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Agrobacterium rhizogenes mediated hairy root induction in endangered Berberis aristata DC.

Brijwal L, Tamta S - Springerplus (2015)

Bottom Line: The presence of rol A and rol B genes during amplification confirmed the transgenic nature of hairy roots and transformed callus.Transformation frequency of callus was further enhanced (from 61.11 ± 1.60 % to 72.22 ± 1.60 %; when infection time was 1 h) by using acetosyringone (100 µM) during co-cultivation period (48 h) on semisolid MS (Murashige and Skoog) medium.In conclusion, this study describes the protocol for hairy root induction which could further be useful for the production of berberin and may reduce the overharvesting of this endangered species from its natural habitat.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Plant Tissue Culture and Molecular Biology Laboratory, Bhimtal Campus, Kumaun University, Nainital, 263136 Uttarakhand India.

ABSTRACT
An efficient protocol for hairy root induction in Berberis aristata DC. was established using two different strains of Agrobacterium rhizogenes, MTCC 532 and 2364 from IMTECH (Institute of Microbial Technology), Chandigarh, India. The strain 532 was more effective than strain 2364 in hairy root induction and in vitro grown callus (61.11 ± 1.60 % transformation frequency) was found to be suitable explant in comparison to leaves (42.59 ± 0.92 % transformation frequency) and nodal segments (34.25 ± 0.92 % transformation frequency) of in vitro grown microshoots for hairy root induction. The presence of rol A and rol B genes during amplification confirmed the transgenic nature of hairy roots and transformed callus. Transformation frequency of callus was further enhanced (from 61.11 ± 1.60 % to 72.22 ± 1.60 %; when infection time was 1 h) by using acetosyringone (100 µM) during co-cultivation period (48 h) on semisolid MS (Murashige and Skoog) medium. In conclusion, this study describes the protocol for hairy root induction which could further be useful for the production of berberin and may reduce the overharvesting of this endangered species from its natural habitat.

No MeSH data available.


Related in: MedlinePlus

PCR amplification of rol A gene
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Fig2: PCR amplification of rol A gene

Mentions: The presence of rol A and rol B genes confirmed the transgenic nature of hairy roots and transformed callus. DNA extracted from hairy roots and transformed callus after PCR amplification demonstrated the product of expected size approximately 500 bp with used rol A primer and approximately 300 bp product with used rol B primer. They were not observed in PCR product of untransformed root and callus DNA, which were used as negative control. The amplicon size observed from bacterial strain DNA exactly matched with hairy root and transformed callus product (Figs. 2, 3) which conforming the transgenic nature of hairy roots and callus.Fig. 2


Agrobacterium rhizogenes mediated hairy root induction in endangered Berberis aristata DC.

Brijwal L, Tamta S - Springerplus (2015)

PCR amplification of rol A gene
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4546071&req=5

Fig2: PCR amplification of rol A gene
Mentions: The presence of rol A and rol B genes confirmed the transgenic nature of hairy roots and transformed callus. DNA extracted from hairy roots and transformed callus after PCR amplification demonstrated the product of expected size approximately 500 bp with used rol A primer and approximately 300 bp product with used rol B primer. They were not observed in PCR product of untransformed root and callus DNA, which were used as negative control. The amplicon size observed from bacterial strain DNA exactly matched with hairy root and transformed callus product (Figs. 2, 3) which conforming the transgenic nature of hairy roots and callus.Fig. 2

Bottom Line: The presence of rol A and rol B genes during amplification confirmed the transgenic nature of hairy roots and transformed callus.Transformation frequency of callus was further enhanced (from 61.11 ± 1.60 % to 72.22 ± 1.60 %; when infection time was 1 h) by using acetosyringone (100 µM) during co-cultivation period (48 h) on semisolid MS (Murashige and Skoog) medium.In conclusion, this study describes the protocol for hairy root induction which could further be useful for the production of berberin and may reduce the overharvesting of this endangered species from its natural habitat.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Plant Tissue Culture and Molecular Biology Laboratory, Bhimtal Campus, Kumaun University, Nainital, 263136 Uttarakhand India.

ABSTRACT
An efficient protocol for hairy root induction in Berberis aristata DC. was established using two different strains of Agrobacterium rhizogenes, MTCC 532 and 2364 from IMTECH (Institute of Microbial Technology), Chandigarh, India. The strain 532 was more effective than strain 2364 in hairy root induction and in vitro grown callus (61.11 ± 1.60 % transformation frequency) was found to be suitable explant in comparison to leaves (42.59 ± 0.92 % transformation frequency) and nodal segments (34.25 ± 0.92 % transformation frequency) of in vitro grown microshoots for hairy root induction. The presence of rol A and rol B genes during amplification confirmed the transgenic nature of hairy roots and transformed callus. Transformation frequency of callus was further enhanced (from 61.11 ± 1.60 % to 72.22 ± 1.60 %; when infection time was 1 h) by using acetosyringone (100 µM) during co-cultivation period (48 h) on semisolid MS (Murashige and Skoog) medium. In conclusion, this study describes the protocol for hairy root induction which could further be useful for the production of berberin and may reduce the overharvesting of this endangered species from its natural habitat.

No MeSH data available.


Related in: MedlinePlus