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Enhancement of rhamnetin production in Vernonia anthelmintica (L.) Willd. cell suspension cultures by eliciting with methyl jasmonate and salicylic acid.

ABSTRACT: Vernonia anthelmintica (L.) Willd. belongs to the family Asteraceae and has anthelmintic, anti-diabetic, diuretic and anti-asthmatic properties. Present study describes the production of rhamnetin: an O-methylated flavonol with anti-oxidant and anti-inflammatory activities; in cell suspension cultures of V. anthelmintica. Suspension cultures were established from cream coloured friable calli initiated from leaf explants and cultured in liquid MS medium supplemented with combinations of IAA (1.5 mg L^-1) and BAP (1.5 mg L^-1). After 20 days of cell suspension culture, the initial inoculum size of 0.5 g was increased to 1.05 g 100 mL^-1 dry weight at exponential phase. When the culture was treated with 0.8 mM methyl jasmonate for 6 days; 2.2 fold accumulation of rhamnetin production was obtained on a dry weight basis compared to unelicited control. Likewise, the elicitation with salicylic acid 1 mM for 6 days brought about 2.7 fold rhamnetin production compared to unelicited control. The rhamnetin content in dried leaf tissue of field grown plants was found to be 0.143 mg g^-1. Elicitation with methyl jasmonate and salicylic acid brought about 7.7 and 9.4 fold increments in rhamnetin production, compared to leaf tissue of field grown plants on a dry weight basis.

SUBMITTER: Rajan M

PROVIDER: S-EPMC7326840 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Using microarray technology and a set of chickpea (Cicer arietinum L.) unigenes and grasspea (Lathyrus sativus L.) ESTs, chickpea responses to treatments with the defence signalling compounds salicylic acid (SA), methyl jasmonate (MeJA), and aminocyclopropane carboxylic acid (ACC) were studied in four chickpea genotypes with ranging levels of resistance to ascochyta blight (Ascochyta rabiei (Pass.) L.). The experimental system minimized environmental effects and was conducted in reference design, where samples from untreated controls acted as references against post-treatment samples. Robust data quality was achieved through the use of three biological replicates (including a dye-swap), the inclusion of negative controls, and strict selection criteria for differentially expressed genes including a fold change cut-off determined by self-to-self hybridizations, Students t test and multiple testing correction (P<0.05). Microarray observations were also validated by quantitative RT-PCR. The time-course expression patterns of 715 experimental microarray features resulted in differential expression of 425 genes in at least one condition. The A. rabiei resistant chickpea genotypes showed a more substantial range of defence-related gene induction by all treatments, indicating that they may possess stronger abilities to resist infection. Further, the involvement of SA, MeJA, and ACC signalling was identified for the regulation of some important A. rabiei responsive genes, as well as cross-talk between these pathways. This study also found evidence to suggest the involvement of A. rabiei-specific signalling mechanisms for the induction of several genes that were previously implicated in A. rabiei resistance. Overall, this study characterised the regulatory mechanisms of many chickpea genes that may be important in defence against various pathogens, as well as other cellular functions. Although the size of the microarray was limited, the results provided novel insights to the molecular control of chickpea cellular processes, which may assist the understanding of chickpea defence mechanisms and allow enhanced development of disease resistant cultivars. Keywords: time course defence-signalling teatment analysis

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Enhancement of rhamnetin production in Vernonia anthelmintica (L.) Willd. cell suspension cultures by eliciting with methyl jasmonate and salicylic acid.

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