桃響應(yīng)LASIODIPLODIA THEOBROMAE侵染的基因表達(dá)變化及桃流膠病菌的侵染特性研究
發(fā)布時(shí)間：2018-10-05 16:58
【摘要】：桃樹流膠病在我國(guó)桃產(chǎn)區(qū)廣泛分布,尤其以長(zhǎng)江流域及以南地區(qū)發(fā)病嚴(yán)重,可導(dǎo)致樹勢(shì)衰弱甚至樹體死亡,嚴(yán)重危害桃樹的栽培和生產(chǎn)。研究報(bào)道,桃流膠病的病原菌屬于葡萄座腔菌屬(Botryosphaeria spp.)真菌,主要包括Fusicoccum aesculi,Diplodia seriata和Lasiodiplodia theobromae三個(gè)種。桃膠的主要成分為多糖,因此研究認(rèn)為膠體形成與樹體內(nèi)的糖代謝關(guān)系密切。本研究利用RNA-Seq,研究了桃樹響應(yīng)L.theobromae侵染后的基因表達(dá)變化,探討了乙烯和茉莉酸在流膠病發(fā)病過程中作用,同時(shí)從病原菌的分離鑒定和組織病理學(xué)方面研究了皮孔流膠現(xiàn)象,分析了流膠病菌的侵染特性并鑒定了一株新的桃流膠病病原菌。主要結(jié)果如下:1.RNA-Seq分析結(jié)果表明,L.theobromae誘導(dǎo)了桃枝條組織的衰老和一系列防御反應(yīng),促進(jìn)了新的多糖合成并抑制了一些多糖的降解。RNA-Seq共檢出20772個(gè)基因,其中4231、3750、3453和3612個(gè)基因分別在接種后12 h、24 h、48 h和60 h時(shí)差異表達(dá)。共差異表達(dá)基因?yàn)?20個(gè),其中515個(gè)上調(diào)表達(dá),405個(gè)下調(diào)表達(dá)。GO分析顯示,接種L.theobromae后,桃枝條的苯丙素合成代謝、尿苷二磷酸(UDP)糖基轉(zhuǎn)移酶相關(guān)基因顯著上調(diào)表達(dá),而光合作用相關(guān)基因表達(dá)受到明顯抑制。糖代謝途徑中的差異基因表達(dá)分析顯示,細(xì)胞壁降解被促進(jìn),但是果膠和纖維素的降解也伴隨著新的合成;與淀粉和蔗糖分解相關(guān)的基因上調(diào)表達(dá),葡萄糖-6-磷酸轉(zhuǎn)運(yùn)蛋白的表達(dá)增強(qiáng);接種后,UDP-半乳糖、UDP-木糖與UDP-D-甘露糖合成相關(guān)基因的表達(dá)被促進(jìn),這些物質(zhì)都是多糖合成的前體;粘多糖降解相關(guān)基因的表達(dá)被抑制。桃枝條接種L.theobromae后促進(jìn)了乙烯的釋放,乙烯合成與信號(hào)轉(zhuǎn)導(dǎo)相關(guān)基因的表達(dá)水平也顯著提高;接種后12 h,茉莉酸合成相關(guān)基因顯著上調(diào)表達(dá)。2%的茉莉酸甲酯(茉莉酸甲酯/羊毛脂,m/m)可穩(wěn)定誘導(dǎo)桃一年生離體枝條流膠,促進(jìn)了枝條乙烯的釋放,降低了可溶性糖的含量;外源茉莉酸甲酯在早期抑制了桃枝條茉莉酸合成相關(guān)基因的表達(dá),但在后期起促進(jìn)作用。2.從桃樹皮孔組織中可以穩(wěn)定的分離到F.aesculi,F.aesculi在田間表現(xiàn)出潛伏侵染特性,分離到的Neofusicoccum parvum(有性型Botryosphaeria parva)鑒定為一株新的桃流膠病菌。F.aesculi在健康的皮孔組織中也能分離到;F.aesculi產(chǎn)生的分生孢子器位于枝條韌皮部,L.theobromae產(chǎn)生的分生孢子器位于外表皮;N.parvum的分生孢子呈紡錘形或橢圓形,透明,有些孢子有雙隔,隔中間顏色變深。人工接種發(fā)現(xiàn)N.parvum的致病力較強(qiáng),枝條接種后可產(chǎn)生明顯的褐色病斑和流膠反應(yīng)。
[Abstract]:Peach gum disease is widely distributed in peach growing areas in China, especially in the Yangtze River valley and south of China, which can lead to the decline of tree potential and even the death of tree body, thus seriously endangering the cultivation and production of peach trees. It is reported that the pathogen of peach gum disease belongs to (Botryosphaeria spp.). Fungi, mainly including Fusicoccum aesculi,Diplodia seriata and Lasiodiplodia theobromae three species. The main component of peach gum is polysaccharide, so it is considered that colloid formation is closely related to sugar metabolism in tree. In this study, RNA-Seq, was used to study the changes of gene expression in peach trees in response to L.theobromae infection, to explore the role of ethylene and jasmonic acid in the pathogenesis of Glutamosis, and to study the phenomenon of pipette in the aspects of isolation and identification of pathogenic bacteria and histopathology. In this paper, the infection characteristics and a new pathogen of peach gum disease were analyzed. The main results were as follows: 1. RNA-Seq analysis showed that L. theobromae induced senescence and a series of defense responses in peach branches, promoted the synthesis of new polysaccharides and inhibited the degradation of some polysaccharides. RNA-Seq detected a total of 20772 genes. The differential expression of 4231r-3750h-3453 and 3612 genes were observed at 24 h and 60 h after inoculation, respectively. There were 920 differentially expressed bases, of which 515 were up-regulated and 405 were down-regulated. Go analysis showed that after inoculation with L.theobromae, phenylpropanin synthesis and uridine diphosphate (UDP) glycosyltransferase related genes were significantly up-regulated in peach branches. However, the expression of photosynthesis-related genes was significantly inhibited. Analysis of differentially expressed genes in glucose metabolism pathway showed that cell wall degradation was promoted, but degradation of pectin and cellulose was accompanied by new synthesis; genes related to starch and sucrose decomposition were up-regulated. The expression of glucose-6-phosphate transporter was enhanced, and the expression of UDP-galactose-xylose and UDP-D- mannose biosynthesis genes were promoted after inoculation, which were all precursors of polysaccharide synthesis, and the expression of mucopolysaccharide degradation related genes was inhibited. Peach branches were inoculated with L.theobromae to promote ethylene release and the expression level of ethylene synthesis and signal transduction related genes were also significantly increased. 12 h after inoculation, the expression of 0.2% methyl jasmonate (methyl jasmonate / lanolin) was significantly up-regulated, which could stably induce the annual branch flow gum, promote the release of ethylene and decrease the content of soluble sugar. Exogenous methyl jasmonate inhibited the expression of genes related to the biosynthesis of jasmonic acid in peach branches at the early stage, but promoted it at a later stage. It was found that F. aesculium F. aesculi showed latent infection in the field. The Neofusicoccum parvum (sex type Botryosphaeria parva) was identified as a new Peach Streptococcus spp. F. aesculi could also be isolated from healthy lenticular tissue. The conidial organ produced by F. aesculi was located in the trunk phloem L. theobromae. The conidial organ was located in the outer epidermis of N. parvum. The conidia are spindle-shaped or elliptical. Transparent, some spores have double septum, the middle of the septum color darkened. The pathogenicity of N.parvum was found to be strong after inoculation.
【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】：2016
【分類號(hào)】：S436.621
[Abstract]:Peach gum disease is widely distributed in peach growing areas in China, especially in the Yangtze River valley and south of China, which can lead to the decline of tree potential and even the death of tree body, thus seriously endangering the cultivation and production of peach trees. It is reported that the pathogen of peach gum disease belongs to (Botryosphaeria spp.). Fungi, mainly including Fusicoccum aesculi,Diplodia seriata and Lasiodiplodia theobromae three species. The main component of peach gum is polysaccharide, so it is considered that colloid formation is closely related to sugar metabolism in tree. In this study, RNA-Seq, was used to study the changes of gene expression in peach trees in response to L.theobromae infection, to explore the role of ethylene and jasmonic acid in the pathogenesis of Glutamosis, and to study the phenomenon of pipette in the aspects of isolation and identification of pathogenic bacteria and histopathology. In this paper, the infection characteristics and a new pathogen of peach gum disease were analyzed. The main results were as follows: 1. RNA-Seq analysis showed that L. theobromae induced senescence and a series of defense responses in peach branches, promoted the synthesis of new polysaccharides and inhibited the degradation of some polysaccharides. RNA-Seq detected a total of 20772 genes. The differential expression of 4231r-3750h-3453 and 3612 genes were observed at 24 h and 60 h after inoculation, respectively. There were 920 differentially expressed bases, of which 515 were up-regulated and 405 were down-regulated. Go analysis showed that after inoculation with L.theobromae, phenylpropanin synthesis and uridine diphosphate (UDP) glycosyltransferase related genes were significantly up-regulated in peach branches. However, the expression of photosynthesis-related genes was significantly inhibited. Analysis of differentially expressed genes in glucose metabolism pathway showed that cell wall degradation was promoted, but degradation of pectin and cellulose was accompanied by new synthesis; genes related to starch and sucrose decomposition were up-regulated. The expression of glucose-6-phosphate transporter was enhanced, and the expression of UDP-galactose-xylose and UDP-D- mannose biosynthesis genes were promoted after inoculation, which were all precursors of polysaccharide synthesis, and the expression of mucopolysaccharide degradation related genes was inhibited. Peach branches were inoculated with L.theobromae to promote ethylene release and the expression level of ethylene synthesis and signal transduction related genes were also significantly increased. 12 h after inoculation, the expression of 0.2% methyl jasmonate (methyl jasmonate / lanolin) was significantly up-regulated, which could stably induce the annual branch flow gum, promote the release of ethylene and decrease the content of soluble sugar. Exogenous methyl jasmonate inhibited the expression of genes related to the biosynthesis of jasmonic acid in peach branches at the early stage, but promoted it at a later stage. It was found that F. aesculium F. aesculi showed latent infection in the field. The Neofusicoccum parvum (sex type Botryosphaeria parva) was identified as a new Peach Streptococcus spp. F. aesculi could also be isolated from healthy lenticular tissue. The conidial organ produced by F. aesculi was located in the trunk phloem L. theobromae. The conidial organ was located in the outer epidermis of N. parvum. The conidia are spindle-shaped or elliptical. Transparent, some spores have double septum, the middle of the septum color darkened. The pathogenicity of N.parvum was found to be strong after inoculation.
【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】：2016
【分類號(hào)】：S436.621
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