Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein

Godwill M. Chewachong, Sally A. Miller, Joshua J. Blakeslee, David M. Francis, T. Jack Morris, Feng Qu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Mild variants of many viruses are able to protect infected plants from subsequent invasion by more severe variants of the same viruses through a process known as cross-protection. In the past, the cross-protective viral variants were commonly derived from mild field isolates that were sometimes genetically heterogeneous, providing variable levels of crossprotection. Here, we report a novel approach to rapidly generate crossprotective variants of the tomato-infecting Pepino mosaic virus (PepMV) independently of the availability of mild field isolates. Our approach sought to attenuate PepMV by mutating less conserved amino acid residues of the abundantly produced capsid protein (CP). These less-conserved amino acid residues were identified through multiple alignments of CPs of six potexviruses including PepMV, and were altered systematically to yield six PepMV mutants. These mutants were subsequently inoculated onto the model plant Nicotiana benthamiana, as well as tomato, to evaluate their accumulation levels, symptom severities, and cross-protection potentials. The mutant KD, in which the threonine (T) and alanine (A) residues at CP positions 66 and 67 were replaced with lysine (K) and aspartic acid (D), respectively, were found to accumulate to low levels in infected plants, cause very mild symptoms, and effectively protect both N. benthamiana and tomato against secondary infections by wild-type PepMV. These data suggest that our approach represents a simple, fast, and reliable way of generating attenuated viral variants capable of cross-protection.

Original languageEnglish (US)
Pages (from-to)126-134
Number of pages9
JournalPhytopathology
Volume105
Issue number1
DOIs
StatePublished - Jan 1 2015

Fingerprint

Pepino mosaic virus
viral proteins
coat proteins
mutagenesis
Nicotiana benthamiana
tomatoes
mutants
signs and symptoms (plants)
Potexvirus
viruses
amino acids
aspartic acid
threonine
alanine
lysine

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein. / Chewachong, Godwill M.; Miller, Sally A.; Blakeslee, Joshua J.; Francis, David M.; Morris, T. Jack; Qu, Feng.

In: Phytopathology, Vol. 105, No. 1, 01.01.2015, p. 126-134.

Research output: Contribution to journalArticle

Chewachong, Godwill M. ; Miller, Sally A. ; Blakeslee, Joshua J. ; Francis, David M. ; Morris, T. Jack ; Qu, Feng. / Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein. In: Phytopathology. 2015 ; Vol. 105, No. 1. pp. 126-134.
@article{b0b6c63764ba4ab8be5d34333ea3ac76,
title = "Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein",
abstract = "Mild variants of many viruses are able to protect infected plants from subsequent invasion by more severe variants of the same viruses through a process known as cross-protection. In the past, the cross-protective viral variants were commonly derived from mild field isolates that were sometimes genetically heterogeneous, providing variable levels of crossprotection. Here, we report a novel approach to rapidly generate crossprotective variants of the tomato-infecting Pepino mosaic virus (PepMV) independently of the availability of mild field isolates. Our approach sought to attenuate PepMV by mutating less conserved amino acid residues of the abundantly produced capsid protein (CP). These less-conserved amino acid residues were identified through multiple alignments of CPs of six potexviruses including PepMV, and were altered systematically to yield six PepMV mutants. These mutants were subsequently inoculated onto the model plant Nicotiana benthamiana, as well as tomato, to evaluate their accumulation levels, symptom severities, and cross-protection potentials. The mutant KD, in which the threonine (T) and alanine (A) residues at CP positions 66 and 67 were replaced with lysine (K) and aspartic acid (D), respectively, were found to accumulate to low levels in infected plants, cause very mild symptoms, and effectively protect both N. benthamiana and tomato against secondary infections by wild-type PepMV. These data suggest that our approach represents a simple, fast, and reliable way of generating attenuated viral variants capable of cross-protection.",
author = "Chewachong, {Godwill M.} and Miller, {Sally A.} and Blakeslee, {Joshua J.} and Francis, {David M.} and Morris, {T. Jack} and Feng Qu",
year = "2015",
month = "1",
day = "1",
doi = "10.1094/PHYTO-01-14-0018-R",
language = "English (US)",
volume = "105",
pages = "126--134",
journal = "Phytopathology",
issn = "0031-949X",
publisher = "American Phytopathological Society",
number = "1",

}

TY - JOUR

T1 - Generation of an attenuated, cross-protective Pepino mosaic virus variant through alignment-guided mutagenesis of the viral capsid protein

AU - Chewachong, Godwill M.

AU - Miller, Sally A.

AU - Blakeslee, Joshua J.

AU - Francis, David M.

AU - Morris, T. Jack

AU - Qu, Feng

PY - 2015/1/1

Y1 - 2015/1/1

N2 - Mild variants of many viruses are able to protect infected plants from subsequent invasion by more severe variants of the same viruses through a process known as cross-protection. In the past, the cross-protective viral variants were commonly derived from mild field isolates that were sometimes genetically heterogeneous, providing variable levels of crossprotection. Here, we report a novel approach to rapidly generate crossprotective variants of the tomato-infecting Pepino mosaic virus (PepMV) independently of the availability of mild field isolates. Our approach sought to attenuate PepMV by mutating less conserved amino acid residues of the abundantly produced capsid protein (CP). These less-conserved amino acid residues were identified through multiple alignments of CPs of six potexviruses including PepMV, and were altered systematically to yield six PepMV mutants. These mutants were subsequently inoculated onto the model plant Nicotiana benthamiana, as well as tomato, to evaluate their accumulation levels, symptom severities, and cross-protection potentials. The mutant KD, in which the threonine (T) and alanine (A) residues at CP positions 66 and 67 were replaced with lysine (K) and aspartic acid (D), respectively, were found to accumulate to low levels in infected plants, cause very mild symptoms, and effectively protect both N. benthamiana and tomato against secondary infections by wild-type PepMV. These data suggest that our approach represents a simple, fast, and reliable way of generating attenuated viral variants capable of cross-protection.

AB - Mild variants of many viruses are able to protect infected plants from subsequent invasion by more severe variants of the same viruses through a process known as cross-protection. In the past, the cross-protective viral variants were commonly derived from mild field isolates that were sometimes genetically heterogeneous, providing variable levels of crossprotection. Here, we report a novel approach to rapidly generate crossprotective variants of the tomato-infecting Pepino mosaic virus (PepMV) independently of the availability of mild field isolates. Our approach sought to attenuate PepMV by mutating less conserved amino acid residues of the abundantly produced capsid protein (CP). These less-conserved amino acid residues were identified through multiple alignments of CPs of six potexviruses including PepMV, and were altered systematically to yield six PepMV mutants. These mutants were subsequently inoculated onto the model plant Nicotiana benthamiana, as well as tomato, to evaluate their accumulation levels, symptom severities, and cross-protection potentials. The mutant KD, in which the threonine (T) and alanine (A) residues at CP positions 66 and 67 were replaced with lysine (K) and aspartic acid (D), respectively, were found to accumulate to low levels in infected plants, cause very mild symptoms, and effectively protect both N. benthamiana and tomato against secondary infections by wild-type PepMV. These data suggest that our approach represents a simple, fast, and reliable way of generating attenuated viral variants capable of cross-protection.

UR - http://www.scopus.com/inward/record.url?scp=84919372268&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84919372268&partnerID=8YFLogxK

U2 - 10.1094/PHYTO-01-14-0018-R

DO - 10.1094/PHYTO-01-14-0018-R

M3 - Article

C2 - 25496364

AN - SCOPUS:84919372268

VL - 105

SP - 126

EP - 134

JO - Phytopathology

JF - Phytopathology

SN - 0031-949X

IS - 1

ER -