Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (17): 3259-3273.doi: 10.3864/j.issn.0578-1752.2017.17.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Genome-Wide Identification and Expression Analysis of WRKY Transcription Factor Under Abiotic Stress in Beta vulgaris

KONG WeiLong1, YU Kun2, DAN NaiZhen1, YANG ShaoZong1, Bao ManZhu1, HUANG XiangRong1, FU XiaoPeng1   

  1. 1College of Horticulture and Forestry Sciences, Huazhong Agricultural University/Key Laboratory of Horticultural Plant Biology, Ministry of Education, Wuhan 430070; 2College of Agronomy, Shihezi University/Key Laboratory of Oasis Ecological Agriculture, Xinjiang Production and Construction Group, Shihezi 832003, Xinjiang
  • Received:2017-02-23 Online:2017-09-01 Published:2017-09-01

Abstract: 【Objective】 WRKY transcription factor is a kind of transcription factor which plays an important role in plant response to biotic, abiotic stress, plant growth and development. Based on the sugar beet genome information, the WRKY family genes (BvWRKYs) were identified, tissue-specific expression profiles and expression pattern under salt and heat stresses were analyzed. The aim of the study is to provide reference for the WRKY gene function research, and lay a foundation for ornamental beet (B. vulgaris) and other Dianthus ornamental plant genetic engineering research. 【Method】 A total of 75 Arabidopsis WRKY proteins were used as references, according to WRKY conserved protein sequence (PF03106), the WRKY genes of B. vulgaris were identified by hmm and BLAST homology searches, and the chromosome location, phylogeny, gene structure, conserved domain, conserved element were also analyzed by MapInspect, GSDS2.0, MEGA5.0, DNAMAN5.0, WebLogo 3 and MEME bioinformatics tools. The specificity of WRKY genes expression and expression pattern under salt stress and heat stress of B. vulgaris were analyzed by RNA-seq and qRT-PCR analysis. 【Result】 B. vulgaris WRKY gene family contained 40 members, 39 were unevenly distributed on 9 chromosomes, 1 on the random fragment. According to the WRKY conserved domain features and the evolution analysis with Arabidopsis thaliana, 40 members were divided into three classes: Ⅰ, Ⅱ, and Ⅲ. ClassⅠhad 9 members, class Ⅱ had 26 members and class Ⅲ had 5 members. According to the evolutionary relationship, class Ⅱ further divided into five subclass: Ⅱa (1), Ⅱb (4), Ⅱc (9), Ⅱd (5) and Ⅱe (7). Genetic structure analysis showed that exon and intron number of B. vulgaris WRKY genes had high variability (2-7 exons), even within the same subgroup. Conserved element analysis showed that within the same class or subclass had the same conserved elements. WRKY conserved domain analysis revealed two mutations in the WRKY domain: WRKYGKK and WRKYGEK. Each WRKY was expressed in at least two tissues, 30 WRKYs were expressed in leaf, 40 WRKYs were expressed in inflorescence, 36 WRKYs were expressed in young leaf, 38 WRKYs were expressed in root, 39 WRKYs were expressed in seedling, and 36 WRKYs were expressed in seed. The expression levels of WRKY genes were also different. All WRKY genes were divided into two types: low expression genes and high expression genes. Such as: BvWRKY23, BvWRKY3, BvWRKY11, BvWRKY7, BvWRKY6, BvWRKY26, BvWRKY4, BvWRKY40, BvWRKY24, BvWRKY2 and BvWRKY28 were highly expressed in all tissues; BvWRKY38, BvWRKY13, BvWRKY36, BvWRKY35, BvWRKY5 and BvWRKY34 were low expressed in all tissues. BvWRKY21, BvWRKY20, BvWRKY22, BvWRKY32, BvWRKY33 and BvWRKY34 genes were up-regulated under heat stress, and BvWRKY1, BvWRKY6, BvWRKY19, BvWRKY31 and BvWRKY33 were up-regulated under salt stress. In addition, BvWRKY33 had a significant response to both heat and salt stress. 【Conclusion】 B. vulgaris WRKY proteins are highly conserved, the length of the gene sequence and the number of introns varied widely, all WRKY genes showed a variety of expression patterns in different tissues, some WRKY genes responded to heat or salt stress, which play an important role in stress physiological regulation.

Key words: Beta vulgaris, WRKY transcription factor, bioinformatics, stress, expression analysis

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