Journal of Plant Interactions
ISSN: 1742-9145 (Print) 1742-9153 (Online) Journal homepage: http://www.tandfonline.com/loi/tjpi20
Characterization of the promoter sequence of chitinase gene from lima bean plant Koichi Sugimoto , Kenji Matsui , Rika Ozawa & Junji Takabayashi To cite this article: Koichi Sugimoto , Kenji Matsui , Rika Ozawa & Junji Takabayashi (2011) Characterization of the promoter sequence of chitinase gene from lima bean plant, Journal of Plant Interactions, 6:2-3, 163-164, DOI: 10.1080/17429145.2010.542566 To link to this article: http://dx.doi.org/10.1080/17429145.2010.542566
Copyright Taylor and Francis Group, LLC
Published online: 11 Mar 2011.
Submit your article to this journal
Article views: 95
View related articles
Citing articles: 2 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=tjpi20 Download by: [117.136.234.11]
Date: 31 January 2016, At: 01:37
Journal of Plant Interactions Vol. 6, Nos. 23, JuneSeptember 2011, 163164
SHORT COMMUNICATION Characterization of the promoter sequence of chitinase gene from lima bean plant Koichi Sugimotoa,b, Kenji Matsuia*, Rika Ozawab and Junji Takabayashib a
Graduate School of Medicine, Yamaguchi University, Yoshida 1677-1, Yamaguchi 753-8515 Japan; bCenter for Ecological Research, Kyoto University, Hirano 2-509-3, Otsu 520-2113 Japan (Received 19 November 2010; final version received 20 November 2010) Chitinase is involved in induced defense against biotic attacks. We analyzed the promoter of the gene in lima bean plants and found cis-elements important for the induction under stress.
Keywords: chitinase; cis-element; induced defense; lima bean
Downloaded by [117.136.234.11] at 01:37 31 January 2016
Introduction Plants induce the expression of a series of defensive genes to protect against invaders. One of the defensive genes, chitinase, was induced by infestation of the herbivorous mite and jasmonate treatment in lima bean plant (Arimura et al. 2000). However, it is unclear how the gene was induced under such conditions. In this study, we show possible regulatory sequences of chitinase gene in lima bean plant. Materials and methods Genomic DNA extracted from lima bean leaf (Phaseolus lunatus cv. Sieva) was digested with BclI, HincII, KpnI, or HinfI and ligated with T4 DNA ligase. The upstream sequence of the chitinase gene was amplified with the circulated DNA by using Taq DNA polymerase using primer set 1 (5?-CTC AGCGCCCTCATATCCAG-3? and 5?-AGAGTG CACCTCCTGCTTGCCTTC-3?) and 2 (5?-ATGTT TTGCTTTTGTGAAGA-3? and 5?-AATCCAACA ATTTTACATCAA-3?) for first amplification, and set 3 (5?-GCCAGAGGCTTCTACACCTACGATG CC-3? and 5?-CTCCGTAGCTTCCTCCAACCA ACAGCATC-3?) and 4 (5?-TGCAGTTTTAATAG TTGCGATTG-3? and 5?-TTATTCCAAGCCAAG TTCCAA-3?) for second amplification, respectively. The amplicons detected after second amplification were sequenced. Result and discussions The regulatory region on upstream sequence of chitinase gene was obtained from lima bean genomic DNA by nested inverse PCR (Figure 1, accession no. AB572066). Transcription and translation start sites were identified as typical TATA-box and Kozak
*Corresponding author. Email:
[email protected] ISSN 1742-9145 print/ISSN 1742-9153 online # 2011 Taylor & Francis DOI: 10.1080/17429145.2010.542566 http://www.informaworld.com
Figure 1. DNA sequence of upstream region of chitinase gene. The coding and promoter region are shown by closed box and bar, respectively. Relative size of sequence is shown by size marker in the figure. The first methionine position of chitinase protein in Kozak sequence was shown by 1. TATA box: open box, G-box core: circle, W-box core: triangle.
sequences, respectively, on the appropriate positions at the upstream of chitinase gene. At approximately 900 bp upstream from the start codon, G-box, and W-box core promoter sequences responsible for jasmonate signaling and elicitor response, respectively, were identified (Mason et al. 1993; Rushton et al. 1996). This result imply that the cis-elements in the promoter sequence possibly contribute to the induced defense of lima bean plant against invaders such as spider mite and to the eavesdropping phenomenon through jasmonate signaling (Arimura et al. 2000). The promoter sequence is also a useful tool to dissect the molecular mechanism of the induced defense response.
Acknowledgements The cDNA sequence of PR-3 gene of lima bean was provided by G. Arimura. This work was supported in part by grants of priority area (S) from MEXT, Core-toCore project (20004) from JSPS, and Scientific Research for Plant Graduate Students from the Nara Institute of Science and Technology supported by MEXT.
164
K. Sugimoto et al.
References
Downloaded by [117.136.234.11] at 01:37 31 January 2016
Arimura G, Ozawa R, Shimoda T, Nishioka T, Boland W, Takabayashi J. 2000. Herbivore-induced volatiles elicit defence genes in lima bean leaves. Nature. 406:512 515. Rushton PJ, Torres JT, Parniske M, Wernert P, Hahlbrock K, Somssich IE. 1996. Interaction of elicitor-induced
DNA-binding proteins with elicitor response element in the promoters of parsley PR1 genes. EMBO J. 15:56905700. Mason HS, DeWald DB, Mullet JE. 1993. Identification of a methyl jasmonate-responsive domain in the soybean vspB promoter. Plant Cell. 5:241251.
