Acta Agriculturae Boreali-Sinica ›› 2022, Vol. 37 ›› Issue (2): 42-48. doi: 10.7668/hbnxb.20192675

Special Issue: Apple Biotechnology

• Crop Genetics & Breeding·Germplasm Resources·Biotechnology • Previous Articles     Next Articles

Cloning of Gene MdSGR2 from Apple and Construction of Plant Overexpression Vector

ZHAO Huiying1, YANG Guangkai1, GAO Yan1,2, ZHANG Xiaojun1,2, HAO Yanyan1,2   

  1. 1.College of Horticulture,Shanxi Agricultural University,Taigu 030801,China
    2.Shanxi Key Laboratory of Germplasm Improvement and Utilization in Pomology,Taiyuan 030031,China
  • Received:2021-12-13 Published:2022-04-28


Gene STAYGREEN(SGR)plays an important role in the process of plant chlorophyll degradation and metabolism.To explore the biological function of gene SGR2,MdSGR2 gene sequence was cloned from the peel of Granny Smith by RT-PCR.The homology,physicochemical properties,protein structure and cis-acting elements of the promoter were predicted and analyzed by bioinformatics software.The plant overexpression vector was constructed by double enzyme digestion.The results showed that the complete open reading frame(ORF)cDNA of gene MdSGR2 was 840 bp.Encoding 279 amino acids in total,it belonged to the Staygreen superfamily.Physicochemical analysis showed that the total molecular weight of the protein was 31.27 ku,and the isoelectric point pI was 8.52,indicating that the protein was an unstable hydrophilic protein.Homology analysis showed that MdSGR2 encoded amino acid sequence was of the highest homology with Pyrus ussuriensis,reaching 84.12%.The protein structure analysis showed that the secondary structure and tertiary structure of MdSGR2 was determined structurally to be 40.55% α-helix and 44.44% random coil.Promoter analysis showed that the promoter region of the gene contained cryogenic cis-acting elements,photo-responsive elements and induced cis-acting elements such as abscisic acid,salicylic acid and methyl jasmonate response elements.These results suggest that gene MdSGR2 may be regulated by many factors such as environment and hormones.The plant overexpression vector pCAMBIA2301-MdSGR2 was successfully constructed.

Key words: Apple, MdSGR2, Gene cloning, Bioinformatics analysis, Overexpression vector

Cite this article

ZHAO Huiying, YANG Guangkai, GAO Yan, ZHANG Xiaojun, HAO Yanyan. Cloning of Gene MdSGR2 from Apple and Construction of Plant Overexpression Vector[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 42-48. doi: 10.7668/hbnxb.20192675.

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