| Grape is a kind of important economic value fruit, nutrient-rich grapes and wine. Grape grey mildew is one of important diseases in grape cultivation, grape production can lead to 15% to 50%, serious or even never produced. In recent years, the main control measures on the production of chemical pesticides to control, breeding of resistant varieties and biological control, however, due to easy to cause environmental pollution, pesticide residues, drug resistance, time-consuming, low efficiency, poor stability and preventing problems, so by increasing the level of grape autoimmune or inactivate pathogens or inhibit the activity of pathogenic bacteria to resist damage more prominent advantages.Resveratrol is a kind of phytoalexins induced by pathogen attack or unadapted environment stimuli, and has been recognized as major maker of grapevine resistance screening, in addition to resveratrol, and its metabolites are related to disease resistance and grapes, it is reported that glycosylation products piceid resveratrol almost no antibacterial activity, but its oxidation product dimer viniferin antibacterial stronger, but less research on the mechanism of its metabolites viniferin. Botrytis cinerea laccase is produced by an important class of pathogenic proteins, the ability of resveratrol to viniferin oxide, and its production and activity of laccase and gray mold pathogenic strength related. UV radiation is one of the main methods for inducing a lot of resveratrol synthesis and metabolism, the study cinerea in grapes and UV induced resveratrol synthesis and metabolism, as well as B. cinerea laccase production catalytic mechanism of resveratrol for understanding the grape antitoxin resistance mechanism of resveratrol, resveratrol and laccase interaction mechanisms, selection and use of resistant varieties of grapes and gray mold prevention and treatment of some theoretical and practical value.In this study, laccase activity was detected in 53 strains of B. cinerea from different host plants, and contents of resveratrol and its derivatives were analyzed, and meanwhile gene expression involved in the resveratrol biosynthesis pathway were examined by qRT-PCR, upon treatment with UV radition or inoculation with a representative strain of Botrytis cinereausing HPLC in three grapevine cultivars (Xiahei, Jufeng and Fapu). Results were as followes:1. Laccase producing capability of B.cinerea from different host plants, was found that all strains had the ability to produce laccase, which was lowest in YSD2 strain, and highest in SYYK112. Futhermore, there was different laccase activity in different strains, which had a maximium level in YM3 and a minimum in YSD1. It had no significant correlation between laccase production capacity and its corresponding activity. Among them, and the SP4 strain was capable of causing different disease incidence in 3 grape leaves and fruits, and the growth rate of the SP4 strain was 11.8 mm.d-1 when laccase activity was highest after 20 days culture.2. After inoculation with B. cinerea, disease symptoms rapidly appeared and disease lesions subsequently spread on the leaves of Jufeng as compared to Fapu and Xiahei. Similarly, UV treatment resulted in a severe damagement in Jufeng rather than other two grapes. Accumulation of resveratrol and its derivatives was different in three grapes and with highest contents in Fapu and lowest in Jufeng, indicating that Fapu showed a high resistance to B. cinerea and tolerance to UV treatment among three grapes.3. Accumulation of resveratrol and its derivatives by HPLC analysis showed that B. cinerea induced different contents of resveratrol, its glucoside Piceid and dimer ?-viniferin, but not Psterostilbene in all three grapes. There was different change of reseveratrol and its derivatives depending on grape cultivars. UV treatment showed a simiar accumulation trend of resveratrol, piceid, psterostilbene and ?-viniferin, and it had a positive correlation between treatment time and contents, at 48 h or 96 h or 168h with the highest content. In addition, B. cinerea also induced resveratrol accumulation in grape fruits, which was much lower than those in grape leaves. Treatment time of UV showed a positive relation with accumulation of resveratrol, piceid, psterostilbene and ?-viniferin. UV treatment for 60 min accumulated the highest contents of all componds. After induced by B.cinerea and UV-radiation in grapevines in different time, Resveratrol, Piced, Pterostibene and ?-viniferin were while increasing the synthesis, it was positively correlated with UV irradiation time, among them, Res synthesis were significantly higher than Piced, Pterostibene and ?-viniferin. It showed grape leaves Resveratrol accumulation was metabolic synthesis Piced, Pterostibene and E-viniferin prerequisite. Compare the contents of different grape varieties found in the product after UV-radiation, the contents were Resveratrol and E-viniferin:Fapu> Xiahei>Jufeng. Comparison of two treatment found, after induced by UV-radiation in leaf synthetic Resveratrol and Piced were significantly higher than induced by B.cinerea, it showed UV radiation induced synthesis and metabolism of resveratrol was more. At the same, by UV-radiation, but by Fapu, Jufeng and Fapu leaves induced Pterostibene were increased significantly, it said that Pterostibene may be the grape leaves in the stilbene compound UV induced response factor.4. Using qRT-PCR technique to detect the expression of B.cinerea in grapes or UV induction of resistance genes, the results show:After induced by B.cinerea, Xiahei and Jufeng leaves RS, STS, PAL, MYBBF2 and MYB14F gene consistent trend:Xiahei and Jufeng had hardly expression, expression of Fapu leaves for the first increase in a trend of decrease, and at 24 h or 48 h up to the maximum. Three kinds of leaves PGIP? PR5 and PRIO gene consistent trend (except Jufeng leaves PGIP gene had hardly expression), all for the first increase in a trend of decrease, and PRIO gene expression was maximum. Induced grape leaves by UV-radiation, except Jufeng leaves RS and Fapu leaves MYBBF2, MYB14F and PR5 gene expression was maximum at 30 minutes, other resistance gene showed a continuous increase in the expression of the trend, gene expression was maximum at 60 minutes, and PR10 gene expression was maximum. After induced by UV-radiation, grape resistance gene expression Fapu> Xiahei> Jufeng. This result indicates, two treatments could induce defense related genes upregulated. Related genes are upregulated expression at the same time before the large-scale synthesis of resveratrol and its metabolites, and the expression of the highest is Fapu, followed by Xiahei, the lowest is Jufeng, it showed that regulation resveratrol-related genes by anabolic, and Fapu was resistance stronger than Xiahei, stronger than Jufeng. |
PDF Full Text Request