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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.2019.tde-20191220-121007
Document
Author
Full name
Keila Maria Roncato Duarte
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2000
Supervisor
Title in Portuguese
Anticorpos monoclonais e policlonais para identificação do tobamovírus do mosaico do tomateiro (ToMV) em plantas de Tomate (Lycopersicon esculentum Mill.)
Keywords in Portuguese
TOBAMOVIRUS
ANTICORPOS
IDENTIFICAÇÃO
MOSAICO-DO-TOMATEIRO
TOMATE
Abstract in Portuguese
O tomateiro (Lycopersicon esculentum Mill.) é uma olerícola de grande importância econômica e uma das mais suscetíveis a viroses, dentre as quais, o tobamovírus do mosaico do tomateiro (ToMV), que causa mosaicos verde claro-escuro nas folhas, afilamento de folíolos, diminuição da produção, entre outros sintomas. Visando a identificação precoce do ToMV, foram produzidos anticorpos monoclonais (MAbs) e anticorpos policlonais. Dentre os MAbs produzidos, escolheu-se aquele que apresentou melhor título em PTA- ELISA ("plate trapped antigen- enzyme linked immunoassay") sem reação cruzada com tobamovírus do mosaico do tabaco (TMV) e com proteínas do extrato da planta. Este MAb (10.Hl), juntamente com os anticorpos policlonais, foram utilizados para avaliar a capacidade de identificação do ToMV em testes de campo com plantas de tomate infectadas. Destas plantas foi isolado o ToMV, que foi purificado e re-inoculado em plantas de tomateiro e de tabaco, para confirmação dos sintomas. Em imuno blot os anticorpos policlonais permitiram reconhecer duas bandas protéicas (de 17,5 kDA e de 30 kDa) enquanto que o MAb 10.Hl reconheceu somente a proteína referente à capa protéica do ToMV (de 17,5 kDa). A maior especificidade do MAb 10.Hl pode permitir o diagnóstico precoce desta doença na fase de plântulas, ainda em casa de vegetação, evitando assim a disseminação desta virose no campo.
Title in English
Monoclonal and polyclonal antibodies to identify tomato mosaic tobamovirus (ToMV) in tomato plants (Lycopersicon esculentum Mill.)
Abstract in English
The tomato crops are one of the most important vegetables for the world economy and it is very susceptible to vírus diseases, among them the tomato mosaic tobamovirus (ToMV) that causes light and dark green mosaic in the leaves, decreases the production, among other symptoms. With the aim to early identify ToMV in biological material, harvested from crop fields, monoclonal antibodies (MAbs) and polyclonal antibodies were produced against ToMV. The MAb 10.Hl was chosen by PTA-ELISA (Plate Trapped Antigen-Enzyme-Linked immunoassay) and do not crossreact with TMV (tobacco mosaic tobamovirus) or with proteins extracted from plant sap. The MAb and the polyclonal antibodies produced were able to identify ToMV from infected plants collected in field. These plants were used to isolate the virus, which were purified and used to re-inoculate tobacco and tomato plants to confirm the symptoms. ln immunoblots assays the polyclonal antibodies recognize two protein bands (17.5 kDA and the 30 kDa bands) and the MAb recognizes only the band corresponding to the coat protein of the ToMV (17.5 kDa). The MAb 10.Hl opens the possibility to identify ToMV in tomato seedlings avoiding its dissemination in the crop fields.
 
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Publishing Date
2019-12-20
 
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