Procedure for detecting tobamovirus in tomato and pepper seeds decreases the cost analysis

João Eduardo Melo Almeida,Mauricio Antônio Lucas,Priscilla De Sousa Geraldino Duarte,Nara Edreira Alencar,Antonia Dos Reis Figueira

doi:10.1590/1678-4499.2017317

João Eduardo Melo Almeida, Mauricio Antônio Lucas + Show 3 more

Open Access

https://doi.org/10.1590/1678-4499.2017317

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Journal: Bragantia	Publication Date: Oct 18, 2018
Citations: 4	License type: cc-by

Affiliation: Universidade Federal de Lavras

Abstract

ABSTRACT The transmission of tobamovirus by tomato and pepper seeds is an important mean of virus introduction in crops. Therefore, detecting its presence in the seed becomes essential for the preventive control of virus diseases. In this study, a method was proposed for the detection of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in tomatoes (Solanum lycopersicum) and Pepper mild mottle virus (PMMoV) in pepper (Capsicum annum) seeds. Seed lots with different levels of incidence were analyzed by biological, serological, and molecular methods. Using DAS-ELISA technique, it was possible to detect TMV up to the limit rate of 1:170 (1 contaminated seed: total seeds) and the ToMV up to 1:200 in tomato seeds; PMMoV in pepper seeds was detected up to 1:140. The IC-RT-PCR detected the TMV and ToMV up to the limit of 1:400 and PMMoV up to the limit of 1:300. The assembled lots containing only 1 contaminated seed in 1000 (1:1000) were combined into 30 sub lots for DAS-ELISA analysis and 10 sub lots for IC-RT-PCR analysis. Both techniques, DAS-ELISA and IC-RT-PCR, were efficient to detect the three viruses in all analyzed samples, but the detection of tobamoviruses with RT-PCR and biological tests was not reliable. Based on the results of this study, in which a combination of seeds in sub lots was made to reduce the number of tests performed, it is possible to make significant savings in the cost of the diagnostic methods routinely conducted in official laboratories, with high efficiency and reliability.

Highlights

The virus species of Tobamovirus genus have worldwide distribution and can occur in different environments, in tropical as well as in temperate climates
In Brazil the most frequently found of tobamoviruses species are Tobacco mosaic virus (TMV), type species of the genus; Tomato mosaic virus (ToMV), which is present in solanaceous crops, including vegetable and ornamentals plants; and Pepper mild mottle virus (PMMoV CP) in pepper (Moreira et al 2003; Eiras et al 2004; Duarte et al 2007; Cezar et al 2009; Silva et al 2008; 2011)
Santa Clara inoculated with ToMV, and pepper cultivar Ikeda inoculated with PMMoV showed typical mosaic symptoms on leaves and fruits and produced 100% of contaminated seeds, as confirmed by DAS-ELISA

Summary

Introduction

The virus species of Tobamovirus genus have worldwide distribution and can occur in different environments, in tropical as well as in temperate climates. The tobamoviruses have no vectors in nature; because of their high stability, they are transmitted mechanically during the cultivation process, and can remain viable in soil for long periods in crop residues (Gülser et al 2008; Candemir et al 2012). They are capable of infecting newly established plants through possible roots injuries, which are common during root growth and expansion in the soil. The majority of tobamoviruses do not infect the embryo or endosperm, viral particles remain externally attached to the integument and can infect plants during the germination process (Chitra et al 1999; Genda et al 2011; Sevik and Kose-Tohumcu 2011; Liu et al 2014)

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