花生抗黃曲霉菌產(chǎn)毒機制的研究
發(fā)布時間：2019-02-16 21:50
【摘要】：花生(Arachis hypogaea L.)是我國重要的油料與經(jīng)濟作物之一,總產(chǎn)量和總產(chǎn)值均已躍居國產(chǎn)油料作物的首位,且其單位面積產(chǎn)量和產(chǎn)油量高、種植效益好、國際競爭力強,發(fā)展花生生產(chǎn)對于保障植物油脂和蛋白質(zhì)供給、提高農(nóng)業(yè)生產(chǎn)效益、增加農(nóng)民收入具有重要作用。然而,黃曲霉毒素污染嚴重危及花生及其制品的食用安全性、消費者健康和國際貿(mào)易,是制約花生產(chǎn)業(yè)持續(xù)發(fā)展的關鍵風險因子。培育抗黃曲霉花生品種是防控毒素污染最為經(jīng)濟有效的措施,而系統(tǒng)開展花生抗黃曲霉菌產(chǎn)毒機制的研究則是深化這一抗性遺傳改良的基礎。花生對黃曲霉菌產(chǎn)毒的抗性(抗產(chǎn)毒)是應答黃曲霉菌侵害的主動防御反應,探討和揭示抗產(chǎn)毒機制具有重要的理論意義和實用價值。本研究利用典型的抗產(chǎn)毒花生品種中花6號(抗)和高產(chǎn)毒品種中花12(感),利用轉(zhuǎn)錄組測序技術,首次比較了抗、感花生應答黃曲霉菌產(chǎn)毒的基因表達差異,分析了黃曲霉菌與抗、感花生互作的基因表達差異,探討了白藜蘆醇與花生抗產(chǎn)毒的關系及其抑制黃曲霉毒素合成的分子機制。主要結論如下。1、明確了典型抗、感花生成熟種子應答黃曲霉菌產(chǎn)毒的基因表達變化及差異。受黃曲霉菌侵染后,抗、感花生種子中均有大量基因的表達發(fā)生了改變,其中表現(xiàn)上調(diào)表達的差異基因比下調(diào)表達的多。與高產(chǎn)毒花生相比,抗產(chǎn)毒花生能迅速啟動較多防衛(wèi)反應來抵御黃曲霉菌的產(chǎn)毒,其中病程相關蛋白(PR-1、PR-2、PR-5、PR-10)、轉(zhuǎn)錄因子(WRKY、bZIP、ERF)和類苯基丙烷(PAL、C4H、4CL、STS、CHS)等抗性相關基因上調(diào)表達水平高,且維持高水平表達的時間較長。2、首次以寄生于花生種子組織內(nèi)的黃曲霉菌為研究對象,明確了抗產(chǎn)毒花生能有效抑制黃曲霉毒素合成相關基因的表達。黃曲霉菌在侵染高產(chǎn)毒花生中花12的過程中,與菌絲穿透力(faeC、abfB、mndA、pgxC)、分生孢子形成(RodA/RolA、AtfA、Con-6、Con-10、Pks P)和黃曲霉毒素合成(aflD、aflX、aflNa)等相關基因表現(xiàn)出顯著上調(diào)表達,而在侵染抗產(chǎn)毒中花6號的過程中,這些基因的表達都受到了顯著抑制,從而有效抑制了其定殖與攝取營養(yǎng)物質(zhì)的能力及毒素合成前體物質(zhì)的供應,表明中花6號具有抑制黃曲霉菌定殖、生長和黃曲霉毒素合成的主動抗性反應。3、通過離體處理,發(fā)現(xiàn)白藜蘆醇能直接抑制黃曲霉毒素合成關鍵基因aflA和aflB的表達,限制毒素合成前體物質(zhì)的供應;能提高超氧化物歧化酶基因的表達,減少氧脅迫對毒素合成的促進作用;能使stuA、fluG和flbC等分生孢子形成相關基因顯著下調(diào)表達,減少分生孢子的形成和再侵染機率。結合白藜蘆醇與花生產(chǎn)毒抗性的相關性分析,證實了中花6號白藜蘆醇含量高是其產(chǎn)毒抗性強的重要機制之一。
[Abstract]:Peanut (Arachis hypogaea L.) It is one of the important oil crops and cash crops in China. The total yield and total output value have leapt to the first place of the domestic oil crops, and its unit area yield and oil yield are high, the planting benefit is good, and the international competitiveness is strong. The development of peanut production plays an important role in ensuring the supply of vegetable oil and protein, improving the benefit of agricultural production and increasing farmers' income. However aflatoxin pollution seriously endangers the edible safety of peanut and its products consumer health and international trade are the key risk factors restricting the sustainable development of peanut industry. Cultivation of peanut varieties resistant to Aspergillus flavus is the most economical and effective measure to prevent and control toxin pollution, and systematic study on the mechanism of resistance to Aspergillus flavus is the basis for further genetic improvement of this resistance. Peanut resistance to Aspergillus flavus is an active defense response in response to Aspergillus flavus. It is of great theoretical significance and practical value to explore and reveal the mechanism of resistance to Aspergillus flavus. In this study, the difference of gene expression in resistance to and response to Aspergillus flavus production in peanut was first compared by using transcription sequencing technique, using typical varieties Zhonghua 6 (resistance) and Zhonghua 12 (sense), which were resistant to toxin production in peanut and Zhonghua 12, respectively. The difference of gene expression between Aspergillus flavus and resistant and susceptible peanut was analyzed. The relationship between resveratrol and antitoxic effect of peanut and the molecular mechanism of inhibiting aflatoxin synthesis were discussed. The main conclusions are as follows: 1. The variation and difference of gene expression in response to Aspergillus flavus in typical resistant and susceptible peanut seeds were identified. After being infected by Aspergillus flavus, a large number of genes expressed in resistant and susceptible peanut seeds changed, among which more differentially expressed genes showed up-regulated than down-regulated ones. Compared with high-yielding peanut, resistant toxigenic peanut can initiate more defense response to resist aflatoxin, including course related protein (PR-1,PR-2,PR-5,PR-10) and transcription factor (WRKY,bZIP,). ERF) and phenylpropanoid (PAL,C4H,4CL,STS,CHS) genes were up-regulated and maintained for a long time. 2. Aspergillus flavus, which was parasitic in peanut seed tissue, was studied for the first time. It is clear that antitoxic peanut can effectively inhibit the expression of aflatoxin related genes. Aspergillus flavus is associated with hyphal penetration (faeC,abfB,mndA,pgxC), conidial formation (RodA/RolA,AtfA,Con-6,Con-10,Pks P) and aflatoxin synthesis (aflD,aflX,) in the process of infecting high-yielding peanut Flower 12 with mycelium penetration (faeC,abfB,mndA,pgxC), conidia formation (RodA/RolA,AtfA,Con-6,Con-10,Pks P) and aflatoxin synthesis (aflD,aflX,). AflNa and other related genes showed a significant up-regulation of expression, and the expression of these genes was significantly inhibited during the infection of the drug-resistant Zhonghua 6. Therefore, the ability of colonization and uptake of nutrients and the supply of precursor substances for toxin synthesis were effectively inhibited, indicating that Zhonghua 6 had an active resistance response to inhibit the colonization, growth and aflatoxin synthesis of Aspergillus flavus. It was found that resveratrol could directly inhibit the expression of aflA and aflB, and restrict the supply of precursor to aflatoxin synthesis. It can increase the expression of superoxide dismutase gene and reduce the effect of oxygen stress on toxin synthesis, and can significantly down-regulate the expression of conidial formation related genes such as stuA,fluG and flbC, and reduce the probability of conidia formation and re-infection. Combined with the correlation analysis between resveratrol and floral toxicity resistance, it was proved that the high content of resveratrol in Zhonghua 6 was one of the important mechanisms of its strong toxic resistance.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S565.2
本文編號：2424869
[Abstract]:Peanut (Arachis hypogaea L.) It is one of the important oil crops and cash crops in China. The total yield and total output value have leapt to the first place of the domestic oil crops, and its unit area yield and oil yield are high, the planting benefit is good, and the international competitiveness is strong. The development of peanut production plays an important role in ensuring the supply of vegetable oil and protein, improving the benefit of agricultural production and increasing farmers' income. However aflatoxin pollution seriously endangers the edible safety of peanut and its products consumer health and international trade are the key risk factors restricting the sustainable development of peanut industry. Cultivation of peanut varieties resistant to Aspergillus flavus is the most economical and effective measure to prevent and control toxin pollution, and systematic study on the mechanism of resistance to Aspergillus flavus is the basis for further genetic improvement of this resistance. Peanut resistance to Aspergillus flavus is an active defense response in response to Aspergillus flavus. It is of great theoretical significance and practical value to explore and reveal the mechanism of resistance to Aspergillus flavus. In this study, the difference of gene expression in resistance to and response to Aspergillus flavus production in peanut was first compared by using transcription sequencing technique, using typical varieties Zhonghua 6 (resistance) and Zhonghua 12 (sense), which were resistant to toxin production in peanut and Zhonghua 12, respectively. The difference of gene expression between Aspergillus flavus and resistant and susceptible peanut was analyzed. The relationship between resveratrol and antitoxic effect of peanut and the molecular mechanism of inhibiting aflatoxin synthesis were discussed. The main conclusions are as follows: 1. The variation and difference of gene expression in response to Aspergillus flavus in typical resistant and susceptible peanut seeds were identified. After being infected by Aspergillus flavus, a large number of genes expressed in resistant and susceptible peanut seeds changed, among which more differentially expressed genes showed up-regulated than down-regulated ones. Compared with high-yielding peanut, resistant toxigenic peanut can initiate more defense response to resist aflatoxin, including course related protein (PR-1,PR-2,PR-5,PR-10) and transcription factor (WRKY,bZIP,). ERF) and phenylpropanoid (PAL,C4H,4CL,STS,CHS) genes were up-regulated and maintained for a long time. 2. Aspergillus flavus, which was parasitic in peanut seed tissue, was studied for the first time. It is clear that antitoxic peanut can effectively inhibit the expression of aflatoxin related genes. Aspergillus flavus is associated with hyphal penetration (faeC,abfB,mndA,pgxC), conidial formation (RodA/RolA,AtfA,Con-6,Con-10,Pks P) and aflatoxin synthesis (aflD,aflX,) in the process of infecting high-yielding peanut Flower 12 with mycelium penetration (faeC,abfB,mndA,pgxC), conidia formation (RodA/RolA,AtfA,Con-6,Con-10,Pks P) and aflatoxin synthesis (aflD,aflX,). AflNa and other related genes showed a significant up-regulation of expression, and the expression of these genes was significantly inhibited during the infection of the drug-resistant Zhonghua 6. Therefore, the ability of colonization and uptake of nutrients and the supply of precursor substances for toxin synthesis were effectively inhibited, indicating that Zhonghua 6 had an active resistance response to inhibit the colonization, growth and aflatoxin synthesis of Aspergillus flavus. It was found that resveratrol could directly inhibit the expression of aflA and aflB, and restrict the supply of precursor to aflatoxin synthesis. It can increase the expression of superoxide dismutase gene and reduce the effect of oxygen stress on toxin synthesis, and can significantly down-regulate the expression of conidial formation related genes such as stuA,fluG and flbC, and reduce the probability of conidia formation and re-infection. Combined with the correlation analysis between resveratrol and floral toxicity resistance, it was proved that the high content of resveratrol in Zhonghua 6 was one of the important mechanisms of its strong toxic resistance.
【學位授予單位】：中國農(nóng)業(yè)科學院
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S565.2
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