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Bulletin of Botanical Research ›› 2018, Vol. 38 ›› Issue (1): 125-131.doi: 10.7525/j.issn.1673-5102.2018.01.015

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Expression and Function Analysis of Walnut JrGRAS2 Gene under Heat Stress

SU Li-Yuan1,2, LI Xiao-Zhe1, CHEN Shu-Wen1,2, ZHANG Fang-Fang1,2, ZHAO Ting-Ting1,2, YANG Gui-Yan1,2   

  1. 1. Walnut and Chestnut Experiment Station of Shanyang, College of Forestry, Northwest A&F University, Yangling 712100;
    2. Key Laboratory of Economic Plant Resources Development and Utilization in Shaanxi Province, College of Forestry, Northwest A&F University, Yangling 712100
  • Received:2017-05-24 Online:2018-01-15 Published:2018-01-06
  • Supported by:
    Fundamental Research Funds for the Central Universities(2452016057/2452015171);China Postdoctoral Science Foundation Project(2016M590979);The Program of Innovative Entrepreneurship Training Programs for Northwest Agriculture & Forestry University Students(1201610712031)

Abstract: The GRAS transcription factor is important for plant response to abiotic stress. To well understand the adaptive mechanism to adverse environment of walnut tree, a GRAS gene was cloned from the transcriptome of Juglans regia(Named as JrGRAS2). The expression of JrGRAS2 was analyzed under heat stress, and JrGRAS2 was inserted into the yeast expression vector pYES2 for constructing recombinant plasmid pYES2-JrGRAS2, which was transformed into a Saccharomyces cerevisiae strain(INVSCI). The yeast transformed with empty pYES2 was used as a negative control. The yeast expression system was used for analysis the heat stress tolerance. The full length open reading frame(ORF) of JrGRAS2 was 1 296 bp, the deduced protein was 47 405.83 Da with 431 amino acids, and the theoretical isoelectric point(pI) was 5.66. Under heat stress, JrGRAS2 was highly induced, especially exposed to 36℃ for 0.5 h in the stems, it was induced to 335.5-fold of control. When both recombinant yeasts were treated with 53℃, the JrGRAS2 expressed yeast displayed higher vitality and survival rate than the control yeast. Therefore, JrGRAS2 gene could effectively response to heat stress and improve heat tolerance of transgenic yeasts, JrGRAS2 may be an important candidate gene for walnut response to adverse stimulus.

Key words: Juglans regia, heat stress, GRAS transcription factor, expression analysis, yeast expression system

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