發(fā)布時(shí)間：2018-04-16 02:06

  本文選題：半野生棉 + 鹽堿�。� 參考：《華中農(nóng)業(yè)大學(xué)》2017年博士論文

【摘要】：棉花是重要的纖維和能源作物,也是鹽堿地種植的先鋒作物。然而高濃度的鹽脅迫依然會(huì)對(duì)其產(chǎn)量和質(zhì)量造成影響。近年來(lái),土壤鹽漬化越來(lái)越嚴(yán)重,極大的制約了棉花的產(chǎn)量和質(zhì)量。半野生棉和野生棉是經(jīng)過嚴(yán)苛的自然界環(huán)境篩選進(jìn)化的材料,其中半野生棉(Semi-wild cotton)是陸地棉的野生類型,又稱陸地棉野生種系(Gossypium hirsutum race),遺傳多樣性豐富,同時(shí)半野生棉與栽培品種之間具有遺傳親和性,是研究棉花抗鹽機(jī)理并進(jìn)行抗性改良的良好材料。本研究制定了室內(nèi)穩(wěn)定鑒定半野生棉萌發(fā)期和苗期復(fù)合鹽堿抗性的方法,并初步探討了ROS在半野生棉對(duì)復(fù)合鹽堿響應(yīng)中的機(jī)制。采用轉(zhuǎn)錄組分析方法分析了克勞茨基棉中SOS、ROS、ABA相關(guān)基因參與高濃度鹽脅迫的響應(yīng)機(jī)制。主要研究結(jié)果如下:1、確立了室內(nèi)鑒定半野生棉復(fù)合鹽堿抗感性的方法:通過對(duì)溫室鑒定結(jié)果與課題多年在新疆鹽堿地塊半野生棉的鑒定結(jié)果的一致性分析,確立了合適土樣;通過對(duì)土樣中主要離子成分的測(cè)定,明確了新疆次生鹽堿地塊中的主要成分,并實(shí)現(xiàn)了溫室重現(xiàn),在進(jìn)行多次驗(yàn)證后,確認(rèn)了方法的可行性;成功的篩選到穩(wěn)定抗性材料瑪利加朗特棉85和穩(wěn)定敏感材料闊葉棉40,抗性對(duì)照材料中16和敏感對(duì)照材料中12。2、苗期半野生棉材料復(fù)合鹽堿脅迫響應(yīng):表型與生理生化顯示以上四種材料的抗性具有生理學(xué)差異,同時(shí)發(fā)現(xiàn)抗氧化相關(guān)的酶在抗感材料之間的活性差異。通過Illumina HiSeq?2500平臺(tái)測(cè)序,對(duì)復(fù)合鹽堿脅迫0h、3h、12h和48h的半野生棉瑪利加朗特棉85、闊葉棉40與陸地棉對(duì)照材料中12和中16的葉片與根的轉(zhuǎn)錄組測(cè)序及生物信息學(xué)分析,發(fā)現(xiàn)64,737個(gè)基因在全部的樣品中表達(dá)。差異基因分析發(fā)現(xiàn)在遭受復(fù)合鹽堿脅迫后下調(diào)基因數(shù)目大于上調(diào)基因數(shù)目,GO富集分析發(fā)現(xiàn),差異基因在響應(yīng)氧化還原進(jìn)程的條目中顯著富集。3、活性氧相關(guān)酶類與基因在半野生棉復(fù)合鹽堿脅迫響應(yīng)中的機(jī)制:通過基因功能注釋,發(fā)現(xiàn)共有265個(gè)ROS產(chǎn)生與清除相關(guān)基因。對(duì)4個(gè)材料的各自的共同差異基因與活性氧代謝相關(guān)的基因進(jìn)行檢索發(fā)現(xiàn)有中12(152),中16(124),闊葉棉40(144),瑪利加朗特棉85(131)個(gè)活性代謝通路相關(guān)的基因在各個(gè)材料中參與ROS調(diào)控。通過SOD、POD、CAT和RBOH四個(gè)基因家族的表達(dá)譜分析,我們確定了主要參與活性氧通路的基因。4、野生棉克勞茨基棉鹽脅迫響應(yīng)機(jī)制與轉(zhuǎn)錄組分析:根據(jù)多個(gè)鹽濃度處理后克勞茨基棉與戴維遜氏棉的表型比對(duì),確定了克勞茨基棉的抗鹽性。在對(duì)300 mM NaCl處理后的克勞茨基棉葉片進(jìn)行生理生化指標(biāo)測(cè)定時(shí),發(fā)現(xiàn)ABA、GSSH、H2O2等含量迅速升高。進(jìn)而通過Illumina HiSeq?2500測(cè)序平臺(tái),對(duì)300 mM NaCl脅迫0h,3h,12h,48h的葉片與根的轉(zhuǎn)錄組測(cè)序及生物信息學(xué)分析,發(fā)現(xiàn)37,278條基因和797條新基因表達(dá)。差異基因分析發(fā)現(xiàn)根部差異基因在3個(gè)時(shí)間節(jié)點(diǎn)都顯著富集。通過基因表達(dá)模式分析,發(fā)現(xiàn)葉片中的差異基因大部分下調(diào)表達(dá)。GO注釋發(fā)現(xiàn)差異基因主要富集在響應(yīng)脅迫與離子轉(zhuǎn)運(yùn)的分類上,KEGG注釋發(fā)現(xiàn)差異主要富集在激素信號(hào)轉(zhuǎn)導(dǎo)與合成通路。5、克勞茨基棉高鹽脅迫響應(yīng)通路:通過對(duì)離子轉(zhuǎn)運(yùn)、活性氧、激素相關(guān)基因的表達(dá)譜分析發(fā)現(xiàn),SOS、ROS、ABA在棉花相應(yīng)鹽脅迫時(shí)起到非常重要的作用,結(jié)合前人研究,初步繪制棉花在高鹽脅迫早期的響應(yīng)通路。
[Abstract]:Cotton is an important fiber and energy crops, saline land planting pioneer crops. However, high concentration of salt stress will affect its yield and quality. In recent years, more and more serious soil salinization, which greatly restricts the yield and quality of cotton. Cotton and semi wild wild cotton is through the harsh nature the evolution of environmental screening materials, including semi wild cotton (Semi-wild cotton) is a type of wild cotton, also known as wild species of Upland Cotton (Gossypium hirsutum race), genetic diversity, and between semi wild cotton and cultivars with genetic affinity, is to study the mechanism of salt tolerance of cotton and improve the resistance of the good material. This study developed a method of indoor identification of stable semi wild cotton at Germination and seedling stage of mixed salt alkali resistance, and to explore the ROS in semi wild cotton on composite saline in the response by rotating mechanism. Record group analysis Klotz Ki cotton SOS, ROS, ABA genes involved in the response mechanism of high concentration of salt stress. The main results are as follows: 1. The established method of indoor identification of semi wild cotton composite salt anti perceptual: Based on the results of greenhouse identification and consistency in the identification of the subject for many years in Xinjiang semi wild saline land cotton analysis, established a suitable soil sample; the determination of main ions in soil samples, the main components of Xinjiang secondary salinization land, and realize the greenhouse in return, after repeated verification, confirm the feasibility of the method; successful screening to stable resistance materials and stable cotton Mary gallante 85 the sensitive material of broad-leaved 40 cotton, 12.2 resistant materials and 16 control sensitive control material, semi wild cotton seedling composite salt stress response: phenotypic and physiological and biochemical display more than four kinds of material resistance With physiological differences, also found that the antioxidant enzyme activity in the resistant and susceptible differences between the materials. Through the Illumina HiSeq 2500 platform? Sequencing of the composite salt stress 0h, 3h, 12h and 48h semi wild cotton Mary gallante 85 cotton, cotton 40 broad-leaved and upland cotton were analyzed in 16 and 12 and leaves root transcriptome sequencing and bioinformatics materials, found that the expression of 64737 genes in all the samples. Analysis of differentially expressed genes found in the suffering of mixed salt alkali stress is greater than the number of up-regulated genes down regulated genes after GO enrichment analysis showed that the number of genes in response to redox process entries were enriched in.3, enzymes related to active oxygen with the gene in semi wild cotton composite salt stress response mechanism in the annotation of gene function, found a total of 265 genes related to ROS production and scavenging. Based on 4 common different materials with live The gene of oxygen metabolism related searches have been found in 12 (152), 16 (124), 40 (144), broad-leaved cotton Mary gallante cotton 85 (131) active metabolic pathway related genes involved in the regulation of ROS in all materials. Through SOD, POD, CAT and RBOH expression analysis four gene family, we identified the.4 gene involved in ROS pathway, Wild Cotton Gossypium klotzschanium Anderss salt stress response mechanism analysis and transcriptome: according to phenotypic comparison of Gossypium klotzschanium Anderss and Davison's cotton multiple salt concentration treatment, determine the salt resistance of Gossypium klotzschanium Anderss. In G. klotzschianum leaves 300 mM after NaCl treatment were determined by physiological and biochemical indexes, ABA, GSSH, H2O2 content increased rapidly. And then through the Illumina HiSeq? 2500 to 300 mM NaCl sequencing platform, 3h, 12h, 0h stress, 48h analysis of the leaf and root transcriptome sequencing and bioinformatics, 37278 797 new genes and gene expression. The difference of gene analysis showed that root genes in 3 time nodes are significantly enriched. Through the analysis of gene expression patterns, differentially expressed genes were found in the leaves of most down regulated expression of.GO annotation found differentially expressed genes were mainly enriched in response to stress and the classification of transport, KEGG notes were found mainly enriched in hormone signal transduction and.5 synthesis pathway, G. klotzschianum high salt stress response pathway: Based on ion transport, reactive oxygen species, spectrum analysis showed that the expression of hormone related genes SOS, ROS, ABA play a very important role in the cotton corresponding to salt stress, combined with previous research, preliminary draw the response pathway of early cotton under high salt stress.

【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：S562

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