【摘要】：根系分泌物在調(diào)控植物與環(huán)境相互作用過程中發(fā)揮著至關(guān)重要的作用。雖然研究者對不同植物在不同生長期、不同生境中根系分泌物組成的變化及其對根際微生物的影響進行了大量的研究,但是對于根系分泌物如何影響根際微生物群落及微生物群落的改變給植物帶來的影響卻知之甚少。本文將植物與根際細菌作為一個系統(tǒng),研究了經(jīng)化學(xué)誘抗劑ASO(3-丙酮基-3-羥基吲哚的水分散粒劑,能誘導(dǎo)煙草對普通花葉病(Tobacco mosaic virus, TMV)和煙草白粉病(Erysiphe cichoracearum)的抗性)誘導(dǎo)處理后,煙草K326根系分泌物的組成變化及其對根際細菌群落組成和功能的影響,并通過對一株分離自植煙土壤的細菌的分類學(xué)地位、抗病性和趨化性等的研究,證明植物能通過根系分泌物吸引特定的根際細菌集聚。取得的主要結(jié)果如下：1、通過GC-MS和離子色譜分析了化學(xué)誘抗劑ASO誘導(dǎo)后煙草K326的根系分泌物中揮發(fā)性組份和有機酸的組成變化。結(jié)果表明,ASO誘導(dǎo)對根系分泌物中小分子物質(zhì)的含量和有機酸組成無顯著影響,但其顯著改變了根系分泌物中小分子物質(zhì)的組成。其中具有抗生素活性的脂肪酸3-甲基丁酸、2-甲基丁酸及具有殺蟲活性的煙堿類衍生物麥斯明、新煙草靈和信號分子乙酰香草酮、乙酰丁香酮和靛紅含量顯著改變；這是根系分泌物直接影響根際微生物群落組成的物質(zhì)基礎(chǔ)和理論依據(jù),同時也表明植物產(chǎn)生系統(tǒng)獲得性抗性后會引發(fā)一系列的、廣泛性的保護性反應(yīng)。2、采用16S rDNA擴增子測序技術(shù),對施用ASO處理的煙草K326根際細菌群落多樣性及其功能的動態(tài)變化進行了研究。結(jié)果表明,K326根際微生物群落的多樣性隨植物的生長和ASO的施用減少。植物的生長時期對根際微生物群落的結(jié)構(gòu)產(chǎn)生顯著影響；ASO的施用則對K326根際微生物群落的結(jié)構(gòu)、種群數(shù)量和功能產(chǎn)生影響,但是這一影響會隨施用時間逐漸減弱。對根際細菌優(yōu)勢群落的組成及功能分析表明,在對照處理組中,根際優(yōu)勢細菌群落的已知功能主要是各類復(fù)雜有機物的降解和參與物質(zhì)循環(huán)；而在ASO處理后,根際優(yōu)勢細菌群落的主要功能則轉(zhuǎn)變?yōu)榉置诳股睾驼T導(dǎo)植物系統(tǒng)抗性；從優(yōu)勢菌群的組成上看,ASO處理后,具有物質(zhì)轉(zhuǎn)換功能的菌群被具有防御功能的相關(guān)菌群所取代,從而使根際微生物群落的功能與植物的生存策略保持一致,進而得出根際微生物是植物適應(yīng)環(huán)境的一個重要組成部分的結(jié)論。3、將ASO誘導(dǎo)后的K326根系分泌物作為選擇性培養(yǎng)物,篩選得到一株具有二孢白粉菌(E. cichoracearum)抗性的細菌菌株,經(jīng)形態(tài)學(xué)、生理生化、細胞化學(xué)和分子生物學(xué)鑒定該菌株為溶桿菌屬的新種,命名為Lysobacter erysipheiresistens (RS-LYSO-3)。該菌株對ASO誘導(dǎo)后的K326根系分泌物表現(xiàn)出較強的趨化性,并在根際土壤中迅速繁殖,豐富度增加；生測實驗表明RS-LYSO-3對煙草白粉病的防治效果明顯。這些實驗結(jié)果都提示著植物能通過根系分泌物成分的改變,招徠功能性的微生物,共同構(gòu)筑防御系統(tǒng),即煙草與其根際微生物能組成一個“共生體”以適應(yīng)環(huán)境變化。
[Abstract]:Root exudates play a vital role in the regulation of the interaction between plants and the environment. Although the changes of root exudates in different habitats and different habitats of different plants and their effects on the rhizosphere microorganisms were studied, However, little is known about how the root exudates affect the microbial community and the change of the microbial community to the plants. In this paper, the plant and the rhizosphere bacteria were used as a system to study the water-dispersible granules of the chemical-induced ASO (3-acetone-3-hydroxy-1-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-3-hydroxy-1- The composition of the root exudates of tobacco K326 and its effect on the composition and function of the bacterial community in the rhizosphere and the taxonomic status, disease resistance and resistance of the bacteria isolated from the tobacco soil were studied, and it was proved that the plant was able to attract specific rhizosphere bacteria by root exudates. The main results were as follows:1. The composition of the volatile component and the organic acid in the root exudates of the tobacco K326 was analyzed by GC-MS and ion chromatography. The results showed that ASO induction had no significant effect on the content of small and medium-sized molecules in root exudates and the composition of small and medium-sized molecules in root exudates. wherein the fatty acid 3-methylbutyric acid,2-methylbutyric acid and the nicotine-like derivative with the insecticidal activity of the antibiotic activity are obviously changed; This is the material base and the theoretical foundation of the root exudates directly affecting the microbial community composition of the rhizosphere, and also shows that the acquired resistance of the plant will trigger a series of protective reactions, and the 16S rDNA amplicon sequencing technology is adopted. The diversity of bacterial community and its function in the rhizosphere of tobacco K326 treated with ASO were studied. The results showed that the diversity of the microbial community in the Rhizosphere of K326 decreased with the growth of plants and the application of ASO. The growth period of the plant has a significant effect on the structure of the rhizosphere microbial community; the application of the ASO has an effect on the structure, the number and function of the microbial community in the rhizosphere of the K326, but the effect will gradually decrease with the time of application. The composition and functional analysis of the bacterial dominant community in the rhizosphere indicated that in the control treatment group, the known function of the bacterial community in the rhizosphere is mainly the degradation of various complex organic matters and the participation of the material circulation, and after the ASO treatment, the main function of the bacterial community in the rhizosphere is transformed into the secretion of antibiotics and the induction of plant systemic resistance; after the ASO treatment, the bacteria group with the substance conversion function is replaced by the related flora with the defense function after the ASO treatment, so that the function of the rhizosphere microorganism community is consistent with the survival strategy of the plant, and then the rhizosphere microorganism is a conclusion of an important part of the environment of the plant, A bacterial strain with the resistance of E. cicum earum was selected, and the strain was identified as a new species of the genus Bacillus by morphological, physiological, biochemical, cytochemical and molecular biology, and was named Lysobacter sphaeirescens (RS-LYSO-3). The results showed that the control effect of RS-LYSO-3 on powdery mildew of tobacco was obvious. The results of these experiments show that the plants can change the components of the root exudates and attract the functional microorganisms to construct the defense system, that is, the tobacco and its rhizosphere microorganisms can make up a "symbiont" to adapt to the change of the environment.
【學(xué)位授予單位】：云南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S572

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