Development of an antibody-based diagnostic method for the identification of Bemisia tabaci biotype B.
The whitefly Bemisia tabaci is a very destructive pest. B. tabaci is composed of various morphologically undistinguishable biotypes, among which biotypes B and Q, in particular, draw attention because of their wide distribution in Korea and differential potentials for insecticide resistance development. To develop a biotype-specific protein marker that can readily distinguishes biotypes B from other biotypes in the field, we established an ELISA protocol based on carboxylesterase 2 (COE2), which is more abundantly expressed in biotypes B compared with Q. Recombinant COE2 was expressed, purified and used for antibody construction. Polyclonal antibodies specific to B. tabaci COE2 [anti-COE2 pAb and deglycosylated anti-COE2 pAb (DG anti-COE2 pAb)] revealed a 3-9-fold higher reactivity to biotype B COE2 than biotype Q COE2 by Western blot and ELISA analyses. DG anti-COE2 pAb exhibited low non-specific activity, demonstrating its compatibility in diagnosing biotypes. Western blot and ELISA analyses determined that one of the 11 field populations examined was biotype B and the others were biotype Q, suggesting the saturation of biotype Q in Korea. DG anti-COE2 pAb discriminates B. tabaci biotypes B and Q with high specificity and accuracy and could be useful for the development of a B. tabaci biotype diagnosis kit for on-site field applications.
College of Pharmacy and Research Institute of Pharmaceutical Science, Gyeongsang National University, 501 Jinju-daero, Jinju 660-701, Republic of Korea.
Ji Hyeong
JH
Baek
Graduate Program for Nanomedical Science, Yonsei University, 134 Shinchon-dong, Seodaemun-Gu, Seoul 120-110, Republic of Korea.
Hye Jung
HJ
Lee
Department of Entomology, Kansas State University, Manhattan, 66506, KS, USA.
Young Ho
YH
Kim
Proteometech Inc., 15-1 Yonhee-dong, Seodaemun-gu, Seoul 120-110, Republic of Korea.
Kook Jin
KJ
Lim
Department of Agricultural Biotechnology, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-921, Republic of Korea; Research Institute for Agriculture and Life Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-921, Republic of Korea.
Si Hyeock
SH
Lee
Proteometech Inc., 15-1 Yonhee-dong, Seodaemun-gu, Seoul 120-110, Republic of Korea. Electronic address: bjkim@proteometech.com.
Bum Joon
BJ
Kim
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D043182:Carboxylesterase / Q000276:immunology; D003001:Cloning, Molecular; D006430:Hemiptera / Q000201:enzymology / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D000906:Antibodies; D011994:Recombinant Proteins; D043182:Carboxylesterase
Bemisia tabaci; Biotype; Carboxylesterase; Protein biomarker; Whitefly