Open AccessAn Immunological System for the Detection of Pepper mild mottle virus in Soil from Green Pepper Fields
Author(s) -
Y Ikegashira,
Takehiro Ohki,
U T Ichiki,
Tsuneyuki Higashi,
Kyoji Hagiwara,
Toshihiro Omura,
Y. Honda,
Shinya Tsuda
Publication year - 2004
Publication title -
plant disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.663
H-Index - 108
eISSN - 1943-7692
pISSN - 0191-2917
DOI - 10.1094/pdis.2004.88.6.650
Subject(s) - pepper , biology , inoculation , mottle , horticulture , virology , tobamovirus , polymerase chain reaction , virus , soil test , extraction (chemistry) , plant virus , soil water , veterinary medicine , agronomy , chromatography , chemistry , gene , medicine , ecology , biochemistry
A reliable method, based on the double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), was developed for the extraction of viruses from soil and optimized for the detection of Pepper mild mottle virus (PMMoV) in soil taken from green pepper (Capsicum annuum) fields. When added to phosphate buffer, Tween 20 increases extraction efficiency and skim milk increases the specificity for PMMoV. Samples positive by DAS-ELISA were verified by inhibition testing using specific anti-PMMoV antibody, immuno-electron microscopy, reverse transcription-polymerase chain reaction, and inoculation tests on assay plants. Our system for detecting PMMoV in soil was successfully tested on samples from 22 infected and uninfected fields in Japan. When used before seedlings are planted, this method allows for the prediction of possible damage to cultivated plants by soil-borne PMMoV.