title: Constitutive heterologous overexpression of a TIR-NB-ARC-LRR gene encoding a putative disease resistance protein from wild Chinese Vitis pseudoreticulata in Arabidopsis and tobacco enhances resistance to phytopathogenic fungi and bacteria.
abstract: Plants use resistance (R) proteins to detect pathogen effector proteins and activate their innate immune response against the pathogen. The majority of these proteins contain an NB-ARC (nucleotide-binding adaptor shared by APAF-1, R proteins, and CED-4) domain along with a leucine-rich repeat (LRR), and some also bear a toll interleukin 1 receptor (TIR) domain. In this study, we characterized a gene encoding a TIR-NB-ARC-LRR R protein (VpTNL1) (GenBank accession number KX649890) from wild Chinese grapevine Vitis pseudoreticulata accession "Baihe-35-1", which was identified previously from a transcriptomic analysis of leaves inoculated with powdery mildew (PM; Erysiphe necator (Schw.)). The VpTNL1 transcript was found to be highly induced in V. pseudoreticulata following inoculation with E. necator, as well as treatment with salicylic acid (SA). Sequence analysis demonstrated that the deduced amino acid sequence contained a TIR domain at the N-terminus, along with an NB-ARC and four LRRs domains within the C-terminus. Constitutive expression of VpTNL1 in Arabidopsis thaliana resulted in either a wild-type or dwarf phenotype. Intriguingly, the phenotypically normal transgenic lines displayed enhanced resistance to Arabidopsis PM, Golovinomyces cichoracearum, as well as to the virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Similarly, constitutive expression of VpTNL1 in Nicotiana tabacum was found to confer enhanced resistance to tobacco PM, Erysiphe cichoacearum DC. Subsequent isolation of the VpTNL1 promoter and deletion analysis indicated that TC-rich repeats and TCA elements likely play an important role in its response to E. necator and SA treatment, respectively. Taken together, these results indicate that VpTNL1 contributes to PM resistance in grapevine and provide an interesting gene target for the future amelioration of grape via breeding and/or biotechnology.
authors.affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China; College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China. Electronic address: zhifengwen@126.com.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Wen
authors.affiliation: Horticultural Plant Biology and Metabolomics Center, Haixia Institute of Science and Technology, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: lpy106080@163.com.
authors.forename: Liping
authors.initials: L
authors.lastname: Yao
authors.affiliation: Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta T6G 2P5, Canada. Electronic address: stacysinger@hotmail.com.
authors.forename: Stacy D
authors.initials: SD
authors.lastname: Singer
authors.affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China. Electronic address: mdhanif1716@yahoo.com.
authors.forename: Hanif
authors.initials: H
authors.lastname: Muhammad
authors.affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China. Electronic address: lizhi@nwsuaf.edu.cn.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China; Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China. Electronic address: wangxiping@nwsuaf.edu.cn.
authors.forename: Xiping
authors.initials: X
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000382:microbiology*; D001203:Ascomycota / Q000187:drug effects / Q000502:physiology*; D001483:Base Sequence; D003001:Cloning, Molecular; D060467:Disease Resistance / Q000187:drug effects / Q000235:genetics*; D017343:Genes, Plant; D000090263:Leucine-Rich Repeat Proteins; D010802:Phylogeny; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011549:Pseudomonas / Q000187:drug effects / Q000502:physiology*; D020156:Salicylic Acid / Q000494:pharmacology; D016415:Sequence Alignment; D014026:Nicotiana / Q000187:drug effects / Q000235:genetics / Q000382:microbiology*; D027843:Vitis / Q000235:genetics*
chemical_list: D000090263:Leucine-Rich Repeat Proteins; D010940:Plant Proteins; D011506:Proteins; D020156:Salicylic Acid
keywords: Disease resistance; Powdery mildew; Promoter analysis; VpTNL1; Wild Chinese vitis