Identification of cDNAs for jasmonic acid-responsive genes in Polygonum minus roots by suppression subtractive hybridization

Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor Malaysia
Acta Physiologiae Plantarum (Impact Factor: 1.52). 01/2011; 33(2):283-294. DOI: 10.1007/s11738-010-0546-2

ABSTRACT Elicitation, the plant-based biotechnology approach that utilizes the ability of plant roots to absorb and secrete a vast
variety of bioactive compounds, was studied on Polygonum minus using jasmonic acid (JA) as an elicitor. To understand the overall molecular responses of P. minus roots to JA induction, a subtracted cDNA library was constructed using the suppression subtractive hybridization (SSH) method.
From a total of 1,344 randomly selected colonies, 190 clones were shown to be differentially expressed using Reverse Northern
hybridization. BLAST analysis revealed that clones were similar to genes associated with the biosynthesis of aromatic compounds
through the oxylipin pathway, such as alcohol dehydrogenase and lipoxygenase. Putative clones involved in the shikimate pathway,
including S-adenosyl-l-methionine synthetase and S-adenosyl-l-homocysteine hydrolase, were identified with predicted roles in phenylpropanoids’ biosynthesis. Genes responding to abiotic
stress unique to JA elicitation, such as ELI3-1, glutathione S-transferase and peroxidase 1, were also identified. The kelch-repeat containing F-box family protein, a possible transcription
factor in response to JA elicitation was also found. The results of the RT-PCR showed that the eight selected clones were
strongly up-regulated, except for lipoxygenase, which showed a slightly higher expression of the transcript levels in response
to the JA elicitation.

KeywordsDifferentially expressed genes–Elicitor–Secondary metabolites–Aromatic compound–Abiotic stress

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