叶斑病对苦水玫瑰品质的影响及其病原菌鉴定

发布时间：2017-12-28 00:06

本文关键词：叶斑病对苦水玫瑰品质的影响及其病原菌鉴定　出处：《甘肃农业大学》2016年博士论文　论文类型：学位论文

【摘要】：苦水玫瑰(R.setate×R.rugosa)是钝齿蔷薇和我国传统玫瑰的自然杂交种,在甘肃省永登县苦水镇已有200多年的栽植历史,是甘肃省主要香料作物之一,玫瑰精油产量约占全国总产量的80%左右。随着市场需求的急剧增加,其经济价值和社会效益日渐凸显,已成为香料用、药用、酿酒等多种用途的特殊经济作物。由于苦水玫瑰特殊的经济价值,促使其种植栽培面积急剧增加,由此引起种植管理粗放、与其他农作物间作等问题,导致病虫害滋生蔓延,最终使得苦水玫瑰的产量严重下降。近年来,田间种植栽培过程中发现,苦水玫瑰叶斑病发病率较高,危害也越来越严重。为了探明叶斑病对苦水玫瑰品质的影响、揭示叶斑病病原菌发病机制,实验就叶斑病对苦水玫瑰活性物质积累的影响、植株抗病过程中生理生化变化、病原菌分离鉴定、以及病原菌的防治等方面进行了研究。结果如下:1)通过采用GC-MS、分光光度法、DPPH和FRAP等方法,对开花旺季苦水玫瑰精油中活性物质进行分离与检测,结果发现,叶斑病引起20种主要特征组分的变化,其中8种组分呈现出不同程度的增加,8种则呈现出不同程度的减少,比如,香茅醇、芳樟醇和异戊醇的相对含量分别降低了62.83%、63.08%和60.18%。随着叶斑病感病程度的增加,可溶性糖、总黄酮、酚类化合物含量均显著减少,Ⅱ级和Ⅲ级感病程度相对于对照(Ⅰ级)分别降低了17.64%和28.72%、49.15%和76.10%、46.60%和67.90%;抗氧化能力也显著减少,I%、FRAP值分别降低了57.24%和73.45%、53.64%和74.80%。2)通过采用HPLC法、qPCR等方法,对开花旺季苦水玫瑰叶片进行生理生化特性的测定,结果发现,随着感病程度的增加,净光合速率、气孔导度、胞间CO2浓度、以及叶绿素含量显著减少,而蒸腾速率、抗氧化酶SOD和PPO活性显著增加;次生代谢关键酶PAL活性、以及关键基因PAL和CHS的表达量也显著增加,其中,PAL、CHS基因的表达量在Ⅱ级和Ⅲ级感病程度,相对于对照分别提高了1.8和4.41倍、2.67和4.23倍。3)通过对苦水玫瑰叶斑病病原进行分离与鉴定,以及室内药剂筛选,结果发现,形态学和致病性测定初步鉴定为链格孢属(Alternaria),叶斑病菌丝DNA经过ITS1F-ITS4引物扩增、测序分析、同源性比对和系统进化树位置进一步证实,引起苦水玫瑰叶斑病的病原菌为链格孢属(Alternaria)芦荟黑斑病(Alternaria arborescens,JQ676197)致病变种。喷施笨醚甲环唑和戊唑醇抑菌效果较好,EC50分别为0.0017μg/mL和0.0041μg/mL。综上所述,叶斑病降低了活性物质的积累和苦水玫瑰的品质。在植株自身抗病过程中,叶斑病刺激了叶片次生代谢过程中关键酶活性以及关键基因的高效表达。苦水玫瑰叶斑病病原菌属于芦荟黑斑病(Alternaria arborescens)变种,利用笨醚甲环唑和戊唑醇可有效防治苦水玫瑰叶斑病的发生。这些研究将为苦水玫瑰大面积种植栽培、叶斑病病原菌防治等方面提供理论基础和技术参考。
[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.
【学位授予单位】：甘肃农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S436.8

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