Genetically transformed root cultures — generation, properties and application in plant sciences

Кузовкина, И. Н.; Schneider, Bernd Uwe

doi:10.1007/3-540-27998-9_13

Cited by 20 publications

(8 citation statements)

References 206 publications

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“…A . rhizogenes A4 is reportedly most potent for dicotyledonous plants (Kuzovkina and Schneider 2006 ). In leaf explants of Pelargonium sp., 100% transformation frequency was reported with strain A4 (Saxena et al 2007 ).…”

Section: Resultsmentioning

confidence: 99%

Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L.

2017

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Semecarpus anacardium L. is a tree species which produces secondary metabolites of medicinal importance. Roots of the plant have been traditionally used in folk medicines. Different strains of Agrobacterium rhizogenes (A4, ATCC15834 and LBA 9402) were used for induction of hairy roots in in vitro grown tissues of the plant. Hairy root initiation was observed after 25–30 days of infection. Optimum transformation frequency of 61% was achieved on leaf explants with ATCC15834 strain. Infection time of 30 min resulted in greater transformation frequency compared to 10 and 20 min, respectively. The hairy roots cultured in growth regulator-free semi-solid woody plant medium differentiated into callus. Whole shoots infected with ATCC 15834 were found to produce more transformants upon co-cultivation for 4 (65%) and 5 (67%) days. Induction of hairy roots in stem explants infected with ATCC 15834 was lower (52%) compared to leaves (62%) after 4 days of co-cultivation. In A4 and LBA9402 strains transformation efficiency was 49 ± 2.8% and 36 ± 5.7% in shoots after 4 days of co-cultivation. Transformation frequency was higher in ATCC15834 strain, irrespective of explants. The hairy roots of S. anacardium elongated slowly upon transfer to half-strength liquid medium. After 3–4 passages in liquid medium slender hairy roots started differentiating which were separated from the original explants. Visible growth of the roots was observed in hormone-free liquid medium after 2–3 months of culturing. Polymerase chain reaction with gene-specific primers from rol A, B and C genes confirms the positive transformation events.

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Section: Resultsmentioning

confidence: 99%

Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L.

2017

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“…During transformation, one or two T-DNA regions (TL and TR) are transferred to the plant genome (Kuzovkina & Schneider 2006). PCR analysis using primers for the virC gene provided no bands corresponding to the size of virC from the hairy root lines, thus eliminating the possibility of A. rhizogenes contamination (Fig.…”

Section: Pcr Analysis Of Transgenic Hairy Rootsmentioning

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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“…The molecular machinery involved in the transfer and integration of A. rhizogenes root-inducing DNA into the plant genome can act in trans on T-DNA regions of binary vectors to yield recombinant hairy roots featuring genes of interest (Ono et al, 2012 ). This approach has been successfully applied to manipulate metabolism occurring in transformed roots of several plant species (Kuzovkina and Schneider, 2006 ). Plants have been engineered to influence soil microorganism signaling as exemplified by transgenic efforts to quench the quorum-sensing-dependent virulence of pathogens belonging to the genus Pectobacterium (Czajkowski et al, 2011 ).…”

Section: Resultsmentioning

confidence: 99%

Plant hairy root cultures as plasmodium modulators of the slime mold emergent computing substrate Physarum polycephalum

et al. 2015

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Roots of the medicinal plant Valeriana officinalis are well-studied for their various biological activities. We applied genetically transformed V. officinalis root biomass to exert control of Physarum polycephalum, an amoeba-based emergent computing substrate. The plasmodial stage of the P. polycephalum life cycle constitutes a single, multinucleate cell visible by unaided eye. The plasmodium modifies its network of oscillating protoplasm in response to spatial configurations of attractants and repellents, a behavior that is interpreted as biological computation. To program the computing behavior of P. polycephalum, a diverse and sustainable library of plasmodium modulators is required. Hairy roots produced by genetic transformation with Agrobacterium rhizogenes are a metabolically stable source of bioactive compounds. Adventitious roots were induced on in vitro V. officinalis plants following infection with A. rhizogenes. A single hairy root clone was selected for massive propagation and the biomass was characterized in P. polycephalum chemotaxis, maze-solving, and electrical activity assays. The Agrobacterium-derived roots of V. officinalis elicited a positive chemotactic response and augmented maze-solving behavior. In a simple plasmodium circuit, introduction of hairy root biomass stimulated the oscillation patterns of slime mold's surface electrical activity. We propose that manipulation of P. polycephalum with the plant root culture platform can be applied to the development of slime mold microfluidic devices as well as future models for engineering the plant rhizosphere.

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Genetically transformed root cultures — generation, properties and application in plant sciences

Cited by 20 publications

References 206 publications

Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L.

Optimizing culture conditions for establishment of hairy root culture of Semecarpus anacardium L.

Induction of Gentiana cruciata hairy roots and their secondary metabolites

Plant hairy root cultures as plasmodium modulators of the slime mold emergent computing substrate Physarum polycephalum

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