發(fā)布時(shí)間：2018-03-10 03:01

  本文選題：同源四倍體泡桐　切入點(diǎn)：轉(zhuǎn)錄組　出處：《河南農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：泡桐是我國速生用材和綠化樹種,用途多,適應(yīng)性強(qiáng),分布廣,具有重要的經(jīng)濟(jì)和生態(tài)價(jià)值。為滿足人們的生產(chǎn)需要,我們利用秋水仙素成功誘導(dǎo)出了同源四倍體泡桐。之前研究顯示同源四倍體泡桐具有很多優(yōu)良特性比如抗性。干旱是影響泡桐的生物產(chǎn)量的非生物因素之一。泡桐的干旱應(yīng)答是多水平的復(fù)雜調(diào)控,但分子機(jī)制還不清楚。本研究以白花泡桐二四倍體、南方泡桐二四倍體、毛泡桐二四倍體和豫雜一號泡桐二四倍體盆栽苗為試驗(yàn)材料,對其分別進(jìn)行15天的25%和75%土壤含水量干旱處理。通過高通量轉(zhuǎn)錄組測序、s RNA測序和降解組測序,以白花泡桐基因組為參考序列,基于生物信息學(xué)分析mRNA和miRNA的表達(dá)變化,鑒定出差異表達(dá)的轉(zhuǎn)錄本和miRNA,同時(shí)利用降解組預(yù)測差異表達(dá)miRNA的靶基因。本研究主要結(jié)果如下:1、四種泡桐二四倍體構(gòu)建的16個(gè)文庫都具有較高的測序質(zhì)量。clean read的GC含量都大于45.68%,Q20都大于94.94%,Q30都大于86.65%,N比例都為0%;clean read與泡桐基因組的比對率都大于79.83%,與泡桐基因比對率都高于44.89%。26個(gè)基因的實(shí)時(shí)熒光定量PCR表達(dá)趨勢和高通量測序獲得的表達(dá)趨勢大致一致,說明測序的可靠性高。2、從四倍體白花泡桐、毛泡桐、南方泡桐和豫雜一號泡桐中鑒定出了302、453、441和372個(gè)與其二倍體的干旱應(yīng)答差異基因。這些基因的Pathway富集在植物信號轉(zhuǎn)導(dǎo)通路、淀粉和糖的代謝通路、苯丙烷的生物合成通路、谷胱甘肽的代謝過程、次生代謝物的合成代謝、過氧化物酶代謝途徑和乙醇酸的代謝。兩種水通道蛋白(PIP和TIP)和脫水蛋白在干旱脅迫后上調(diào)表達(dá)有利于泡桐根部對于水分的吸收。GO富集分析顯示下調(diào)的轉(zhuǎn)錄本作用的細(xì)胞組分主要集中在光合單位的膜系統(tǒng),發(fā)揮的分子功能富集在結(jié)合作用和催化功能,參與的生物過程多富集在了光合作用的光反應(yīng)和暗反應(yīng)過程。它們保護(hù)了泡桐的光合體系免受來自還原力和氧化電子的破壞,但減少了泡桐生物量的積累,其中尤為顯著的就是碳水化合物的合成。但與二倍體相比較,這些基因在同源四倍體泡桐中下調(diào)的幅度較小,能夠在干旱脅迫條件下積累較多的光合產(chǎn)物。3、通過sRNA測序,從不同倍性不同干旱處理的白花泡桐、毛泡桐、南方泡桐和豫雜一號泡桐分別鑒定出miRNA有180、169、178和246條,四個(gè)庫中19、27、11、25個(gè)mi RNA預(yù)測到了28、23、16和46靶基因。s RNA測序水平上,MIR398、MIR171、MIR172、MIR159、Pau-mir-10、Ptf-mir68和Ptf-mir116差異表達(dá),它們靶定的基因包括鉀離子通道KAT1、Ca2+結(jié)合蛋白、脯氨酸-t RNA連接酶、葡聚糖合成酶、氨基酸透性酶、ABC轉(zhuǎn)運(yùn)家族、ABA受體和脯氨酸氨肽酶,參與逆境激素ABA的信號轉(zhuǎn)導(dǎo),鐵氧合酶的活性調(diào)節(jié),脯氨酸的合成代謝以及細(xì)胞內(nèi)可溶性糖、可溶性蛋白的合成、運(yùn)輸和積累過程。維持泡桐體內(nèi)滲透平衡和氧化平衡,最終提高泡桐在干旱逆境下根部的吸水能力,減少地上部分的失水量。綜上所述,以泡桐基因組為背景,通過高通量測序和生物信息學(xué)鑒定和分析大量干旱應(yīng)答相關(guān)的基因。植物激素ABA上調(diào)與干旱耐性相關(guān),可溶性蛋白和糖類上調(diào)有利于維持細(xì)胞膨壓和滲透壓平衡與干旱耐性相關(guān),抗氧化酶和非抗氧化酶(抗壞血酸酶和谷胱甘肽酶)上調(diào)維持細(xì)胞的氧化平衡和離子平衡相關(guān)。從不同層次、不同角度,研究和探討了泡桐的干旱耐性相關(guān)的分子機(jī)制,為將來泡桐的分子育種奠定理論基礎(chǔ)。
[Abstract]:Paulownia is a fast-growing timber tree species in China and, multiple purposes, strong adaptability, wide distribution, has important economic and ecological value. In order to meet the production needs of people, we successfully induced by colchicine in Autotetraploid Paulownia. Previous studies indicated that tetraploid Paulownia foam has many good characteristics such as resistance. The drought is one of the the non biological factors of Paulownia biomass. The drought response of Paulownia is a complex multi level regulation, but the molecular mechanism is not clear. In this study, p.fortunei 24 times, 24 times south of Paulownia, Paulownia 24 times and 24 times of Paulownia tomentosa * p.fortune body potted seedlings as the experimental materials, the were 15 days of drought treatment of water containing 25% and 75% soil. Through high-throughput transcriptome sequencing, s sequencing and RNA sequencing to Degradome, Paulownia genome reference sequence, based on Bioinformatics Analysis of the changes of the expression of mRNA and miRNA, identified differentially expressed transcripts and miRNA, at the same time the degradation group predicted the differentially expressed target genes of miRNA. The main results of this study are as follows: 1. The GC content of four species of Paulownia was 24 times as much as the construction of the 16 libraries have high quality.Clean read sequencing is higher than 45.68%. Q20 is greater than 94.94%, Q30 greater than 86.65%, the proportion of N is 0%; clean and read ratio of Paulownia genome rate is greater than 79.83%, and the gene expression trend of Paulownia comparison rate is higher than the real time fluorescence quantitative PCR 44.89%.26 gene expression trends and high-throughput sequencing was roughly consistent, indicating a high reliability.2 sequencing. From the tetraploid Paulownia, Paulownia, Paulownia australis and Paulownia tomentosa * p.fortune identified genetic differences between the 302453441 and 372 drought response and its diploid. These genes Pathway enrichment in plant signal transduction Pathway, metabolic pathway of starch and sugar, phenylpropanoid biosynthesis pathway, metabolism of glutathione, biosynthesis of secondary metabolites, the metabolic pathway of peroxidase and glycolate metabolism. Two kinds of water channel proteins (PIP and TIP) and dehydration protein after drought stress expression is conducive to root for the absorption of.GO enrichment analysis of Paulownia display cell group down transcripts function focused on membrane system of photosynthetic unit water, molecular function enrichment in binding and catalytic function, biological process in the enrichment in the light reaction of photosynthesis and dark reaction process. They protect the photosynthetic system from Paulownia reduction stress and oxidative damage of electronic, but reduced the biomass of Paulownia accumulation, which is particularly significant is the synthesis of carbohydrates. But compared with the diploid, the homologous gene in Small scale down of tetraploid Paulownia, will accumulate more in drought stress conditions of photosynthetic products in.3, through sRNA sequencing, from different times of different drought treatment of Paulownia, Paulownia, Paulownia australis and Paulownia tomentosa * p.fortune were identified with 180169178 miRNA and 246 19,27,11,25, MI RNA four a library to 28,23,16 46 and.S RNA sequencing of target gene level, MIR398, MIR171, MIR172, MIR159, Pau-mir-10, Ptf-mir68 and Ptf-mir116 expression differences, their target genes including potassium channel KAT1, Ca2+ binding protein, proline -t RNA ligase, glucan synthase, amino acid permease, ABC transporter family. ABA receptor and proline aminopeptidase, signal transduction involved in the stress hormone ABA, regulation of iron oxide synthase activity, proline metabolism and intracellular synthesis of soluble sugar, soluble protein, transport and accumulation Cheng. Maintain the osmotic balance of Paulownia and oxidative balance, and ultimately improve the water absorbing ability of Paulownia in roots under drought stress, reduce the amount of water on the ground part. To sum up, in the background of Paulownia genome, by high-throughput sequencing and bioinformatics identification and analysis of a large number of drought responsive genes related to plant hormone. ABA increased with drought tolerance correlation of soluble protein and carbohydrate increase helps to maintain cell turgor and osmotic balance and drought tolerance, antioxidant enzymes and non antioxidant enzymes (ascorbate peroxidase and glutathione peroxidase) up-regulated maintain oxidative balance and ion balance in cells. Different from the different levels and angles, studied and discussed the drought tolerance of Paulownia related molecular mechanism and lay the theoretical foundation for the molecular breeding of Paulownia in the future.

【學(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S792.43
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本文編號：1591448