Agrobacterium-Mediated Genetic Transformation of Pogostemon cablin (Blanco) Benth. Using Leaf Explants: Bactericidal Effect of Leaf Extracts and Counteracting Strategies

Paul, Anamika; Bakshi, S. K.; Sahoo, Debee Prasad; Kalita, Mohan Chandra; Sahoo, Lingaraj

doi:10.1007/s12010-012-9612-0

Cited by 23 publications

(16 citation statements)

References 77 publications

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“…Preconditioning also stimulates the plant cells to enter into the regeneration pathway, resulting in improved transformation efficiency (Ainsley et al, 2001). Our results are in agreement with previous findings where high transformation frequency was achieved in 2-day precultured leaf explants of medicinally important Scoparia dulcis L. (Aileni et al, 2011) and in Pogostemon cablin (Blanco) Benth (Paul et al, 2012). The density of bacterial suspension used for infection of the explant was also found to be a key factor affecting the transformation frequency (Mishra et al, 2013).…”

Section: Discussionsupporting

confidence: 92%

Direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation of Solanum trilobatum L.

Sakthivel¹,

Jayashre²,

Largia³

et al. 2016

Turk J Biol

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Solanum trilobatum L. is one of the most valued medicinal plants of Indian Ayurveda and Siddha medicine. An efficient protocol for direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation using in vitro derived leaf explants of S. trilobatum was established for the first time. High frequency direct shoot regeneration (89%) was achieved in MS medium containing 2 mg/L 6-benzylaminopurine and 0.25 mg/L indole-3-acetic acid. The Agrobacterium strain EHA 105 harboring the binary vector pCAMBIA 1301, which contained hygromycin phosphotransferase (hpt) as a selectable marker gene and β-glucuronidase (gus) as a reporter gene, was used for the transformation. The factors that influence the transformation frequency were optimized as follows: age of explants (40 days old), acetosyringone concentration (150 µM), duration of preculture (2 days), bacterial optical density (O.D. 0.4), Tween 20 concentration (0.01%), infection time (15 min), pH of cocultivation medium (5.4), and cocultivation duration (3 days). The putative transformants were screened on selection medium containing 15 mg/L hygromycin. The transgenic plants were confirmed by histochemical gus analysis, PCR, and RT-PCR analysis. A stable transformation efficiency of 64% was achieved under optimized transformation conditions. Thus, the protocol can be used for genetic improvement of this plant in the future.

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Section: Discussionsupporting

confidence: 92%

Direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation of Solanum trilobatum L.

Sakthivel¹,

Jayashre²,

Largia³

et al. 2016

Turk J Biol

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“…Wounding is an integral step in the Agrobacterium -mediated transformation because acetosyringone released from injured part plays chemotactic role and induces the vir genes to initiate T-DNA transfer [36, 37]. Figure 4(b) shows that wounding with scalpel produced the highest percentage of transient GUS expression.…”

Section: Discussionmentioning

confidence: 99%

Agrobacterium-Mediated Transformation of the RecalcitrantVandaKasem's Delight Orchid with Higher Efficiency

Gnasekaran

Antony

Uddain

et al. 2014

The Scientific World Journal

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The presented study established Agrobacterium-mediated genetic transformation using protocorm-like bodies (PLBs) for the production of transgenic Vanda Kasem's Delight Tom Boykin (VKD) orchid. Several parameters such as PLB size, immersion period, level of wounding, Agrobacterium density, cocultivation period, and concentration of acetosyringone were tested and quantified using gusA gene expression to optimize the efficiency of Agrobacterium-mediated genetic transformation of VKD's PLBs. Based on the results, 3-4 mm PLBs wounded by scalpel and immersed for 30 minutes in Agrobacterium suspension of 0.8 unit at A 600nm produced the highest GUS expression. Furthermore, cocultivating infected PLBs for 4 days in the dark on Vacin and Went cocultivation medium containing 200 𝜇M acetosyringone enhanced the GUS expression. PCR analysis of the putative transformants selected in the presence of 250 mg/L cefotaxime and 30 mg/L geneticin proved the presence of wheatwin1, wheatwin2, and nptII genes.

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“…Very efficient Agrobacterium-mediated transformation of the liverwort Marchantia polymorpha thalli was developed with the help of L-glutamine used for co-cultivation medium (Kubota et al 2013). Additional L-glutamine to Agrobacterium infection medium counteracted the bactericidal effect of Pogostemon cablin and significantly increased the T-DNA delivery to its leaf explants (Paul et al 2012). In our studies, L-glutamine appeared an essential factor for obtaining callus tissue and plants presenting stable transgenesis.…”

Section: The Effect Of B-lactam Antibiotics (Timentin and Cefatoxime)mentioning

confidence: 73%

Genetic transformation of gentian Gentiana tibetica (King) leaf explants with Agrobacterium tumefaciens strain C58C1

Wójcik

Rybczyński

2016

Acta Physiol Plant

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Experiments were carried out on transformation of leaf explants derived from the axenic culture of Gentiana tibetica (King) using a co-culture of Agrobacterium tumefaciens. A. tumefaciens octopine strain C58C1 carried neomycine phosphotransferase (nptII) and b-glucuronidase (uidA) genes. The influence of plasmid helper pCH32 on transformation efficiency was stressed. After co-cultivation, explant was cultured on three different regeneration media (RM1-3) at the presence of the timentin and callus formation and plants regeneration occurred. The transgenic character of the selected tissue and transformants T0 in the presence of kanamycin has been confirmed by the histochemical analysis of reporter enzyme activity (ß-glucuronidase), polymerase chain reaction, and southern hybridization detecting uidA and nptII genes. L-glutamine used during inoculation period had a significant influence on the transformation efficiency in relation to its concentration and time of treatment.

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Agrobacterium-Mediated Genetic Transformation of Pogostemon cablin (Blanco) Benth. Using Leaf Explants: Bactericidal Effect of Leaf Extracts and Counteracting Strategies

Cited by 23 publications

References 77 publications

Direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation of Solanum trilobatum L.

Direct shoot organogenesis and Agrobacterium tumefaciens mediated transformation of Solanum trilobatum L.

Agrobacterium-Mediated Transformation of the RecalcitrantVandaKasem's Delight Orchid with Higher Efficiency

Genetic transformation of gentian Gentiana tibetica (King) leaf explants with Agrobacterium tumefaciens strain C58C1

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