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Bulletin of Botanical Research ›› 2023, Vol. 43 ›› Issue (3): 412-420.doi: 10.7525/j.issn.1673-5102.2023.03.011

• Molecular biology • Previous Articles     Next Articles

Cloning of WRKY40 Gene from Leymus chinensis and Analysis of Disease Resistance in Transgenic Tobacco

Danni LI1, Jiali LIU1, Jitao ZHANG2, Baoxiang GU1, Fengjin ZHU1, Qingjie GUAN1()   

  1. 1.Key Laboratory of Restoration and Reconstruction of Northeast Saline-alkali Vegetation,Ministry of Education,College of Life Sciences,Northeast Forestry University,Harbin 150040
    2.North-east Institute of Geography and Agroecology,Chinese Academy of Sciences,Changchun 130000
  • Received:2022-08-21 Online:2023-05-20 Published:2023-05-11
  • Contact: Qingjie GUAN E-mail:guanqingjie@nefu.edu.cn
  • About author:LI Danni(1999—),female,master,major in molecular biology research.
  • Supported by:
    National Natural Science Foundation of China(32171989);Science and Technology basic Resources Survey Project(2021FY100403)


In order to cultivate rice varieties resistant to Xanthomonas oryzae pv. oryzaeRhizoctonia solaniMagnaporthe oryzae and Fusarium fujikuroi, the method of mining resistance genes is an important way to select resistant varieties. RT-PCR method was used to clone the LcWRKY40 gene (MN187915) from the leaves of Leymus chinensis. The result showed that the CDS was 1 053 bp in length and encoded 350 amino acids with a molecular weight of 38.1 kDa. The results of bioinformatics analysis showed that the primary structure of LcWRKY40 contained WRKY domain, zinc finger protein domain and nuclear localization sequence. Phylogenetic tree construction and motif analysis showed that the phylogenetic relationship between LcWRKY40 and HvWRKY38 was closed. The results of subcellular localization in tobacco showed that the LcWRKY40 protein was located in the nucleus, verified by the software prediction. qRT-PCR tissue specific expression analysis showed that LcWRKY40 gene was expressed in root, stem, leaf, leaf sheath, Lemma and anther of Leymus chinensis respectively, but the expression level was the highest in leaf but the lowest in Lemma. Transgenic LcWRKY40 tobacco plants and wild-type tobacco plants were inoculated with X. oryzae pv. Oryzae, R. solani, M. oryzae and F. fujikuroi, respectively, which showed that transgenic LcWRKY40 tobacco plants could alleviate the four pathogens in different degrees, and showed high resistance to M. oryzae and F. fujikuroi. Therefore, it was speculated that LcWRKY40 protein played a key regulatory role in signal pathways such as resisting disease stress and improved the resistance of plant pathogens, which layed a molecular foundation for the study of LcWRKY40 function and abiotic stress.

Key words: Leymus chinensis, LcWRKY40 gene, transgenic tobacco, disease resistance

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