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

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.

Arabidopsis thalianaSKP1 interacting protein 11 (At2g02870) negatively regulates the release of green leaf volatiles

“…This gene was found to respond to jasmonic acid elicitation and co-up-regulated with PmADH1 and PmLOX1 upon exogenous treatment with jasmonic acid. 30,31 BLAST searches were run to nd a PmF-box1 homolog in Arabidopsis thaliana. The results returned by the BLAST search show that SKP1 interacting protein 11 (SKIP11) possesses the highest (69%) sequence similarity with PmF-box1.…”

Expression profile of gene encoding Kelch repeat containing F-box protein (PmF-box1) in relation to the production of green leaf volatiles

“…In the previous study, the expression of PmF-box1 was found to be increased in Persicaria minor roots after JA treatment (Gor et al 2010). Similar observations were also obtained for several other KFB proteins, such as chickpea (C. arietinum L.) grape (V. vinifera L.) and mung bean (P. vulgaris L.) (Jia et al 2011;Maldonado-Calderón et al 2011;Paquis et al 2010).…”

“…A previous study in our lab has revealed that in Persicaria minor, a gene encoding a kelch repeat containing F-box protein named PmF-box1, an ADH gene named PmADH1 and a LOX gene named PmLOX were found differentially expressed in P. minor after the plants were treated with JA (Gor et al 2010). P. minus or commonly known as kesum is very famous among Malaysians as a plant that exerts sweet and pleasant aroma.…”

Identification of sequences expressed during compatible black pepper—Fusarium solani f. sp. piperis interaction

“…piperis infection by suppression subtractive hybridization (SSH) (Diatchenko et al 1996), which has been used with success to identify differentially expressed genes (Aslam et al 2010;Fan et al 2010;Gor et al 2010). Our results revealed the identification of sequences coding for putative proteins related to oxidative burst and defense response processes, such as superoxide dismutase, cytochrome p450, cysteine proteinase inhibitor, and pathogenesis-related protein 4.…”