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ACTA HORTICULTURAE SINICA ›› 2015, Vol. 42 ›› Issue (2): 289-300.doi: 10.16420/j.issn.0513-353x.2014-0686

• Ornamental Plants • Previous Articles     Next Articles

Cloning and Expression Analysis of Coronatine Insensitive Gene Encoding a Jasmonate Receptor from Gladiolus hybridus

WU Chen-yu1,HE Jun-na1,ZHONG Xiong-hui1,SENG Shan-shan1,WU Jian1,SUI Juan-juan1,2,LIU Chen1,WU Ze1,LIU Chao1,and YI Ming-fang1,*   

  1. 1Department of Ornamental Horticulture and Landscape Architecture,China Agricultural University,Beijing 100193,China;2College of Biology,Fuyang Normal College,Fuyang,Anhui 236037,China
  • Online:2015-02-25 Published:2015-02-25

Abstract: A full length cDNA of jasmonate receptor CORONATINE INSENSITIVE1(COI1)was amplified from the leaves of Gladiolus hybridus‘Rose Supreme’by the technique of RACE,and it was named GhCOI1. The gene had a length of 2 603 bp,and it included a 5′-untranslated region(UTR)of 484 bp,a 3′-UTR of 277 bp and an open reading frame that encoded a protein of 613 amino acid residues with a predicted molecular mass of 69.46 kD. The results of multiple alignments revealed that the deduced amioacid sequence of GhCOI1 had a high similarity(64.3%)to LeCOI1. Phylogenetic analysis showed that GhCOI1 shared a close relationship with the OsCOI1. The results of qRT-PCR demonstrated that the highest level of GhCOI1 mRNA was detected in leaves which followed by roots,while the extreme low level was observed in stolon,cormel,corm,petal,stamen and pistil,suggesting that the homologous genes of GhCOI1 expressed specifically in organs. Leaves were treated with the solution of 0.05,0.1 and 0.5 mmol ? L-1 methyl jasmonate(MeJA)respectively,the transcript level of GhCOI1 was up-regulated significantly by 0.1 mmol ? L-1 MeJA. The expression of GhCOI1 showed an alternating pattern of rise and fall by treatment during 12 hours with 0.1 mmol ? L-1 MeJA. Following transient expression of GhCOI1 in onion epidermal cells,GhCOI1 was found to be localized in both the cytoplasm and the plastid. The research suggested that GhCOI1 has specific function in jasmonate signal pathway of Gladiolus hybridus.

Key words: Gladiolus hybridus, CORONATINE INSENSITIVE1, jasmonic acid, cloning, expression analysis

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