In recent years, acid rot is one of the main diseases found in fruit ripening and storage and transportation. It not only causes rotten fruit in late harvest, but also seriously affects the quality of wine grape and its products. Grape acid rot is caused by acid rot fungus. At present, chemical fungicides are mainly used to control acid rot fungus, but this chemical method will not only lead to the production of drug-resistant strains, but also the fungicides left on the surface of fruits will seriously harm human health. In recent years, more attention has been paid to phytoalexin. Pterostilbene is an important phytoalexin produced by grapes under the pressure of resisting external biological or non biological stress. In the early stage, it was found that pterostilbene can effectively inhibit the activity of acid rot fungi through in vitro antibacterial experiment. However, the defense mechanism of red pterostilbene against acid rot fungi in grape is not clear, so this experiment focused on the mechanism of it against acid rot fungi, isolated and identified the secondary metabolites of grape defense acid rot fungi, cloned and analyzed the key genes defense acid rot fungi, and The research results include the following aspects: 1.
1. The inhibitory effect of different concentrations of pterostilbene on acid rot was studied by in vitro plate bacteriostasis experiment. The results showed that with the increase of the concentration of it, the inhibitory effect was significantly enhanced. When the concentration of red pterostilbene was 400ug / ml, the inhibitory rate reached 79.91%. In order to explore the defense against acid rot fungus in grape, the grape was infected by acid rot fungus, and the grape decay was observed from the physiological level. It was found that 48 hours after infection, the grape began to rot, accompanied by rotten sour smell; 72 hours after infection, the grape seriously rotted. In order to resist disease infection, grape will produce secondary metabolites. Two main defense products, resveratrol and pterostilbene, were isolated by uhplc-qqq-ms2. The content of these metabolites before and after infection was detected to explore the changes in the defense process. The results showed that it was significantly improved by acid rot fungus infection. High content of resveratrol and pterostilbene in grape.
2. According to the related reports and the existing grape EST database, we cloned the key gene of pterostilbene defense, resveratrol oxygen methyltransferase gene (wgreom). Sequence analysis showed that the open reading frame of wgreom was 1116bp, encoding 305 amino acids, molecular weight was 34.4kda, isoelectric point was 6.52.The phylogenetic analysis showed that wereom and vvromt belong to the same subfamily, and they are closely related to rcoomt. The results of amino acid sequence homology comparison showed that wgreom had 95.86% homology with European grape vvromt, and 65.61%, 62.10% and 30.57% homology with Chinese rose rcoomt1, castor rsomt and Dutch iris homt, respectively.
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