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Resistance Monitoring and cDNA Cloning of Cytochrome P450 in Panonychus Citri (McGregor) (Acari: Tetranychidae)

Author: LiuYongHua
Tutor: WangJinJun
School: Southwestern University
Course: Agricultural Entomology and Pest Control
Keywords: Panonychus citri Sensitivity monitoring Acaricides Synergist Clone Cytochrome P450
CLC: S436.66
Type: Master's thesis
Year: 2010
Downloads: 81
Quote: 2
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Abstract


Citrus red mite Panonychus citri (McGregor) is a serious harm of citrus pest mites worldwide. Attaches great importance to the mite resistance problem has caused scientists and management departments worldwide. This thesis by slide dipping Determination Panonychus citri field populations of the sensitivity of the commonly used acaricides, A Comparative Study of the difference in sensitivity to acaricides Panonychus mites feeding on different citrus germplasm resources determination of triphenyl phosphate esters (TPP), maleic acid diethyl ethyl ester (DEM) and piperonyl butoxide (PBO) 3 the synergist on acaricides synergies research aims to grasp citrus full mite acaricide resistance level and the formation of drug-resistant physiological and biochemical mechanisms, to provide a basis for the citrus red mite resistance management. In addition, using RT-PCR and RACE clones obtained Panonychus mites cytochrome P450 gene full-length cDNA fragment of each one, provided the basic data for further study P450 gene-mediated citrus Panonychus mites molecular mechanisms of resistance. The main findings are as follows: 1. Citrus whole mite field populations of sensitivity to commonly used acaricides 1.1 citrus red mite sensitivity monitoring of four kinds of acaricide sensitive monitoring results for the biennium 2008-2009, the citrus fenpropathrin (Panonychus acaricides tested four kinds of pyrethroid pesticides) decreased sensitivity fastest LCso value of 4.93 mg · L-1 and 25.17 mg · L-1, respectively, in the two-year period relative up to 5.1 times confrontational; followed pyridaben (Heterocyclic pesticide) LCso value of 3.78 mg · L-1 and 8.38 mg L-1 were relatively resistant up to 2.2 times; Panonychus citri abamectin (biological source pesticides) sensitivity basic remain unchanged; increased sensitivity of emamectin benzoate (semi-synthetic antibiotics, pesticides), 2009 Panonychus citri emamectin benzoate relative The sensitivity is 1.4 times that of 2008. There is a certain sensitivity 1.2 feeding on different citrus germplasm resources citrus red mite sensitivity to commonly used acaricides determination were collected from different citrus germplasm resources Panonychus citri fenpropathrin and pyridaben differences. Panonychus citri of different food stocks on the fenpropathrin the sensitivity descending order of grapefruit, orange and trifoliate which Aurantii populations of fenpropathrin phase confrontational is 2.34 times the population of the grapefruit; on da pyridaben sensitivity descending order of the grapefruit, Citrus aurantium and orange populations, Orange populations pyridaben relatively resistant grapefruit population of 3.92 times; feeding citrus germplasm resources Panonychus citri avermectin sensitivity did not differ significantly. Three kinds of synergist synergist commonly used acaricides synergies synergies measurement results, TPP fenpropathrin, abamectin and emamectin benzoate significant synergies, efficiency multiple of 3.2 times, 2.0 times and 3.8 times, respectively. DEM pyridaben and emamectin benzoate has significant synergies, efficiency multiple of 3.3 times and 2.6 times, respectively. The PBO of the fenpropathrin Pyridaben and emamectin benzoate have significant synergies, efficiency multiple of 2.7 times, 2.4 times and 7.2 times. PBO avermectin synergistic effect, its efficiency factor of 1.7 times. TPP on pyridaben DEM fenpropathrin and abamectin had no synergistic effect. Citrus full mite cytochrome P450 gene cloning use of RT-PCR, and cloned two Panonychus citri P450 gene fragments N1 and N2, GenBank accession number FJ480403 and FJ480404. By RACE technology, cloning N2 full-length sequence, the full-length gene was named CYP4CF1 its sequence of this 1880 bp, including the open reading frame of 1527 bp encoding 508 amino acids, 87 bp 5 'non-coding region 286 bp 3 'non-coding region of the initiation codon ATG and stop codon TAA. In its nucleic acid sequence 3 'terminal poly-A structure of a typical polyadenylation signal (ATTAAA). Its deduced amino acid containing the P450 gene classic heme binding site consensus sequence F × xGxxxcxc located the the helix K involved in the stable core structure of a completely conserved sequence Ex × R and CYP4 family characteristic sequence EVDTFMFEGHDTT.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Horticultural Crops Pest and Disease Control > Fruit tree pests and diseases > Citrus pests and diseases
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