Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (10): 1852-1861.doi: 10.3864/j.issn.0578-1752.2017.10.010

• HORTICULTURE • Previous Articles     Next Articles

Cloning and Expression Analysis of Cadmium Tolerance Related Gene CsNAC019 in Cucumber

LI HuiYuan, TIAN ChunYu, ZHENG YuYing, WU Tao   

  1. Horticultural and Landscape Architecture College, Northeast Agricultural University/Key Laboratory of Biology and Germplasm Creation for Horticultural Crops, Harbin 150030
  • Received:2016-11-25 Online:2017-05-16 Published:2017-05-16

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Abstract: 【Objective】 To understand the molecular mechanism of Cd tolerance in cucumber, a NAC transcription factor CsNAC019 was obtained by using bioinformatics analysis. CsNAC019 was cloned and its expression was analyzed under Cd stress. The results of the present study will provide an experimental foundation for breeding of cucumber cultivars with high capacity for Cd tolerance.【Method】The full length sequence of CsNAC019 was cloned by PCR. Sequence comparison and conserved domain were analyzed by NCBI and DNAMAN. Amino acid composition, stability coefficient, and hydrophilic coefficient were analyzed by online software Expasy and TMHMM. The promoter region was analyzed by online software PlantCARE. Phylogenic tree was constructed by MEGA 6.0 according to the NJ method. The real-time PCR (qRT-PCR) was used to analyze the expression pattern of CsNAC019 under Cd treatments. The variance and significance were analyzed by DPS 7.05.【Result】CsNAC019 contained a typical conserved NAC domain. BLAST analysis revealed that CsNAC019 had 60% homology with NAC019 in the amino acid sequence, and the five relatively conserved regions of its amino acid sequences, A, B, C, D and E, were highly consistent between them from the N-terminal to the C-terminal. The full-length CDS of CsNAC019 was 960 bp, which encoded 319 amino acids with a molecular weight of 35.66 kD. The theoretical isoelectric point is 8.72, the instability coefficient is 68.18, and the average hydrophilic coefficient is -0.483. Besides the eukaryotic promoter TATA and CAAT natural elements, promoter analysis showed that the promoter sequence of CsNAC019 also had cis elements in response to stress, such as G-box, ABRE, W-box, P-box, TCA-element, TC-richrepeats, and HSE, etc. Phylogenetic analysis showed that CsNAC019 had 64%-96% homology with NAC transcription factors of other 15 plants (melon, peach, apple, citrus, grape and soybean, etc). CsNAC019 had the highest homology, 96%, with the NAC sequence of melon. qRT-PCR analysis revealed that the expression of CsNAC019 was significantly increased (8.2 folds) compared with the control under Cd tolerance condition.【Conclusion】CsNAC019 is a Cd tolerance induced gene. It was speculated that CsNAC019 may regulate the cucumber to response to Cd tolerance by regulating the expression of downstream genes.

Key words: cucumber, Cd, CsNAC019, gene clone, expression analysis

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