本文選題：紅小豆 + 銹病��； 參考：《黑龍江八一農(nóng)墾大學(xué)》2017年碩士論文

【摘要】：紅小豆銹病是紅小豆生產(chǎn)中主要病害之一,嚴(yán)重影響紅小豆的產(chǎn)量和品質(zhì)。利用抗病品種是防治紅小豆銹病最經(jīng)濟(jì)有效的措施,然而目前紅小豆種質(zhì)資源對(duì)銹病的抗性評(píng)價(jià)尚無(wú)研究報(bào)道,主栽品種均不抗銹病,生產(chǎn)上難有可利用的抗病資源,紅小豆對(duì)銹病的抗病機(jī)制亦不明確。因此,本研究從紅小豆資源對(duì)紅小豆銹病的抗性評(píng)價(jià)入手,篩選抗病品種,研究紅小豆銹病菌(Uromyces vignae)與不同抗性紅小豆品種互作后,病原菌在葉片上侵染過(guò)程的差異,以及與抗病相關(guān)的CAT、SOD、POD、PPO活性和抗病相關(guān)基因的變化情況,為紅小豆抗銹病資源利用奠定基礎(chǔ)。研究取得主要結(jié)果如下:1.采用人工接種方法對(duì)85份紅小豆資源進(jìn)行抗銹性評(píng)價(jià),獲得2個(gè)免疫品種,16個(gè)高抗品種、10個(gè)中抗品種、26個(gè)中感品種和31個(gè)高感品種。抗病品種與感病品種相比,其對(duì)銹病抗性主要表現(xiàn)為接種銹病菌后顯癥及產(chǎn)孢的時(shí)間推遲2-3 d,并且產(chǎn)生夏孢子堆的密度小于與感病品種。2.利用熒光顯微技術(shù)研究了銹病菌在不同抗性品種葉片上侵染過(guò)程的差異。銹病菌夏孢子在感病品種上于接種后6 h即可萌發(fā)并形成附著胞,24 h可產(chǎn)生氣孔下囊和吸器母細(xì)胞,48 h形成胞內(nèi)吸器,接種后8 d,葉片表面即可產(chǎn)生夏孢子堆�？共∑贩N與感病品種比較,夏孢子附著胞形成時(shí)間推遲,接種后12 h才形成附著胞,夏孢子萌發(fā)率和侵染率降低。在抗病品種中,接種后48 h,菌絲靠近的葉肉細(xì)胞出現(xiàn)抑制菌絲擴(kuò)展的胞壁沉積物,產(chǎn)孢時(shí)間明顯推遲。感病品種葉肉細(xì)胞不出現(xiàn)抑制菌絲擴(kuò)展的組織結(jié)構(gòu)。以上結(jié)果表明抗病品種對(duì)銹病菌的抗性表現(xiàn)為抗侵入和抗擴(kuò)展兩種特性。3.利用生理生化技術(shù)研究了與抗銹性相關(guān)的防御酶系活性的變化和抗病性的關(guān)系。結(jié)果表明:抗病和感病品種與紅小豆銹病菌互作后,葉片中CAT、POD、SOD、PPO活性均有不同程度的升高�？共∑贩N和感病品種葉片中CAT和POD活性在接種后48 h達(dá)到第一個(gè)高峰,并且抗病品種中CAT和POD活性要高于感病品種,分別高30.2%和35.7%;抗病品種中,接種后12 h,葉片中的SOD活性比感病品種低25.9%,而接種48 h后,抗病品種中SOD活性要高于感病品種;抗病品種和感病品種中PPO活性在接種后12 h即開(kāi)始升高,并且在整個(gè)互作過(guò)程中,抗病品種葉片中的PPO活性要始終高于感病品種。以上結(jié)果表明CAT、POD、SOD、PPO活性變化與品種抗病性呈正相關(guān)。4.利用qRT-PCR技術(shù)研究了幾丁質(zhì)酶(CHI)基因和β-1,3-葡聚糖酶(GLU)基因在不同抗性品種中的表達(dá)情況。結(jié)果表明:抗病品種接種紅小豆銹病菌后24 h,CHI基因出現(xiàn)上調(diào)表達(dá),并在接種后48 h表達(dá)量達(dá)到最大,此時(shí)相對(duì)表達(dá)量是對(duì)照處理的24.5倍。在感病品種中,CHI基因的表達(dá)量在接種后12 h開(kāi)始上調(diào)表達(dá),之后表達(dá)量開(kāi)始下調(diào),并與對(duì)照相比無(wú)明顯變化;抗病品種中GLU基因相對(duì)表達(dá)量在接種后0-48 h內(nèi)無(wú)明顯變化,在接種后120 h明顯上調(diào),并且此時(shí)的表達(dá)量達(dá)到峰值,上調(diào)7.1倍。感病品種中GLU基因的相對(duì)表達(dá)量與抗病品種的變化趨勢(shì)相似,但是感病品種中GLU基因的表達(dá)量上調(diào)幅度要低于抗病品種。以上結(jié)果說(shuō)明CHI基因和GLU基因與品種抗銹性密切相關(guān)。
[Abstract]:Red bean rust is one of the main diseases in the production of red bean, which seriously affects the yield and quality of red bean. The use of resistant varieties is the most economical and effective measure to prevent red bean rust. However, there is no research report on the resistance evaluation of red bean germplasm resources to rust, and the main varieties are not resistant to rust, and it is difficult to make use of disease resistance in production. Resources, the resistance mechanism of red bean to rust is also not clear. Therefore, this study, starting with the evaluation of the resistance of red bean resources to red bean rust, screened the resistant varieties, and studied the difference of the infection process on the leaves of the pathogen of red bean rust fungus (Uromyces vignae) and the different resistant red bean varieties, as well as the CAT, S related to disease resistance. The changes of OD, POD, PPO activity and disease resistance related genes were used to lay the foundation for the utilization of red bean rust resistant resources. The main results were as follows: 1. using artificial inoculation method to evaluate the rust resistance of 85 red bean resources, obtained 2 immune varieties, 16 high resistant varieties, 10 medium resistant varieties, 26 medium varieties and 31 high sensitive varieties. Compared with susceptible varieties, their resistance to rust was mainly manifested by the delay of 2-3 d after the inoculation of rust fungus and the time of sporulation, and the density of the spore heap was less than that of the susceptible variety.2. using fluorescence microscopy to study the difference in the infection process of the rust fungus on the leaves of different resistant varieties. The spore of the rust fungus was susceptible to the disease. 6 h after inoculation can germinate and form attachments, 24 h can produce the inferior stoma and sucker mother cells, 48 h form the intracellular sucker, 8 d after inoculation, and the summer spore heap can be produced on the leaf surface. In the resistant varieties, 48 h after inoculation, the cell wall sediment which inhibited the mycelial expansion of the mycelium cells, the sporulation time was delayed obviously. The susceptible variety leaf cells did not have the tissue structure to inhibit the mycelium expansion. The results showed that the resistance of the resistant varieties to the rust fungus was two kinds of anti invasion and anti expansion. Sexual.3. uses physiological and biochemical techniques to study the relationship between the changes in the activity of defense enzymes related to rust resistance and the relationship between the resistance to disease. The results show that the activities of CAT, POD, SOD, and PPO in the leaves of the resistant and susceptible varieties and the red bean rust fungus increase in varying degrees. The activity of CAT and POD in the leaves of the resistant and susceptible varieties is 48 h after inoculation. The activity of CAT and POD in the resistant varieties was higher than that of susceptible varieties, 30.2% and 35.7%, respectively, and 12 h after inoculation, the SOD activity in leaves was 25.9% lower than that of susceptible varieties. After inoculation 48 h, the activity of SOD in the resistant varieties was higher than that of susceptible varieties, and the activity of PPO in the resistant and susceptible varieties was 12 h after inoculation. The PPO activity in the leaves of the disease resistant varieties was always higher than that of susceptible varieties during the whole process. The above results showed that the changes of CAT, POD, SOD, PPO activity and disease resistance were positively correlated.4. using qRT-PCR technology to study the table of the chitinase (CHI) gene and the beta -1,3- glucan enzyme (GLU) gene in different resistant varieties. The results showed that 24 h, CHI gene was up-regulated after inoculation of the red bean rust fungus, and the expression of 48 h reached the maximum after inoculation, and the relative expression was 24.5 times as high as that of the control treatment. In susceptible varieties, the expression of CHI gene was up to be expressed at 12 h after inoculation, and then the expression began to be downregulated and compared with the control. There was no obvious change in the relative expression of GLU gene in 0-48 h after inoculation, the 120 h increased obviously after inoculation, and the expression amount reached the peak and up to 7.1 times. The relative expression of the GLU gene in the susceptible variety was similar to that of the disease resistant varieties, but the expression of GLU gene in the susceptible varieties was expressed. The results showed that the CHI gene and GLU gene were closely related to the rust resistance of cultivars.
【學(xué)位授予單位】：黑龍江八一農(nóng)墾大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S435.21

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