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Identification of the genetic determinants of tobacco etch virus responsible for the wilting response of tabasco pepper
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Identification of the genetic determinants of tobacco etch virus responsible for the wilting response of tabasco pepper
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Identification of the genetic determinants of tobacco etch virus responsible for the wilting response of tabasco pepper
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Dissertation
Identification of the genetic determinants of tobacco etch virus responsible for the wilting response of tabasco pepper
1995
Overview
Infection of Tabasco pepper (Capsicum frutescens L. cv. Tabasco) by tobacco etch virus (TEV) typically causes wilting due to root necrosis. A recently isolated strain of TEV, designated TEV non-wilting (TEV NW), causes mosaic symptoms but not wilting in Tabasco pepper. It also induces larger local lesions on Chenopodium amaranticolor than the type strain of TEV, TEV HAT. In order to find the mutations responsible for the inability of TEV NW to cause wilting, the complete nucleotide sequence of TEV NW was determined. TEV NW has 9496 nucleotides and a polyadenylated tail; translation of a single putative polyprotein can be deduced. Compared to the published sequence of TEV HAT, there are 767 nucleotide and 126 amino acid changes scattered throughout the TEV NW genome. By using the available TEV NW sequence and its derived restriction map, a TEV NW infectious clone, pTEVNW, was constructed based on the TEV HAT infectious clone, pTEV7DA. The progeny virus generated from pTEVNW caused mosaic but not wilting symptoms on Tabasco pepper, and produced the same size local lesions on C. amaranticolor as TEV NW. The location of the wilting determinants was pursued by exchanging DNA fragments between pTEVNW and pTEV7DA. Substitutions of the individual P1, HC, P3, NIb and CP coding regions of TEV NW into TEV HAT did not alter the wilting response. When the pTEV7DA EcoR I (4264)-BamH I (5916) or Eco47 III (4967)-BamH I (5916) fragments, containing the C-terminal portion of CI and the entire 6-kDa coding sequence, were inserted into pTEVNW, the resulting chimeric viruses elicited wilting in Tabasco pepper. Therefore, the EcoR I (4264)-BamH I (5916) region contains sufficient sequence for eliciting the wilting response. With further exchange experiments, another wilting determinant was located within the EcoR I fragment (2383-4264) of TEV HAT. This fragment encompasses the entire P3 and the N-terminal CI coding sequence.
