Abstract
The 3-hydroxy-3-methyl glutaryl CoA reductase (HMGR) is the key enzyme of mevalonate pathway in plants. A partial genomic DNA fragment encoding HMGR conserved domain (denoted as AbHMGR) is isolated from Aconitum balfourii Stapf. It comprises 871 bp encoding 290 amino acids. In silico analysis reveals that it had extensive similarities to other plant HMGR gene. Domain analysis of AbHMGR showed two highly conserved NADPH and HMG CoA domains. Docking study predicted inhibitor, substrate and cofactor binding sites in the protein. Expression analysis revealed that AbHMGR is similarly expressed in all tested tissues with differential pattern. The highest expression was found in leaf tissue. However, fold expression in root and shoot tissue was almost similar. Enzyme activity of HMGR was found to be much higher in leaf tissue as compared to other tissues. The highest aconitine content (0.015 %) was obtained in root tissues. Our data laid a foundation for further investigation of HMGR role in Aconitum balfourii. Electronic supplementary materialThe online version of this article (doi:10.1007/s13205-016-0405-y) contains supplementary material, which is available to authorized users.
Highlights
Aconitum balfourii (A. balfourii) Stapf is one of the endangered herbs of the genus Aconitum
isopentyl pyrophosphate (IPP) formation follows two distinct routes, the classical mevalonate pathway (MVA) which is effective in cytosol and another one is 2-C-methyl-D-erythritol 4-phosphate (MEP) in plastids (McGarvey and Croteau 1995)
The putative sequence was subjected to BLAST search in GenBank database and homology search confirmed the identity of hydroxy-3-methyl glutaryl CoA reductase (HMGR) sequence
Summary
Aconitum balfourii (A. balfourii) Stapf is one of the endangered herbs of the genus Aconitum. The value of aconitine as a medicine has been recognized in modern times, and it ranks as one of the most useful drugs, in homeopathy, Ayurveda and Unani systems of medicine (Kirtikar and Basu 1965). It cures several ailments like rheumatism, arthritis, gout, neuralgia, sciatica, migraine and cancer. Gene mining of rate-limiting enzymes of aconitine biosynthesis pathway and their studies is one of the strategies which needs to be focused. In the view of above, we successfully attempted the isolation of HMG CoA reductase gene from A. balfourii Stapf. Expression pattern analysis and HMGR enzyme activities and aconitine content analysis at tissue level were performed
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