Resistance monitoring and target gene cloning of Bemisia tabaci Q-biotype from Jiangsu Province

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1 Chinese Journal of Applied Entomology * Q Q Bemisia tabaci Gennadius 5 Q RCR 287 bp 184 bp ace1 para- Q F331W L925I T929V Resistance monitoring and target gene cloning of Bemisia tabaci Q-biotype from Jiangsu Province YUAN Lin-Ze 1 WANG Shao-Li 2 CHEN Xue-Lin 1 DU Yu-Zhou 1 ZHANG You-Jun 2 WANG Jian-Jun 1 1. College of Horticulture and Plant Protection Yangzhou University Yangzhou 2. Institute of Vegetables and Flowers Chinese Academy of Agricultural Sciences Beijing China China Abstract The levels of resistance in three field populations of the Bemisia tabaci Q-biotype from Yangzhou Dongtai and Wuxi in Jiangsu Province to five representative insecticides were determined using the leaf dipping method. The result show that these three populations had low to moderate levels of resistance to cypermethrin and imidacloprid but remain largely susceptible to abamectin compared to a relatively susceptible population. Two fragments of the B. tabaci acetylcholinesterase enzyme ace1 and para-type voltage gated sodium channel gene with respective lengths of 287 bp and 184 bp were amplified by PCR. Sequence analysis suggested the presence of the F331W mutation in acetylcholinesterase ace1 and L925I and T929V mutations in the sodium channel gene mutations associated with resistance to organophosphate and pyrethroid insecticides respectively. Key words Bemisia tabaci resistance monitoring acetylcholinesterase sodium channel target-site associated resistance Bemisia tabaci Gennadius Q Homoptera Aleyrodidae Bemisia 2005 Ⅰ COⅠ 110 Jones 2003 Q Q B Q Chu et al Chu et al * drjianjun@ yahoo. com. cn

2 1 Q 49 Q B Table1 Origins and host plants of B. tabaci populations Hostplant collection Date of Population Location Biotype YZ Q Yangzhou Gerbera WX Q Wuxi Cucumber Elbert and Nauen 2000 Kady and Devine Roditakis et al BJ B Beijing Cabbage S- The letters of population are same for Table 2. Byrne et al Wang and Wu 2007 Alon et al Karunker et al Tsagkarakou et al Wang et al cm 3 Q 10 s Q ace1 2 ml 15 g / L 20 ~ 25 4 Q 25 L D = h LC 50 mg / L DNA DNA μl 1 l% SDS 500 mmol / L Tris-HCI 20 mmol / 3 1 L NaCI 1mmol / L EDTA 200 mg / ml K Q ~ 3 h min % Dichlorvos DNA 20% Carbosulfan PCR Tsagkarakou 10% 2009 Cypermethrin ace1 BtAcHE1F1 5-10% Imidacloprid TAGGGATCTGCGACTTCCC % Abamectin DT Dongtai Capsicum Q Feng 2010 K - 20 BtAcHE1R1 5 - GTTCAGCCAGTCCG

3 50 Chinese Journal of Applied Entomology 48 TGTACT Bt-Kdr-F1 5 - GCCAAATCCTGGCCAACT B 3 Bt-kdr-RIntr1 5 - LC 50 GAGACAAAAGTCCTGTAGC μL PCR μlddh 2 O 2. 5μL10 raction buffer 2 μl 25 mmol / L MgCl μl 10mmol / L dntp 0. 5 μl10 μmol / L / 1 μl 10 Taq DNA 1μL gdna 94 5 min s 60 ~ s s s s 72 PCR mg / L 30 s min PCR ~ pgem-t vector 2. 2 PCR 287 bp Q LC ~ mg / L ~ LC mg / L Q LC ~ mg / L ace1 1 Q ace1 Alon SUD-S 3 GenBank EF Q T C G G Table 2 2 Q 5 Resistance in field populations of Q type B. tabaci to five insecticides Populations Insectcides ± SE Slope ± SE LC 50 mg / L 95% 95% FL RF BJ YZ WX DT Dichlorvos ± ~ Carbosulfan ± ~ Cypermethrin ± ~ Imidacloprid ± ~ Abamectin ± ~ Dichlorvos ± ~ Carbosulfan ± ~ Cypermethrin ± ~ Imidacloprid ± ~ Abamectin ± ~ Dichlorvos ± ~ Carbosulfan ± ~ Cypermethrin ± ~ Imidacloprid ± ~ Abamectin ± ~ Dichlorvos ± Carbosulfan ± ~ Cypermethrin ± ~ Imidacloprid ± ~ Abamectin ± ~

4 1 Q 51 Torpedo californica AJ Q F331W 1 A C G T PCR 184 bp Q T929V Q IIS Morin 2002 T A 925 SUD-S GenBank L925I Fig. 1 1 Q Nucleotide and deduced amino acid sequences of the acetylcholinesterase gene fragment amplified from field populations of B. tabaci Q-biotype. The resistance-associated mutation is in bold face. 3 Fig. 2 2 Q Nucleotide and deduced amino acid sequences of the sodium channel gene fragment amplified from field populations of B. tabaci Q-biotype The resistance-associated mutation is in bold face

5 52 Chinese Journal of Applied Entomology 48 B ⅡS4-6 cdna L925I Roditakis ~ ~ ~ ~ L925I 22 Luo T929V 3 Q 3 Q 2 B ace1 Q B Q F331W B Q L925I T929V 3 20 ~ 170 Q 3 Q References Alon M Alon F Nauen R Morin S Organophosphate Q resistance in the B biotype of Bemisia tabaci Hemiptera LC 50 Aleyrodidae is associated with a point mutation in an ace ~ mg / L ~ mg / L type acetylcholinesterase and overexpression of LC ~ carboxylesterase Insect. Biochem. Mol. Biol mg / L LC ~ mg / L Byrne FJ Gorman KJ Cahill M Denholm I Devonshire AL The role of B-type esterases in conferring resistance in the tobacco whitefly. Pest. Manag. Sci Chu D Jiang T Liu GX Jiang DF Tao YL Fan ZX Zhou HX Bi YP Biotype status and distribution of Bemisia tabaci Hemiptera Aleyrodidae in Shandong province of China based on mitochondrial DNA markers. Environmental Alon 2008 B Entomology ace1 Chu D Zhang YJ Brown JK Cong B Xu BY Wu QJ Zhu 331 F331W GR The introduction of the exotic Q biotype of PCR-RFLP 15 Q Bemisia tabaci Gennadius from the Mediterranean region into China on ornamental crops. Florida Entomologist 89 F331W Q Q Tsagkarakou et al Morin 2002 B Q B Elbert A Nauen R Resistance of Bemisia tabaci IIS Homoptera Aleyrodidae to insecticide s in southern Spain L925I 88 with special reference to neonicotinoids. Pest. Manag. Sci. 9 L925I L925I B Feng Y Wu Q Wang S Chang X Xie W Xu B Zhang Y 2004 RT-PCR Cross-resistance study and biochemical mechanisms of B thiamethoxam resistance in B-biotype Bemisia tabaci

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