Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (7): 1277-1288.doi: 10.3864/j.issn.0578-1752.2016.07.006

• PLANT PROTECTION • Previous Articles     Next Articles

Expression and Functional Analysis of CsWRKY30 in Cucumber Under Propamocarb Stress

LI Sheng-nan, QIN Zhi-wei, XIN Ming, ZHOU Xiu-yan   

  1. College of Horticulture, Northeast Agricultural University/Key Laboratory of Biology and Germplasm Creation for Horticultural Crops, Ministry of Agriculture, Harbin 150030
  • Received:2015-12-31 Online:2016-04-01 Published:2016-04-01

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Abstract: 【Objective】The objective of this study is to screen, clone and analyze the function of the cucumber transcription factor gene CsWRKY30, which was up-regulated under propamocarb stress in previous transcriptome analyses, CsWRKY30, and to understand the molecular mechanism of low propamocarb residue in cucumbers.【Method】The full length of CsWRKY30 was cloned by PCR, and protein sequences and promoter region of the gene were analyzed with NCBI and PlantCARE separately. The real-time PCR was used to analyze the expression pattern of CsWRKY30 under different treatments. The subcellular localization of CsWRKY30 was conducted through fusion with GFP protein. CsWRKY30 was over-expressed in Arabidopsis by the inflorescence infection method and got pure transgenic lines to identify gene function.【Result】The full length of CsWRKY30 was 1 014 bp and encoded 337 amino acids, which contained a WRKY conservative domain consisting of 60 amino acids. Expression pattern analysis showed that CsWRKY30 was induced by propamocarb stress in the low propamocarb residue D0351, while there was no differences in the high propamocarb residue D9320. The expression level of CsWRKY30 in D0351 was significantly higher than that in the control during 0.5-9 h, while the expression of this gene was no longer significantly up-regulated after 24 h. Tissue specific expression analysis showed that CsWRKY30 was mainly expressed in cucumber fruits. Subcellular localization indicated that CsWRKY30 was mainly located on the nucleus in plant cells. Over-expression CsWRKY30 Arabidopsis under propamocarb stress test showed that there were no significant phenotypic differences between transgenic Arabidopsis and wild types under the untreated condition, while under 2 mmol?L-1 propamocarb stress the transgenic Arabidopsis germination rate and root length were apparently higher than that of the wild type. Expression of CsWRKY30 also showed a trend of increasing under Corynespora cassiicola treatment and was induced by abscisic acid (ABA), while showing no effects in drought and high salt stresses.【Conclusion】CsWRKY30 played an important role under propamocarb stress in cucumber, and over-expression of CsWRKY30 enhanced transgenic plants tolerance.

Key words: cucumber, CsWRKY30, propamocarb, function analysis

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