title: Molecular cloning and functional analysis of GbRVd, a gene in Gossypium barbadense that plays an important role in conferring resistance to Verticillium wilt.
abstract: Most of the disease resistance genes already characterized in plants encode nucleotide-binding site-leucine rich repeat (NBS-LRR) proteins that have key roles in resistance to Verticillium dahliae. Using a cDNA library and RACE protocols, we cloned a coiled-coil (CC)-NBS-LRR-type gene, GbRVd, from a resistant tetraploid cotton species, Gossypium barbadense (RVd=Resistance to V. dahliae). We also applied RT-qPCR and VIGS technologies to analyze how expression of GbRVd was induced upon attack by V. dahliae. Its 2862-bp ORF encodes a predicted protein containing 953 amino acid residues, with a predicted molecular weight of 110.17kDa and an isoelectric point of 5.87. GbRVd has three domains - CC, NBS, and LRR - and is most closely related to Gossypium raimondii RVd (88% amino acid identity). Profiling demonstrated that GbRVd is constitutively expressed in all tested tissues, and transcript levels are especially high in the leaves. In plants inoculated with V. dahliae, GbRVd was significantly up-regulated when compared with the control, with expression peaking at 48h post-inoculation. Silencing of GbRVd in cotton through VIGS dramatically down-regulated SA, NO, and H2O2 production, resulting in greater susceptibility to V. dahliae. Taken together, these results suggest that GbRVd has an important role in protecting G. barbadense against infection by V. dahliae.
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Ma
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China.
authors.forename: Xingfen
authors.initials: X
authors.lastname: Wang
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China.
authors.forename: Guiyin
authors.initials: G
authors.lastname: Zhang
authors.affiliation: North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding 071001, China. Electronic address: mzhy@hebau.edu.cn.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Ma
mesh_terms: D003001:Cloning, Molecular; D060467:Disease Resistance; D018506:Gene Expression Regulation, Plant; D003368:Gossypium / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots; D014018:Tissue Distribution; D020172:Verticillium / Q000502:physiology*
chemical_list: D010940:Plant Proteins; D009569:Nitric Oxide; D006861:Hydrogen Peroxide
keywords: GbRVd; Gossypium barbadense; Resistance; Verticillium wilt; Virus-induced gene silence