Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor

The study was undertaken to induce hairy roots in Glycyrrhiza glabra in leaf explants and to optimize the nutritional requirement for its growth kinetics at shake flask and bioreactor level. Pathogenecity of Agrobacterium depends upon transformation ability of strain and age, type, and physiological...

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Autores principales:	Mehrotra,Shakti, Kumar Kukreja,Arun, Singh Khanuja,Suman Preet, Nath Mishra,Bhartendu
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	Agrobacterium bioreactor Glycyrrhiza scale-up transformed roots
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200009
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spelling	oai:scielo:S0717-345820080002000092009-01-21Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactorMehrotra,ShaktiKumar Kukreja,ArunSingh Khanuja,Suman PreetNath Mishra,Bhartendu Agrobacterium bioreactor Glycyrrhiza scale-up transformed roots The study was undertaken to induce hairy roots in Glycyrrhiza glabra in leaf explants and to optimize the nutritional requirement for its growth kinetics at shake flask and bioreactor level. Pathogenecity of Agrobacterium depends upon transformation ability of strain and age, type, and physiological state of explants. Agrobacterium rhizogenes strain K599 was used to infect leaf explants of G. glabra. Explants of different age groups were obtained from 2 to 5 weeks old in vitro grown cultures. Bacterial strain K599 could induce hairy roots in 3 and 4 weeks old leaf explants cultured on B5, MS, NB and WP basal semi-solid medium. Leaf explants of 2 and 5 weeks old culture were not responsive to bacterial infection in terms of hairy root induction. Maximum transformation frequency (TF) of tested bacterial strain was 47% obtained in 3 weeks old explants after 25 days of incubation on MS basal semi solid medium. NB and B5 both media composition showed 20% of transformation frequency after 28 and 38 days respectively. WP medium did not support induction of roots in cultured leaf explants infected with A. rhizogenes strain K599even after 50 days of incubation. Further, when all the four media combinations were tested for root growth it was found that though WP was not responsive for hairy root induction, yet all four basal media supported hairy root growth and a gradual increase in fresh weight biomass was observed with an increase in culture duration. However amongst all, the NB medium composition supported best growth of hairy roots followed by MS, B5 and WP media. About 20 times increase in root biomass on fresh weight basis was recorded after 45days of culture in NB medium. Initial inoculum of roots (0.18 g. F.wt./ flask) containing 50 ml of liquid culture medium produced 3.59 g (F. wt.) biomass. A fast growing hairy root clone G6 was grown in a 5 l capacity mechanically agitated bioreactor provided with a nylon mesh septum. After 30 days of sterile run, 310 g of root biomass was harvested from the bioreactor culture vessel, recording about 20 times increase over initial inoculum (16.0 g).info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.2 20082008-04-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200009en10.4067/S0717-34582008000200009
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Agrobacterium bioreactor Glycyrrhiza scale-up transformed roots
spellingShingle	Agrobacterium bioreactor Glycyrrhiza scale-up transformed roots Mehrotra,Shakti Kumar Kukreja,Arun Singh Khanuja,Suman Preet Nath Mishra,Bhartendu Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
description	The study was undertaken to induce hairy roots in Glycyrrhiza glabra in leaf explants and to optimize the nutritional requirement for its growth kinetics at shake flask and bioreactor level. Pathogenecity of Agrobacterium depends upon transformation ability of strain and age, type, and physiological state of explants. Agrobacterium rhizogenes strain K599 was used to infect leaf explants of G. glabra. Explants of different age groups were obtained from 2 to 5 weeks old in vitro grown cultures. Bacterial strain K599 could induce hairy roots in 3 and 4 weeks old leaf explants cultured on B5, MS, NB and WP basal semi-solid medium. Leaf explants of 2 and 5 weeks old culture were not responsive to bacterial infection in terms of hairy root induction. Maximum transformation frequency (TF) of tested bacterial strain was 47% obtained in 3 weeks old explants after 25 days of incubation on MS basal semi solid medium. NB and B5 both media composition showed 20% of transformation frequency after 28 and 38 days respectively. WP medium did not support induction of roots in cultured leaf explants infected with A. rhizogenes strain K599even after 50 days of incubation. Further, when all the four media combinations were tested for root growth it was found that though WP was not responsive for hairy root induction, yet all four basal media supported hairy root growth and a gradual increase in fresh weight biomass was observed with an increase in culture duration. However amongst all, the NB medium composition supported best growth of hairy roots followed by MS, B5 and WP media. About 20 times increase in root biomass on fresh weight basis was recorded after 45days of culture in NB medium. Initial inoculum of roots (0.18 g. F.wt./ flask) containing 50 ml of liquid culture medium produced 3.59 g (F. wt.) biomass. A fast growing hairy root clone G6 was grown in a 5 l capacity mechanically agitated bioreactor provided with a nylon mesh septum. After 30 days of sterile run, 310 g of root biomass was harvested from the bioreactor culture vessel, recording about 20 times increase over initial inoculum (16.0 g).
author	Mehrotra,Shakti Kumar Kukreja,Arun Singh Khanuja,Suman Preet Nath Mishra,Bhartendu
author_facet	Mehrotra,Shakti Kumar Kukreja,Arun Singh Khanuja,Suman Preet Nath Mishra,Bhartendu
author_sort	Mehrotra,Shakti
title	Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
title_short	Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
title_full	Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
title_fullStr	Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
title_full_unstemmed	Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor
title_sort	genetic transformation studies and scale up of hairy root culture of glycyrrhiza glabra in bioreactor
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200009
work_keys_str_mv	AT mehrotrashakti genetictransformationstudiesandscaleupofhairyrootcultureofglycyrrhizaglabrainbioreactor AT kumarkukrejaarun genetictransformationstudiesandscaleupofhairyrootcultureofglycyrrhizaglabrainbioreactor AT singhkhanujasumanpreet genetictransformationstudiesandscaleupofhairyrootcultureofglycyrrhizaglabrainbioreactor AT nathmishrabhartendu genetictransformationstudiesandscaleupofhairyrootcultureofglycyrrhizaglabrainbioreactor
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Genetic transformation studies and scale up of hairy root culture of Glycyrrhiza glabra in bioreactor

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