Nutrient requirements of suspension cultures of soybean root cells

Gamborg, O. L.; Miller, Rebecca; Ojima, Kunihiko

doi:10.1016/0014-4827(68)90403-5

Cited by 8,615 publications

(3,324 citation statements)

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“…2 cm in length), isolated from ca. 2-week-old root cultures, was pre-propagated in liquid B5 medium [27] containing 1% sucrose for 2 weeks. The medium was then exchanged for either a Fe-deficient B5 medium or a Fe-containing B5 medium, each with 1% sucrose, followed in each case by further culture for up to 1 week.…”

Section: Root Materials and Culture Conditionsmentioning

confidence: 99%

“…The medium was then exchanged for either a Fe-deficient B5 medium or a Fe-containing B5 medium, each with 1% sucrose, followed in each case by further culture for up to 1 week. Only in the case of observation by confocal microscopy, a root tip was directly inoculated and cultured on solid Fe-deficient B5 medium [27] containing 1% sucrose. Fe-deficient B5 medium was prepared by omission of Fe-EDTA from B5 basal medium, prior to autoclaving at 121 ˚C for 15 min.…”

Section: Root Materials and Culture Conditionsmentioning

confidence: 99%

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Increased de novo riboflavin synthesis and hydrolysis of FMN are involved in riboflavin secretion from Hyoscyamus albus hairy roots under iron deficiency

Higa

Khandakar

Mori

et al. 2012

Plant Physiology and Biochemistry

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Riboflavin secretion by Hyoscyamus albus hairy roots under Fe deficiency was examined to determine where riboflavin is produced and whether production occurs via an enhancement of riboflavin biosynthesis or a stimulation of flavin mononucleotide (FMN) hydrolysis. Confocal fluorescent microscopy showed that riboflavin was mainly localized in the epidermis and cortex of the root tip and, at the cellular level, in the apoplast. The expressions of three genes involved in the de novo biosynthesis of riboflavin (GTP cyclohydrolase II/3,6, riboflavin synthase) were compared between Fe-starved and Fe-replete roots over a time course of 7 days, using RT-PCR. All three genes were found to be highly expressed over the period 1-7 days in the roots cultured under Fe deficiency. Since riboflavin secretion began to be detected only from 3 days, there was a lag phase observed between the increased transcript accumulations and riboflavin secretion. To determine whether FMN hydrolysis might contribute to the riboflavin secretion in Fe-deficient root cultures, FMN hydrolase activity was determined and was found to be substantially increased after 3 days, when riboflavin secretion became detectable. These results suggested that not only de novo riboflavin synthesis but also the hydrolysis of FMN contributes to riboflavin secretion under conditions of Fe deficiency. Respiration activity was assayed during the time-course, and was also found to be enhanced after 3 days under Fe deficiency, suggesting a possible link with riboflavin secretion. On the other hand, several respiratory inhibitors were found not to affect riboflavin synthase transcript accumulation.Key words: Hyoscyamus albus; hairy roots; iron deficiency; riboflavin secretion; riboflavin biosynthesis; FMN hydrolase; respiration. RibA: GTP cyclohydrolase II · RibB: 3,4-dihydroxy-2-butanone 4-phosphate synthase · RibC: 6,7-dimethyl-8-ribityllumazine synthase · RibD: riboflavin synthase

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Section: Root Materials and Culture Conditionsmentioning

confidence: 99%

Section: Root Materials and Culture Conditionsmentioning

confidence: 99%

Increased de novo riboflavin synthesis and hydrolysis of FMN are involved in riboflavin secretion from Hyoscyamus albus hairy roots under iron deficiency

Higa

Khandakar

Mori

et al. 2012

Plant Physiology and Biochemistry

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“…The hard nodular cell masses termed as 'nodular calli' were induced on the BAP preconditioned explants cultured on the Gamborg's B 5 medium (Gamborg et al 1968) …”

Section: Callus Inductionmentioning

confidence: 99%

Genetic transformation of Indian isolate of Lemna minor mediated by Agrobacterium tumefaciens and recovery of transgenic plants

Chhabra

Chaudhary

Sainger

et al. 2011

Physiol Mol Biol Plants

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Transgenic plants of an Indian isolate of Lemna minor have been developed for the first time using Agrobacterium tumefaciens and hard nodular cell masses 'nodular calli' developed on the BAP -pretreated daughter frond explants in B 5 medium containing sucrose (1.0 %) with 2,4-D (5.0 μM) and 2-iP (50.0 μM) or 2,4-D (50.0 μM) and TDZ (5.0 μM) under light conditions. These calli were co-cultured with A. tumefaciens strain EHA105 harboring a binary vector that contained genes for β-glucuronidase with intron and neomycin phosphortransferase. Transformed cells selected on kanamycin selection medium were regenerated into fronds whose transgenic nature was confirmed by histochemical assay for GUS activity, PCR analysis and Southern hybridization. The frequency of transformation obtained was 3.8 % and a period of 11-13 weeks was required from initiation of cultures from explants to fully grown transgenic fronds. The pretreatment of daughter fronds with BAP, use of non-ionic surfactant, presence of acetosyringone in co-cultivation medium, co-culture duration of 3 d and 16 h photoperiod during culture were found crucial for callus induction, frond regeneration and transformation of L. minor. This transformation system can be used for the production of pharmaceutically important protein and in bioremediation.

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“…Effect of antioxidants on the transformation of garlic calli To optimize the efficiency of gene transfer into garlic, we examined the influence of basal salts such as N6 (Chu et al, 1975), MS (Murashige and Skoog, 1962), and B5 (Gamborg et al, 1968) on this process. Our preliminary experiment using calli derived from the roots of garlic cv.…”

Section: Resultsmentioning

confidence: 99%

Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

Ahn

Yoon

Jeon

2013

Molecules and Cells

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The genetic improvement of garlic plants (Allium sativum L.) with agronomical beneficial traits is rarely achieved due to the lack of an applicable transformation system. Here, we developed an efficient Agrobacterium-mediated transformation procedure with Danyang, an elite Korean garlic cultivar. Examination of sGFP (synthetic green fluorescence protein) expression revealed that treatment with 2-(N-morpholino) ethanesulfonic acid (MES), L-cysteine and/or dithiothreitol (DTT) gives the highest efficiency in transient gene transfer during Agrobacterium co-cultivation with calli derived from the roots of in vitro plantlets. To increase stable transformation efficiency, a two-step selection was employed on the basis of hygromycin resistance and sGFP expression. Of the hygromycin-resistant calli initially produced, only sGFP-expressing calli were subcultured for selection of transgenic calli. Transgenic plantlets produced from these calli were grown to maturity. The transformation efficiency increased up to 10.6% via our optimized procedure. DNA and RNA gel-blot analysis indicated that transgenic garlic plants stably integrated and expressed the phosphinothricin acetyltransferase (PAT) gene. A herbicide spraying assay demonstrated that transgenic plants of garlic conferred herbicide resistance, whilst nontransgenic plants and weeds died. These results indicate that our transformation system can be efficiently utilized to produce transgenic garlic plants with agronomic benefits.

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Nutrient requirements of suspension cultures of soybean root cells

Cited by 8,615 publications

References 10 publications

Increased de novo riboflavin synthesis and hydrolysis of FMN are involved in riboflavin secretion from Hyoscyamus albus hairy roots under iron deficiency

Increased de novo riboflavin synthesis and hydrolysis of FMN are involved in riboflavin secretion from Hyoscyamus albus hairy roots under iron deficiency

Genetic transformation of Indian isolate of Lemna minor mediated by Agrobacterium tumefaciens and recovery of transgenic plants

Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

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