本文關(guān)鍵詞： 南方型紫花苜蓿 葉片 鹽脅迫 轉(zhuǎn)錄組　出處：《農(nóng)業(yè)生物技術(shù)學(xué)報(bào)》2017年10期 　論文類型：期刊論文

【摘要】：紫花苜蓿(Medicago sativa)是世界上被廣泛種植的一種優(yōu)質(zhì)牧草。鹽脅迫對(duì)紫花苜蓿的生長(zhǎng)和產(chǎn)量具有明顯抑制作用。為了理解南方型紫花苜蓿(M.sativa'Millennium')受鹽脅迫的內(nèi)在分子機(jī)制,挖掘其與耐鹽密切相關(guān)的功能基因。以250 mmol/L Na Cl處理72 h的南方型紫花苜蓿耐鹽突變體葉片進(jìn)行Illumina Hi SeqTM2000高通量轉(zhuǎn)錄組測(cè)序,并對(duì)所獲得的差異表達(dá)基因進(jìn)行基因本體(Gene Ontology,GO)和京都基因與基因組百科全書(shū)(Kyoto Encyclopedia of Genes and Genomes,KEGG)pathway生物信息學(xué)分析,獲得可能耐鹽潛在靶標(biāo)基因。同時(shí),挑選8個(gè)差異表達(dá)基因驗(yàn)證測(cè)序結(jié)果的可靠性。結(jié)果表明,過(guò)濾后對(duì)照(control,CK)和鹽處理(salt stress,ST)樣本分別保留了60 395 324和60 303 692對(duì)reads,其中54.18%和53.77%的reads能精確比對(duì)到參考序列蒺藜苜蓿(M.truncatula)上。差異表達(dá)基因(differentially expressed genes,DEGs)結(jié)果顯示,在樣品中共檢測(cè)到30 900個(gè)基因表達(dá)發(fā)生改變,其中4 187上調(diào)表達(dá),3 507下調(diào)表達(dá)。GO功能分析顯示,差異表達(dá)基因主要表現(xiàn)在結(jié)合、催化活性、細(xì)胞組分和細(xì)胞等。KEGG Pathway分析顯示,差異表達(dá)基因廣泛涉及次生代謝、代謝途徑及苯丙素的生物合成。另外,篩選了與紫花苜蓿鹽脅迫應(yīng)答相關(guān)的基因谷胱甘肽硫轉(zhuǎn)移酶、超氧化物歧化酶Cu/Zn蛋白、L-抗壞血酸過(guò)氧化物酶、類受體蛋白激酶、誘導(dǎo)類受體蛋白激酶、蔗糖非發(fā)酵型蛋白激酶、類鈣調(diào)素蛋白、膽堿單加氧酶、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類乙烯響應(yīng)的轉(zhuǎn)錄因子、b HLH36轉(zhuǎn)錄因子、NAI1轉(zhuǎn)錄因子、b ZIP轉(zhuǎn)錄因子、C3H鋅指蛋白、核酸結(jié)合轉(zhuǎn)錄因子活性、Myb轉(zhuǎn)錄因子、NAC轉(zhuǎn)錄因子蛋白、序列特異性DNA結(jié)合轉(zhuǎn)錄因子蛋白和WRKY轉(zhuǎn)錄因子等。本研究為揭示紫花苜蓿耐鹽分子機(jī)制提供了基礎(chǔ)資料。
[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.
【作者單位】： 浙江農(nóng)林大學(xué)風(fēng)景園林與建筑學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(No.31272494) 浙江省自然科學(xué)基金(No.LY16C170003)
【分類號(hào)】：S541.9
【正文快照】： 土壤鹽堿化已成為最主要的非生物脅迫之一,對(duì)農(nóng)作物的生長(zhǎng)和生產(chǎn)量等方面造成不利影響。解析農(nóng)作物的耐鹽分子機(jī)制有助于創(chuàng)制耐鹽能力提高的作物種質(zhì)。被稱為“牧草皇后”的紫花苜蓿(Medicago sativa)在生態(tài)環(huán)境修復(fù)和奶產(chǎn)業(yè)方面具有具有重要價(jià)值。我國(guó)紫花苜蓿種植,主要以北

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