Functional analysis of the tomato Ve resistance locus against Verticillium wilt

E.F. Fradin

Research output: Thesisinternal PhD, WU

Abstract

Verticillium dahliae, V. albo-atrum and V. longisporum are soil-borne plant pathogens that are responsible for Verticillium wilt diseases in temperate and subtropical regions. Collectively they can infect over 200 hosts, including many economically important crops. Chapter 1 is a “pathogen profile” which describes the most important aspects of the biology of the Verticillium wilt pathogens. They colonize the xylem vessels of their host plants and cause symptoms such as wilting, chlorosis, stunting, necrosis and vein clearing. Verticillium species are notoriously difficult to control as there are no fungicides available to cure plants once they are infected. Therefore, genetic resistance is the preferred method for disease control.

Chapter 2 describes the functional characterization of the tomato (Solanum lycopersicum) Ve locus. This locus is responsible for resistance against race 1 strains of V. dahliae and V. albo-atrum and comprises two closely linked inversely oriented genes, Ve1 and Ve2. Both genes encode cell surface receptor proteins of the extracellular leucine-rich repeat (eLRR) receptor-like protein (RLP) class of disease resistance proteins. In chapter 2, it is demonstrated that Ve1, but not Ve2, provides resistance in tomato against race 1 but not against race 2 strains of V. dahliae and V. albo-atrum. Using virus-induced gene silencing in tomato, the signaling cascade downstream of Ve1 was shown to require both EDS1 (enhanced disease susceptibility1) and NDR1 (non-race-specific disease resistance1). In addition, also NRC1 (NB-LRR protein required for hypersensitive response-associated cell death1), ACIF (Avr9/Cf-9–induced F-box1), MEK2 (MAP/ERK kinase2), and SERK3/BAK1 (somatic embryogenesis receptor kinase 3/brassinosteroid-associated kinase 1) act as positive regulators of Ve1 in tomato. In conclusion, Ve1-mediated resistance signaling only partially overlaps with signaling mediated by Cf proteins, type members of the eLRR-RLP-class of resistance proteins.

In chapter 3 an attempt to introduce Nicotiana benthamiana as a model to facilitate the study of Ve1-mediated resistance is described. Challenge of wild type plants with several race 1 and race 2 strains of V. dahliae and V. albo-atrum demonstrated that N. benthamiana is susceptible to both Verticillium species. To obtain Verticillium wilt resistant plants, N. benthamiana was engineered to express the tomato Ve1 coding sequence. However, out of thirteen transgenic lines, six showed clear phenotypic aberrancies that included severe stunting and malformed leaves when compared to wild type plants. The seven Ve1-transgenic lines that did not show any phenotypic alterations were challenged with race 1 and race 2 strains. Although the pathogenicity assays indicated that in few lines Ve1 expression temporarily reduced disease development, most lines were as susceptible as wild type parental line. In conclusion, in chapter 3 it is demonstrated that the Ve1-transgenic N. benthamiana lines could not be used to study Ve1-mediated resistance signaling.

In chapter 4, the use of Arabidopsis (Arabidopsis thaliana) as model to facilitate the study of Ve1-mediated resistance is presented. To this end, tomato Ve1 was expressed in susceptible Arabidopsis plants. Upon challenge with race 1 strains of V. dahliae or V. albo-atrum, Ve1-expressing plants were found to be resistant. In contrast, Ve1-expressing plants were susceptible to race 2 strains of both V. dahliae and V. albo-atrum. Furthermore, expression of Ve1 in Arabidopsis plants did not prevent colonization by V. longisporum strains. Through Ve1-expression in Arabidopsis defense signaling mutants, it was demonstrated that signaling downstream of Ve1 is highly conserved between tomato and Arabidopsis.

In previous chapters it was shown that the receptor kinase SERK3/BAK1 is required for Ve1-mediated resistance in tomato as well as in Arabidopsis. In Arabidopsis, SERK3/BAK1 belongs to a gene family consisting of five members. In chapter 5, the requirement of the different SERK family members in Ve1-mediated resistance in Arabidopsis is investigated, revealing the requirement of SERK1 and, although to a lesser extent, SERK4 for resistance. Using virus-induced gene silencing, it was subsequently shown that SERK1 is also required for Ve1-mediated resistance in tomato. In conclusion, the results of chapter 5 demonstrate that Arabidopsis can be used as model to unravel the molecular mechanisms of Ve1-mediated resistance.

In chapter 6, the recognition specificity of Ve1 was further investigated by performing domain-swaps with Ve2 and expressing the chimeric Ve proteins in Arabidopsis. Various domain swaps in which eLRRs from Ve1 were replaced by those of Ve2 suggest that the region between eLRR22 and eLRR35 is required for full Ve1-mediated resistance. However, plants expressing a Ve chimera in which eLRR1 to eLRR30 of Ve1 was replaced with those of Ve2 were resistant against Verticillium. Overall, these results suggest that Ve2 may still bind the elicitor in the eLRR domain, but its C-terminal domain is not able to activate a successful defense response.

Finally in Chapter 7, highlights of this thesis are discussed and placed in a broader perspective. 

Original languageEnglish
QualificationDoctor of Philosophy
Awarding Institution
  • Wageningen University
Supervisors/Advisors
  • de Wit, Pierre, Promotor
  • Thomma, Bart, Co-promotor
Award date19 Sep 2011
Place of Publication[s.l.]
Print ISBNs9789085859611
Publication statusPublished - 2011

Keywords

  • verticillium dahliae
  • verticillium albo-atrum
  • solanum lycopersicum
  • tomatoes
  • disease resistance
  • genes
  • resistance breeding
  • functional analysis
  • vertical resistance
  • wilts

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