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为探究棉酚对热带假丝酵母(Candida tropicalis)ZD-3 MAPK信号通路相关基因的影响，加入棉酚至200μg/mL胁迫C.tropicalis ZD-3,分别在棉酚胁迫后0 h、 12 h和24 h对C.tropicalis ZD-3 cDNA进行转录组测序。转录组测序结果发现棉酚胁迫下C.tropicalis ZD-3 12 h相较于0 h筛选出差异表达基因(different expression genes, DEGs)1 391个，DEGs中有678个基因表达量上调，有713个基因表达量下调。24 h相对于0 h DEGs 1 715个，其中765个基因表达量上调，有950个基因表达量下调。对韦恩分析后基因集中的基因进行KEGG代谢途径富集分析发现，DEGs主要富集于MAPK通路。其中44个MAPK相关基因表达量差异显著。经Stem分析发现MAPK相关显著下调基因中有7个基因表达量在12 h和24 h呈递减趋势。在44个MAPK通路基因中选取了Hog1、Fks、Far1、Clb、Gpd1、KatE以及Hot1进行RT-qPCR验证，另对未加棉酚胁迫的0 h、 12 h和24 h热带假丝酵母ZD-3 Clb、Hog1和Fks基因进行定量。结果显示，Hog1、Fks、Far1、Clb、Gpd1、KatE和Hot1表达量与RNA-seq的检测结果一致，未加棉酚的热带假丝酵母ZD-3 Clb、Hog1以及Fks基因在0 h、 12 h、 24 h的表达量变化差异不显著。综上，棉酚可引起C.tropicalis ZD-3 MAPK通路基因的响应，本研究为后续C.tropicalis ZD-3耐受性分子机制的研究提供科学依据。
Abstract：To investigate the effects of gossypol on MAPK signaling pathway-related genes of Candida tropicalis ZD-3, C. tropicalis ZD-3 was stressed by adding gossypol to 200 μg/mL. Transcriptome sequencing of C. tropicalis ZD-3 cDNA was performed at 0 h, 12 h, and 24 h after gossypol stress. Transcriptome sequencing found that 1 391 differentially expressed genes(DEGs) of C. tropicalis ZD-3 were screened out under gossypol stress for 12 h compared with 0 h. The expression quantity of 678 genes in the DEGs showed an up-regulated trend and 713 genes showed a down-regulated trend. Compared with 0 h the 1 715 DEGs screened at 24 h, the expression quantity of 765 genes in the DEGs were up-regulated, and the expression quantity of 950 genes were down-regulated. KEGG metabolic pathway enrichment analysis of the genes in the gene set after Venn analysis revealed that DEGs were mainly enriched in the MAPK pathway. Among them, the expression quantity of 44 MAPK related genes were significantly different. Stem analysis revealed that the expression quantity of 7 genes in the significantly down-regulated MAPK-related genes showed a decreasing trend at 12 h and 24 h. Hog1, Fks, Far1, Clb, Gpd1, KatE, and Hot1 were selected in 44 MAPK pathway genes for RT-qPCR validation and even the Clb, Hog1 and Fks genes of C. tropicalis ZD-3 at 0 h, 12 h and 24 h without gossypol stress were also quantified. The results showed that the expression quantity of Hog1, Fks, Far1, Clb, Gpd1, KatE and Hot1 were consistent with the results of RNA-seq and the changes in the expression quantity of Clb, Hog1 and Fks genes were not significant at 0 h, 12 h and 24 h in C. tropicalis ZD-3 without gossypol stress. To sum up, gossypol can induce the response of MAPK pathway gene in C. tropicalis ZD-3, and this study provides a scientific evidence for the subsequent research on the molecular mechanism of gossypol tolerance in C. tropicalis ZD-3.
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基本信息:
DOI：10.13417/j.gab.042.000318
中图分类号:TQ925;Q78
引用信息:
[1]杨晓龙,张莉,陈程,等.棉酚对热带假丝酵母ZD-3 MAPK信号通路响应的影响[J].基因组学与应用生物学,2023,42(03):318-326.DOI:10.13417/j.gab.042.000318.
基金信息:
国家自然科学基金项目(32060770); 兵团重点领域科技攻关计划项目“肉羊健康养殖模式关键技术集成创新与示范”(2021AB014); 青年创新拔尖人才项目(KX01500301); 石河子大学高层次人才科研项目(RCZK201901)共同资助
2023-07-31
2023-07-31
2023-07-31