GbWRKY1, a novel cotton (Gossypium barbadense) WRKY gene isolated from a bacteriophage full-length cDNA library, is induced by infection with Verticillium dahliae.
WRKY transcription factor proteins play important roles in diverse stress responses. In this study, we first cloned a novel WRKY from our constructed bacteriophage full-length cDNA library for cotton (Gossypium barbadense). The plants were stressed by exposure to a defoliating strain of Verticillium dahliae. The capacity of primary cDNA library was 1.28 x 10(6) PFU and the titer of the amplified cDNA library was > 10(10) PFU mL(-1). The recombination rate of the library was 94% and average insert size was about 1.1 kb. This novel gene, named GbWRKY1 was 1971 bp long and encodes a protein of 489 amino acids. It contains two characteristic WRKY domains and two zinc finger motifs. The sub-cellular assay indicated that GbWRKY1-GFP fusion protein was localized in the nucleus. Furthermore, Northern blot analysis showed that expression pattern of GbWRKY1 was similar among tissue types (roots, stems and leaves), but differed between pathogen-infiltrated and Czapek medium-infiltrated (untreated control) plants. Quantitative real-time PCR showed that GbWRKY1 could also be induced by salicylic acid (SA), methyl jasmonate (MeJA) and 1-aminocyclopropane-1-carboxylic acid (ACC). These findings clearly suggest that as a pathogen-inducible transcription factor GbWRKY1 plays an important role in plant defense responses.
North China Key Laboratory for Crop Germplasm Resources, Education Ministry of China, Agricultural University of Hebei, Baoding 071001, China.
Zhang
Z
Shu-Ling
Wang
W
Xing-Fen
Zhang
Z
Yan
Liu
L
Jian-Feng
Wu
W
Li-Zhu
Zhang
Z
Dong-Mei
Ma
M
Zhi-Ying
D000595:Amino Acid Sequence; D000598:Amino Acids, Cyclic / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D001435:Bacteriophages / Q000235:genetics*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D017343:Genes, Plant / Q000187:drug effects / Q000502:physiology*; D003368:Gossypium / Q000235:genetics* / Q000382:microbiology; D008969:Molecular Sequence Data; D018696:Neuroprotective Agents / Q000494:pharmacology; D010802:Phylogeny; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020156:Salicylic Acid / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D020172:Verticillium / Q000187:drug effects / Q000502:physiology*; D016335:Zinc Fingers
D000598:Amino Acids, Cyclic; D000935:Antifungal Agents; D018696:Neuroprotective Agents; D012333:RNA, Messenger; C023863:1-aminocyclopropane-1-carboxylic acid; D020156:Salicylic Acid