A Cytochrome p450 and Its Application in Degrading Pesticide Residues
A cytochrome and population technology, applied in the field of biotechnology and environmental protection, can solve the problems of dead, white leaves, yellow and dry leaves, etc.
Active Publication Date: 2022-06-24
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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Problems solved by technology
Tetranychus urticae like to cluster on the back of plant leaves, and do damage by sucking plant juice in three developmental stages: adults, larvae, and nymphs. First, white spots are formed around the main veins of leaves, which in turn causes leaves to turn white and affects plant photosynthesis. In severe cases, the leaves turn yellow and dry, and even the whole plant dies, which greatly affects the yield and quality of crops.
After Tetranychus urticae developed resistance to organophosphorus insecticides, the P450 genes involved in detoxification and metabolism with definite physiological functions were rarely disclosed. The lack of these key genes involved in the formation of resistance also hindered the development of new pesticides and Development of pest resistance management strategies
Based on this, the present invention has discovered that a P450 gene CYP392D8 is overexpressed in the population of resistant spider mite, and the gene can be metabolized in vitro after heterologous expression and protein purification. The P450 gene involved in the resistance of Tetranychus urticae to organophosphorus insecticides has been reported. At the same time, it also provides a theoretical basis for the design of new pesticide targets
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[0018] The present invention is further described below in conjunction with specific embodiments, but does not constitute a limitation of the present invention.
[0019] Materials and Methods
[0020] Test insects
[0021] Two-spotted spider mite indoor sensitive population (Lab-IPP) was provided by the Natural Enemy Insect Research Group of the Institute of Plant Protection, Chinese Academy of Agricultural Sciences. Phaseolus vulgaris Linn) has been raised on leaves until now, and has never been exposed to any chemicals during this period. The rearing conditions were: temperature 26±1°C, humidity 60±5% and photoperiod of L:D=16h:8h. The leaves used for raising the test mites were "Bifeng" kidney bean leaves.
[0022] The three test field populations were collected from eggplant and strawberry hosts in Changping (CP), Beijing, Miyun (MY), and Shouguang, Shandong (SG), respectively. The two-spotted spider mite population collected from the field was reared on clean and in...
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Abstract
The invention discloses a cytochrome P450, the amino acid sequence of which is shown in SEQ ID NO: 2. The P450 gene is overexpressed in the populations with low to medium level resistance to organic phosphorus in the field. The gene can be used as a molecular Marker-assisted early detection and early warning of resistance to organophosphorus insecticides in the field Tetranychus urticae population, and the detection process is simple and fast. The gene can metabolize chlorpyrifos and pirimiphos-methyl in vitro, and is expected to metabolize other organophosphorus pesticides. Therefore, the engineered bacteria expressed by this gene can degrade organic phosphorus pesticide residues in fruits and vegetables, waste water and sewage It has the potential for industrial development and application. The cloned P450 gene involved in drug resistance can also be used as a target to design inhibitors of specific detoxification enzymes and provide ideas for the development of new drugs in the future.
Description
technical field [0001] The invention belongs to the fields of biotechnology and environmental protection, and particularly relates to a cytochrome P450 and its application in degrading pesticide residues. Background technique [0002] Two-spotted spider mite Tetranychus urticae It belongs to the order Arachnida and is a major agricultural pest with a worldwide distribution. The two-spotted spider mite has a diverse diet and can eat more than 1100 kinds of host plants including vegetables, fruit trees, flowers, peanuts, etc. Two-spotted spider mites like to gather on the back of plant leaves, and suck plant sap in the three developmental stages of adult mites, larvae, and nymphs. They first form white spots around the main veins of leaves, which in turn cause the leaves to turn white and affect plant photosynthesis. In severe cases, the leaves will turn yellow and wither, or even the whole plant will die, which greatly affects the yield and quality of crops. At present, in...
Claims
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IPC IPC(8): C12N9/02C12N15/53C12Q1/6888B09C1/10C02F3/34A23B7/155C12N15/87A01K67/033C12N15/866C12N5/10C02F101/38
CPCC12N9/0071C12Q1/6888B09C1/10C02F3/342A23B7/155C12N15/87A01K67/0337C12N15/86C12N5/0601C12Q2600/158C12Q2600/106C02F2101/306C02F2101/40A01K2207/05A01K2227/70C12N2710/14143C12N2800/105C12N2510/02
Inventor 王少丽徐丹丹张友军谢文吴青君郭兆将
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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