本文選題：茶樹 + cDNA-AFLP��； 參考：《福建農(nóng)林大學(xué)》2016年碩士論文

【摘要】：茶葉是世界上最流行的健康飲料之一,具有抗氧化、利尿、抗突變強(qiáng)心解痙等功效,茶葉需求量也節(jié)節(jié)攀升。為保證茶葉的品質(zhì)和產(chǎn)量,茶樹病害的有效防治顯得越發(fā)重要。茶樹炭疽病遍及各大主要茶區(qū),嚴(yán)重影響茶葉的品質(zhì)和產(chǎn)量,為此,有必要探究炭疽菌-茶樹的互作機(jī)理,為有效防治茶炭疽病提供一些理論依據(jù)。本研究通過對不同茶樹品種葉片分別接種不同炭疽病病原菌菌株,茶炭疽病病斑多為圓形、半圓形,有的會被葉脈隔斷,形狀不規(guī)則,有的病斑則串聯(lián)到一起,病害嚴(yán)重的葉片蜷縮成卷形；顏色以褐色、暗褐色為主,有的為暗綠色,從病斑中心位置至病斑邊緣漸漸轉(zhuǎn)化為灰白色,病斑中心密生小黑點(diǎn)并呈現(xiàn)隆起狀.。不同茶樹品種對炭疽病病原菌的抗病能力強(qiáng)弱也有差異,鐵觀音、肉桂、大葉烏龍屬于抗性較弱品種,毛蟹、福鼎大白茶抗性較強(qiáng),本研究結(jié)合茶樹形態(tài)學(xué)為基礎(chǔ)對炭疽病病原菌的致病能力分化進(jìn)行了探索,篩選獲得強(qiáng)致病菌株ZRG,較弱致病力菌株ATG、FSX,從形態(tài)上分析強(qiáng)弱菌株的附著胞、分生孢子等結(jié)構(gòu)差異特性。針對強(qiáng)致病力菌株ZRG侵染茶樹葉片后部分生理生化反應(yīng)進(jìn)行了探究,以期為揭開炭疽菌-茶樹互作機(jī)制提供理論依據(jù),并通過qRT-PCR對茶樹受病原菌侵染主要幾種抗氧化酶類表達(dá)進(jìn)行分析,旨在為茶樹生理生化研究提供一種新的模式和茶樹分子手段選育抗病品種的開展提供借鑒。利用炭疽病病原菌強(qiáng)致病菌株ZRG侵染茶樹品種毛蟹后,對茶樹葉片基因差異表達(dá)譜,即cDNA-AFLP體系進(jìn)行了構(gòu)建。對146條差異條帶功能進(jìn)行分析歸類,其中參與碳水化合物和能量代謝11.6%、脂質(zhì)代謝2.7%、核酸代謝6.8%、蛋白質(zhì)代謝15.1%、應(yīng)激反應(yīng)21.9%、生物調(diào)控與信號轉(zhuǎn)導(dǎo)17.8%、運(yùn)輸4.8%、細(xì)胞壁與細(xì)胞骨架代謝3.4%和其他代謝過程12.3%。對其中部分差異條帶進(jìn)行進(jìn)一步的qRT-PCR驗(yàn)證cDNA-AFLP圖譜,驗(yàn)證設(shè)立三次重復(fù)實(shí)驗(yàn),一致率為92.6%,證明了cDNA-AFLP體系的可靠性。在其中發(fā)現(xiàn)了WEKY轉(zhuǎn)錄因子、乙烯轉(zhuǎn)錄因子和過氧化物酶等與植物抗逆境密切相關(guān)基因表達(dá)上調(diào)�？傊�,本研究篩選出茶樹被炭疽菌侵入后差異相關(guān)基因,并對這些基因進(jìn)行分析,但未對其在茶樹抗炭疽病中的功能進(jìn)行研究,這些工作有待在還需在茶樹中進(jìn)行進(jìn)一步轉(zhuǎn)基因驗(yàn)證。
[Abstract]:Tea is one of the most popular health drinks in the world, with antioxidant, diuretic, anti-mutation, strong spasmolysis and other functions, tea demand is also rising. In order to guarantee the quality and yield of tea, the effective control of tea plant disease becomes more and more important. Anthracnose of tea plants is widespread in major tea areas, which seriously affects the quality and yield of tea leaves. Therefore, it is necessary to explore the interaction mechanism of anthrax and tea plants in order to provide some theoretical basis for effective control of tea anthracnose. In this study, different strains of anthracnose were inoculated into different leaves of different tea varieties. Most of the anthracnose spots were round and semicircular, some of them were separated by leaf veins, some of them were irregular in shape, and some of them were connected together. The leaves with severe disease were curled up into coils. The color was brown, dark brown, and some of them were dark green. From the center of the spot to the edge of the spot, the leaf gradually changed into gray and white, and the center of the disease was dense with small black spots and appeared as a bulge. The resistance of different tea varieties to anthracnose pathogens was also different. Tieguanyin, cinnamon and Oolong were among the less resistant varieties, hairy crab and Fuding Dabai tea had stronger resistance to anthracnose. Based on the morphology of tea plant, the pathogenicity differentiation of anthracnose pathogen was studied, and the strong pathogenic strain ZRG, the weak pathogenic strain ATGG FSX were obtained, and the attachment cells of the strong and weak strains were analyzed from morphological point of view. Conidia and other structural differences. The physiological and biochemical responses of the strongly pathogenic strain ZRG to tea leaves were studied in order to provide theoretical basis for uncovering the interaction mechanism of anthrax and tea plants. The expression of several antioxidant enzymes in tea plants infected by pathogens was analyzed by qRT-PCR in order to provide a new model for the physiological and biochemical studies of tea plants and to provide reference for the selection of resistant varieties by molecular means of tea plants. The gene differential expression profile of tea leaves, or cDNA-AFLP system, was constructed after infecting the tea variety hairy crab with ZRG, a strong pathogen of anthracnose. The function of 146 different bands was analyzed and classified. Among them, 11.6 involved in carbohydrate and energy metabolism, 2.7 in lipid metabolism, 6.8 in nucleic acid metabolism, 15.1in protein metabolism, 21.9in stress reaction, 17.8in biological regulation and signal transduction, 4.8in transport, 3.4% in cell wall and cytoskeleton metabolism and 12.3% in other metabolic processes. Some of the differential bands were further verified by qRT-PCR to verify the cDNA-AFLP map, and three repeated experiments were established, and the consistency rate was 92.6, which proved the reliability of the cDNA-AFLP system. The expression of WEKY transcription factors, ethylene transcription factors and peroxidase were up-regulated. In conclusion, the differentially related genes of tea plants after invasion by anthrax were screened and analyzed, but their functions in anthracnose resistance of tea plants were not studied. These efforts need to be further verified in tea plants.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S435.711

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