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Discerning picroside-I biosynthesis via molecular dissection of in vitro shoot regeneration in Picrorhiza kurroa

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Expression analysis of primary and secondary metabolic pathways genes vis-à-vis shoot regeneration revealed developmental regulation of picroside-I biosynthesis in Picrorhiza kurroa.

Abstract

Picroside-I (P-I) is an important iridoid glycoside used in several herbal formulations for treatment of various disorders. P-I is synthesized in shoots of Picrorhiza kurroa and Picrorhiza scrophulariiflora. Current study reports on understanding P-I biosynthesis in different morphogenetic stages, viz. plant segment (PS), callus initiation (CI), callus mass (CM), shoot primordia (SP), multiple shoots (MS) and fully developed (FD) stages of P. kurroa. Expression analysis of genes involved in primary and secondary metabolism revealed that genes encoding HMGR, PMK, DXPS, ISPE, GS, G10H, DAHPS and PAL enzymes of MVA, MEP, iridoid and shikimate/phenylpropanoid pathways showed significant modulation of expression in SP, MS and FD stages in congruence with P-I content compared to CM stage. While HK, PK, ICDH, MDH and G6PDH showed high expression in MS and FD stages of P. kurroa, RBA, HisK and CytO showed high expression with progress in regeneration of shoots. Quantitative expression analysis of secondary metabolism genes at two temperatures revealed that 7 genes HMGR, PMK, DXPS, GS, G10H, DAHPS and PAL showed high transcript abundance (32–87-folds) in FD stage derived from leaf and root segments at 15 °C compared to 25 °C in P. kurroa. Further screening of these genes at species level showed high expression pattern in P. kurroa (6–19-folds) vis-à-vis P. scrophulariiflora that was in corroboration with P-I content. Therefore, current study revealed developmental regulation of P-I biosynthesis in P. kurroa which would be useful in designing a suitable genetic intervention study by targeting these genes for enhancing P-I production.

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Abbreviations

26S:

26S rRNA

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

HMGR:

3-Hydroxy-3-methylglutaryl-CoA reductase

PMK:

Phosphomevalonate kinase

DXPS:

1-Deoxy-d-xylulose-5-phosphate synthase

ISPD:

2-C-methylerythritol 4-phosphate cytidyl transferase

ISPE:

4-(Cytidine-5-diphospho)-2-C-methylerythritol kinase

GS:

Geraniol synthase

G10H:

Geraniol-10-hydroxylase

10-HGO:

10-Hydroxygeraniol dehydrogenase

IS:

Iridoid synthase

DAHPS:

3-Deoxy-d-arabino-heptulosonate 7-phosphate synthase

PAL:

Phenylalanine ammonia lyase

HK:

Hexokinase

PK:

Pyruvate kinase

ICDH:

Isocitrate dehydrogenase

MDH:

Malate dehydrogenase

G6PDH:

Glucose-6-phosphate dehydrogenase

RBA:

RuBisCO activase

ARP:

Auxin response protein

ARF7:

Auxin response factor 7

HisK:

Histidine kinase

CytO:

Cytokinin oxidase

MVA:

Mevalonate

MEP:

Non-mevalonate

PS:

Plant segment stage

CI:

Callus initiation stage

CM:

Callus mass stage

SP:

Shoot primordia stage

MS:

Multiple shoots stage

FD:

Fully developed stage

MS medium:

Murashige and Skoog medium

P-I:

Picroside-I

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Acknowledgments

The authors are thankful to Jaypee University of Information Technology for providing the necessary research facilities and Department of Biotechnology, Ministry of Science and Technology, Govt. of India for providing research grant in the form of a programme support on high value medicinal plants to RSC and HS. We are also thankful to the Himalayan Forest Research Institute (HFRI), Manali, India and National Bureau of Plant Genetic Resources, New Delhi, India for providing plant material of P. kurroa and P. scrophulariiflora, respectively.

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  1. Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, HP, India

    Neha Sharma, Rajinder Singh Chauhan & Hemant Sood

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  1. Neha Sharma
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  2. Rajinder Singh Chauhan
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  3. Hemant Sood
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Correspondence to Hemant Sood.

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Communicated by H. Ebinuma.

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Sharma, N., Chauhan, R.S. & Sood, H. Discerning picroside-I biosynthesis via molecular dissection of in vitro shoot regeneration in Picrorhiza kurroa . Plant Cell Rep 35, 1601–1615 (2016). https://doi.org/10.1007/s00299-016-1976-0

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