Journal of Nuclear Agricultural Sciences ›› 2015, Vol. 29 ›› Issue (4): 651-662.DOI: 10.11869/j.issn.100-8551.2015.04.0651

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Digital Expression Profile of Different Cotton (Gossypium hirsutum) Cultivars Resistance to Fusarium Wilt Disease by Solexa Sequencing

HAN Zegang1, ZHAO Zengqiang1, HE Lanlan1, CHAI Mengliang2, LI Huihui1, ZHANG Wei1   

  1. 1. College of Agronomy, Shihezi University, Shihezi, Xinjiang 832000;
    2. College of Life Science, Shihezi University, Shihezi, Xinjiang 832000
  • Received:2014-04-14 Revised:2014-10-21 Online:2015-04-13 Published:2015-04-16

应用Solexa测序技术分析棉花不同抗病品种的数字表达谱

韩泽刚1, 赵曾强1, 何兰兰1, 柴蒙亮2, 李会会1, 张薇1   

  1. 1. 石河子大学农学院, 新疆 石河子 832000;
    2. 石河子大学生命科学学院, 新疆 石河子 832000
  • 通讯作者: 张薇, 教授, 主要从事棉花分子育种研究.E-mail:zhw_agr@shzu.edu.cn.
  • 作者简介:韩泽刚, 男, 主要从事棉花分子育种研究.E-mail:hanzegang501706851@126.com
  • 基金资助:

    国家自然基金(31260358),农业部转基因生物新品种培育重大专项(2011ZX08005-005)

Abstract:

Fusarium wilt is a worldwide serious disease and constructing digital gene expression profiles of various varieties resistant to fusarium wilt disease will provide a theoretical basis for further study on molecular regulation mechanism and screening related genes resistant to the disease High resistance to fusarium wilt disease upland cotton cultivar‘Zhongmiansuo 12’and susceptible upland cotton cultivar‘Xinluzao 7’were used to analysis the gene expression of cotton seedling root tissue by Solexa sequencing technology. After Solexa sequencing, we built a tag library as many as 3.5 million reads, then selected and identified 1 080 significantly differentially expressed genes, which included 302 up-regulated genes and 778 down-regulated genes. The gene function analysis showed that these annotated genes could be divided into three functional annotation onrology, molecular function, biological processes, and cellular components, and then further be subdivided into 37 functional categories. 112 metabolic pathways and annotated 1 238 significantly differentially expressed genes were identified, which involved amino acid metabolism, glycan biosynthesis and metabolism, lipid metabolism, energy metabolism and signal transduction, etc. There were 26 known functional genes involving in plant hormone signal transduction pathway, and 4 genes encoding AP2/ERF transcription factors, 4 genes relating to LRR receptor-like serine/threonine-protein kinase or protein kinase, 5 genes associated with protein phosphatase 2c. All of these genes have been shown to be involved in disease resistance of plants. Finally, by RT-PCR validation of random selecting 5 differentially expressed genes, the results were consistent with Solexa sequencing resluts. The study preliminary revealed the differential expression of cotton cultivars resistant and sensitive to Cotton Fusarium wilt at the transcriptome level, and hen a large number of differentially expressed genes were obtained for a wealth of data resources for the further study.

Key words: Gossypium hirsutum, fusarium wilt, solexa, expression profiling

摘要:

棉花枯萎病是一种危害严重的世界性病害, 构建棉花枯萎病不同抗病品种的基因表达谱将为进一步研究棉花抗枯萎病的分子调控机制及筛选抗枯萎病相关基因奠定基础.本研究选用高抗枯萎病的陆地棉品种中棉所12和高感枯萎病的陆地棉品种新陆早7号为材料, 利用 Solexa高通量测序技术分析棉花幼苗根部组织中的基因表达情况.Solexa高通量测序后构建了多达350万个reads的标签文库, 根据已知序列信息, 显著差异表达基因1 080个, 其中上调基因302个, 下调基因778个.基因功能分析表明, 已注释的显著差异表达基因可划分为分子功能、生物过程和细胞组件3个功能注释本体, 并进一步细分为37个功能类别.鉴定出112条代谢通路, 注释了1 238个显著差异表达基因, 共涉及氨基酸代谢、多糖的生物合成和代谢、脂类代谢、能量代谢及信号转导等方面.在植物激素信号转导通路分析中, 涉及到26个已知功能基因, 4个基因编码AP2/ERF转录因子, 4个基因与LRR类受体丝氨酸/苏氨酸蛋白激酶或蛋白激酶有关, 5个基因与蛋白磷酸酶2C有关, 而这些基因都参与植物的抗病反应.随机抽取5个基因进行实时荧光定量PCR验证, 其结果与测序结果一致.本研究初步揭示了棉花枯萎病抗感品种在转录组水平上的表达差异, 获得了大量差异表达基因.

关键词: 陆地棉, 枯萎病, Solexa, 基因表达谱