The application of root cultures to problems of biological chemistry

Robins, Richard J.

doi:10.1039/a815549y

Cited by 14 publications

(3 citation statements)

References 230 publications

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“…45 Robins reported a complete loss of nicotine in the root clones of Nicotiana rustica treated with synthetic auxins. 46 Strikingly, the roots with TK morphology were 100% positive for the TL-and TR-DNA genes. In contrast, the TN roots showed 0% presence of the aux1 and ags genes, and only 16.7% of them were positive for mas1 integration into their genome.…”

Section: Tl-and Tr-dna Integration Into the Genome Of Transgenic Hairmentioning

confidence: 99%

Effect of pRi T-DNA genes and elicitation on morphology and phytoecdysteroid biosynthesis in Ajuga bracteosa hairy roots

et al. 2017

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The medicinal plant Ajuga bracteosa is a rich source of biologically active and metabolism-enhancing phytoecdysteroids. Transgenic hairy roots from A. bracteosa were obtained by infection with Agrobacterium rhizogenes strains A4, LBA-9402 and ARqua1. The 59 selectively established and rolCpositive hairy root lines increased in size up to 6.6-fold (L42) after one month of in vitro culturing and produced a phytoecdysteroid content ranging from 69.3 to 4449 mg g À1 (L3 and A2, respectively). The clones of these transgenic roots were maintained for successive subcultures on hormone-free medium to obtain a stabilized morphology. Hairy roots displayed four different morphologies: typical hairy root (THR) (59%), callus-like (CM) (17%), thick (TK) (14%) and thin (TN) (10%). The growth rate of the transgenic hairy root lines varied according to their morphology, with CM showing the highest rate (3.93-fold per month). However, THR exhibited the highest phytoecdysteroid content (1538.5 mg g À1 ). Considering the hairy root morphology in relation to the pRi TR-DNA genes, 100% of the hairy root lines harbored the mas1 and ags genes and 90% of CM lines the aux1 gene. Strikingly, the clones with TK morphology were all positive for the TL-and TR-DNA genes. In contrast, 0% of TN clones harbored the aux1 and ags genes, and only 16.7% the mas1 gene. All CM and TK hairy roots were positive for the ags gene, while mas1 was present in all hairy root types except TN. As expected, all the root lines considered in this work were positive for the rol C gene. Eleven hairy root lines displaying a high growth rate and phytoecdysteroid content were elicited with methyl jasmonate (MeJ) and coronatine (Cor). MeJ doubled the phytoecdysteroid content after 14 days of elicitation (8356 mg g À1 in L2) compared with unelicited control hairy roots, and in in vitro-grown untransformed roots the increase was 5.6-fold. a Growth capacity of transgenic clones of different root lines expressed as (harvested FW/inoculum FW) aer 28 days of culture in hormone-free half strength MS medium. Each value is the average of 3 replicates. THR: typical hairy root, CM: callus like morphology, TK: hairy root with thick morphology, TN: hairy root with thin morphology, PE: phytoecdysteroid content.47948 | RSC Adv., 2017, 7, 47945-47953This journal is Fig. 3 HPLC chromatographs showing the elution of phytoecdysteroids. (a) Standard ecdysteroids (100 mg ml À1 of each standard), (b) phytoecdysteroid elution pattern in one of the hairy root extract.This journal is

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Section: Tl-and Tr-dna Integration Into the Genome Of Transgenic Hairmentioning

confidence: 99%

Effect of pRi T-DNA genes and elicitation on morphology and phytoecdysteroid biosynthesis in Ajuga bracteosa hairy roots

et al. 2017

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“…In recent years, the use of root organ cultures was found to be efficient for the production of natural compounds, e.g., tropane alkaloids [35]. Thus, hairy root cultures of Datura candida x D. aurea were established after transformation with Agrobacterium rhizogenes strains.…”

Section: Applicationsmentioning

confidence: 99%

Capillary electrophoresis-diode array detection — electrospray mass spectrometry for the analysis of selected tropane alkaloids in plant extracts

et al. 1999

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Capillary zone electrophoresis, coupled to UV and interfaced with electrospray ionization mass spectrometry (ESI‐MS), is described for the simultaneous analysis of hyoscyamine and scopolamine. On‐line UV detection occurred at 22 cm from the inlet of the capillary and ESI‐MS monitoring was performed along the entire length of the capillary (85 cm). An alkaline solution of 40 mM ammonium acetate at pH 8.5 was suitable for the analysis of the alkaloids under consideration. Under the optimized conditions, including CE and ESI‐MS parameters, the two alkaloids were resolved within a short time and with very high sensitivity. The differentiation of hyoscyamine and its positional isomer littorine, commonly encountered in plant material, is also presented using up‐front collision‐induced dissociation. Finally, the developed method was applied to the analysis of these alkaloids in Belladonna leaf extract and in Datura candida x D. aurea hairy root extract.

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“…Several studies using different transformed roots have shown genetic and biochemical stabilities over 10 years (Baiza et al, 1999;Lanoue et al, 2002;Maldonado-Mendoza et al, 1993). Since the mid-1980s, hairy roots have been used as an experimental system for plant root metabolism (Robins, 1998) and also as an alternative to plant cell suspension cultures for the production of fine chemicals (Shanks and Morgan, 1999) on a large scale (Choi et al, 2000;Rhodes et al, 1992). More than 80 plant species have already been transformed with A. rhizogenes (Giri and Narasu, 2000), providing a wide spectrum of applications.…”

Section: Introductionmentioning

confidence: 99%

Occurrence of circadian rhythms in hairy root cultures grown under controlled conditions

Lanoue

Shakourzadeh

Marison

et al. 2004

Biotech & Bioengineering

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Hairy roots obtained by transformation via Agrobacterium rhizogenes provide an artificial plant material devoid of aerial parts with high growth on hormone-free media. Fundamental knowledge of hairy root physiology is essential to develop and control its culture. In contrast to shake-flask cultures, a bioreactor set-up combined with on-line data logging provides an efficient tool to study rapid physiological variations in hairy root cultures. Datura innoxia hairy roots were grown in a bioreactor equipped with on-line data analyses of pH, dissolved oxygen (pO2), conductivity, oxygen, and carbon dioxide. The experiments were done at a constant temperature and in the absence of light cues. The results obtained showed that the carbon dioxide evolution rate (CER) presented regular oscillations during the culture. Similar oscillations were also observed for the oxygen uptake rate (OUR). These signals were treated mathematically to look for the existence of a rhythm. An autocorrelation function was used to detect any periodic components. The results demonstrate that hairy root respiration exhibited peaks of 1 day. These oscillations, having a period of about 24 h, were also observed in pH and conductivity signals, although not for the pO2 signal. The data acquired in the absence of hairy roots showed that the observed periodic behavior was not an artifact. No effect on rhythms was observed by the imposition of an external "day/night" cycle. The fact that oscillations persisted in the absence of external stimuli, with a free-running period of 24 h, suggests that a circadian rhythm exists in hairy roots of D. innoxia.

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The application of root cultures to problems of biological chemistry

Cited by 14 publications

References 230 publications

Effect of pRi T-DNA genes and elicitation on morphology and phytoecdysteroid biosynthesis in Ajuga bracteosa hairy roots

Effect of pRi T-DNA genes and elicitation on morphology and phytoecdysteroid biosynthesis in Ajuga bracteosa hairy roots

Capillary electrophoresis-diode array detection — electrospray mass spectrometry for the analysis of selected tropane alkaloids in plant extracts

Occurrence of circadian rhythms in hairy root cultures grown under controlled conditions

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