Functional analysis and mode of action of phytotoxic Nep1-like proteins of Botrytis cinerea

Arenas, Y. Cuesta; Kalkman, Eric R.; Schouten, Alexander; Dieho, Mirjam; Vredenbregt, P.; Uwumukiza, B.; Ruiz, Miriam Osés; Kan, Jan A.L. van

doi:10.1016/j.pmpp.2010.06.003

Cited by 71 publications

(47 citation statements)

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“…This suggests that individuals PcNLP may have an effect on its ability to establish infection on plant. Several studies have considered the function of NLP genes, and most conclude that several NLPs are indispensable for fungal infection [33,65]. Our study concluded that PcNLP 2, PcNLP 6, and PcNLP 14 contribute greatly to the induction of necrosis during infection by P. capsici , and suggested that the simultaneous presence of PcNLP 2, PcNLP 6, and PcNLP 14 may be required for a complete necrotic response.…”

Section: Discussionmentioning

confidence: 56%

“…On the other hand, the NLP genes in pathogens such as F. oxysporum f. sp. erythroxyli and Mycosphaerella graminicola do not appear to affect their virulence [31,32], and the NLPs, Bcnep 1 and Bcnep 2 are apparently not related to virulence during Botrytis cinerea infection [33]. Thus, the members of the NLP families from different pathogens encode functionally different phytotoxic proteins that appear to perform a variety of functions during infection and produce variable extended phenotypes.…”

Section: Discussionmentioning

confidence: 99%

“…erythroxyli and Mycosphaerella graminicola does not reduce their virulence [31,32]. Similarly, mutants of Bcnep 1 or Bcnep 2 in pathogenic strains of Botrytis cinerea result in virulence similar to that of the wild type strains [33]. However, there is strong evidence that NLPs function as virulence factors that accelerate disease and pathogen growth in host plants.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of necrosis-inducing NLP proteins in Phytophthora capsici

Feng

Zhu

et al. 2014

BMC Plant Biol

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BackgroundEffector proteins function not only as toxins to induce plant cell death, but also enable pathogens to suppress or evade plant defense responses. NLP-like proteins are considered to be effector proteins, and they have been isolated from bacteria, fungi, and oomycete plant pathogens. There is increasing evidence that NLPs have the ability to induce cell death and ethylene accumulation in plants.ResultsWe evaluated the expression patterns of 11 targeted PcNLP genes by qRT-PCR at different time points after infection by P. capsici. Several PcNLP genes were strongly expressed at the early stages in the infection process, but the expression of other PcNLP genes gradually increased to a maximum at late stages of infection. The genes PcNLP2, PcNLP6 and PcNLP14 showed the highest expression levels during infection by P. capsici. The necrosis-inducing activity of all targeted PcNLP genes was evaluated using heterologous expression by PVX agroinfection of Capsicum annuum and Nicotiana benthamiana and by Western blot analysis. The members of the PcNLP family can induce chlorosis or necrosis during infection of pepper and tobacco leaves, but the chlorotic or necrotic response caused by PcNLP genes was stronger in pepper leaves than in tobacco leaves. Moreover, PcNLP2, PcNLP6, and PcNLP14 caused the largest chlorotic or necrotic areas in both host plants, indicating that these three genes contribute to strong virulence during infection by P. capsici. This was confirmed through functional evaluation of their silenced transformants. In addition, we further verified that four conserved residues are putatively active sites in PcNLP1 by site-directed mutagenesis.ConclusionsEach targeted PcNLP gene affects cells or tissues differently depending upon the stage of infection. Most PcNLP genes could trigger necrotic or chlorotic responses when expressed in the host C. annuum and the non-host N. benthamiana. Individual PcNLP genes have different phytotoxic effects, and PcNLP2, PcNLP6, and PcNLP14 may play important roles in symptom development and may be crucial for virulence, necrosis-inducing activity, or cell death during infection by P. capsici.

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Section: Discussionmentioning

confidence: 56%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of necrosis-inducing NLP proteins in Phytophthora capsici

Feng

Zhu

et al. 2014

BMC Plant Biol

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“…carotovora is required for full virulence of bacteria on potato tuber but not in potato stem or on other tested host plants 14 . Moreover, other studies show that deletion of a member of NLP family in B. cinerea or F. oxysporum does not impair the fungal virulence on tomato, N. benthamiana 23 , or coca plants 29 , suggesting that NLPs may be dispensable for the virulence of pathogens. It is also possible that in these studies additional NLP genes may compensate the loss of a member of the family, which may mask the potential phenotype of mutant strains.…”

Section: Discussionmentioning

confidence: 98%

“…The up-regulation of NLP gene expression during infection has been described in other plant-pathogen interactions as well. In necrotrophic fungal pathogen Botrytis cinerea , the induction of BcNep1 and BcNep2 is at early and late stage of infection, respectively 23 . In addition, transcript levels of most of the expressed PsNLPs in P. sojae 12, 24 and two VdNLPs in V. dahliae 13 are highly induced at the late stage of infection; in contrast, the induction of four HaNLPs in the obligate biotroph oomycete Hyaloperonospora arabidopsidis occurs at the early stage of infection 16 .…”

Section: Discussionmentioning

confidence: 99%

The Nep1-like protein family of Magnaporthe oryzae is dispensable for the infection of rice plants

Fang

Peng

Fan

2017

Sci Rep

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The necrosis- and ethylene-inducing protein 1 (Nep1)-like proteins (NLPs) are a class of microbe-associated molecular patterns widely distributed across diverse groups of plant-associated microorganisms. In spite of the cytotoxic activity in dicot plants, the role of most NLPs in the virulence of plant pathogens is still largely unknown. We showed that the MoNLP family of rice blast fungus varied very little in amino acid sequence, transient expression of three MoNLPs induced cell death and the production of reactive oxygen species in Nicotiana benthamiana, and the expression of MoNLPs was induced during infection of susceptible rice plants. To further investigate the biological role of the MoNLP family, a marker-free gene replacement vector was developed and used to knock out the whole family in Magnaporthe oryzae. Results showed no significant difference in disease levels caused by wild type and the quadruple ΔMoNLP mutant strains. Likewise, the sporulation and radial growth of the two strains were similar under various unfavorable cultural conditions including malnutrition and abiotic stresses. These observations demonstrated that the MoNLP family is dispensable for the fungal tolerance to the tested adverse cultural conditions, and more importantly, for the virulence of blast fungus on susceptible rice plants.

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