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CHINESE JOURNAL OF OIL CROP SCIENCES ›› 2020, Vol. 42 ›› Issue (1): 85-.doi: 10.19802/j.issn.1007-9084.2019085

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Cloning and functional analysis of CtDHN1 gene related to abiotic stress in Carthamus tinctorius

  

  1. 1. College of Life Sciences,Jilin Agricultural University,Changchun 130118,China; 2. College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China
  • Online:2020-02-28 Published:2020-04-22

Abstract:

 Dehydrates was a member of the group 2 late embryogenesis abundant protein family, which were
high disorder proteins and responded to adversity stress. Based on the transcriptome data of safflower under drought
stress, CtDHN1, a dehydrin gene respondimg to drought stress was identified and cloned by using RT-PCR tech⁃
niques, which encoded 318 amino acids with the molecular weight of 31.1 kDa and the pI value of 6.36. Bioinformat⁃
ics analysis indicated that CtDHN1 protein belonged to YnSKn-type dehydrin. Prokaryotic expression analysis re⁃
vealed that CtDHN1 protein was a soluble protein. The protein had the highest expression level when the concentra⁃
tion of IPTG was 0.4 mmol/L, the OD600 value was 0.8, the induction time was 4 h, and the induction temperature
was 37℃. The E. coli stress tolerance analysis demonstrated that CtDHN1 gene tolerated a drought and a high salt
environment of 1.5 mol/L sorbitol and 1.3 mol/L sodium chloride. In vitro enzyme activity assays indicated that Ct⁃
DHN1 protein protected LDH enzyme activity in 500 mmol/L and sorbitol 300 mmol/L sodium chloride. This re⁃
search provided a theoretical basis for the molecular mechanism of CtDHN1 gene in safflower resistance to abiotic
stress.

Key words: dehydrins, abiotic stress, prokaryotic expression, LDH activity

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