Monoclonal Antibody-Based Enzyme Linked Immunosorbent Assay for the Analysis of Jasmonates in Plants

作     者：Aixing Deng Weiming Tan Suping He Wei Liu Tiegui Nan Zhaohu Li Baomin Wang Qing X. Li 

作者机构：College of Agronomy and Biotechnology China Agricultural University Beijing 100094 China Department of Molecular Biosciences and Bioengineering University of Hawaii Honolulu Hawaii 96822 USA 

出 版 物：《Journal of Integrative Plant Biology》 (植物学报（英文版）)

年 卷 期：2008年第50卷第8期

页      面：1046-1052页

核心收录：

学科分类：0710[理学-生物学] 071001[理学-植物学] 07[理学] 

主　　题：enzyme-linked immunosorbent assay immunoassay methyl jasmonate plant growth regulators 

摘      要：Methyl jasmonate （MeJA） and its free-acid form, jasmonic acid （JA） are naturally occurring plant growth regulators widely distributed in higher plants. In order to improve the sensitivity for the analysis of MeJA at low levels in small amounts of plant samples, a monoclonal antibody （MAb） （designated as MAb 3E5D7C4B6） against MeJA was derived from a JA- bovine serum albumin （BSA） conjugate as an immunogen. The antibody belongs to the IgG1 subclass with a κ type light chain and has a dissociation constant of approximately 6.07 ×10^-9 M. MAb3E5D7C4B6 is very specific to MeJA. It was used to develop a direct competitive enzyme-linked immunosorbent assay （dcELISA）, conventional and simplified indirect competitive ELISAs （icELISA）. JA was derivatized into MeJA for the ELISA analysis. The IC50 value and detection range for MeJA were, respectively, 34 and 4-257 nglmL by the conventional icELISA, 21 and 3-226 nglmL by the simplified icELISA and 5.0 and 0.7-97.0 nglmL by the dcELISA. The dcELISA was more sensitive than either the conventional or simplified icELISA. The assays were used to measure the content of jasmonates as MeJA in tobacco leaves under drought stress or inoculated with tobacco mosaic virus and tomato leaves inoculated with tomato mosaic virus or Lirioinyza sativae Blanchard as compared with the corresponding healthy leaves. The increased jasmonates content indicated its role in response to the drought stress and pathogens.