RESEARCH ARTICLE

Cloning and pharmaceutical analysis of CaMK gene of Botrytis cinerea

  • Yuxia DONG 1 ,
  • Jihong XING 1 ,
  • Jiao JIA 1 ,
  • Qiaoyun WENG 2 ,
  • Zhimin HAO 1 ,
  • Jingao DONG , 1
Expand
  • 1. Molecular Plant Pathology Lab, Agricultural University of Hebei, Baoding 071001, China
  • 2. Department of Agricultural and Forest Technology, Hebei North University, Zhangjiakou 075131, China

Received date: 08 Nov 2010

Accepted date: 03 Dec 2010

Published date: 05 Sep 2011

Copyright

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg

Abstract

Using a PCR homology approach, DNA and cDNA sequences of calcium/calmodulin-dependent protein kinase (CaMK) gene of Botrytis cinerea were obtained. Southern blotting result displayed that CaMK was single copy in the genome of B. cinerea. The cDNA sequence of CaMK revealed an open reading frame of 2190 nucleotides encoding a 730 amino acid protein with predicted molecular weight of 81.8748 kDa. The genomic sequence of CaMK revealed the same ORF interrupted by six introns. Bioinformatics analysis showed that this protein had the distinctive features that characterize CaMK ATP binding region signature and serine/threonine protein kinase active-site signature. Pharmaceutical analysis displayed that the CaMK specific inhibitor, KN-62, could inhibit conidial germination, pathogenicity and herbicidal activity of B. cinerea BC4 strain. It was suggested that CaMK played an important role in regulating conidial germination, pathogenicity and herbicidal activity of B. cinerea.

Cite this article

Yuxia DONG , Jihong XING , Jiao JIA , Qiaoyun WENG , Zhimin HAO , Jingao DONG . Cloning and pharmaceutical analysis of CaMK gene of Botrytis cinerea[J]. Frontiers of Agriculture in China, 2011 , 5(3) : 299 -304 . DOI: 10.1007/s11703-011-1085-3

1
Anand T, Chandrasekaran A, Kuttalam S, Senthilraja G, Samiyappan R (2010). Integrated control of fruit rot and powdery mildew of chili using the biocontrol agent Pseudomonas fluorescens and a chemical fungicide. Biol Control, 52(1): 1–7

DOI

2
Chen H, Xiao X, Wang J, Wu L J, Zheng Z M, Yu Z L (2008). Antagonistic effects of volatiles generated by Bacillus subtilis on spore germination and hyphal growth of the plant pathogen, Botrytis cinerea. Biotechnol Lett, 30(5): 919–923

DOI PMID

3
Drenth A, Goodwin S B, Fry W E, Davidse L C (1993). Genotypic diversity of Phytophthora infestans in the Netherlands revealed by DNA polymorphisms. Phytopathology, 83(10):1087–1092

4
Hanks S K, Quinn A M (1991). Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. Methods Enzymol, 200: 38–62

DOI PMID

5
Jayashree T, Praveen Rao J, Subramanyam C (2000). Regulation of aflatoxin production by Ca2+/calmodulin-dependent protein phosphorylation and dephosphorylation. FEMS Microbiol Lett, 183(2): 215–219

DOI PMID

6
Joseph J D, Means A R (2000). Identification and characterization of two Ca2+/CaM-dependent protein kinases required for normal nuclear division in Aspergillus nidulans. J Biol Chem, 275(49): 38230–38238

DOI PMID

7
Joseph J D, Means A R (2002). Calcium binding is required for calmodulin function in Aspergillus nidulans. Eukaryot Cell, 1(1): 119–125

DOI PMID

8
Kameshita I, Yamada Y, Nishida T, Sugiyama Y, Sueyoshi N, Watanabe A, Asada Y (2007). Involvement of Ca2+/calmodulin-dependent protein kinases in mycelial growth of the basidiomycetous mushroom, Coprinus cinereus. Biochim Biophys Acta, 1770(9): 1395–1403

DOI

9
Li C G (2003). Studies on herbicidal activity of metabolites from Botrytis cinerea and isolation of active substance. Dissertation for the Master Degree. Hebei: Agricultural University of Hebei (in Chinese)

10
Liu X H, Lu J P, Dong B, Gu Y, Lin F C (2010). Disruption of MoCMK1, encoding a putative calcium/calmodulin-dependent kinase, in Magnaporthe oryzae. Microbiol Res, 165(5): 402–410

DOI PMID

11
Ma J (2006). Mutation and differential display of herbicidal-related genes of Botrytis Cinerea. Dissertation for the Master Degree. Hebei: Agricultural University of Hebei (in Chinese)

12
Ma L, Liang S P, Jones R L, Lu Y T (2004). Characterization of a novel calcium/calmodulin-dependent protein kinase from tobacco. Plant Physiol, 135(3): 1280–1293

DOI PMID

13
Mukherjee P K, Kenerley C M (2010). Regulation of morphogenesis and biocontrol properties in Trichoderma virens by a VELVET protein, Vel1. Appl Environ Microbiol, 76(7): 2345–2352

14
Praskova M, Kalenderova S, Miteva L, Poumay Y, Mitev V (2002). Ca2+ /calmodulin-dependent protein kinase (CaM-kinase) inhibitor KN-62 suppresses the activity of mitogen-activated protein kinase (MAPK), c-myc activation and human keratinocyte proliferation. Arch Dermatol Res, 294(4): 198–202

DOI PMID

15
Timmins J M, Ozcan L, Seimon T A, Li G, Malagelada C, Backs J, Backs T, Bassel-Duby R, Olson E N, Anderson M E, Tabas I (2009). Calcium/calmodulin-dependent protein kinase II links ER stress with Fas and mitochondrial apoptosis pathways. J Clin Invest, 119(10): 2925–2941

DOI PMID

16
Tokumitsu H, Chijiwa T, Hagiwara M, Mizutani A, Terasawa M, Hidaka H (1990). KN-62, 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine, a specific inhibitor of Ca2+ /calmodulin-dependent protein kinase II. J Biol Chem, 265(8): 4315–4320

PMID

17
Tsai P J, Tu J, Chen T H (2002). Cloning of a Ca2+€€/calmodulin-dependent protein kinase gene from the filamentous fungus Arthrobotrys dactyloides. FEMS Microbiol Lett, 212(1): 7–13

DOI PMID

18
Valle-Aviles L, Valentin-Berrios S, Gonzalez-Mendez R R, Rodriguez-Del Valle N (2007). Functional, genetic and bioinformatic characterization of a calcium/calmodulin kinase gene in Sporothrix schenckii. BMC Microbiol, 7(1): 107

DOI PMID

19
Yang Y, Cheng P, Zhi G, Liu Y (2001). Identification of a calcium/calmodulin-dependent protein kinase that phosphorylates the Neurospora circadian clock protein FREQUENCY. J Biol Chem, 276(44): 41064–41072

DOI PMID

20
Zheng M, Xu K, Dong J G (2008). Purification and structural identification of herbicides from Botrytis cinerea. Acta Microbiologica Sinica, 48(10): 1362–1366 (in Chinese)

PMID

Outlines

/