A reliable and efficient protocol for inducing genetically transformed roots in medicinal plant Nepeta pogonosperma

Valimehr, Sepideh; Sanjarian, Forough; Sohi, Haleh Hashemi; Sharafi, Ali; Sabouni, Farzaneh

doi:10.1007/s12298-014-0235-5

Cited by 31 publications

(25 citation statements)

References 17 publications

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“…Prior studies indicated that vir G expression can be triggered by low levels of PO 4 and proposed that PO 4 deficiency could be a helpful signal to prompt genetic transformation. Similar results were described in transformation by A. tumefaciens in Lilium (36) and genetically transformed root generation of Papaver bracteatum, Nepeta pogonosperma, Dracocephalum kotschyi, and Artemisia aucheri (26,31,34,37). 4 Jundishapur J Nat Pharm Prod.…”

Section: Discussionsupporting

confidence: 80%

“…Secondary metabolites, probably apigenin, were released in ½ MS liquid culture medium after inoculation with hairy roots compared to control as indicated by the color of the medium (Figure 3). Former reports showed that A. rhizogenes strain and form of explants could influence hairy root initiation (26,32,34). In some explants, tumorigenic calli (galls) were induced.…”

Section: Discussionmentioning

confidence: 99%

“…Total Genomic DNA was extracted from putative transgenic hairy root lines and control root line with the CTAB protocol (26). The PCR analysis of hairy roots and controls resulted in the amplification of expected fragment in hairy root lines similar to that of the positive control, while there was no extension in the DNA isolated from normal roots Figure 6.…”

Section: Polymerase Chain Reaction Analysismentioning

confidence: 99%

“…The biosynthesis of secondary metabolites is induced by increasing plant defense genes expression. Fast growth of hairy roots offers an additional benefit to use as a continuous source for valuable secondary metabolite production in the absence of growth regulators (which are usually expensive) in a sterile culture medium (24)(25)(26). For increasing the production of pharmaceutical compounds in hairy root cultures, the identification of key genes, which are involved in regulating secondary metabolic pathway stages can aid in secondary metabolic engineering.…”

Section: Introductionmentioning

confidence: 99%

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Genetically Transformed Root-Based Culture Technology in Medicinal Plant Cosmos bipinnatus

Jaberi¹,

Sharafi²,

Sharafi³

et al. 2018

Jundishapur J Nat Pharm Prod

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Background: Cosmos bipinnatus is an important medicinal plant with antioxidative, antigenotoxic, anti-inflammatory, and antiproliferative effects on several cancer cell lines. It has great potential for development as a promising cancer chemo-preventive agent. Hairy root-based culture technology is a new sustainable production platform for producing specific pharmaceutical secondary metabolites. Objectives: The current study developed and introduced a reliable transformation system for C. bipinnatus by optimization of aspects important in transformation frequency using Agrobacterium rhizogenes. Methods: Five bacterial strains, including ATCC 15834, ATCC 31798, A7, MAFF-02-10266, and MSU440, 2 explant types (leaf and stem), and 2 co-cultivation media (full MS and ½ MS) were examined. Genomic DNA was extracted using a modified CTAB protocol from putative transgenic root lines and the control root. Transgenic hairy root lines were approved by means of Polymerase Chain Reaction (PCR) using specific rolB gene primers. Results:The highest ratio of genetically transformed root induction was found from leaf explants using A. rhisogenes strains ATCC15834 and MSU440 (72% to 73%). When ½ MS medium was used as a co-cultivation medium, a significant increase in transformation frequency (84%) was observed. Conclusions: The MSU440 Agrobacterium strain and ½ MS co-cultivation medium could significantly improve genetic transformation efficiency for establishment of hairy root-based cultures for C. bipinnatus.

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Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Section: Polymerase Chain Reaction Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genetically Transformed Root-Based Culture Technology in Medicinal Plant Cosmos bipinnatus

Jaberi¹,

Sharafi²,

Sharafi³

et al. 2018

Jundishapur J Nat Pharm Prod

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“…A. aucheri leaves were susceptible to infection by all A. rhizogenes strains, as shown by the percentage of hairy roots emerged. Valimehr et al (2014) reported that stem explants were the best explants for A. rhizogenes mediated transformation in Nepeta pogonosperma while leaf explants showed a low rate of hairy root induction. The highest rate of hairy root induction was obtained using strain ATCC15834 and MSU440 (Fig.…”

Section: Hairy Roots Inductionmentioning

confidence: 99%

In vitro regeneration and Agrobacterium mediated genetic transformation of Artemisia aucheri Boiss

Sharafi

Sohi

Mirzaee

et al. 2014

Physiol Mol Biol Plants

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In the present study, we developed an efficient protocol for in vitro plant regeneration and genetically transformed root induction in medicinal plant Artemisia aucheri Boiss. Leaf explants were cultivated in MS medium supplemented by combination of plant growth regulators including α-naphthalene-acetic acid, 6-benzyl-aminopurine, indole-3-acetic acid and 2, 4-dichlorophenoxyaceticacid. The highest frequency of shoot organogenesis occurred on MS medium supplemented with 0.05 mg/l NAA plus 2 mg/l BA (96.3 %) and MS medium supplemented with 0.5 mg/l IAA plus 2 mg/l BA (88.3 %). Root induction was obtained on MS medium supplemented with 0.5 mg/l IBA. This is a simple, reliable, rapid and high efficient regeneration system for A. aucheri Boiss in short period via adventitious shoot induction approach. Also, an efficient genetically transformed root induction for A. aucheri was developed through Agrobacterium rhizogenes-mediated transformation by four bacterial strains, A4, ATCC15834, MSU440, and A13 (MAFF-02-10266). The maximum frequency of hairy root induction was obtained using MSU440 (93 %) and ATCC15834 (89 %) bacterial strains. Hairy root lines were confirmed by PCR using the rolB gene specific primers and Southern blot analysis.

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Potential of the genetically transformed root cultures of Plumbago europaea for biomass and plumbagin production

Beigmohamadi

Movafeghi

Jafari

et al. 2019

Biotechnology Progress

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Plumbago europaea L. is the main source of plumbagin which is a well‐known pharmacological active compound. In this investigation, genetically transformed roots of P. europaea were obtained by improving some factors affecting the efficiency of Agrobacterium rhizoigenes‐mediated transformation such as explant type, A. rhizoigenes strain, bacterial infection period, co‐cultivation period and acetosyringone concentration. The leaf, hypocotyl and stem explants from in vitro grown plantlets were infected with bacterial strains (A4, ATCC15834, MSU440 and A13). The highest transformation rate of 69.3% was achieved after 7–9 days by inoculating A. rhizogenes MSU440 strain onto the 3‐week‐old stem explants followed by a co‐cultivation period of 2 days on a medium containing 100 μM acetosyringone. To investigate the existence of the rolB gene, polymerase chain reaction was carried out using specific primers. Effects of growth media (MS, 1/2 MS, MS‐B5 and ½ MS‐B5), different sucrose concentrations and illumination on biomass production and plumbagin biosynthesis in P. europaea hairy root cultures were analyzed using stem explants after infection with MSU440 strain. ½ MS‐B5 liquid medium containing 30 g L−1 sucrose incubated in the dark resulted in the efficient biomass production of transformed hairy roots (12.5 g fresh weight, 1.8 g dry weight) with 3.2 mg g−1 DW plumbagin accumulation. This procedure provides a framework for large‐scale cultivation of hairy roots for plumbagin production. This is the first report describing the establishment of P. europaea hairy root culture with special emphasis on plumbagin production.

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A reliable and efficient protocol for inducing genetically transformed roots in medicinal plant Nepeta pogonosperma

Cited by 31 publications

References 17 publications

Genetically Transformed Root-Based Culture Technology in Medicinal Plant Cosmos bipinnatus

Genetically Transformed Root-Based Culture Technology in Medicinal Plant Cosmos bipinnatus

In vitro regeneration and Agrobacterium mediated genetic transformation of Artemisia aucheri Boiss

Potential of the genetically transformed root cultures of Plumbago europaea for biomass and plumbagin production

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