Analysis of knowdown resistance gene mutation in Culex tritaeniorhynchus resistant to DDT and deltamethrin in Yunnan Province, China

Abstract

Abstract:

Objective To detect the knockdown resistance（kdr） gene mutations in Culex tritaeniorhynchus resistant or susceptible to DDT and deltamethrin, in order to elucidate the relationship between the resistant phenotype and kdr mutations.Methods C.tritaeniorhynchus mosquitoes were captured in Zhaoyang, Mang Shi, Jiang Cheng and Meng Lian in Yunnan Province, and exposed to DDT and deltamethrin, respectively.The resulting alive and dead ones were counted.Genomic DNA was extracted by individual mosquitoes.The kdr gene was amplified by PCR and analyzed.The frequency of kdr genotypes was estimated in resistant and susceptible samples.The relationship between kdr genotypes and resistant phenotypes was analyzed by Chi-square test.Results A total of 411 mosquitoes were genotyped for the kdr gene.Mutation was detected at codon 1014, yielding two alleles, the wild-type TTA/L and the mutant TTT/F, and three genotypes, the wild-type homozygote L/L （frequency, 96.35%; 396/411）, the mutant homozygote F/F （0.73%; 3/411） and the wild/mutant heterozygous L/F （2.92%; 12/411）.The frequencies of mutant genotype in DDT-resistant and -susceptible individuals were 3.42% （4/117） and 3.88% (4/103), respectively; while those in deltamethrin-resistant and -susceptible individuals were 3.96% (4/101) and 3.33% （3/90）, respectively.Chi-square test revealed that there was no correlation between kdr genotype frequency and the resistance phenotypes to either DDT （χ² = 0.034, P > 0.05） or deltamethrin （χ² = 0.053, P > 0.05）.Conclusion We found a novel L1014F mutant allele for kdr, albeit with a low frequency.There is no correlation between kdr gene mutation and the resistance phenotypes to either DDT or deltamethrin.

Key words: Culex tritaeniorhynchus, Knockdown resistance, Gene mutation

CLC Number:

R384.112

Jin-yong JIANG, Hui-ying CHEN, Hong-ning ZHOU, Ya-jun MA. Analysis of knowdown resistance gene mutation in Culex tritaeniorhynchus resistant to DDT and deltamethrin in Yunnan Province, China[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2017, 35(6): 536-540.

Figures/Tables 3

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引物名 Primers	序列 (5′→3′) Sequences (5′→3′)	片段长度/bp Product length/bp
CT3	TGCTGTGCGGAGAGTGGATC	800
CT6	TTTCATTCAACTTGGCTCTG
TRI5	CTTCACCGACTTCATGCACTC	200
TRI6	GATTTTGGACAAAAG CAAGGC

杀虫剂 Insecticide	地点 Sampling site	表型 Phenotype	蚊虫数 No.	野生型纯合子(L/L) Wild type (L/L)			突变型杂合子(L/F) Heterozygote (L/F)			突变型纯合子(F/F) Mutant genotype (F/F)
杀虫剂 Insecticide	地点 Sampling site	表型 Phenotype	蚊虫数 No.	蚊虫数 No.mosquitoes	频率/% Frequency/%		蚊虫数 No.mosquitoes		频率/% Frequency/%	蚊虫数 No.mosquitoes		频率/% Frequency/%
DDT	昭阳Zhaoyang	R	45	41		91.11	4	8.89		0	0
	昭阳Zhaoyang	S	55	51		92.73	3	5.45		1	1.82
	芒市Mangshi	R	26	26		100	0	0		0	0
	芒市Mangshi	S	35	35		100	0	0		0	0
	江城Jiangcheng	R	13	13		100	0	0		0	0
	江城Jiangcheng	S	6	6		100	0	0		0	0
	孟连Menglian	R	33	33		100	0	0		0	0
	孟连Menglian	S	7	7		100	0	0		0	0
	小计Subtotal	R	117	113		96.58	4	3.42		0	0
	小计Subtotal	S	103	99		96.12	3	2.91		1	0.97
溴氰菊酯 Deltamethrin	昭阳Zhaoyang	R	23	21		91.30	1	4.35		1	4.35
	昭阳Zhaoyang	S	24	21		87.5	2	8.33		1	8.32
	芒市Mangshi	R	36	35		97.22	1	2.78		0	0
	芒市Mangshi	S	44	44		100	0	0		0	0
	江城Jiangcheng	R	17	17		100	0	0		0	0
	江城Jiangcheng	S	9	9		100	0	0		0	0
	孟连Menglian	R	25	24		96.00	1	4.00		0	0
	孟连Menglian	S	13	13		100	0	0		0	0
	小计Subtotal	R	101	97		96.04	3	2.97		1	0.99
	小计Subtotal	S	90	87		96.67	2	2.22		1	1.11