title: Proteomics-based identification of midgut proteins correlated with Cry1Ac resistance in Plutella xylostella (L.).
abstract: The diamondback moth, Plutella xylostella (L.), is a worldwide pest of cruciferous crops and can rapidly develop resistance to many chemical insecticides. Although insecticidal crystal proteins (i.e., Cry and Cyt toxins) derived from Bacillus thuringiensis (Bt) have been useful alternatives to chemical insecticides for the control of P. xylostella, resistance to Bt in field populations of P. xylostella has already been reported. A better understanding of the resistance mechanisms to Bt should be valuable in delaying resistance development. In this study, the mechanisms underlying P. xylostella resistance to Bt Cry1Ac toxin were investigated using two-dimensional differential in-gel electrophoresis (2D-DIGE) and ligand blotting for the first time. Comparative analyses of the constitutive expression of midgut proteins in Cry1Ac-susceptible and -resistant P. xylostella larvae revealed 31 differentially expressed proteins, 21 of which were identified by mass spectrometry. Of these identified proteins, the following fell into diverse eukaryotic orthologous group (KOG) subcategories may be involved in Cry1Ac resistance in P. xylostella: ATP-binding cassette (ABC) transporter subfamily G member 4 (ABCG4), trypsin, heat shock protein 70 (HSP70), vacuolar H(+)-ATPase, actin, glycosylphosphatidylinositol anchor attachment 1 protein (GAA1) and solute carrier family 30 member 1 (SLC30A1). Additionally, ligand blotting identified the following midgut proteins as Cry1Ac-binding proteins in Cry1Ac-susceptible P. xylostella larvae: ABC transporter subfamily C member 1 (ABCC1), solute carrier family 36 member 1 (SLC36A1), NADH dehydrogenase iron-sulfur protein 3 (NDUFS3), prohibitin and Rap1 GTPase-activating protein 1. Collectively, these proteomic results increase our understanding of the molecular resistance mechanisms to Bt Cry1Ac toxin in P. xylostella and also demonstrate that resistance to Bt Cry1Ac toxin is complex and multifaceted.
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China; Department of Biocontrol, Institute of Plant Protection, Heilongjiang Academy of Agricultural Sciences, Harbin, 150080, China. Electronic address: jixingxia@126.com.
authors: Jixing
authors: J
authors: Xia
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: zjguo1988@163.com.
authors: Zhaojiang
authors: Z
authors: Guo
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: yyll88523@126.com.
authors: Zezhong
authors: Z
authors: Yang
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: zhuxun99@gmail.com.
authors: Xun
authors: X
authors: Zhu
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: kangshi0718@163.com.
authors: Shi
authors: S
authors: Kang
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: xinyang3856@163.com.
authors: Xin
authors: X
authors: Yang
authors: Key Laboratory of Molecular Biology of Heilongjiang Province, College of Life Sciences, Heilongjiang University, Harbin 150080, China. Electronic address: yangfshan@126.com.
authors: Fengshan
authors: F
authors: Yang
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: wuqingjun@caas.cn.
authors: Qingjun
authors: Q
authors: Wu
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: wangshaoli@caas.cn.
authors: Shaoli
authors: S
authors: Wang
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: xiewen@caas.cn.
authors: Wen
authors: W
authors: Xie
authors: Department of Biocontrol, Institute of Plant Protection, Heilongjiang Academy of Agricultural Sciences, Harbin, 150080, China. Electronic address: xu.weijun@163.com.
authors: Weijun
authors: W
authors: Xu
authors: Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: zhangyoujun@caas.cn.
authors: Youjun
authors: Y
authors: Zhang
mesh_terms: D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D004731:Endotoxins; D041981:Gastrointestinal Tract; D006460:Hemolysin Proteins; D019476:Insect Proteins / Q000187:drug effects / Q000378:metabolism*; D007305:Insecticide Resistance; D007306:Insecticides; D007814:Larva; D009036:Moths / Q000187:drug effects / Q000378:metabolism*; D040901:Proteomics / Q000379:methods
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D019476:Insect Proteins; D007306:Insecticides; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: 2D-DIGE; Bacillus thuringiensis; Cry1Ac resistance; Ligand blotting; Plutella xylostella