Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 17-25.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0077

Previous Articles     Next Articles

Cloning and Expression Analysis of GjMnSOD Gene in Gerbera jamesonni

WU Huan1(), LU Zhen-hong2,3, HAO Xiang-yang1, WANG Bin1, JIAO Yuan-chen1, YANG Chun-mei2,3, CHENG Chun-zhen1,4()   

  1. 1. College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002
    2. Flower Research Institute,Yunnan Academy of Agricultural Sciences,Kunming 650100
    3. Yuxi Yunxing Biotechnology Co.,Ltd.,Yuxi 652604
    4. College of Horticulture,Shanxi Agricultural University,Taigu 030801
  • Received:2021-01-18 Online:2021-10-26 Published:2021-11-12
  • Contact: CHENG Chun-zhen E-mail:wuhuan980422@163.com;ld0532cheng@126.com

Abstract:

Having the ‘Linglong’ Gerbera jamesonni as experimental material,RT-PCR technology was used to successfully clone a SOD gene(GenBank accession number:MH708534.1)with CDS length of 519 bp. Then,bioinformatics analysis was to analyze the sequences of the gene and its encoded protein,and quantitative real time PCR(qRT-PCR)was adapted to investigate the gene expression patterns under treatments of 3 phytohormones(100 µmol/L IAA,GA3 and SA)and 3 abiotic stresses(salt stress using 200 mmol/L NaCl,mimicked drought stress using 30% PEG,and cold stress using 4℃). Results showed that the SOD gene encoded a stable hydrophilic protein with no signal peptide and transmembrane structure and its molecular weight was 19 203.85 and pI was 7.93. The protein contained typical Sod-Fe-C conserved domain and a MnSOD characteristic sequence(DVWEHAYY)ranging from 141 th to 148th amino acids,thus it was named as GjMnSOD. Wolf Psort prediction revealed that GjMnSOD was located in mitochondria. GjMnSOD shared the highest similarity(93.86%)with Cynara cardunculus MnSOD,and they were in the closest genetic relationship. qRT-PCR result showed that the expression of GjMnSOD was significantly inhibited by IAA,SA and GA3 treatments,salt and 4℃ low temperature stresses. Under drought treatment,it showed ‘fall-rise’ expression pattern and its expression level was found to be significantly higher than control from 4 h post-treatment. The above results indicate that GjMnSOD is widely involved in the stress responses of G. jamesonni,especially in drought response.

Key words: Gembera jamesonni, manganese superoxide dismutase, gene cloning, expression analysis, stress response