本文關(guān)鍵詞：葉斑病對(duì)苦水玫瑰品質(zhì)的影響及其病原菌鑒定　出處：《甘肅農(nóng)業(yè)大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 苦水玫瑰 葉斑病 活性物質(zhì)積累 生理生化特性 PAL和CHS 病原菌分離鑒定 藥劑防治

【摘要】：苦水玫瑰(R.setate×R.rugosa)是鈍齒薔薇和我國傳統(tǒng)玫瑰的自然雜交種,在甘肅省永登縣苦水鎮(zhèn)已有200多年的栽植歷史,是甘肅省主要香料作物之一,玫瑰精油產(chǎn)量約占全國總產(chǎn)量的80%左右。隨著市場需求的急劇增加,其經(jīng)濟(jì)價(jià)值和社會(huì)效益日漸凸顯,已成為香料用、藥用、釀酒等多種用途的特殊經(jīng)濟(jì)作物。由于苦水玫瑰特殊的經(jīng)濟(jì)價(jià)值,促使其種植栽培面積急劇增加,由此引起種植管理粗放、與其他農(nóng)作物間作等問題,導(dǎo)致病蟲害滋生蔓延,最終使得苦水玫瑰的產(chǎn)量嚴(yán)重下降。近年來,田間種植栽培過程中發(fā)現(xiàn),苦水玫瑰葉斑病發(fā)病率較高,危害也越來越嚴(yán)重。為了探明葉斑病對(duì)苦水玫瑰品質(zhì)的影響、揭示葉斑病病原菌發(fā)病機(jī)制,實(shí)驗(yàn)就葉斑病對(duì)苦水玫瑰活性物質(zhì)積累的影響、植株抗病過程中生理生化變化、病原菌分離鑒定、以及病原菌的防治等方面進(jìn)行了研究。結(jié)果如下:1)通過采用GC-MS、分光光度法、DPPH和FRAP等方法,對(duì)開花旺季苦水玫瑰精油中活性物質(zhì)進(jìn)行分離與檢測,結(jié)果發(fā)現(xiàn),葉斑病引起20種主要特征組分的變化,其中8種組分呈現(xiàn)出不同程度的增加,8種則呈現(xiàn)出不同程度的減少,比如,香茅醇、芳樟醇和異戊醇的相對(duì)含量分別降低了62.83%、63.08%和60.18%。隨著葉斑病感病程度的增加,可溶性糖、總黃酮、酚類化合物含量均顯著減少,Ⅱ級(jí)和Ⅲ級(jí)感病程度相對(duì)于對(duì)照(Ⅰ級(jí))分別降低了17.64%和28.72%、49.15%和76.10%、46.60%和67.90%;抗氧化能力也顯著減少,I%、FRAP值分別降低了57.24%和73.45%、53.64%和74.80%。2)通過采用HPLC法、qPCR等方法,對(duì)開花旺季苦水玫瑰葉片進(jìn)行生理生化特性的測定,結(jié)果發(fā)現(xiàn),隨著感病程度的增加,凈光合速率、氣孔導(dǎo)度、胞間CO2濃度、以及葉綠素含量顯著減少,而蒸騰速率、抗氧化酶SOD和PPO活性顯著增加;次生代謝關(guān)鍵酶PAL活性、以及關(guān)鍵基因PAL和CHS的表達(dá)量也顯著增加,其中,PAL、CHS基因的表達(dá)量在Ⅱ級(jí)和Ⅲ級(jí)感病程度,相對(duì)于對(duì)照分別提高了1.8和4.41倍、2.67和4.23倍。3)通過對(duì)苦水玫瑰葉斑病病原進(jìn)行分離與鑒定,以及室內(nèi)藥劑篩選,結(jié)果發(fā)現(xiàn),形態(tài)學(xué)和致病性測定初步鑒定為鏈格孢屬(Alternaria),葉斑病菌絲DNA經(jīng)過ITS1F-ITS4引物擴(kuò)增、測序分析、同源性比對(duì)和系統(tǒng)進(jìn)化樹位置進(jìn)一步證實(shí),引起苦水玫瑰葉斑病的病原菌為鏈格孢屬(Alternaria)蘆薈黑斑病(Alternaria arborescens,JQ676197)致病變種。噴施笨醚甲環(huán)唑和戊唑醇抑菌效果較好,EC50分別為0.0017μg/mL和0.0041μg/mL。綜上所述,葉斑病降低了活性物質(zhì)的積累和苦水玫瑰的品質(zhì)。在植株自身抗病過程中,葉斑病刺激了葉片次生代謝過程中關(guān)鍵酶活性以及關(guān)鍵基因的高效表達(dá)。苦水玫瑰葉斑病病原菌屬于蘆薈黑斑病(Alternaria arborescens)變種,利用笨醚甲環(huán)唑和戊唑醇可有效防治苦水玫瑰葉斑病的發(fā)生。這些研究將為苦水玫瑰大面積種植栽培、葉斑病病原菌防治等方面提供理論基礎(chǔ)和技術(shù)參考。
[Abstract]:R.setate R.rugosa is a natural hybrid of Rosa rugosa and Chinese traditional rose. It has been planted for more than 200 years in Yongdeng, Gansu province. It has been planted for more than 200 years. It is one of the main spice crops in Gansu province. The yield of rose essential oil is about 80% of the total output of the country. With the rapid increase of market demand, its economic and social benefits are becoming increasingly prominent. It has become a special economic crop for many uses, such as spices, medicine and wine making. Due to the special economic value of bitter water rose, its cultivation and cultivation area increased sharply, which caused extensive planting management and intercropping with other crops, which led to the spread of disease and insect pests, and ultimately led to a serious decline in yield of bitter water rose. In recent years, the incidence of rose leaf spot in bitter water rose is higher and the harm is more and more serious in the field of cultivation and Cultivation in the field. In order to explore, to the quality of leaf spot disease Kushui reveal the pathogenesis of leaf spot disease pathogens, and experiments on effect of leaf spot disease Kushui accumulation of active substances in the process of physiological and biochemical changes of plant disease resistance, pathogen isolation and identification of pathogenic bacteria, and the prevention and other aspects of the study. The results are as follows: 1) by using GC-MS, spectrophotometry, DPPH and FRAP, were separated and detected on active substances in the flowering season of rose essential oil, leaf spot disease caused by 20 kinds of main features of the change of the composition, the 8 components showed different degrees of increase, while the 8 show different degrees of reduction, for example, the relative content of geraniol, linalool and isoamyl alcohol were decreased by 62.83%, 63.08% and 60.18%. With the increasing degree of sense of leaf spot disease, soluble sugar, total flavonoids, phenolic compounds were significantly reduced, grade II and III disease level compared to the control (grade I) were decreased by 17.64% and 28.72%, 49.15% and 76.10%, 46.60% and 67.90%; the antioxidant capacity also decreased significantly, I% and FRAP respectively. Decreased by 57.24% and 73.45%, 53.64% and 74.80%. 2) by using HPLC method, qPCR method, determination of physiological and biochemical characteristics of flowering season Kushui rose leaves were found, with the increased sense of disease degree, net photosynthetic rate, stomatal conductance, intercellular CO2 concentration, and chlorophyll content decreased significantly, and the transpiration rate, antioxidant enzymes SOD and PPO the activity increased significantly; key enzyme activity, PAL secondary metabolism and expression of key genes PAL and CHS were significantly increased, the expression of PAL and CHS gene disease level in grade II and III, compared to the control respectively increased 1.8 and 4.41 times, 2.67 times and 4.23. 3) through the isolation and identification of the pathogen of leaf spot disease and Kushui rose, fungicides, results show that the morphology and pathogenicity identification of Alternaria leaf spot (Alternaria), after ITS1F-ITS4 amplification, DNA wire sequencing, homology and phylogenetic tree of position further confirmed that the pathogen causing bacteria Kushui rose leaf spot for Alternaria Leaf spot (Alternaria) (Alternaria arborescens, aloe JQ676197) PV. Good at stupid Difenoconazole and tebuconazole antibacterial effect, EC50 were 0.0017 g/mL and 0.0041 g/mL. To sum up, leaf spot disease reduces the accumulation of active substances and the quality of bitter water rose. In the process of plant disease resistance, leaf spot disease stimulates the activity of key enzymes and the efficient expression of key genes during the secondary metabolism of leaves. The pathogen of leaf spot disease rose grievances (Alternaria arborescens) belongs to the aloe leaf spot varieties, use stupid Difenoconazole and tebuconazole can effectively prevent the occurrence of Kushui rose leaf spot. These studies will provide theoretical basis and technical reference for large area planting and cultivation of bitter water rose, prevention and cure of pathogen of leaf spot disease.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S436.8

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