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Title of Journal: Protoplasma

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Abbravation: Protoplasma

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Springer Vienna

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1615-6102

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Metabolic shift from withasteroid formation to phe

Authors: Bipradut Sil Chiranjit Mukherjee Sumita Jha Adinpunya Mitra
Publish Date: 2014/12/23
Volume: 252, Issue: 4, Pages: 1097-1110
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Abstract

Cotransformed hairy roots containing a gene that encodes a fungal elicitor protein βcryptogein were established in Withania somnifera a medicinal plant widely used in Indian systems of medicine To find out whether βcryptogein protein endogenously elicits the pathway of withasteroid biosynthesis withaferin A and withanolide A contents along with transcript accumulation of farnesyl pyrophosphate FPP synthase 3hydroxy3methylglutarylCoA reductase HMGR and sterol glycosyltransferase SGT were analyzed in both cryptogeincotransformed and normal hairy roots of W somnifera It was observed that the withaferin A and withanolide A contents were drastically higher in normal hairy roots than cryptogeincotransformed ones Similar trends were also observed on the levels of transcript accumulation Subsequently the enzyme activity of phenylalanine ammonia lyase PAL one of the key enzymes of phenylpropanoid pathway was measured in both cryptogeincotransformed and normal hairy roots of W somnifera along with the levels of PAL transcript accumulation Upliftment of PAL activity was observed in cryptogeincotransformed hairy roots as compared to the normal ones and the PAL expression also reflected a similar trend ie enhanced expression in the cryptogeincotransformed lines Upliftment of wallbound ferulic acid accumulation was also observed in the cryptogeincotransformed lines as compared to normal hairy root lines Thus the outcome of the above studies suggests a metabolic shift from withanolide accumulation to phenylpropanoid biosynthesis in cryptogeincotransformed hairy roots of W somniferaS Jha thanks David Tepfer of INRA Versailles France for providing the Agrobacterium rhizogenes 9402 strain harboring a binary vector pBIN19 containing the βcryptogein gene B Sil thanks the Council of Scientific and Industrial Research CSIR India for the award of an individual senior research fellowship SRFNET Facilities created from a research grant SERB/SR/S0/PS/18/2011 to A Mitra and S Jha of the Science and Engineering Research Board Department of Science and Technology India for a related project were utilized in this workAM BS and CM conceived and designed research BS did all the experiments BS and CM analyzed the data SJ guided the genetic transformation experiments and took the overall responsibility AM wrote the final manuscript All authors read and approved the manuscriptPCR amplification of cryptogein gene in normal hairy roots A and cryptcotransformed B hairy roots of W somnifera C Positive control A rhizogenes strain LBA9402 harboring pBIN19CRYPT binary vector M Molecular marker Gene Ruler 100 bp DNA ladder GIF 56 kb

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