Agrobacterium rhizogenes mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment

Kumar, Vinod; Sharma, Ashwani; Prasad, Bellur Chayapathy Narasimha; Gururaj, H. B.; Ravishankar, G. A.

doi:10.2225/vol9-issue4-fulltext-4

Cited by 48 publications

(30 citation statements)

References 18 publications

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“…Primers for detecting the virC gene (5'-ATCATTTGTAGC GACT-3' and 5'-AGCTCAAACCTGCTTC-3') (Kumar et al 2006) outside the T-DNA of Ri plasmid were also used to eliminate the possibility of A. rhizogenes contamination of the hairy root lines. A. rhizogenes cells, in addition to primers and reaction buffer, were incubated in a DNA thermal cycler (MJ Research Inc., Nevada, USA) at 94 • C. PCR products were examined by electrophoresis on 2% agarose gels (w/v) and staining with ethidium bromide.…”

Section: Growth Measurementmentioning

confidence: 99%

Induction of Gentiana cruciata hairy roots and their secondary metabolites

et al. 2011

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Gentiana cruciata L. (cross gentian) is a medicinal and ornamental plant. The root extracts of this species are known to exhibit many curative properties. The natural Gentiana populations are exposed to great danger because of their uncontrolled usage. In this study, hairy roots from Gentiana cruciata L. stem and leaf explants belonging to three different clones were induced by inoculation with four different Agrobacterium rhizogenes wild strains namely A4, 15834, 8196 and R1000. Induction of the root transformation was significantly dependent on the explant type used. On the other hand, the genotype and bacterial strain had no significant effect on hairy root formation. Hairy root formation percentages of the explants varied between 5.6-33.3% in the stem explants, and between 0.0-6.7% in the leaf explants. Transformations of the hairy roots were confirmed by PCR using rolC specific primers, and revealed the absence of contaminating A. rhizogenes with virC primers. Total of twelve hairy root clones were obtained, and their secondary metabolite content was also analyzed by HPLC. Quantitative results exhibited that gentiopicroside was the most abundant compound in all root samples. Furthermore, metabolites such as loganic acid, swertiamarin, and sweroside were also identified and quantified in the samples.

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Section: Growth Measurementmentioning

confidence: 99%

Induction of Gentiana cruciata hairy roots and their secondary metabolites

et al. 2011

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“…rhizogenes strain interaction and light intensity. The effects of physical factors were also reported in many other crops like pumpkin (Katavic et al, 1991), pine (McAfee et al, 1993) and tobacco (Kumar et al, 2006). The variation in virulence gene activity in stem and petiole is different.…”

Section: Discussionmentioning

confidence: 81%

Enhanced Virulence Gene Activity of Agrobacterium in Muskmelon (Cucumis melo L.) cv. ‘Birdie’

Mohiuddin¹,

Abdullah²,

Chowdhury

et al. 2011

Not Sci Biol

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Muskmelon (Cucumis melo L.) cultivar 'Birdie' , was evaluated for its response to the tumorigenic Agrobacterium tumefaciens and the oncogenic A. rhizogenes strains. Stem and petiole of three week-old in vitro-grown muskmelon plants were inoculated with five strains of A. tumefaciens and A. rhizogenes each and observed phenotypic expressions i.e. induction of crown galls and hairy roots. This phenotypic expression was efficaciously increased when virulence gene activity of different strains of two Agrobacterium species was enhanced. Intensive studies on enhancement of virulence gene activity of Agrobacterium found to be correlated to the appropriate light intensity (39.3 µmol m -2 s -1) with a specific concentration of monocyclic phenolic compound, acetosyringone (20 µM). The gene activity was also influenced by several other physical factors e.g. plant tissue type, Agrobacterium species and their strains, and plant tissue-Agrobacterium interaction. Among the different A. tumefaciens strains, LBA4404 showed the best virulence gene activity in both stem and petiole through the formation of higher rate of crown galls. On the other hand, strain 15834 of A. rhizogenes showed better gene activity in stem and 8196 in petiole through the formation of higher rate of hairy roots as well as higher average number of hairy roots. Among the two different types of explants, petiole was more susceptible to both Agrobacterium species. Thus it was concluded that future muskmelon transformation study can efficiently be carried out with LBA4404, 15834 and 8196 strains using petiole explants by adding 20 µM of acetosyringone in the medium.

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“…Two fragments, defined as T L -DNA and T R -DNA, can be transferred and integrated independently into the plant genome during the infection process. However, the integration capacity of T L -DNA was much higher than T R -DNA Costantion et al 1984;David et al 1984;Grant et al 1991;Phelep et al 1991;Nilsson and Olsson 1997;Holefors et al 1998;Sevon and Oksman-Caldentey 2002;Kumar et al 2006;Navarrete et al 2006;Bensaddek et al 2008). Furthermore, the present findings imply that a higher number of Ri-T-DNA copies integrated into the plant genome increase the phenotypic effect in the Ri-line (Christensen et al 2008).…”

Section: Rit-dnagenesmentioning

confidence: 99%

“…Certain bacterial species are capable of transferring some of their genes to higher plants ending up with insertion and permanent integration in the nuclear genome (Broothaerts et al 2005;Kumar et al 2006). Members of genus Agrobacterium are widely known for their ability of forming a wide variety of different neoplastic diseases, including crown gall ( A. tumefaciens and A. vitis), hairy root ( A. rhizogenes) and cane gall ( A. rubi) (Gelvin 2009;Ozyigit 2012).…”

Section: Characteristics Of Agrobacteriumrhizogenesmentioning

confidence: 99%

“…The bacterium moves toward the wounded sites by chemotaxis and infect plant cells. Subsequent infection at wound site followed by transfer of a particular DNA segment (T-DNA) from the root-inducing (Ri) plasmid (pRi) of the bacteria (Kumar et al 2006). A. rhizogenes-induced roots have the unique property of being able to grow in vitro without exogenous plant growth regulators (Lee et al 2001;Rao and Ravishankar 2002).…”

Section: Hairy Root Diseasementioning

confidence: 99%

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Agrobacterium rhizogenes-Mediated Transformation and Its Biotechnological Applications in Crops

2013

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The history of Agrobacterium-related plant biotechnology goes back for more than three decades with the discovery of molecular mechanisms of crown gall disease in plants. After 1980s, gene technologies began developing rapidly and today, related with the improved gene transfer methods, plant biotechnology has become one of the most important branches in science. Till now, the most important genes related with agricultural affairs have been utilized for cloning of plants with the deployment of different techniques used in genetic engineering. Especially, Agrobacterium tumefaciens was used extensively for transferring desired genetic materials to plants rapidly and effectively by the researchers to create transgenic plants. Recognition of the biology of Agrobacterium species and newly developed applications of their T-DNA systems has been a great step in plant biotechnology. This chapter provides the reader with extensive information on A. rhizogenes which is responsible for the development of hairy root disease in a wide range of dicotyledonous plants and its T-DNA system. This knowledge will be useful in improving utilization of crops and the formulation of new and up-graded transgenic based food products.

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Agrobacterium rhizogenes mediated genetic transformation resulting in hairy root formation is enhanced by ultrasonication and acetosyringone treatment

Cited by 48 publications

References 18 publications

Induction of Gentiana cruciata hairy roots and their secondary metabolites

Induction of Gentiana cruciata hairy roots and their secondary metabolites

Enhanced Virulence Gene Activity of Agrobacterium in Muskmelon (Cucumis melo L.) cv. ‘Birdie’

Agrobacterium rhizogenes-Mediated Transformation and Its Biotechnological Applications in Crops

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