙江大学学报(农业与生命科学版), 2016, 42(3): 313-320.

CHEN Renliang, LI Xiao, LONG Ying, ZHANG Lei, YAN Junjie, HUANG Qianhui, XIONG Hongmin, XIE Baogui. Analysis on the co-expression of hydrophobin Fv-Hyd1 and its potential transcription factor Fv-Rtg3 in Flammulina velutipes. Journal of Zhejiang University (Agriculture and Life Sciences), 2016, 42(3): 313-320.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2015.10.091    或    http://www.zjujournals.com/agr/CN/Y2016/V42/I3/313

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