南方型紫花苜蓿耐盐突变体叶片盐胁迫应答差异基因鉴定与分析

发布时间：2018-01-23 23:20

本文关键词： 南方型紫花苜蓿叶片盐胁迫转录组　出处：《农业生物技术学报》2017年10期 　论文类型：期刊论文

【摘要】：紫花苜蓿(Medicago sativa)是世界上被广泛种植的一种优质牧草。盐胁迫对紫花苜蓿的生长和产量具有明显抑制作用。为了理解南方型紫花苜蓿(M.sativa'Millennium')受盐胁迫的内在分子机制,挖掘其与耐盐密切相关的功能基因。以250 mmol/L Na Cl处理72 h的南方型紫花苜蓿耐盐突变体叶片进行Illumina Hi SeqTM2000高通量转录组测序,并对所获得的差异表达基因进行基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)pathway生物信息学分析,获得可能耐盐潜在靶标基因。同时,挑选8个差异表达基因验证测序结果的可靠性。结果表明,过滤后对照(control,CK)和盐处理(salt stress,ST)样本分别保留了60 395 324和60 303 692对reads,其中54.18%和53.77%的reads能精确比对到参考序列蒺藜苜蓿(M.truncatula)上。差异表达基因(differentially expressed genes,DEGs)结果显示,在样品中共检测到30 900个基因表达发生改变,其中4 187上调表达,3 507下调表达。GO功能分析显示,差异表达基因主要表现在结合、催化活性、细胞组分和细胞等。KEGG Pathway分析显示,差异表达基因广泛涉及次生代谢、代谢途径及苯丙素的生物合成。另外,筛选了与紫花苜蓿盐胁迫应答相关的基因谷胱甘肽硫转移酶、超氧化物歧化酶Cu/Zn蛋白、L-抗坏血酸过氧化物酶、类受体蛋白激酶、诱导类受体蛋白激酶、蔗糖非发酵型蛋白激酶、类钙调素蛋白、胆碱单加氧酶、1-吡咯啉-5-羧酸合成酶、蛋白磷酸酶2C、海藻糖磷酸酯酶等和AP2类乙烯响应的转录因子、b HLH36转录因子、NAI1转录因子、b ZIP转录因子、C3H锌指蛋白、核酸结合转录因子活性、Myb转录因子、NAC转录因子蛋白、序列特异性DNA结合转录因子蛋白和WRKY转录因子等。本研究为揭示紫花苜蓿耐盐分子机制提供了基础资料。
[Abstract]:Medicago sativa. Salt stress has a significant inhibitory effect on the growth and yield of alfalfa. M. sativaa Millennium) the intrinsic molecular mechanism of salt stress. The functional genes closely related to salt tolerance were excavated. The southern alfalfa salt-tolerant mutant leaves treated with 250 mmol/L NaCl for 72 h were treated with Illumina Hi. SeqTM2000 high-throughput transcriptome sequencing. The differentially expressed genes were analyzed by gene Ontology. ) and the Kyoto Encyclopedia of Genes and Genomes. KEGG)pathway bioinformatics analysis showed that the potential target genes for salt tolerance were obtained. At the same time, 8 differentially expressed genes were selected to verify the reliability of the sequencing results. After filtration, the control group (CK) and salt treated salt salt (stress) were used. ST) samples retained 60 395,324 and 60 303,692 pairs of reads, respectively. The reads of 54.18% and 53.77% can be accurately compared to the reference sequence of M. truncatula. Differentially expressed genes. The results of DEGs showed that a total of 30 900 genes were detected in the samples, among which 4 187 upregulated and 3 507 down-regulated expression. Go functional analysis showed. KEGG Pathway analysis showed that differentially expressed genes were extensively involved in secondary metabolism, such as binding, catalytic activity, cell composition and cell. In addition, genes related to salt stress response of alfalfa, glutathione S-transferase and superoxide dismutase (Cu/Zn) protein were screened. L-ascorbic acid peroxidase, receptor-like protein kinase, inducible receptor-like protein kinase, sucrose non-fermentative protein kinase, calmodulin-like protein, choline monooxygenase 1-pyrrolidin-5-carboxylic acid synthase. Protein phosphatase 2C, trehalose phosphatase and AP2 ethylene responsive transcription factor B HLH36 transcription factor Nai 1 transcription factor B ZIP transcription factor C 3H zinc finger protein. Nucleic acid binding transcription factor activity Myb transcription factor NAC transcription factor protein. Sequence-specific DNA binding transcription factor protein and WRKY transcription factor. This study provides basic information for revealing the molecular mechanism of salt tolerance in alfalfa.
【作者单位】：浙江农林大学风景园林与建筑学院;
【基金】：国家自然科学基金(No.31272494) 浙江省自然科学基金(No.LY16C170003)
【分类号】：S541.9
【正文快照】： 土壤盐碱化已成为最主要的非生物胁迫之一,对农作物的生长和生产量等方面造成不利影响。解析农作物的耐盐分子机制有助于创制耐盐能力提高的作物种质。被称为“牧草皇后”的紫花苜蓿(Medicago sativa)在生态环境修复和奶产业方面具有具有重要价值。我国紫花苜蓿种植,主要以北

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