高温胁迫下坛紫菜的数字基因表达谱研究

赖晓娟; 严小军; 杨锐; 骆其君; 陈海敏

高温胁迫下坛紫菜的数字基因表达谱研究

宁波大学海洋生物工程重点实验室, 应用海洋生物技术教育部重点实验室, 浙江宁波 315211

基金项目: 浙江省重大科技专项（2012C12907-6）；国家公益性行业（海洋）科研专项经费项目（201105023）；浙江省创新团队项目（2012R10025-07）；宁波市创新团队项目（2011B81007）；宁波市科技攻关项目（201201C1011016）；宁波大学研究生科研创新基金项目。

计量
- 文章访问数: 2360
- HTML全文浏览量: 7
- PDF下载量: 1661
- 被引次数: 0
出版历程
- 收稿日期: 2013-06-05

Digital gene expression profiling analysis of Pyropia haitanensis under high temperature stress

Marine Biotechnology Laboratory, Key Laboratory of Applied Marine Biotechnology, Ministry of Educaltion, Ningbo University, Ningbo 315211, China

摘要

摘要: 坛紫菜是潮间带重要的经济藻种，对高温、渗透压等逆境具有独特的调控机制。本文采用基于高通量测序的数字基因表达谱（DGE）技术研究了坛紫菜在高温胁迫下的基因表达差异，并分析其相应的响应方式；利用实时定量PCR技术对DGE部分数据进行验证；检测了其中较有代表性的应答基因hsp70的差异表达。结果显示，高温胁迫下坛紫菜中有256个unigene上调表达，以HSP、核糖体蛋白L12、延伸因子EF-Tu及部分光合作用相关基因为代表，3 820个unigene下调表达，主要为核酸、蛋白以及糖类等合成代谢相关基因。Gene Ontology分析表明，差异表达基因主要定位于质体等有膜细胞器，参与繁殖和发育过程，行使催化和连接酶活性的功能。Pathway分析显示，这些基因分布于107条pathway中。其中，下调表达基因最显著富集于mRNA监督和RNA转运途径，而上调表达基因部分富集于内质网的蛋白加工、RNA降解及光合作用途径。验证表明此次DGE结果具有较高准确性，hsp70基因对高温响应积极。综上所述，DGE结果反应出，在高温胁迫时，坛紫菜出现基础代谢减慢、合成速度下降、能量合成受阻、碳同化降低等现象，但光合作用前期未受影响，同时补救途径启动。
- 坛紫菜 /
- 高温胁迫 /
- 数字基因表达谱(DGE) /
- 差异表达基因 /
- 响应
Abstract: Pyropia haitanensis is an economically important red alga cultivated in China. As an intertidal alga,P. haitanensis developed various and highly effective strategies to overcome those environmental stressors,such as temperature fluctuation and osmotic shock. In this research,digital gene expression profiling (DGE) technique based on high throughput sequencing was used to analysis the gene expression difference of P. haitanensis under high temperature stress. To validate the DGE data,eight genes were selected for real-time quantitative PCR (QRT-PCR) analysis. The expression level of high-temperature response gene hsp70 was particularly investigated. Results indicated that 256 unigenes including HSP,ribosomal protein L12,elongation factor EF-Tu and some photosynthesis-related genes were up-regulated,3 820 unigenes involved in anabolism and metabolism of nucleic acid,protein and carbohydrate were down-regulated. Gene Ontology analysis indicated that differentially expressed genes mainly involved in reproduction and developmental process,exercised the functions of catalytic activity and ligase activity,and located in membrane-bounded organelle such as plastid. Pathway enrichment analysis showed that these genes distributed in 107 pathways. Down-regulated genes were significantly enriched in mRNA surveillance and RNA transport pathway,while up-regulated genes were partially enriched in protein processing in endoplasmic reticulum,RNA degradation and photosynthesis pathway. QRT-PCR confirmed the accuracy of DGE,and the positive response of hsp70 to high temperature was also demonstrated. In summary,DGE results indicated that under high temperature stress,primary metabolism,biosynthesis,energy synthesis,and carbon assimilation of P. haitanensis were weakened,whereas,the rescue strategies were activated.
- Pyropia haitanensis /
- high temperature stress /
- digital gene expression profiling (DGE) /
- differentially expressed gene /
- response

HTML全文

参考文献(22)

张学成,秦松,马家海,等. 海藻遗传学[M]. 北京:中国农业出版社,2005.

杨锐,张晓龙,徐丽宁,等. 坛紫菜耐高温胁迫机理之初步研究//中国海洋湖沼学会藻类学分会第七届会员大会暨第十四次学术讨论会论文摘要集,2007.

Chen S,Jiang J,Li HY,et al. The salt-responsive transcriptome of Populus simonii × Populus nigra via DGE[J]. Gene,2012,504(2):203-212.

Choi S,Hwang M S,Im S,et al. Transcriptome sequencing and comparative analysis of the gametophyte thalli of Pyropia tenera under normal and high temperature conditions[J]. J Appl Phycol,2013,25(4):1237-1264.

杨惠. 条斑紫菜功能基因组及重复序列特征研究. 青岛:中国海洋大学,2011.

Mortazavi A,Williams B A,McCue K,et al. Mapping and quantifying mammalian transcriptomes by RNA-Seq[J]. Nat Methods,2008,5(7): 621-628.

Li R,Yu C,Li Y,et al. SOAP2: an improved ultrafast tool for short read alignment[J]. Bioinformatics,2009,25 (15): 1966-1967.

Audic S,Claverie J M. The significance of digital gene expression profiles[J]. Genome Res,1997,7(10): 986-995.

Kanehisa M,Araki M,Goto S,et al. KEGG for linking genomes to life and the environment[J]. Nucleic Acids Res, 2008,36: 480-484.

朱竹君,骆其君,严小军,等. 琼胶寡糖聚合度对坛紫菜抗性诱导的影响[J].海洋学报,2012,34(6):205-209.

Bray E A,Bailey-Serres J,Weretilnyk E. Responses to abiotic stresses//Buchanan B,Gruissem W,Jones R. Biochemistry and molecular biology of plants. American Society of Plant Biologists,Rockville,2000: 1158-1203.

Wang W,Vinocur B,Shoseyov O,et al. Role of plant heat-shock proteins and molecular chaperons in the abiotic stress response[J]. Trends Plant Sci,2004,9(5): 244-252.

Allakhverdiev S I,Kreslavski V D,Klimov V V,et al. Heat stress: an overview of molecular responses in photosynthesis[J]. Photosynth Res,2008,98(1-3): 541-550.

McKersie B D,Leshem Y Y. Stress and stress coping in cultivated plants[M]. Dordrecht: Kluwer Academic Publishers,1994:181-193.

Daugaard M,Rohde M. The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions[J]. FEBS Lett,2007,581(19): 3702-3710.

Kim E,Park H S,Jung Y,et al. Identification of the high-temperature response genes from Porphyra seriata(rhodophyta) expression sequence tags and enhancement of heat tolerance of Chlamydomonas(chlorophyta) by expression of the porphyra HTR2 gene[J]. J Phycol,2011,47(4): 821-828.

Kim Y K,Jang K. Continuous heat shock enhances translational initiation directed by internal ribosomal entry site[J]. Biochem Biophys Res Comun,2002,297(2): 224-231.

Warner J R,McIntosh K B. How common are extraribosomal functions of ribosomal proteins[J]. Mol Cell,2009,34(1): 3-11.

Fu J,MomciloviéI,Clemente T E,et al. Heterologous expression of a plastid EF-Tu reduces protein thermal aggregation and enhances CO₂ fixation in wheat (Triticum aestivum) following heat stress[J]. Plant Mol Biol,2008,68(3):277-288.

Urano K,Kurihara Y,Seki M,et al. "Omics" analyses of regulatory networks in plant abiotic stress responses[J]. Curr Opin Plant Biol,2010,13(2): 132-138.

Mittler R,Flinka A,Goloubinoff P. How do plant feel the heat？[J]. Trends Biochem Sci,2012,37(3): 118-125.

Qin D,Wu H,Peng H,et al. Heat stress-responsive transcriptome analysis in heat susceptible and tolerant wheat (Triticum aestivum L.) by using Wheat Genome Array[J]. BMC Genomic,2008,9: 432.

施引文献

资源附件(0)

访问统计