Culture methods and detection of glucanases in suspension cultures of wheat and barley

Gamborg, O. L.; Eveleigh, D. E.

doi:10.1139/o68-063

Cited by 308 publications

(91 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In experiments to test the separate effects of growth regulators on shoot and callus initiation, 6-benzyladenine (BA), kinetin, 6-(γ,γ-dimethylallylamino)-purine (2iP), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), -naphthalene acetic acid (NAA) or 2,4-diclorophenoxyacetic acid (2,4-D) were added at either 0, 0.31, 0.62, 1.25, 2.5 or 5.0 µM to MS medium supplemented with 2% (w/v) sucrose and 6% (w/v) agar (Agar Type, Sigma Co.). In evaluations on the abilities of different basal media to support shoot culture establishment, MS (total and half strength), B5 (Gamborg et al 1968), AR (Anderson 1978), W (White 1963), SH (Schenk & Hildebrandt 1972), KM (Kao & Michayluk 1975) and WPM (Lloyd & McCown 1981) salts were supplemented with 0.62 µM BA. For root culture initiation, root segments were inoculated in 125 mL erlenmeyer flasks containing 20 mL of MS liquid medium supplemented with 2% (w/v) sucrose, 1.1 µM NAA and the cultures maintained on a rotatory shaker at 100 rpm.…”

Section: Methodsmentioning

confidence: 99%

In vitro culture of Phyllanthus stipulatus (Euphorbiaceae)

Catapan¹,

Otuki²,

Viana³

2001

Rev. bras. Bot.

View full text Add to dashboard Cite

-(In vitro culture of Phyllanthus stipulatus (Euphorbiaceae)). An efficient micropropagation protocol was developed for the medicinal plant Phyllanthus stipulatus (Euphorbiaceae) using nodal segments for axillary shoot proliferation. Maximum multiplication rates (8-9 shoots per explant) was achieved on MS media supplemented with either 2.5-5.0 µM IBA. The best basal media for axillary shoot proliferation when 0.62 µM BA was supplemented were MS, MS/2 and AR (4-5 shoots per explant). Rooting was achieved with 100% of the microshoots on MS medium without growth regulators. Regenerated plants were successfully acclimatized and about 88% of plantlets survived under ex vitro conditions. Flowering was observed in 81% of the ex vitro grown plantlets after 12 weeks of acclimatization. High frequency callus initiation and growth was achieved when nodal segment explants were inoculated either in the vertical position, in the light on MS medium supplemented with 5.0 µM NAA or horizontally oriented, in the dark on MS supplemented with 5.0 µM NAA or 1.25-5.0 µM BA or 2iP. Root cultures were successfully established on MS medium containing 1.1 µM NAA. The optimized micropropagation, callus and root culture protocols offer the possibility to use cell/organ culture techniques for vegetative propagation and secondary metabolism studies.RESUMO -(Cultura in vitro de Phyllanthus stipulatus (Euphorbiaceae)). Um eficiente protocolo de micropropagação foi desenvolvido para a espécie medicinal Phyllanthus stipulatus (Euphorbiaceae) através da utilização de segmentos nodais para a proliferação de ramos. Taxas máximas de multiplicação (8-9 ramos por explante) foram obtidas em meio MS suplementado com 2,5-5,0 µM de IBA. As melhores formulações de meios de cultura quando 0,62 µM de BA foi suplementado foram MS, MS/2 e AR (4-5 ramos por explante). O enraizamento foi obtido em 100% das microestacas em meio MS sem reguladores de crescimento. As microplantas regeneradas foram aclimatadas com successo e ao redor de 88% delas sobreviveram às condições ex vitro. A floração foi observada em 81% das microplantas crescidas ex vitro 12 semanas após a aclimatação. Altas freqüências de iniciação e crescimento de calos foram obtidas quando os segmentos nodais foram inoculados na posição vertical, na luz, no meio MS suplementado com 5,0 µM NAA ou na posição horizontal, no escuro em meio MS suplementado com 5,0 µM NAA or 1,25-5,0 µM de BA ou 2iP. Culturas de raízes foram estabelecidas em meio MS suplementado com 1,1 µM NAA. Os protocolos otimizados de micropropagação, cultura de calos e de raízes possibilitam a utilização das técnicas de cultura de células/órgãos para a propagação vegetativa e estudos sobre o metabolismo secundário.

show abstract

Section: Methodsmentioning

confidence: 99%

In vitro culture of Phyllanthus stipulatus (Euphorbiaceae)

Catapan¹,

Otuki²,

Viana³

2001

Rev. bras. Bot.

View full text Add to dashboard Cite

show abstract

“…,-1 , 3-Glucanases have been reported to be widespread in higher plants (6). Among these are two reports of ,B-1 ,3-glucanase activity in suspension-cultured plant cells (10,18 growth (19,20,28,29). Cell wall-degrading enzymes have been reported to be present in crude preparations of cell walls (12)(13)(14).…”

mentioning

confidence: 99%

“…,-1 , 3-Glucanases have been reported to be widespread in higher plants (6). Among these are two reports of ,B-1 ,3-glucanase activity in suspension-cultured plant cells (10,18). Recently, Heyn reported a number of glucanase activities in Avena coleoptiles, including a very active 0-1,3-glucanase (11).…”

mentioning

confidence: 99%

The Involvement of Glycosidases in the Cell Wall Metabolism of Suspension-cultured Acer pseudoplatanus Cells

Keegstra

Albersheim

1970

Plant Physiol.

View full text Add to dashboard Cite

Several glycosidases have been isolated from suspensioncultured sycamore (Acer pseudoplatanus) cells. These include an a-galactosidase, an a-mannosidase, a f-N-acetylglucosaminidase, a ,B-glucosidase, and two ,3-galactosidases.The pH optimum of each of these enzymes was determined. Glycosidases have been reported in a variety of bacteria, fungi, animals, and higher plants. In plants, glycosidases have long been known to be present in seeds and storage material (1,2,4,8,9,15,17,(24)(25)(26)(27). Recently, there has been considerable interest in these glycosidases. One of the major reasons for the increased interest in these enzymes has been their usefulness as tools for studying the structure of oligo-and polysaccharides. Li has studied an a-mannosidase from jack bean meal and has used it to study a-mannosides present in glycoproteins (15-17). Agrawal and Bahl have studied the glycosidases from the cotyledons of Phaseolus vulgaris (1, 4). They have used these enzymes to study the structure of the saccharide portion of a glycoprotein hormone (3). An a-galactosidase from coffee beans has been used to study various galactomannans (7).Further interest in these enzymes stems from the possibility that they may be involved in growth of microbial and plant cells. ,-1 , 3-Glucanases have been reported to be widespread in higher plants (6). Among these are two reports of ,B-1 ,3-glucanase activity in suspension-cultured plant cells (10,18 growth (19,20,28,29). Cell wall-degrading enzymes have been reported to be present in crude preparations of cell walls (12)(13)(14). These findings suggest that glycosidases partially degrade cell walls and, in so doing, facilitate cell growth.In this report we describe a number of glycosidases of sycamore cells (Acer pseudoplatanus) grown in suspension culture. Some of the glycosidases are found associated with the cell walls. The activity of some of these hydrolytic enzymes is greater during log phase growth than prior to or after the growth period of these suspension-cultured cells.MATERIAL AND METHODS Sycamore cells (A. pseudoplatanus) were grown on M-6 media as described earlier (5) except that 20 g/liter of sucrose was used rather than 40 g/liter.For the growth experiments, cells were grown in 2800-ml Fernbach flasks with a three-way stopcock attached near the bottom. The stopcock was sterilized by passing 70% ethanol through it before and after each use. Cells were withdrawn while the culture was agitated by a magnetic stirrer. Representative samples of the cell population could be obtained in this manner.The cells were collected on a coarse scintered glass funnel. The collected cells were washed with 5 volumes of 0.5 M potassium phosphate, pH 7.0, and the wash solution was saved. The washed cells were suspended in additional phosphate buffer (1.0 ml of buffer for each g of cells) and disrupted by forcing the suspension through a French pressure cell at 2,000 p.s.i. The resulting preparation was centrifuged for 60 min at 40,000g.The enzymes in the 0.5 M wash from the whole ...

show abstract

“…radiobacter carrying a nopaline Ti plasmid forms the subject of this paper. citrate, 0.5 g; sodium glutamate, 1.0 g; K2HP04, 1.05 g; KH2P04, 0.45 g; Fe(N03)3, 5 mg; biotin, 200 pg; trace elements (Gamborg & Eveleigh, 1968), 1.0 ml; and (added as separately sterilized solutions) glucose, 2.0 g; MgS04. 7H20, 0.2 g; thiamin, 5 mg; the final pH was 7.0.…”

Section: Introductionmentioning

confidence: 99%

A Basis for Agrocin 84 Sensitivity in Agrobacterium radiobacter

Murphy¹,

Roberts²

1979

Journal of General Microbiology

View full text Add to dashboard Cite

~ ~~~Agrocin 84 inhibits a virulent strain of Agrobacterium radiobacter containing a nopaline Ti plasmid, but not the same strain lacking the Ti plasmid; this specificity was investigated. Sensitivity to agrocin 84 was correlated with its active uptake by a high-affinity transport system with a K, of 5-88 x M. Unmetabolized agrocin 84 was transported inside the sensitive strain and was associated with the soluble fraction of ruptured bacteria and not with the outer walls or cytoplasmic membrane. Agrocin 84 bound to a protein fraction isolated from the periplasmic space of the sensitive strain; there was no equivalent binding activity in the insensitive strain. It is proposed that sensitivity to agrocin 84 is due to the presence of one or more plasmid-coded binding proteins which are associated with transport of agrocin 84 into sensitive strains.

show abstract

Culture methods and detection of glucanases in suspension cultures of wheat and barley

Cited by 308 publications

References 0 publications

In vitro culture of Phyllanthus stipulatus (Euphorbiaceae)

In vitro culture of Phyllanthus stipulatus (Euphorbiaceae)

The Involvement of Glycosidases in the Cell Wall Metabolism of Suspension-cultured Acer pseudoplatanus Cells

A Basis for Agrocin 84 Sensitivity in Agrobacterium radiobacter

Contact Info

Product

Resources

About