發(fā)布時(shí)間：2018-05-02 16:33

  本文選題：煙粉虱 + 棉花蛋白組��； 參考：《華中農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：棉花作為我國(guó)最重要的經(jīng)濟(jì)作物之一,在生產(chǎn)過(guò)程中長(zhǎng)期遭到各種蟲(chóng)害的嚴(yán)重威脅,近些年來(lái)很多棉花種植區(qū)煙粉虱頻繁發(fā)生,給棉花生產(chǎn)帶來(lái)嚴(yán)重危害,使得棉花產(chǎn)量和纖維品質(zhì)大幅度下降。煙粉虱刺吸脅迫后,棉花植株響應(yīng)機(jī)制的研究還沒(méi)有詳細(xì)報(bào)道,其自身的生理機(jī)制的變化還不清楚。主要乳狀蛋白(MLPs)是Bet v1蛋白亞家族之一,MLPs與另一個(gè)亞家族病理機(jī)制相關(guān)蛋白10(PR10)非常相似,然而目前關(guān)于MLP家族蛋白在植物響應(yīng)脅迫方面的機(jī)制還不清楚,我們希望通過(guò)相關(guān)的分子生物學(xué)技術(shù)探索MLP蛋白在棉花響應(yīng)煙粉虱脅迫過(guò)程中的作用。我們通過(guò)蛋白組手段對(duì)煙粉虱抗性較強(qiáng)的JH棉花品種和抗性較弱的ZS棉花品種進(jìn)行蛋白組表達(dá)差異分析,將表達(dá)差異的蛋白質(zhì)進(jìn)行質(zhì)譜鑒定分析,成功鑒定出263個(gè)棉花蛋白質(zhì),去除重復(fù)冗余后有176個(gè)可供分析的特異蛋白質(zhì)。通過(guò)功能富集分析和代謝通路分析發(fā)現(xiàn)這些蛋白質(zhì)有相當(dāng)一部分參與了氧化還原反應(yīng),而且部分是與活性氧信號(hào)路徑相關(guān)的酶。為了進(jìn)一步了解煙粉虱侵染棉花后活性氧水平變化情況,我們對(duì)抗性材料JH進(jìn)行酶活力測(cè)定發(fā)現(xiàn)超氧化物歧化酶活性比對(duì)照顯著升高,過(guò)氧化氫含量初期升高后逐漸降低,而過(guò)氧化物酶活力顯著降低,表明活性氧信號(hào)路徑可能參與了棉花抵御煙粉虱侵染的過(guò)程。煙粉虱侵染后棉花的GhMLP423蛋白表達(dá)上升。為了探索棉花MLP蛋白在棉花響應(yīng)煙粉虱脅迫中的作用,我們選取了棉花GhMLP423基因進(jìn)行后續(xù)的生物學(xué)驗(yàn)證。通過(guò)VIGS技術(shù)成功干涉了GhMLP423基因并獲得了干涉植株。同時(shí),我們對(duì)干涉植株進(jìn)行了酶活力和過(guò)氧化氫含量測(cè)定。棉花干涉系GhMLP423的過(guò)氧化物酶活力低于對(duì)照,超氧化物歧化酶活力高于對(duì)照,將超氧根離子轉(zhuǎn)化為過(guò)氧化氫,因此初期過(guò)氧化氫含量高于對(duì)照,隨著時(shí)間點(diǎn)延長(zhǎng)而逐漸降低。另外,我們對(duì)干涉植株進(jìn)行了激素水平測(cè)定發(fā)現(xiàn)GhMLP423干涉系植株葉片SA含量低于正常植株,煙粉虱侵染后明顯高于非侵染植株,同時(shí)GhMLP423干涉系植株葉片中過(guò)氧化氫含量也低于正常植株,煙粉虱侵染后都有所升高,干涉系也明顯低于對(duì)照,可能由于煙粉虱侵染棉花時(shí)SA積累抑制了過(guò)氧化氫酶的活性使得過(guò)氧化氫積累。與對(duì)照植株相比,GhMLP423干涉系沒(méi)有積累ABA和JA而且對(duì)煙粉虱表現(xiàn)出更感的現(xiàn)象,初步推斷GhMLP423可能參與了JA信號(hào)路徑的傳導(dǎo)過(guò)程。我們通過(guò)對(duì)煙粉虱不同抗性的棉花蛋白組進(jìn)行表達(dá)差異分析,篩選對(duì)抗煙粉虱的棉花抗性相關(guān)的基因,同時(shí)通過(guò)對(duì)煙粉虱侵染后的棉花進(jìn)行生理指標(biāo)分析,進(jìn)一步驗(yàn)證在棉花防御煙粉虱過(guò)程中相關(guān)蛋白的作用。
[Abstract]:Cotton, as one of the most important cash crops in China, has been seriously threatened by various insect pests for a long time in the production process. In recent years, whitefly occurs frequently in many cotton growing areas, which brings serious harm to cotton production. The cotton yield and fiber quality decreased significantly. The study of cotton plant response mechanism has not been reported in detail, and the change of its physiological mechanism is not clear. MLPs, one of the subfamilies of Bet v1 protein, is very similar to that of another subfamily. However, the mechanism of MLP family proteins in response to plant stress is still unclear. We hope to explore the role of MLP protein in cotton response to whitefly stress through molecular biological techniques. We analyzed the protein expression difference of Bemisia Tabaci resistance JH cotton varieties and ZS cotton varieties with weak resistance by proteomic analysis, and identified 263 cotton proteins by mass spectrometry. There are 176 specific proteins that can be analyzed after repetitive redundancy is removed. Through functional enrichment analysis and metabolic pathway analysis, it was found that some of these proteins were involved in the redox reaction, and some were enzymes related to the signal pathway of reactive oxygen species (Ros). In order to further understand the changes of reactive oxygen species (Ros) level in cotton infected with Bemisia Tabaci, we found that the activity of superoxide dismutase (SOD) of resistant material JH was significantly higher than that of the control, and that the content of hydrogen peroxide decreased gradually after the initial increase of the content of hydrogen peroxide. However, the activity of peroxidase decreased significantly, suggesting that the signal pathway of reactive oxygen species may be involved in the resistance of cotton to whitefly infection. The expression of GhMLP423 protein increased in cotton infected with Bemisia Tabaci. In order to explore the role of cotton MLP protein in cotton response to Bemisia Tabaci stress, we selected cotton GhMLP423 gene for further biological verification. The GhMLP423 gene was successfully interfered with by VIGS technique and the interference plants were obtained. At the same time, we determined the enzyme activity and hydrogen peroxide content of interference plants. The activity of peroxidase and superoxide dismutase (SOD) of cotton interference line GhMLP423 was lower than that of the control, and the superoxide dismutase activity was higher than that of the control, so the content of hydrogen peroxide in the initial stage was higher than that of the control, and gradually decreased with the extension of time. In addition, we determined the hormone level of the interference plants and found that the SA content in the leaves of GhMLP423 interference lines was lower than that in normal plants, and that in Bemisia Tabaci infected plants was significantly higher than that in non-infected plants. At the same time, the content of hydrogen peroxide in the leaves of GhMLP423 interference lines was lower than that in normal plants, and the content of hydrogen peroxide in the leaves of Bemisia Tabaci increased after infection. It was possible that SA accumulation inhibited the activity of catalase and resulted in hydrogen peroxide accumulation in cotton infected by Bemisia Tabaci. Compared with the control plant, GhMLP423 interference line did not accumulate ABA and JA and showed more sensitivity to Bemisia Tabaci. It was preliminarily concluded that GhMLP423 might be involved in the transmission of JA signal pathway. By analyzing the difference of expression of cotton protein groups with different resistance to Bemisia Tabaci, we screened the genes related to cotton resistance to Bemisia Tabaci, and analyzed the physiological indexes of cotton infected by Bemisia Tabaci. To further verify the role of related proteins in cotton defense against whitefly.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：S435.622

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相關(guān)碩士學(xué)位論文 前1條

1 李素麗;棉花與煙粉虱互作的蛋白組學(xué)分析和GhMLP423基因功能分析[D];華中農(nóng)業(yè)大學(xué);2016年

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本文編號(hào)：1834661