Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in Glycyrrhiza glabra
Identifieur interne : 000167 ( Pmc/Corpus ); précédent : 000166; suivant : 000168Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in Glycyrrhiza glabra
Auteurs : Suphla Gupta ; Pankaj Pandotra ; Ajai P. Gupta ; M. K. Verma ; Ashok Ahuja ; Ram A. VishwakarmaSource :
- Acta Physiologiae Plantarum [ 0137-5881 ] ; 2013.
Abstract
Direct rhizogenesis from leaf explants and establishment of an in vitro stolon culture system and subsequent plant regeneration for
Url:
DOI: 10.1007/s11738-013-1302-1
PubMed: NONE
PubMed Central: 7089307
Links to Exploration step
PMC:7089307Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in <italic>Glycyrrhiza glabra</italic></title><author><name sortKey="Gupta, Suphla" sort="Gupta, Suphla" uniqKey="Gupta S" first="Suphla" last="Gupta">Suphla Gupta</name><affiliation><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Plant Biotechnology Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Pandotra, Pankaj" sort="Pandotra, Pankaj" uniqKey="Pandotra P" first="Pankaj" last="Pandotra">Pankaj Pandotra</name><affiliation><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Plant Biotechnology Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Gupta, Ajai P" sort="Gupta, Ajai P" uniqKey="Gupta A" first="Ajai P." last="Gupta">Ajai P. Gupta</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Quality Control and Assurance Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Verma, M K" sort="Verma, M K" uniqKey="Verma M" first="M. K." last="Verma">M. K. Verma</name><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Instrumentation Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Ahuja, Ashok" sort="Ahuja, Ashok" uniqKey="Ahuja A" first="Ashok" last="Ahuja">Ashok Ahuja</name><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Biodiversity and Applied Botany Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Vishwakarma, Ram A" sort="Vishwakarma, Ram A" uniqKey="Vishwakarma R" first="Ram A." last="Vishwakarma">Ram A. Vishwakarma</name><affiliation><nlm:aff id="Aff5"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Medicinal Chemistry Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmc">7089307</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7089307</idno><idno type="RBID">PMC:7089307</idno><idno type="doi">10.1007/s11738-013-1302-1</idno><idno type="pmid">NONE</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">000167</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000167</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in <italic>Glycyrrhiza glabra</italic></title><author><name sortKey="Gupta, Suphla" sort="Gupta, Suphla" uniqKey="Gupta S" first="Suphla" last="Gupta">Suphla Gupta</name><affiliation><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Plant Biotechnology Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Pandotra, Pankaj" sort="Pandotra, Pankaj" uniqKey="Pandotra P" first="Pankaj" last="Pandotra">Pankaj Pandotra</name><affiliation><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Plant Biotechnology Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Gupta, Ajai P" sort="Gupta, Ajai P" uniqKey="Gupta A" first="Ajai P." last="Gupta">Ajai P. Gupta</name><affiliation><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Quality Control and Assurance Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Verma, M K" sort="Verma, M K" uniqKey="Verma M" first="M. K." last="Verma">M. K. Verma</name><affiliation><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Instrumentation Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Ahuja, Ashok" sort="Ahuja, Ashok" uniqKey="Ahuja A" first="Ashok" last="Ahuja">Ashok Ahuja</name><affiliation><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Biodiversity and Applied Botany Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author><author><name sortKey="Vishwakarma, Ram A" sort="Vishwakarma, Ram A" uniqKey="Vishwakarma R" first="Ram A." last="Vishwakarma">Ram A. Vishwakarma</name><affiliation><nlm:aff id="Aff5"><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Medicinal Chemistry Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</nlm:aff></affiliation></author></analytic><series><title level="j">Acta Physiologiae Plantarum</title><idno type="ISSN">0137-5881</idno><idno type="eISSN">1861-1664</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Direct rhizogenesis from leaf explants and establishment of an in vitro stolon culture system and subsequent plant regeneration for <italic>Glycyrrhiza glabra</italic> have been described. MS liquid medium supplemented with 0.01 mg l<sup>−1</sup> of NAA was most effective for stolon proliferation. Extensive proliferation of stolon and shoot regeneration was achieved on medium containing 3 % sucrose with 0.01 mg l<sup>−1</sup> NAA. Stolons with nodes showing growth was transferred under light for plantlet regeneration in the same medium. This paper is the first report in <italic>G. glabra</italic> describing a complete regeneration procedure via in vitro stolon proliferation along with quantitative data for glycyrrhizin and genetic fidelity of plant regenerated in vitro there from. In vitro stolon proliferation described here would be an efficient way for regeneration of plants for functional genomics studies and better understanding of glycyrrhizin (GA) metabolism.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Ayabe, S" uniqKey="Ayabe S">S Ayabe</name></author><author><name sortKey="Takano, H" uniqKey="Takano H">H Takano</name></author><author><name sortKey="Fujita, T" uniqKey="Fujita T">T Fujita</name></author><author><name sortKey="Furuya, T" uniqKey="Furuya T">T Furuya</name></author><author><name sortKey="Hirota, H" uniqKey="Hirota H">H Hirota</name></author><author><name sortKey="Takahashi, T" uniqKey="Takahashi T">T Takahashi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cinatl, J" uniqKey="Cinatl J">J Cinatl</name></author><author><name sortKey="Morgenstern, B" uniqKey="Morgenstern B">B Morgenstern</name></author><author><name sortKey="Bauer, G" uniqKey="Bauer G">G Bauer</name></author><author><name sortKey="Chandra, P" uniqKey="Chandra P">P Chandra</name></author><author><name sortKey="Rabenau, H" uniqKey="Rabenau H">H Rabenau</name></author><author><name sortKey="Doerr, Hw" uniqKey="Doerr H">HW Doerr</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Doyle, Jj" uniqKey="Doyle J">JJ Doyle</name></author><author><name sortKey="Doyle, Jl" uniqKey="Doyle J">JL Doyle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Finney, Rsh" uniqKey="Finney R">RSH Finney</name></author><author><name sortKey="Somers, Gf" uniqKey="Somers G">GF Somers</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fiore, C" uniqKey="Fiore C">C Fiore</name></author><author><name sortKey="Eisenhut, M" uniqKey="Eisenhut M">M Eisenhut</name></author><author><name sortKey="Krausse, R" uniqKey="Krausse R">R Krausse</name></author><author><name sortKey="Ragazzi, E" uniqKey="Ragazzi E">E Ragazzi</name></author><author><name sortKey="Pellati, D" uniqKey="Pellati D">D Pellati</name></author><author><name sortKey="Armanini, D" uniqKey="Armanini D">D Armanini</name></author><author><name sortKey="Bielenberg, J" uniqKey="Bielenberg J">J Bielenberg</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Franklin, G" uniqKey="Franklin G">G Franklin</name></author><author><name sortKey="Sheeba, Cj" uniqKey="Sheeba C">CJ Sheeba</name></author><author><name sortKey="Laksmisita, G" uniqKey="Laksmisita G">G Laksmisita</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gupta, V" uniqKey="Gupta V">V Gupta</name></author><author><name sortKey="Kak, A" uniqKey="Kak A">A Kak</name></author><author><name sortKey="Singh, Bb" uniqKey="Singh B">BB Singh</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Hayashi, H" uniqKey="Hayashi H">H Hayashi</name></author><author><name sortKey="Fukui, H" uniqKey="Fukui H">H Fukui</name></author><author><name sortKey="Tabata, M" uniqKey="Tabata M">M Tabata</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Ito, M" uniqKey="Ito M">M Ito</name></author><author><name sortKey="Sato, A" uniqKey="Sato A">A Sato</name></author><author><name sortKey="Hirabayashi, K" uniqKey="Hirabayashi K">K Hirabayashi</name></author><author><name sortKey="Tanabe, F" uniqKey="Tanabe F">F Tanabe</name></author><author><name sortKey="Shigeta, S" uniqKey="Shigeta S">S Shigeta</name></author><author><name sortKey="Baba, M" uniqKey="Baba M">M Baba</name></author><author><name sortKey="Clercq, Ed" uniqKey="Clercq E">ED Clercq</name></author><author><name sortKey="Nakashima, H" uniqKey="Nakashima H">H Nakashima</name></author><author><name sortKey="Yamamoto, N" uniqKey="Yamamoto N">N Yamamoto</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jatav, Vs" uniqKey="Jatav V">VS Jatav</name></author><author><name sortKey="Singh, Sk" uniqKey="Singh S">SK Singh</name></author><author><name sortKey="Khatri, P" uniqKey="Khatri P">P Khatri</name></author><author><name sortKey="Sharma, Ak" uniqKey="Sharma A">AK Sharma</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kojoma, M" uniqKey="Kojoma M">M Kojoma</name></author><author><name sortKey="Ohyama, K" uniqKey="Ohyama K">K Ohyama</name></author><author><name sortKey="Seki, H" uniqKey="Seki H">H Seki</name></author><author><name sortKey="Hiraoka, Y" uniqKey="Hiraoka Y">Y Hiraoka</name></author><author><name sortKey="Asazu, Sn" uniqKey="Asazu S">SN Asazu</name></author><author><name sortKey="Sawa, S" uniqKey="Sawa S">S Sawa</name></author><author><name sortKey="Sekizaki, H" uniqKey="Sekizaki H">H Sekizaki</name></author><author><name sortKey="Yoshida, S" uniqKey="Yoshida S">S Yoshida</name></author><author><name sortKey="Muranaka, T" uniqKey="Muranaka T">T Muranaka</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Li, W" uniqKey="Li W">W Li</name></author><author><name sortKey="Asad, Y" uniqKey="Asad Y">Y Asad</name></author><author><name sortKey="Yoshikawa, T" uniqKey="Yoshikawa T">T Yoshikawa</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mehrotra, S" uniqKey="Mehrotra S">S Mehrotra</name></author><author><name sortKey="Kukreja, Ak" uniqKey="Kukreja A">AK Kukreja</name></author><author><name sortKey="Khanuja, Sps" uniqKey="Khanuja S">SPS Khanuja</name></author><author><name sortKey="Mishra, Bn" uniqKey="Mishra B">BN Mishra</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mehrotra, S" uniqKey="Mehrotra S">S Mehrotra</name></author><author><name sortKey="Khwaja, O" uniqKey="Khwaja O">O Khwaja</name></author><author><name sortKey="Kukreja, Ak" uniqKey="Kukreja A">AK Kukreja</name></author><author><name sortKey="Rahman, L" uniqKey="Rahman L">L Rahman</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mousa, N" uniqKey="Mousa N">N Mousa</name></author><author><name sortKey="Siagura, P" uniqKey="Siagura P">P Siagura</name></author><author><name sortKey="Wiryowidagdo, S" uniqKey="Wiryowidagdo S">S Wiryowidagdo</name></author><author><name sortKey="Wagih, Me" uniqKey="Wagih M">ME Wagih</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Nagat, A" uniqKey="Nagat A">A Nagat</name></author><author><name sortKey="Mousa, Tr" uniqKey="Mousa T">TR Mousa</name></author><author><name sortKey="Siaguru, I" uniqKey="Siaguru I">I Siaguru</name></author><author><name sortKey="Wiryowidagdo, S" uniqKey="Wiryowidagdo S">S Wiryowidagdo</name></author><author><name sortKey="Wagih, Me" uniqKey="Wagih M">ME Wagih</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Park, Hy" uniqKey="Park H">HY Park</name></author><author><name sortKey="Park, Sh" uniqKey="Park S">SH Park</name></author><author><name sortKey="Yoon, Hk" uniqKey="Yoon H">HK Yoon</name></author><author><name sortKey="Han, Mj" uniqKey="Han M">MJ Han</name></author><author><name sortKey="Kim, Dh" uniqKey="Kim D">DH Kim</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Saito, K" uniqKey="Saito K">K Saito</name></author><author><name sortKey="Kaneko, H" uniqKey="Kaneko H">H Kaneko</name></author><author><name sortKey="Yamazaki, M" uniqKey="Yamazaki M">M Yamazaki</name></author><author><name sortKey="Yoshida, M" uniqKey="Yoshida M">M Yoshida</name></author><author><name sortKey="Murakoshi, I" uniqKey="Murakoshi I">I Murakoshi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shah, Rr" uniqKey="Shah R">RR Shah</name></author><author><name sortKey="Dalal, Kc" uniqKey="Dalal K">KC Dalal</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shirazi, Z" uniqKey="Shirazi Z">Z Shirazi</name></author><author><name sortKey="Piri, K" uniqKey="Piri K">K Piri</name></author><author><name sortKey="Asl, Am" uniqKey="Asl A">AM Asl</name></author><author><name sortKey="Hasanloo, T" uniqKey="Hasanloo T">T Hasanloo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Thengane, R" uniqKey="Thengane R">R Thengane</name></author><author><name sortKey="Kulkarni, Dk" uniqKey="Kulkarni D">DK Kulkarni</name></author><author><name sortKey="Krishnamurthy, Kv" uniqKey="Krishnamurthy K">KV Krishnamurthy</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Toivonen, L" uniqKey="Toivonen L">L Toivonen</name></author><author><name sortKey="Rosenqvist, H" uniqKey="Rosenqvist H">H Rosenqvist</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Wongwicha, W" uniqKey="Wongwicha W">W Wongwicha</name></author><author><name sortKey="Tanaka, H" uniqKey="Tanaka H">H Tanaka</name></author><author><name sortKey="Shoyama, Y" uniqKey="Shoyama Y">Y Shoyama</name></author><author><name sortKey="Tuvshintogtokh, I" uniqKey="Tuvshintogtokh I">I Tuvshintogtokh</name></author><author><name sortKey="Putalun, W" uniqKey="Putalun W">W Putalun</name></author></analytic></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Zhang, Mz" uniqKey="Zhang M">MZ Zhang</name></author><author><name sortKey="Jie, X" uniqKey="Jie X">X Jie</name></author><author><name sortKey="Bing, Y" uniqKey="Bing Y">Y Bing</name></author><author><name sortKey="Huiyong, Y" uniqKey="Huiyong Y">Y Huiyong</name></author><author><name sortKey="Qiuyin, C" uniqKey="Qiuyin C">C Qiuyin</name></author><author><name sortKey="Martha, J" uniqKey="Martha J">J Martha</name></author><author><name sortKey="Shrubsole, Xc" uniqKey="Shrubsole X">XC Shrubsole</name></author><author><name sortKey="Valentina, K" uniqKey="Valentina K">K Valentina</name></author><author><name sortKey="Wei, Z" uniqKey="Wei Z">Z Wei</name></author><author><name sortKey="Ambra, P" uniqKey="Ambra P">P Ambra</name></author><author><name sortKey="Harris, Rc" uniqKey="Harris R">RC Harris</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Zietkiewicz, E" uniqKey="Zietkiewicz E">E Zietkiewicz</name></author><author><name sortKey="Rafalski, A" uniqKey="Rafalski A">A Rafalski</name></author><author><name sortKey="Labuda, D" uniqKey="Labuda D">D Labuda</name></author></analytic></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Acta Physiol Plant</journal-id><journal-id journal-id-type="iso-abbrev">Acta Physiol. Plant</journal-id><journal-title-group><journal-title>Acta Physiologiae Plantarum</journal-title></journal-title-group><issn pub-type="ppub">0137-5881</issn><issn pub-type="epub">1861-1664</issn><publisher><publisher-name>Springer Berlin Heidelberg</publisher-name><publisher-loc>Berlin/Heidelberg</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmc">7089307</article-id><article-id pub-id-type="publisher-id">1302</article-id><article-id pub-id-type="doi">10.1007/s11738-013-1302-1</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Paper</subject></subj-group></article-categories><title-group><article-title>Direct rhizogenesis, in vitro stolon proliferation and high-throughput regeneration of plantlets in <italic>Glycyrrhiza glabra</italic></article-title></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name><surname>Gupta</surname><given-names>Suphla</given-names></name><address><fax>+91-0191-2569019</fax><email>suphlabg@gmail.com</email><email>sgupta@iiim.ac.in</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Pandotra</surname><given-names>Pankaj</given-names></name><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Gupta</surname><given-names>Ajai P.</given-names></name><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Verma</surname><given-names>M. K.</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Ahuja</surname><given-names>Ashok</given-names></name><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author"><name><surname>Vishwakarma</surname><given-names>Ram A.</given-names></name><xref ref-type="aff" rid="Aff5">5</xref></contrib><aff id="Aff1"><label>1</label><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Plant Biotechnology Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</aff><aff id="Aff2"><label>2</label><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Quality Control and Assurance Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</aff><aff id="Aff3"><label>3</label><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Instrumentation Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</aff><aff id="Aff4"><label>4</label><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Biodiversity and Applied Botany Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</aff><aff id="Aff5"><label>5</label><institution-wrap><institution-id institution-id-type="GRID">grid.418225.8</institution-id><institution-id institution-id-type="ISNI">0000000418026428</institution-id><institution>Medicinal Chemistry Division,</institution><institution>CSIR-Indian Institute of Integrative Medicine,</institution></institution-wrap>Jammu, 180 001 India</aff></contrib-group><author-notes><fn fn-type="com"><p>Communicated by S. Werbrouck.</p></fn></author-notes><pub-date pub-type="epub"><day>21</day><month>5</month><year>2013</year></pub-date><pub-date pub-type="ppub"><year>2013</year></pub-date><volume>35</volume><issue>9</issue><fpage>2699</fpage><lpage>2705</lpage><history><date date-type="received"><day>24</day><month>1</month><year>2013</year></date><date date-type="rev-recd"><day>25</day><month>3</month><year>2013</year></date><date date-type="accepted"><day>8</day><month>5</month><year>2013</year></date></history><permissions><copyright-statement>© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2013</copyright-statement><license><license-p>This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.</license-p></license></permissions><abstract id="Abs1"><p>Direct rhizogenesis from leaf explants and establishment of an in vitro stolon culture system and subsequent plant regeneration for <italic>Glycyrrhiza glabra</italic> have been described. MS liquid medium supplemented with 0.01 mg l<sup>−1</sup> of NAA was most effective for stolon proliferation. Extensive proliferation of stolon and shoot regeneration was achieved on medium containing 3 % sucrose with 0.01 mg l<sup>−1</sup> NAA. Stolons with nodes showing growth was transferred under light for plantlet regeneration in the same medium. This paper is the first report in <italic>G. glabra</italic> describing a complete regeneration procedure via in vitro stolon proliferation along with quantitative data for glycyrrhizin and genetic fidelity of plant regenerated in vitro there from. In vitro stolon proliferation described here would be an efficient way for regeneration of plants for functional genomics studies and better understanding of glycyrrhizin (GA) metabolism.</p></abstract><kwd-group xml:lang="en"><title>Keywords</title><kwd>Acclimatization</kwd><kwd>Glycyrrhizin</kwd><kwd>ISSR</kwd><kwd>Root induction</kwd></kwd-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2013</meta-value></custom-meta></custom-meta-group></article-meta></front><body><sec id="Sec1" sec-type="introduction"><title>Introduction</title><p><italic>Glycyrrhiza</italic> (<italic>Fabaceae</italic>), commonly known as licorice, is an important source of flavonoids and terpenoids class of compounds, important among them is glycyrrhizin (GA). Roots and stolons of <italic>Glycyrrhiza</italic> plants (<italic>G. uralensis</italic> Fisch. and <italic>G. glabra</italic> L., Fabaceae) are highly valued commercial products used in oriental medicine (Anonymous <xref ref-type="bibr" rid="CR1">2005</xref>). In modern medicine it is considered an important crude drug for its various pharmacological activities (Jatav et al. <xref ref-type="bibr" rid="CR12">2011</xref>) including anti-diabetic, anti-inflammatory (Finney and Somers <xref ref-type="bibr" rid="CR5">1958</xref>), hepato-protective, anti-ulcer and anti-allergic (Park et al. <xref ref-type="bibr" rid="CR19">2004</xref>). Glycyrrhizin also has antiviral activity (Fiore et al. <xref ref-type="bibr" rid="CR6">2008</xref>) against various DNA and RNA viruses, including HIV (Ito et al. <xref ref-type="bibr" rid="CR11">1988</xref>) and severe acute respiratory syndrome (SARS)-associated <italic>corona virus</italic> (Cinatl et al. <xref ref-type="bibr" rid="CR3">2003</xref>). In recent studies it is shown to selectively block the tumor COX-2 pathway and suppress colon carcinogenesis in mice and humans (Zhang et al. <xref ref-type="bibr" rid="CR27">2009</xref>).</p><p>Glycyrrhizin, an oleanane type terpenoid saponin, and its derivative compounds are exclusively obtained from the dried roots and stolons of licorice. Chemical analyses have failed to detect them in the aerial part (Hayashi et al. <xref ref-type="bibr" rid="CR9">1988</xref>); the plants have been indiscriminately exploited to meet the high demand, resulting in desertification of the habitat. The plant is naturally found in areas restricted to Central and South China, Russia and Mediterranean regions. Under cultivation, the plant is rarely propagated by seeds due to its hard testa (Gupta et al. <xref ref-type="bibr" rid="CR8">1997</xref>).Vegetative propagation of the plant is annual and germination percentage is low, being highly under the influence of environmental conditions.</p><p>Developing in vitro regeneration methods of <italic>Glycyrrhiza glabra</italic> utilizing different explants viz, stem segments (Thengane et al. <xref ref-type="bibr" rid="CR23">1998</xref>) and axillary bud (Shah and Dalal <xref ref-type="bibr" rid="CR21">1980</xref>) to produce multiple shoots (Henry et al. <xref ref-type="bibr" rid="CR10">1991</xref>), shoot regeneration from callus (Wongwicha et al. <xref ref-type="bibr" rid="CR25">2008</xref>) shoots differentiation and clonal propagation (Mousa et al. <xref ref-type="bibr" rid="CR17">2006</xref>) has been the subject of several studies. Recent study by Mehrotra et al. (<xref ref-type="bibr" rid="CR16">2012</xref>) has demonstrated the potential of encapsulated in vitro-grown axillary micro-shoots of <italic>Glycyrrhiza glabra</italic> in long-term storage and year round availability of germplasm for commercial cultivation. Hairy root induction using <italic>Agrobacterium</italic><italic>rhizogenes</italic> have been recently developed in <italic>G. glabra</italic> (Li et al. <xref ref-type="bibr" rid="CR14">2000</xref>; Shirazi et al. <xref ref-type="bibr" rid="CR22">2012</xref>) showing the feasibility to cultivate these transformed roots in air lift bioreactors (Mehrotra et al. <xref ref-type="bibr" rid="CR15">2008</xref>). Tissue culture techniques have raised expectation regarding possibility of their application for propagation and understanding of glycyrrhizin biosynthesis. Tissue culture method to propagate <italic>G. glabra</italic> on commercial scale has also been reported by Shah and Dalal (<xref ref-type="bibr" rid="CR21">1980</xref>). Recent studies regarding glycyrrhizin production by in vitro cultures have shown contrasting and conflicting results. Most of the attempts to detect glycyrrhizin from cultured tissues of <italic>G. glabra</italic> have been unsuccessful (Hayashi et al. <xref ref-type="bibr" rid="CR9">1988</xref>; Saito et al. <xref ref-type="bibr" rid="CR20">1990</xref>; Ayabe et al. <xref ref-type="bibr" rid="CR2">1990</xref>; Toivonen and Rosenqvist <xref ref-type="bibr" rid="CR24">1995</xref>). Callus and cell suspension cultures of selected <italic>G. glabra</italic> clones have shown production of glycyrrhizin, detected using TLC-tentative analysis (Nagat et al. <xref ref-type="bibr" rid="CR18">2007</xref>). Similarly, Wongwicha et al. (<xref ref-type="bibr" rid="CR25">2008</xref>) reported glycyrrhizin production in callus cultures of <italic>G. uralensis</italic> (36.52 mg g<sup>−1</sup>), <italic>G. glabra</italic> (14.39 mg g<sup>−1</sup>), and <italic>G. inflata</italic> (16.14 mg g<sup>−1</sup>).</p><p>Although development of in vitro regeneration procedures utilizing various explants in <italic>G. glabra</italic> have been the subject of many studies, most of these reports lacked information on the molecular stability and biochemistry of the plant regenerated using tissue culture procedures. Inter-simple sequence repeats (ISSR) was chosen as molecular marker in the present study because this technique relies on the high polymorphism and can detect low differentiation levels having more stringent primer annealing conditions than RAPDs leading to higher accuracy and reproducibility (Zietkiewicz et al. <xref ref-type="bibr" rid="CR28">1994</xref>).</p><p>The present study focuses on utilization of root culture, stolon proliferation for high-throughput plant regeneration and analyze glycyrrhizin and related terpenoids accumulation in root cultures and in vitro-proliferated stolons. Additionally, molecular analysis of regenerated plants using ISSR markers have also been undertaken to assess genetic uniformity.</p></sec><sec id="Sec2" sec-type="materials|methods"><title>Materials and methods</title><sec id="Sec3"><title>Plant material</title><p>A selected clone of <italic>G. glabra</italic> plants collected from agricultural farms of Himachal Pradesh Agricultural University, Solan, located in the north Himalayan region (30.92°N and 77.15°E) of Indian subcontinent was utilized for the present study.</p></sec><sec id="Sec4"><title>Direct rhizogenesis from foliar explants</title><p>Fully expanded young leaves of the <italic>G. glabra</italic> plant were collected and washed under tap water. Whole leaf was surface sterilized with 0.1 % HgCl<sub>2</sub> for 5 min. The leaves were further rinsed with sterile water five times and inoculated on culture medium. Plantlets were regenerated in vitro using shooting and rooting mediums. The leaves of the in vitro raised plants were used for studying the prospect of direct root initiation and plant regeneration. Young, soft leaves measuring 0.5–1 cm<sup>2</sup> were obtained from 5-week-old in vitro raised plantlets along with petiole under aseptic conditions and cultured on MS (Murashige and Skoog 1962) medium containing 3 % sucrose and supplemented with cascade of growth regulators (IBA, Kinetin, IAA and 2,4-D) as shown in Table <xref rid="Tab1" ref-type="table">1</xref>. The pH of the medium was adjusted to 5.8 and autoclaved at 121 °C under pressure of 1.2 kg cm<sup>−1</sup> for 15 min. First observation was recorded after 14 days of culturing and subsequent observations were made every week thereon. Primary cultures showing rhizogenesis (Fig. <xref rid="Fig1" ref-type="fig">1</xref>a, b) were transferred on to stolen proliferation liquid MS medium (3 % sucrose, 0.01 mg l<sup>−1</sup> NAA), incubated in dark and grown at 28° ± 2 °C on a rotary shaker at 100 rpm.
<table-wrap id="Tab1"><label>Table 1</label><caption><p>Effect of various plant growth regulators in MS medium from leaf explants of <italic>Glycyrrhiza glabra</italic> (after 3 weeks)</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left">PGR/(mg ml<sup>−1</sup>)</th><th align="left">NAA</th><th align="left">IAA</th><th align="left">IBA</th><th align="left">2,4 D</th><th align="left">BAP</th><th align="left">Kinetin</th><th align="left">Mean no. of roots</th><th align="left">Response (%)</th><th align="left">Morphology</th></tr></thead><tbody><tr><td align="left" rowspan="15"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">5</td><td align="left">100</td><td align="left">R + C</td></tr><tr><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">3</td><td align="left">60</td><td align="left">C</td></tr><tr><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">3</td><td align="left">60</td><td align="left">C</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">l</td><td align="left"></td><td align="left">2</td><td align="left">60</td><td align="left">NR</td></tr><tr><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">4</td><td align="left">80</td><td align="left">R</td></tr><tr><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">5</td><td align="left">100</td><td align="left">R</td></tr><tr><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">5</td><td align="left">100</td><td align="left">R</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">1</td><td align="left">2</td><td align="left">40</td><td align="left">C</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left">–</td><td align="left">–</td><td align="left">–</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">–</td><td align="left">–</td><td align="left">–</td></tr><tr><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left">20</td><td align="left">R</td></tr><tr><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">3</td><td align="left">60</td><td align="left">R + C</td></tr><tr><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">3</td><td align="left">60</td><td align="left">R</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">1</td><td align="left"></td><td align="left"></td><td align="left">2</td><td align="left">40</td><td align="left">C</td></tr><tr><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left"></td><td align="left">–</td><td align="left">–</td><td align="left">NR</td></tr></tbody></table><table-wrap-foot><p><italic>R</italic> roots, <italic>C</italic> callus, <italic>NR</italic> no response</p></table-wrap-foot></table-wrap><fig id="Fig1"><label>Fig. 1</label><caption><p>In vitro stolon organogenesis and plant regeneration from cultured stolons in <italic>Glychyrriza glabra</italic>. <bold>a</bold> Adventitious root induction from leaf explants; <bold>b</bold> root cultures and stolon proliferation from cultured roots in MS medium containing 3 % sucrose, 0.01 mg l<sup>−1</sup> NAA; <bold>c</bold>, <bold>d</bold> stolon segments and shoot differentiation with plantlet regeneration; <bold>e</bold>, <bold>f</bold> regenerated plantlets after acclimatization (3 months); <bold>g</bold> ISSR profile of ten randomly selected regenerated plants from roots; <bold>h</bold> LC–MS chromatogram showing peaks of glycyrrhizin analysis of cultured stolons</p></caption><graphic xlink:href="11738_2013_1302_Fig1_HTML" id="MO1"></graphic></fig></p></sec><sec id="Sec5"><title>De novo shoot proliferation from cultured stolons</title><p>The stolon cultures grown in dark at 28 ± 2 °C under shaking conditions were transferred to light under a 16-h photoperiod provided by cool white fluorescent tubes (Phillips, India) having a light intensity of 40 μmol m<sup>−2</sup> s<sup>−1</sup> (Fig. <xref rid="Fig1" ref-type="fig">1</xref>c, d)</p></sec><sec id="Sec6"><title>Transfer of plant to soil</title><p>Plantlets with well developed shoot and root systems were transferred to the green house for hardening. Roots of plantlets regenerated in vitro were washed thoroughly to remove agar and planted into plastic pots containing sterilized sand–soil mixture (2:1). They were kept covered for the first 2 weeks and watered twice a week in the first week using ½ MS stock solution followed by sterile water in the subsequent week on the alternative days (Fig. <xref rid="Fig1" ref-type="fig">1</xref>e, f)</p></sec></sec><sec id="Sec7"><title>Molecular and biochemical analysis</title><sec id="Sec8"><title>Molecular analysis</title><p>The plants regenerated via stolons were selected randomly for assessing genetic stability using five ISSR markers. Young fresh leaves of the ten selected in vitro regenerated plants, hardened under glass house conditions, were utilized for molecular analysis. High-quality DNA was extracted following CTAB method (Doyle and Doyle <xref ref-type="bibr" rid="CR4">1987</xref>) with slight modifications. The quality and quantity of the DNA was ascertained by spectrophotometer (Nanodrop, Thermo scientific) and agarose gel electrophoresis. PCR reactions were carried out in a 10× reaction buffer with 1.5 mM MgCl<sub>2</sub>, 1 mM dNTP, 10 pmol of primer, 25 ng of genomic DNA for 20 μl reaction volume with 1 Unit of <italic>Taq</italic> DNA polymerase. Amplification was performed in a thermal cycler (Eppendorf, Vapo protect), with one cycle of initial denaturation at 95 °C for 5 min, followed by 34 cycles at 94 °C for 30 s, annealing at their respective temperature (40–45 °C) for 2 min and extension at 72 °C for 1 min followed by final extension at 72 °C for 8 min. PCR products were separated on 1.5 % agarose gel containing 1× TAE and stained with ethidium bromide (0.5 μg ml<sup>−1</sup>). A 100 bp DNA ladder (Fermentas) was used as a standard molecular weight marker. Banding pattern was photographed and stored as digital picture in gel documentation system (Syngene, USA). Each experiment was repeated thrice (Fig. <xref rid="Fig1" ref-type="fig">1</xref>g).</p></sec><sec id="Sec9"><title>Biochemical analysis</title><sec id="Sec10"><title>Chemicals and reagents</title><p>ACS certified HPLC-grade solvents supplied by JT Baker, India, were used in the analysis. Pure glycyrrhizin (GA) was purchased from Sigma, India. Water was purified using Milli-Q-plus filter systems (Millipore, Bedford, MA, USA).</p></sec></sec><sec id="Sec11"><title>Ultrasound-assisted extraction of glycyrrhizin</title><p>Green and brown stolons and root tuft were weighed and crushed using mortar, extracted for 10 min using 2 ml of 50 % ethanol under ultra-sonication (Elma, Germany), for 30 min at 45 MHz, at room temperature, twice. The extract was filtered (Millipore filter 0.45 μm) concentrated on a rotary-evaporator (Buchi, Switzerland) and 2 ml of solvent was added to the sample before analysis.</p></sec><sec id="Sec12"><title>Glycyrrhizin detection and quantification</title><p>For the identification and quantification of glycyrrhizin present in in vitro raised stolons, a UPLC-qTOF-MS system (Synapt, Waters, USA, equipped with MassLynx acquisition software, version 4.1) was used. The system consisted of: column, C-18 (50 mm × 2.1 mm) with particle size 1.7 μm (Acquity, BEH) having mobile phase of acetonitrile and acetic acid (2 %) in water (75:25, v/v), at a flow rate of 200 μl min<sup>−1</sup>. The injection volume was 1 μl. Separation of the analyte was achieved at retention time of 1.17 min. Column and sample temperature were 30 and 10 °C, respectively. Infusion experiments were performed using an in-built syringe pump. A UPLC–qTOF-MS with ESI interface was used for MS/MS analysis, with following parameters: capillary voltage, 2.7 kV for positive mode; source temperature, 100 °C; desolvation temperature, 200 °C; cone gas flow, 50 l h<sup>−1</sup> and desolvation gas flow, 550 l h<sup>−1</sup>. The multiple reaction monitoring (MRM) mode was used to monitor the transition of glycyrrhizin <italic>mz</italic><sup>−1</sup> 845.1873 [M+Na], 867.1644 [M+K] 823.2040 [M+H] (Fig. <xref rid="Fig1" ref-type="fig">1</xref>h).</p></sec></sec><sec id="Sec13"><title>Results and discussion</title><p>Roots could be initiated on MS medium enriched with three different growth hormones (IBA, NAA and Kn) displaying well-grown, white roots after 2 weeks of culturing of leaf explants. Roots mostly induced at the proximal end of the whole leaf cultured on medium containing combination of Kn (1.0 mg l<sup>−1</sup>) with NAA or IBA (1.0 mg l<sup>−1</sup>) and IAA (1.0 mg l<sup>−1</sup>) alone (Fig. <xref rid="Fig1" ref-type="fig">1</xref>a, b). Addition of cytokinin or BAP along with IAA or NAA resulted in synergistic influence on evoking response on the leaf segments. The root culture proliferates very rapidly on transferring roots in liquid medium with 0.01 mg l<sup>−1</sup> NAA, with the passage of time roots transferred to liquid medium assumed a mat like appearance within a period of 3 weeks. Repeated sub-culturing demonstrated autonomy of root meristem which grew vigorously as a whitish cottony mass under dark incubation. Among the five different PGR combinations—(1) NAA (1.0 mg l<sup>−1</sup>) + Kn (1.0 mg l<sup>−1</sup>); (2) IBA (1.0 mg l<sup>−1</sup>) + Kn (1.0 mg l<sup>−1</sup>); (3) NAA (1.0 mg l<sup>−1</sup>); (4) IAA (1.0 mg l<sup>−1</sup>) and (5) 2,4-D (1.0 mg l<sup>−1</sup>)—root development was most vigorous in MS medium supplemented with IAA or IBA with kinetin (Table <xref rid="Tab1" ref-type="table">1</xref>). With the passage of time elongated roots slowly got transformed into thick and stout pale white or light brown stolons in medium under dark conditions. Several shoot primordial could be observed on stolons within 3 weeks on MS medium containing 3 % sucrose with 1.0 mg l<sup>−1</sup> NAA. Medium supplemented with NAA (Fig. <xref rid="Fig1" ref-type="fig">1</xref>c) was found most suitable for shoot differentiation. Root differentiation and whole plantlet regeneration was observed after 2 subculture passages within 3 weeks when kept under light (Fig. <xref rid="Fig1" ref-type="fig">1</xref>c, d). Cultured stolons retained characteristic color similar to field grown stolons. Shoot bud and its differentiation in well developed shoots occurred mostly on middle portion of in vitro stolon which indicated that certain degree of maturity of stolon is required prior to shoot differentiation. On transferring, segments of cultured stolons to MS media treatment containing NAA (0.01 mg l<sup>−1)</sup> resulted in >8 shoots from each stolon segment (3 cm). The shoots obtained were found to root efficiently and resulted in complete plantlets. These were further maintained on the same medium under illumination. Plantlets were successfully acclimatized to soil in pots with 70 % survival, plants grew normal and vigorously under green house conditions (Fig. <xref rid="Fig1" ref-type="fig">1</xref>e, f; Table <xref rid="Tab2" ref-type="table">2</xref>).The present described procedure is being reported for the first time for <italic>G. glabra</italic> where root cultures established in vitro could be effectively utilized to retrieve high-frequency plant regeneration via stolon proliferation. The present results provide protocol for complete in vitro regeneration utilizing excised in vitro regenerated roots leading to stolon proliferation and subsequent in vitro regeneration in <italic>G. glabra</italic>. Using present procedure 20–25 plants/stolon could be produced within 3 months. The root cultures are advantageous over other explants in terms of their easy manipulation and high regenerative potential (Franklin et al. <xref ref-type="bibr" rid="CR7">2004</xref>). However, such report in other species of <italic>Glycyrrhiza</italic> (<italic>G. uralensis</italic>) is available (Kojoma et al. <xref ref-type="bibr" rid="CR13">2010</xref>). The present described method holds promise in being simple, reproducible and high throughput for mass cloning of ‘elite’ genotypes of <italic>G. glabra.</italic> The application of developed procedure could be in providing large number of <italic>G. glabra</italic> plants in comparison to earlier published reports (Shah and Dalal <xref ref-type="bibr" rid="CR21">1980</xref>; Henry et al. <xref ref-type="bibr" rid="CR10">1991</xref>; Mousa et al. <xref ref-type="bibr" rid="CR17">2006</xref>). <table-wrap id="Tab2"><label>Table 2</label><caption><p>Performance of <italic>Glycyrrhiza glabra</italic> plants regenerated from in vitro cultured stolons under acclimatization chamber mist-irrigated conditions</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left">Plant</th><th align="left">Height (cm)</th><th align="left">Nodes (cm)</th><th align="left">Internode (cm)</th><th align="left">Leaves (number)</th></tr></thead><tbody><tr><td align="left">G1</td><td char="±" align="char">9.2 ± 0.010</td><td char="±" align="char">6.0 ± 0.009</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">16.2 ± 0.001</td></tr><tr><td align="left">G2</td><td char="±" align="char">6.5 ± 0.02</td><td char="±" align="char">8.1 ± 0.000</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">23.1 ± 0.030</td></tr><tr><td align="left">G3</td><td char="±" align="char">12.0 ± 0.001</td><td char="±" align="char">8.3 ± 0.000</td><td char="±" align="char">2.2 ± 0.003</td><td char="±" align="char">25.4 ± 0.039</td></tr><tr><td align="left">G4</td><td char="±" align="char">13.3 ± 4.002</td><td char="±" align="char">9.3 ± 0.003</td><td char="±" align="char">2.0 ± 0.003</td><td char="±" align="char">25.3 ± 0.039</td></tr><tr><td align="left">G5</td><td char="±" align="char">17.2 ± 2.021</td><td char="±" align="char">13.2 ± 0.021</td><td char="±" align="char">1.2 ± 0.001</td><td char="±" align="char">33.3 ± 0.075</td></tr><tr><td align="left">G6</td><td char="±" align="char">11.5 ± 0.003</td><td char="±" align="char">8.8 ± 0.002</td><td char="±" align="char">1.2 ± 0.000</td><td char="±" align="char">24.4 ± 0.036</td></tr><tr><td align="left">G7</td><td char="±" align="char">12.0 ± 3.001</td><td char="±" align="char">11.2 ± 0.012</td><td char="±" align="char">2.3 ± 0.003</td><td char="±" align="char">22.3 ± 0.025</td></tr><tr><td align="left">G8</td><td char="±" align="char">15.1 ± 1.012</td><td char="±" align="char">13.3 ± 0.021</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">17.2 ± 0.002</td></tr><tr><td align="left">G9</td><td char="±" align="char">13.3 ± 2.002</td><td char="±" align="char">7.1 ± 0.005</td><td char="±" align="char">1.5 ± 0.001</td><td char="±" align="char">14.1 ± 0.010</td></tr><tr><td align="left">G10</td><td char="±" align="char">19.6 ± 1.030</td><td char="±" align="char">17.5 ± 0.040</td><td char="±" align="char">2.3 ± 0.003</td><td char="±" align="char">24.3 ± 0.034</td></tr><tr><td align="left">G11</td><td char="±" align="char">17.5 ± 3.021</td><td char="±" align="char">11.3 ± 0.012</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">21.2 ± 0.021</td></tr><tr><td align="left">G12</td><td char="±" align="char">15.2 ± 0.012</td><td char="±" align="char">11.8 ± 0.016</td><td char="±" align="char">1.2 ± 0.000</td><td char="±" align="char">19.6 ± 0.014</td></tr><tr><td align="left">G13</td><td char="±" align="char">17.0 ± 1.021</td><td char="±" align="char">8.1 ± 0.000</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">17.3 ± 0.002</td></tr><tr><td align="left">G14</td><td char="±" align="char">12.3 ± 6.001</td><td char="±" align="char">7.1 ± 0.005</td><td char="±" align="char">1.0 ± 0.001</td><td char="±" align="char">12.3 ± 0.019</td></tr><tr><td align="left">G15</td><td char="±" align="char">15.1 ± 5.012</td><td char="±" align="char">13.2 ± 0.021</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">12.3 ± 0.019</td></tr><tr><td align="left">G15</td><td char="±" align="char">17.0 ± 8.021</td><td char="±" align="char">8.1 ± 0.000</td><td char="±" align="char">1.3 ± 0.001</td><td char="±" align="char">17.3 ± 0.002</td></tr><tr><td align="left">G16</td><td char="±" align="char">27.1 ± 6.035</td><td char="±" align="char">8.0 ± 0.000</td><td char="±" align="char">0.5 ± 0.001</td><td char="±" align="char">27.3 ± 0.025</td></tr><tr><td align="left">G17</td><td char="±" align="char">17.6 ± 0.023</td><td char="±" align="char">8.8 ± 0.002</td><td char="±" align="char">0.7 ± 0.002</td><td char="±" align="char">17.0 ± 0.002</td></tr><tr><td align="left">G18</td><td char="±" align="char">7.8 ± 3.024</td><td char="±" align="char">6.5 ± 0.009</td><td char="±" align="char">2.3 ± 0.003</td><td char="±" align="char">13.1 ± 0.015</td></tr><tr><td align="left">G19</td><td char="±" align="char">8.2 ± 4.019</td><td char="±" align="char">6.2 ± 0.009</td><td char="±" align="char">1.5 ± 0.000</td><td char="±" align="char">12.3 ± 0.019</td></tr><tr><td align="left">G20</td><td char="±" align="char">13.4 ± 0.002</td><td char="±" align="char">7.0 ± 0.005</td><td char="±" align="char">1.3 ± 0.001</td><td char="±" align="char">14.2 ± 0.010</td></tr><tr><td align="left">G21</td><td char="±" align="char">5.0 ± 3.033</td><td char="±" align="char">4.1 ± 0.018</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">5.3 ± 0.051</td></tr><tr><td align="left">G22</td><td char="±" align="char">5.8 ± 6.033</td><td char="±" align="char">5.2 ± 0.014</td><td char="±" align="char">0.5 ± 0.003</td><td char="±" align="char">10.1 ± 0.028</td></tr><tr><td align="left">G23</td><td char="±" align="char">7.6 ± 0.021</td><td char="±" align="char">5.6 ± 0.011</td><td char="±" align="char">1.1 ± 0.000</td><td char="±" align="char">10.8 ± 0.025</td></tr><tr><td align="left">G24</td><td char="±" align="char">9.0 ± 3.007</td><td char="±" align="char">7.1 ± 0.002</td><td char="±" align="char">2.6 ± 0.001</td><td char="±" align="char">14.2 ± 0.005</td></tr><tr><td align="left">G25</td><td char="±" align="char">5.6 ± 7.033</td><td char="±" align="char">3.3 ± 0.023</td><td char="±" align="char">1.5 ± 0.000</td><td char="±" align="char">5.3 ± 0.051</td></tr><tr><td align="left">G26</td><td char="±" align="char">14.5 ± 0.007</td><td char="±" align="char">10.2 ± 0.008</td><td char="±" align="char">1.5 ± 0.000</td><td char="±" align="char">12.2 ± 0.019</td></tr><tr><td align="left">G27</td><td char="±" align="char">13.2 ± 0.002</td><td char="±" align="char">6.3 ± 0.009</td><td char="±" align="char">3.3 ± 0.007</td><td char="±" align="char">7.5 ± 0.042</td></tr><tr><td align="left">G28</td><td char="±" align="char">8.2 ± 0.01</td><td char="±" align="char">3.5 ± 0.012</td><td char="±" align="char">3.0 ± 0.004</td><td char="±" align="char">12.3 ± 0.010</td></tr><tr><td align="left">G29</td><td char="±" align="char">9.8 ± 0.011</td><td char="±" align="char">5.8 ± 0.010</td><td char="±" align="char">2.2 ± 0.004</td><td char="±" align="char">10.2 ± 0.028</td></tr></tbody></table><table-wrap-foot><p>RH, 80–90 %; temperature 25 ± 2 °C; potting mixture, sand/soil (2:1); mist-irrigated conditions</p></table-wrap-foot></table-wrap></p><p>Regenerated plantlets derived through stolon proliferation were subjected to molecular analysis in order to evaluate their genetic uniformity and/or detect likely existing variations among in vitro-derived plantlets. Ten ISSR primers were used for evaluating ISSR fingerprinting successfully validated true-to- type of in vitro raised regenerates (Fig. <xref rid="Fig1" ref-type="fig">1</xref>g). As such the described method could be used for mass cloning with minimum possibility of any in vitro-induced variability.</p><p>Glycyrrhizin production in three stages of in vitro raised stolons (young, mature and root tuft) confirmed the presence of glycyrrhizin in cultivated stolons using sensitive LC–MS analytical technique. However, quantitative differences in the glycyrrhizin content were observed in all the three stages. It was interesting to note that green immature stolon produced minimum glycyrrhizin (4.0 μg g<sup>−1</sup>) while maximum content (4.7 μg g<sup>−1</sup>) was detected in mature brown stolon in the present study (Table <xref rid="Tab3" ref-type="table">3</xref>). Previous reports of Kojoma et al. (<xref ref-type="bibr" rid="CR13">2010</xref>) detected the glycyrrhizin production in cultured stolons of <italic>G. uralensis</italic> support our present results. However, earlier published reports of glycyrrhizin production in tissue cultures of <italic>G. glabra</italic> were conflicting. Yoo and Kim (<xref ref-type="bibr" rid="CR26">1976</xref>) reported glycyrrhizin production by callus of <italic>G.</italic><italic>glabra</italic>. Later Hayashi et al. (<xref ref-type="bibr" rid="CR9">1988</xref>), Ayabe et al. (<xref ref-type="bibr" rid="CR2">1990</xref>) and Saito et al. (<xref ref-type="bibr" rid="CR20">1990</xref>) failed to detect any glycyrrhizin in <italic>G. glabra.</italic><table-wrap id="Tab3"><label>Table 3</label><caption><p>Glycyrrhizin content in various culture lines in <italic>Glycyrrhiza glabra</italic></p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left">Experimental material</th><th align="left">Glycyrrhizin content (μg g<sup>−1</sup>)</th></tr></thead><tbody><tr><td align="left">Immature stolon</td><td align="left">4.00 ± 0.03</td></tr><tr><td align="left">Mature stolon</td><td align="left">4.70 ± 0.06</td></tr><tr><td align="left">Root tuft</td><td align="left">4.21 ± 0.07</td></tr></tbody></table><table-wrap-foot><p>Values are expressed as mean ± SE (<italic>n</italic> = 3)</p></table-wrap-foot></table-wrap></p></sec><sec id="Sec14" sec-type="conclusion"><title>Conclusion</title><p>In conclusion, the present described strategies were found to be most effective in induction of stolons which proliferated and extended high-throughput regeneration procedure in <italic>Glycyrrhiza glabra</italic> to obtain ideal true-to-type regenerants. Accumulation of glycyrrhizin in root cultures of <italic>G glabra</italic> and in vitro stolon system extends uniform tissue cultures which could be utilized for functional genomics and understanding glycyrrhizin biosynthetic studies. This would broaden available knowledge regarding the biosynthetic pathway and mechanism responsible for glycyrrhizin metabolism and its regulation. The mechanism and the biosynthetic enzymes responsible for its over production is still an open area for research.</p><sec id="d29e1850"><title><bold>Author contribution</bold></title><p>R.A.V. is responsible for conceptualizing the idea. A.A. designed the experiments. S.G., P.P. were responsible for establishment of tissue culture experiment, chemical analysis was done by A.P.G. and M.K.V. A.A. and S.G. are also responsible for compiling the data in the form of MS.</p></sec></sec></body><back><glossary><title>Abbreviations</title><def-list><def-item><term>2,4-D</term><def><p>2,4-Dichlorophenoxy acetic acid</p></def></def-item><def-item><term>BA</term><def><p>6-Benzyladenine</p></def></def-item><def-item><term>HPLC</term><def><p>High performance liquid chromatography</p></def></def-item><def-item><term>IAA</term><def><p>Indole-3-acetic acid</p></def></def-item><def-item><term>IBA</term><def><p>Indole-3-butyric acid</p></def></def-item><def-item><term>KN</term><def><p>Kinetin</p></def></def-item><def-item><term>MS</term><def><p>Murashige and Skoog medium</p></def></def-item><def-item><term>NAA</term><def><p>1-Naphthaleneacetic acid</p></def></def-item><def-item><term>μM</term><def><p>Micro mole</p></def></def-item><def-item><term>ISSR</term><def><p>Inter-simple sequence repeats</p></def></def-item><def-item><term>LCMS</term><def><p>Liquid chromatography–mass spectrometry</p></def></def-item><def-item><term>GA</term><def><p>Glycyrrhizin</p></def></def-item><def-item><term>DNA</term><def><p>Deoxyribonucleotide triphosphate</p></def></def-item><def-item><term>RNA</term><def><p>Ribonucleic acid</p></def></def-item><def-item><term>SARS</term><def><p>Severe acute respiratory syndrome</p></def></def-item><def-item><term>COX-2</term><def><p>Cyclo-oxygenase-2</p></def></def-item><def-item><term>TLC</term><def><p>Thin layer chromatography</p></def></def-item><def-item><term>PCR</term><def><p>Polymerase chain reaction</p></def></def-item><def-item><term>CTAB</term><def><p>Cetyl trimethylammonium bromide</p></def></def-item><def-item><term>dNTP</term><def><p>Di-nucleotide phosphate</p></def></def-item><def-item><term>TAE</term><def><p>Tris-acetate-ethylenediaminetetraacetic acid</p></def></def-item></def-list></glossary><ack><p>This work was carried out under Research Council approved project MLP-3012 (Work Package 4). Authors are grateful to Members Research Council, for their valuable comments during the course of the study. We thank Mr. Rajender Gupta for HPLC and Mr. Sunil Kumar for photography.</p></ack><ref-list id="Bib1"><title>References</title><ref id="CR1"><mixed-citation publication-type="other">Anonymous (2005) A dictionary of Indian raw materials and industrial products, first supplement series. In: The wealth of India. National Institute of Sciences Communication and Information Resources, CSIR, New Delhi, 3, pp 195–198</mixed-citation></ref><ref id="CR2"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Ayabe</surname><given-names>S</given-names></name><name><surname>Takano</surname><given-names>H</given-names></name><name><surname>Fujita</surname><given-names>T</given-names></name><name><surname>Furuya</surname><given-names>T</given-names></name><name><surname>Hirota</surname><given-names>H</given-names></name><name><surname>Takahashi</surname><given-names>T</given-names></name></person-group><article-title>Triterpenoid biosynthesis in tissue cultures of <italic>Glycyrrhiza glabra</italic> var. <italic>glandulifera</italic></article-title><source>Plant Cell Rep</source><year>1990</year><volume>9</volume><fpage>181</fpage><lpage>184</lpage><pub-id pub-id-type="doi">10.1007/BF00232175</pub-id><pub-id pub-id-type="pmid">24226698</pub-id></element-citation></ref><ref id="CR3"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Cinatl</surname><given-names>J</given-names></name><name><surname>Morgenstern</surname><given-names>B</given-names></name><name><surname>Bauer</surname><given-names>G</given-names></name><name><surname>Chandra</surname><given-names>P</given-names></name><name><surname>Rabenau</surname><given-names>H</given-names></name><name><surname>Doerr</surname><given-names>HW</given-names></name></person-group><article-title>Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus</article-title><source>Lancet</source><year>2003</year><volume>361</volume><fpage>2045</fpage><lpage>2046</lpage><pub-id pub-id-type="doi">10.1016/S0140-6736(03)13615-X</pub-id><pub-id pub-id-type="pmid">12814717</pub-id></element-citation></ref><ref id="CR4"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Doyle</surname><given-names>JJ</given-names></name><name><surname>Doyle</surname><given-names>JL</given-names></name></person-group><article-title>A rapid DNA isolation procedure for small quantities of fresh leaf tissue</article-title><source>Phytochem Bull</source><year>1987</year><volume>19</volume><fpage>11</fpage><lpage>15</lpage></element-citation></ref><ref id="CR5"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Finney</surname><given-names>RSH</given-names></name><name><surname>Somers</surname><given-names>GF</given-names></name></person-group><article-title>The antiinflammatory activity of glycyrrhetinic acid and derivatives</article-title><source>J Pharm Pharmacol</source><year>1958</year><volume>10</volume><fpage>613</fpage><lpage>620</lpage><pub-id pub-id-type="doi">10.1111/j.2042-7158.1958.tb10349.x</pub-id><pub-id pub-id-type="pmid">13588543</pub-id></element-citation></ref><ref id="CR6"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Fiore</surname><given-names>C</given-names></name><name><surname>Eisenhut</surname><given-names>M</given-names></name><name><surname>Krausse</surname><given-names>R</given-names></name><name><surname>Ragazzi</surname><given-names>E</given-names></name><name><surname>Pellati</surname><given-names>D</given-names></name><name><surname>Armanini</surname><given-names>D</given-names></name><name><surname>Bielenberg</surname><given-names>J</given-names></name></person-group><article-title>Antiviral effects of <italic>Glycyrrhiza</italic> species</article-title><source>Phytother Res</source><year>2008</year><volume>22</volume><fpage>141</fpage><lpage>148</lpage><pub-id pub-id-type="doi">10.1002/ptr.2295</pub-id><pub-id pub-id-type="pmid">17886224</pub-id></element-citation></ref><ref id="CR7"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Franklin</surname><given-names>G</given-names></name><name><surname>Sheeba</surname><given-names>CJ</given-names></name><name><surname>Laksmisita</surname><given-names>G</given-names></name></person-group><article-title>Regeneration of eggplant (<italic>Solanum melongena</italic> L.) from root explants</article-title><source>In Vitro Cell Dev Biol Plant</source><year>2004</year><volume>40</volume><fpage>188</fpage><lpage>191</lpage><pub-id pub-id-type="doi">10.1079/IVP2003491</pub-id></element-citation></ref><ref id="CR8"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Gupta</surname><given-names>V</given-names></name><name><surname>Kak</surname><given-names>A</given-names></name><name><surname>Singh</surname><given-names>BB</given-names></name></person-group><article-title>Studies on seed germination and seedling vigour in liquorice (<italic>Glycyrrhiza glabra</italic>)</article-title><source>JMAPS</source><year>1997</year><volume>2</volume><fpage>412</fpage><lpage>413</lpage></element-citation></ref><ref id="CR9"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Hayashi</surname><given-names>H</given-names></name><name><surname>Fukui</surname><given-names>H</given-names></name><name><surname>Tabata</surname><given-names>M</given-names></name></person-group><article-title>Examination of triterpenoids produced by callus and cell suspension cultures of <italic>Glycyrrhiza glabra</italic></article-title><source>Plant Cell Rep</source><year>1988</year><volume>7</volume><fpage>508</fpage><lpage>511</lpage><pub-id pub-id-type="doi">10.1007/BF00272743</pub-id><pub-id pub-id-type="pmid">24240403</pub-id></element-citation></ref><ref id="CR10"><mixed-citation publication-type="other">Henry M, Edy MA, Desmarest P, Manoir DJ (1991) <italic>Glycyrrhiza glabra</italic> L. (Licorice): cell culture, regeneration, and the production of glycyrrhizin. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 15, Medicinal and aromatic plants III. Springer, Berlin, pp 270–282</mixed-citation></ref><ref id="CR11"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Ito</surname><given-names>M</given-names></name><name><surname>Sato</surname><given-names>A</given-names></name><name><surname>Hirabayashi</surname><given-names>K</given-names></name><name><surname>Tanabe</surname><given-names>F</given-names></name><name><surname>Shigeta</surname><given-names>S</given-names></name><name><surname>Baba</surname><given-names>M</given-names></name><name><surname>Clercq</surname><given-names>ED</given-names></name><name><surname>Nakashima</surname><given-names>H</given-names></name><name><surname>Yamamoto</surname><given-names>N</given-names></name></person-group><article-title>Mechanism of inhibitory effect of glycyrrhizin on replication of human immunodeficiency virus (HIV)</article-title><source>Antiviral Res</source><year>1988</year><volume>10</volume><fpage>289</fpage><lpage>298</lpage><pub-id pub-id-type="doi">10.1016/0166-3542(88)90047-2</pub-id><pub-id pub-id-type="pmid">3250333</pub-id></element-citation></ref><ref id="CR12"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Jatav</surname><given-names>VS</given-names></name><name><surname>Singh</surname><given-names>SK</given-names></name><name><surname>Khatri</surname><given-names>P</given-names></name><name><surname>Sharma</surname><given-names>AK</given-names></name></person-group><article-title>Recent pharmacological trends of <italic>Glycyrrhiza glabra</italic> Linn.</article-title><source>Unani Res</source><year>2011</year><volume>1</volume><fpage>1</fpage><lpage>4</lpage><pub-id pub-id-type="doi">10.5530/ur.2.2011.3</pub-id></element-citation></ref><ref id="CR13"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Kojoma</surname><given-names>M</given-names></name><name><surname>Ohyama</surname><given-names>K</given-names></name><name><surname>Seki</surname><given-names>H</given-names></name><name><surname>Hiraoka</surname><given-names>Y</given-names></name><name><surname>Asazu</surname><given-names>SN</given-names></name><name><surname>Sawa</surname><given-names>S</given-names></name><name><surname>Sekizaki</surname><given-names>H</given-names></name><name><surname>Yoshida</surname><given-names>S</given-names></name><name><surname>Muranaka</surname><given-names>T</given-names></name></person-group><article-title><italic>In vitro</italic> proliferation and triterpenoid characteristics of licorice (<italic>Glycyrrhiza uralensis</italic> Fischer, Leguminosae) stolons</article-title><source>Plant Biotechnol</source><year>2010</year><volume>27</volume><fpage>59</fpage><lpage>66</lpage><pub-id pub-id-type="doi">10.5511/plantbiotechnology.27.59</pub-id></element-citation></ref><ref id="CR14"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Li</surname><given-names>W</given-names></name><name><surname>Asad</surname><given-names>Y</given-names></name><name><surname>Yoshikawa</surname><given-names>T</given-names></name></person-group><article-title>Flavonoid constituents from <italic>Glycyrrhiza glabra</italic> hairy root cultures</article-title><source>Phytochemistry</source><year>2000</year><volume>55</volume><fpage>447</fpage><lpage>456</lpage><pub-id pub-id-type="doi">10.1016/S0031-9422(00)00337-X</pub-id><pub-id pub-id-type="pmid">11140606</pub-id></element-citation></ref><ref id="CR15"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Mehrotra</surname><given-names>S</given-names></name><name><surname>Kukreja</surname><given-names>AK</given-names></name><name><surname>Khanuja</surname><given-names>SPS</given-names></name><name><surname>Mishra</surname><given-names>BN</given-names></name></person-group><article-title>Genetic transformation studies and scale up of hairy root culture of <italic>Glycyrrhiza glabra</italic> in bioreactor</article-title><source>Electron J Biotechnol</source><year>2008</year><volume>11</volume><fpage>1</fpage><lpage>7</lpage><pub-id pub-id-type="doi">10.2225/vol11-issue2-fulltext-6</pub-id></element-citation></ref><ref id="CR16"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Mehrotra</surname><given-names>S</given-names></name><name><surname>Khwaja</surname><given-names>O</given-names></name><name><surname>Kukreja</surname><given-names>AK</given-names></name><name><surname>Rahman</surname><given-names>L</given-names></name></person-group><article-title>ISSR and RAPD based evaluation of genetic stability of encapsulated micro shoots of <italic>Glycyrrhiza glabra</italic> following six months of storage</article-title><source>Mol Biotechnol</source><year>2012</year><volume>52</volume><fpage>262</fpage><lpage>268</lpage><pub-id pub-id-type="doi">10.1007/s12033-011-9491-6</pub-id><pub-id pub-id-type="pmid">22215432</pub-id></element-citation></ref><ref id="CR17"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Mousa</surname><given-names>N</given-names></name><name><surname>Siagura</surname><given-names>P</given-names></name><name><surname>Wiryowidagdo</surname><given-names>S</given-names></name><name><surname>Wagih</surname><given-names>ME</given-names></name></person-group><article-title>Rapid clonal propagation of licorice (<italic>Glycyrrhiza glabra</italic>) by in vitro shoot culture</article-title><source>Sugar Tech</source><year>2006</year><volume>8</volume><fpage>292</fpage><lpage>298</lpage><pub-id pub-id-type="doi">10.1007/BF02943571</pub-id></element-citation></ref><ref id="CR18"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Nagat</surname><given-names>A</given-names></name><name><surname>Mousa</surname><given-names>TR</given-names></name><name><surname>Siaguru</surname><given-names>I</given-names></name><name><surname>Wiryowidagdo</surname><given-names>S</given-names></name><name><surname>Wagih</surname><given-names>ME</given-names></name></person-group><article-title>Establishment of regenerative callus and cell suspension system of licorice (<italic>Glycyrrhiza glabra</italic>) for the production of the sweetener glycyrrhizin in vitro</article-title><source>Sugar Tech</source><year>2007</year><volume>9</volume><fpage>72</fpage><lpage>82</lpage><pub-id pub-id-type="doi">10.1007/BF02956917</pub-id></element-citation></ref><ref id="CR19"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Park</surname><given-names>HY</given-names></name><name><surname>Park</surname><given-names>SH</given-names></name><name><surname>Yoon</surname><given-names>HK</given-names></name><name><surname>Han</surname><given-names>MJ</given-names></name><name><surname>Kim</surname><given-names>DH</given-names></name></person-group><article-title>Anti-allergic activity of 18β-glycyrrhetinic acid-3-<italic>O</italic>-beta-<sc>d</sc> glucuronide</article-title><source>Arch Pharm Res</source><year>2004</year><volume>27</volume><fpage>57</fpage><lpage>60</lpage><pub-id pub-id-type="doi">10.1007/BF02980047</pub-id><pub-id pub-id-type="pmid">14969340</pub-id></element-citation></ref><ref id="CR20"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Saito</surname><given-names>K</given-names></name><name><surname>Kaneko</surname><given-names>H</given-names></name><name><surname>Yamazaki</surname><given-names>M</given-names></name><name><surname>Yoshida</surname><given-names>M</given-names></name><name><surname>Murakoshi</surname><given-names>I</given-names></name></person-group><article-title>Stable transfer and expression of chimeric genes in licorice (<italic>Glycyrrhiza uralensis</italic>) using a Ri plasmid binary vector</article-title><source>Plant Cell Rep</source><year>1990</year><volume>8</volume><fpage>718</fpage><lpage>721</lpage><pub-id pub-id-type="doi">10.1007/BF00272102</pub-id><pub-id pub-id-type="pmid">24232925</pub-id></element-citation></ref><ref id="CR21"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Shah</surname><given-names>RR</given-names></name><name><surname>Dalal</surname><given-names>KC</given-names></name></person-group><article-title>In vitro multiplication of <italic>Glycyrrhiza</italic></article-title><source>Curr Sci</source><year>1980</year><volume>49</volume><fpage>69</fpage><lpage>71</lpage></element-citation></ref><ref id="CR22"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Shirazi</surname><given-names>Z</given-names></name><name><surname>Piri</surname><given-names>K</given-names></name><name><surname>Asl</surname><given-names>AM</given-names></name><name><surname>Hasanloo</surname><given-names>T</given-names></name></person-group><article-title>Glycyrrhizin and isoliquiritigenin production by hairy root culture of <italic>Glycyrrhiza glabra</italic></article-title><source>J Med Plants Res</source><year>2012</year><volume>6</volume><fpage>4640</fpage><lpage>4646</lpage><pub-id pub-id-type="doi">10.5897/JMPR11.1651</pub-id></element-citation></ref><ref id="CR23"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Thengane</surname><given-names>R</given-names></name><name><surname>Kulkarni</surname><given-names>DK</given-names></name><name><surname>Krishnamurthy</surname><given-names>KV</given-names></name></person-group><article-title>Micropropagation of licorice (<italic>Glycyrrhiza glabra</italic> l.) through shoot tip and nodal cultures</article-title><source>In Vitro CeIl Dev Biol Plant</source><year>1998</year><volume>34</volume><fpage>331</fpage><lpage>334</lpage><pub-id pub-id-type="doi">10.1007/BF02822743</pub-id></element-citation></ref><ref id="CR24"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Toivonen</surname><given-names>L</given-names></name><name><surname>Rosenqvist</surname><given-names>H</given-names></name></person-group><article-title>Establishment and growth characteristics of <italic>Glycyrrhiza glabra</italic> hairy root cultures</article-title><source>Plant Cell Tiss Org Cult</source><year>1995</year><volume>41</volume><fpage>249</fpage><lpage>258</lpage><pub-id pub-id-type="doi">10.1007/BF00045089</pub-id></element-citation></ref><ref id="CR25"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Wongwicha</surname><given-names>W</given-names></name><name><surname>Tanaka</surname><given-names>H</given-names></name><name><surname>Shoyama</surname><given-names>Y</given-names></name><name><surname>Tuvshintogtokh</surname><given-names>I</given-names></name><name><surname>Putalun</surname><given-names>W</given-names></name></person-group><article-title>Production of glycyrrhizin in callus cultures of licorice</article-title><source>Z Naturforsch</source><year>2008</year><volume>63</volume><fpage>413</fpage><lpage>417</lpage></element-citation></ref><ref id="CR26"><mixed-citation publication-type="other">Yoo S, Kim S (1976) Studies in tissue culture of medicinal plants (II). Tissue culture of <italic>Glycyrrhiza glabra</italic> L. var. <italic>glandulifera</italic> Reg. et Herder. Kor J Pharmacol 7:55D57</mixed-citation></ref><ref id="CR27"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Zhang</surname><given-names>MZ</given-names></name><name><surname>Jie</surname><given-names>X</given-names></name><name><surname>Bing</surname><given-names>Y</given-names></name><name><surname>Huiyong</surname><given-names>Y</given-names></name><name><surname>Qiuyin</surname><given-names>C</given-names></name><name><surname>Martha</surname><given-names>J</given-names></name><name><surname>Shrubsole</surname><given-names>XC</given-names></name><name><surname>Valentina</surname><given-names>K</given-names></name><name><surname>Wei</surname><given-names>Z</given-names></name><name><surname>Ambra</surname><given-names>P</given-names></name><name><surname>Harris</surname><given-names>RC</given-names></name></person-group><article-title>Inhibition of 11β-hydroxysteroid dehydrogenase type II selectively blocks the tumor COX-2 pathway and suppresses colon carcinogenesis in mice and humans</article-title><source>J Clin Invest</source><year>2009</year><volume>119</volume><fpage>876</fpage><lpage>885</lpage><pub-id pub-id-type="doi">10.1172/JCI37398</pub-id><pub-id pub-id-type="pmid">19307727</pub-id></element-citation></ref><ref id="CR28"><element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Zietkiewicz</surname><given-names>E</given-names></name><name><surname>Rafalski</surname><given-names>A</given-names></name><name><surname>Labuda</surname><given-names>D</given-names></name></person-group><article-title>Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification</article-title><source>Genomics</source><year>1994</year><volume>20</volume><fpage>176</fpage><lpage>183</lpage><pub-id pub-id-type="doi">10.1006/geno.1994.1151</pub-id><pub-id pub-id-type="pmid">8020964</pub-id></element-citation></ref></ref-list></back></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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