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Characterization of necrosis-inducing NLP proteins in Phytophthora capsici.
BMC Plant Biol. 2014 May 08; 14:126.BP

Abstract

BACKGROUND

Effector 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.

RESULTS

We 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.

CONCLUSIONS

Each 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.

Authors+Show Affiliations

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableDepartment of Plant Pathology, Shandong Agricultural University, 61, Daizong Street, Tai'an, Shandong 271018, China. zhxg@sdau.edu.cn.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24886309

Citation

Feng, Bao-Zhen, et al. "Characterization of Necrosis-inducing NLP Proteins in Phytophthora Capsici." BMC Plant Biology, vol. 14, 2014, p. 126.
Feng BZ, Zhu XP, Fu L, et al. Characterization of necrosis-inducing NLP proteins in Phytophthora capsici. BMC Plant Biol. 2014;14:126.
Feng, B. Z., Zhu, X. P., Fu, L., Lv, R. F., Storey, D., Tooley, P., & Zhang, X. G. (2014). Characterization of necrosis-inducing NLP proteins in Phytophthora capsici. BMC Plant Biology, 14, 126. https://doi.org/10.1186/1471-2229-14-126
Feng BZ, et al. Characterization of Necrosis-inducing NLP Proteins in Phytophthora Capsici. BMC Plant Biol. 2014 May 8;14:126. PubMed PMID: 24886309.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of necrosis-inducing NLP proteins in Phytophthora capsici. AU - Feng,Bao-Zhen, AU - Zhu,Xiao-Ping, AU - Fu,Li, AU - Lv,Rong-Fei, AU - Storey,Dylan, AU - Tooley,Paul, AU - Zhang,Xiu-Guo, Y1 - 2014/05/08/ PY - 2013/06/01/received PY - 2014/05/02/accepted PY - 2014/6/3/entrez PY - 2014/6/3/pubmed PY - 2015/1/17/medline SP - 126 EP - 126 JF - BMC plant biology JO - BMC Plant Biol. VL - 14 N2 - BACKGROUND: Effector 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. RESULTS: We 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. CONCLUSIONS: Each 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. SN - 1471-2229 UR - https://www.unboundmedicine.com/medline/citation/24886309/Characterization_of_necrosis_inducing_NLP_proteins_in_Phytophthora_capsici_ L2 - https://bmcplantbiol.biomedcentral.com/articles/10.1186/1471-2229-14-126 DB - PRIME DP - Unbound Medicine ER -