ORIGINAL ARTICLE
New insights into the novel and functional promoter sequences of β-1,3-glucanase gene from Hevea brasiliensis
| 1 | Advanced Center for Molecular Biology and Biotechnology, Rubber Research Institute of India, Kottayam, Kerala, India |
| 2 | Department of Biotechnology, University of Kerala, Thiruvananthapuram, Kerala, India |
| 3 | Faculty of Science and Engineering, University of Turku, Turku, Finland |
| 4 | Department of Cell Biology and Molecular Medicine, Rutgers New Jersey Medical School, Newark, New Jersey, United States of America |
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article
CORRESPONDING AUTHOR
Arjunan Thulaseedharan
Advanced Center for Molecular Biology and Biotechnology, Rubber Research Institute of India, Puthuppally, 686 009, Kottayam, India
Advanced Center for Molecular Biology and Biotechnology, Rubber Research Institute of India, Puthuppally, 686 009, Kottayam, India
Submission date: 2020-05-25
Acceptance date: 2020-09-16
Online publication date: 2021-02-25
Journal of Plant Protection Research 2021;61(1):28–40
RELATED ARTICLE
KEYWORDS
abnormal leaf fallHevea brasiliensisPhytophthoraPR-proteinsystemic acquired resistance (SAR)β-13-glucanase
TOPICS
ABSTRACT
β-1,3-glucanases play a major role in combating the abnormal leaf fall disease (ALF) caused
by the oomycete Phytophthora spp. in Hevea brasiliensis, the major commercial source
of natural rubber. In this study, partial sequences of four novel promoters of different
β-1,3-glucanase genomic forms were amplified through inverse PCR from the H. brasiliensis
clone RRII 105 and sequence characterized. This is the first report showing
β-1,3-glucanase genes driven by a different set of promoter sequences in a single clone
of Hevea. The nucleotide sequencing revealed the presence of 913, 582, 553 and 198 bp
promoter regions upstream to the translation initiation codon, ‘ATG’, and contained the
essential cis-elements that are usually present in biotic/abiotic stress-related plant gene promoters
along with other complex regulatory regions. The amplified regions showed strong
nucleosome formation potential and in two of the promoters CpG islands were observed
indicating the tight regulation of gene expression by the promoters. The functional efficiency
of the isolated promoter forms was validated using promoter: reporter gene (GUS)
fusion binary vectors through Agrobacterium mediated transformation in Hevea callus and
tobacco. GUS gene expression was noticed in Hevea callus indicating that all the promoters
are functional. The transgenic tobacco plants showed no GUS gene expression. The implication
of these novel promoter regions to co-ordinate the β-1,3-glucanase gene expression
can be utilized for defense specific gene expression in future genetic transformation attempts
in Hevea and in a wide variety of plant systems.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
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