Showing papers on "Murashige and Skoog medium published in 1995"

Improved plant regeneration from barley callus cultures by increased copper levels

Abstract: Incorporation of cupric sulfate into callus induction, maintenance, and regeneration media significantly enhanced plant regeneration from callus cultures of barley (Hordeum vulgare L.) immature embryos. Embryos from the cultivars ‘Hector’ and ‘Excel’ were cultured on MS medium containing 0, 0.1 (MS level), 0.5, 1.0, 5.0, 10.0, 50.0, or 100.0 μM cupric sulfate. Plants were regenerated beginning at 8 weeks and continuing through 36 weeks. For Hector, medium containing 50 μM copper regenerated significantly more plants than any other medium, with an average of 17 plants per embryo. In comparison, medium with MS copper levels (0.1 μM) regenerated only 5 plants per embryo. For Excel, medium containing 5.0 μM copper was the best, regenerating 1.4 plants per embryo. No Excel regenerants were obtained on medium with MS copper levels. Increased copper levels also increased the percentage of embryos that regenerated at least one plant, in both cultivars. The results indicate that MS copper levels are not optimized for barley callus cultures, and that improved plant regeneration can be obtained at higher copper concentrations.

Establishment of oil palm cell suspensions and plant regeneration

Abstract: Primary globular callus from immature zygotic embryos and friable embryogenic tissue derived from mature zygotic embryos were used to establish suspension cultures. Callus cultures were established either on modified Y3 or MS medium containing 475–500 μM 2,4-D or 250 μM picloram and 0.3% (w/v) activated charcoal. Suspension cultures of both cell lines were established in modified Y3 medium containing 10 μM 2,4-D. The establishment of cell suspensions from friable embryogenic tissue took only 2 months, in contrast with suspensions from primary globular callus which took 3–5 months to establish. Embryo differentiation was observed only in cell suspensions derived from the friable embryogenic tissue after plating aliquots on regeneration medium. Germinated embryos were recovered and plantlets were successfully established under greenhouse conditions.

Use of starch-gelled medium for tissue culture of some fruit crops

Abstract: Six cultivars of apple and two of red raspberry consistently produced equal or significantly better shoot proliferation on modified Murashige and Skoog medium gelled with a mixture of corn starch and Gelrite than on the same medium gelled with agar. Two pear cultivars grown on starch-Gelrite medium produced hyperhydric shoots and almost no growth, but the addition of a polysaccharide hydric control (‘antivitrifying’) agent to the medium eliminated hyperhydricity. The resulting shoot proliferation equaled or exceeded that on the agar-gelled medium. The starch-Gelrite mixture is easy to prepare and gelling agent costs are only 10–15% of agar, or less if starch is purchased in bulk. Although the opaque gray-white medium makes it more difficult to detect internal contaminants, external contaminants are easily discerned.

Cryopreservation of in vitro-grown apical meristems of lily by vitrification

Abstract: Apical meristems from adventitious buds induced by culturing of bulb-scale segments of Japanese Pink Lily (Lilium japonicum Thunb) were successfully cryopreserved by a vitrification The excised apical meristems were precultured on a solidified Murashige & Skoog medium, containing 03 M sucrose, for 1 day at 25°C and then loaded in a mixture of 2 M glycerol plus 04 M sucrose for 20 min at 25°C Cryoprotected meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) at 25°C for 20 min or at 0°C for 110 min prior to a plunge into liquid nitrogen After rapid warming in a water bath at 40°C, the meristems were placed in 18 ml of 12 M sucrose for 20 min and then, placed on filter papers over gellan gum-solidified MS medium The revived meristems resumed growth within 5 days and directly produced shoots The rate of shoot formation was approximately 80% after 4 weeks When bulb-scale segments with adventitious buds were cold-hardened at 0°C for more than 7 days before the procedure, the rates of shoot formation were significantly increased This vitrification method was successfully applied to five other lily cultivars Thus, this vitrification procedure for cryopreservation appears promising as a routine method for cryopreserving meristems of lily

Somatic embryogenesis of Prunus subhirtella autumno rosa and regeneration of transgenic plants after Agrobacterium-mediated transformation.

Abstract: Embryogenic lines of Prunus subhirtella autumno rosa were established on a modified MS medium supplemented with 1 mg/l NAA, 0.06 mg/l IBA and 0.04 mg/l BA from petioles of axenically grown shoots of adult origin. To induce normal development of plantlets we compared a range of approaches on solid culture media as well as in suspension cultures including treatments with ABA, GA3, zeatin, darkness, and cold. A series of experiments were conducted to follow the temporal pattern of somatic embryo development.

In vitro microrhizome production in Zingiber officinale Rosc.

Abstract: Microrhizomes of Zingiber officinale were successfully produced from tissue culture derived shoots by transferring them to liquid MS medium supplemented with 1 mg/l BAP, 2 mg/l calcium pantothenate, 0.2 mg/l GA3 and 0.05 mg/l NAA for shoot proliferation. After 4 weeks of incubation, the medium was replaced with microrhizome induction medium, consisting of MS salts supplemented with 8 mg/l BAP and 75 g/l sucrose. Microrhizome formation started after 20 d of incubation in stationary cultures at 25+1 ° in the dark. Microrhizomes with 1-4 buds and weighing 73.8 to 459 mg each were harvested after 50-60 d. After storage for 2 months in moist sand at room temperature, 80% of the microrhizomes sprouted producing roots and shoots.

Somatic embryogenesis and plant regeneration from callus culture of Acacia catechu - a multipurpose leguminous tree

Abstract: Plant regeneration via somatic embryogenesis was achieved from callus derived from immature cotyledons of Acacia catechu Willd. on Woody Plant Medium (WPM) supplemented with 13.9 μM kinetin and 2.7 μM 1-naphthaleneacetic acid. The addition of 0.9–3.5 mM L-proline to the medium influenced development of somatic embryos and also promoted secondary somatic embryogenesis. The light-green somatic embryos germinated on half-strength MS medium supplemented with 2% (w/v) sucrose. Somatic embryos germinated into plantlets that were acclimatized in the greenhouse and subsequently transferred to the field.

A Novel Method for Rapid Micropropagation of Pineapple

Abstract: A novel micropropagation method for pineapple (Ananas comosus L.), based on shoot elongation induced in vitro, was demonstrated for two cultivars. Decapitated in vitro plantlets were used as explants. Shoot etiolation was induced by placing explants in a Murashige and Skoog (MS) medium containing NAA (10 μM) and incubating in darkness at 28C for 30 to 40 days. The mean number of the regenerated etiolated shoots per explant was 2.6 ± 0.29. The etiolated shoots were placed into N6 medium supplemented with kinetin or BA (25 or 20 μM, respectively). After 4 to 6 weeks, shoots regenerated along the nodes. The highest regeneration rate was 15 and 13 plantlets per node with 25 μM kinetin and 20 μM BA, respectively. Regenerated plantlets were rooted on a growth-regulator-free MS medium. Residual shoots of the initial explants could be recycled by rooting on a growth- regulator-free MS medium. This procedure enables the regeneration of several thousand plantlets per year. Chemical names used: naphthaleneacetic acid (NAA); benzyladenine (BA).

Isolated microspore culture of wheat ( Triticum aestivum L.) in a defined media I. Effects of pretreatment, isolation methods, and hormones

Abstract: Significant improvements were achieved in the production of haploid and doubled haploid plants from isolated microspore culture of wheat c.v. Chris on a defined media. Procedures found to be of benefit included: A 7-day pretreatment of anthers in 0.4M mannitol plus the macronutrients from FHG medium; the inclusion of 4.5 mg/liter abscisic acid in the pretreatment solution; the isolation of microspores from pretreated anthers by vortexing; and the use of phenylacetic acid (PAA) as the auxin source in MS medium. The best response was achieved with 4.0 mg/liter PAA in MS medium containing 90 g/liter maltose as the sugar source. Under these conditions, 68% of viable microspores underwent division, and an average of 93 embryos and 92 green plants were regenerated per 100 anthers used. The root-tip chromosome number and the fertility of 114 regenerating green plants revealed that 75% were completely fertile spontaneously doubled haploids.

Antihepatotoxic agent in micropropagated plantlets of Eclipta alba

Abstract: A protocol for in vitro micropropagation of Eclipta alba (L.) Hassk from nodal segment explants has been established. The maximum number of shoots was obtained after 60 days of culture in Murashige & Skoog (MS) medium supplemented with 4.4 μM benzyladenine. Multiple rooting was achieved using MS medium with 2.4 μM 2-isopentyladenine. Wedelolactone was present in shoots cultured in media containing cytokinins.

Callus induction and plant regeneration from gladiolus

Abstract: A method for the initiation of callus capable of plant regeneration from in vivo grown cormels of gladiolus (Gladiolus x grandiflorus Hort.) is described. Sliced cormels of the large-flowering hybrid, ‘Peter Pears’ were cultured in vitro on a modified Murashige and Skoog medium, supplemented with various auxins. Yellow callus, which was either friable or compact, could be induced on all media tested. Callus induced on media with naphthaleneacetic acid failed to proliferate. Callus induced on media with 9 mM 2,4-dichlorophenoxyacetic acid showed the best growth. Addition of micro-elements and vitamins increased the induction and growth of callus capable of plant regeneration. Explants taken from the middle part of the cormels had the highest competence for callus initiation. Callus was induced on several gladiolus hybrids and the South African species G. garnierii Klatt. Callus induction was genotype dependent and among the cultivars tested, ‘Peter Pears’ and ‘White Prosperity’ were superior with respect to callus production on the media with either 2,4-dichlorophenoxyacetic acid or picloram. Plants were regenerated from yellow compact callus of all genotypes on media containing zeatin and benzyladenine in various concentrations.

Plantlet regeneration through different morphogenic pathways in pommelo tissue culture

Abstract: Pommelo (Citrus grandis Osbeck) plantlets were regenerated through different morphogenic pathways in culture. Multiple shoot regeneration through de novo organogenesis was obtained with epicotyl segments and root cultures. Shoot regeneration was observed in 84% of the midtal epicotyl segments cultured in Murashige and Skoog's medium (MS) with 2.2 μM benzyladenine (BA) and 83% of the middle and proximal epicotyl segments cultured on basal medium. Isolated root segments cultured on medium containing 0.089 μM BA showed best shoot regeneration at 71% with an average of 3.3 shoots per segment. Callus tissues derived from cotyledon and leaf explants regenerated shoots on BA-enriched medium. Shoots were also obtained at high frequencies from shoot-tip and nodal explants. Roots developed when regenerated shoots were excised and cultured on half strength MS medium with 2.5 μM indolebutyric acid.

In vitro culture of switchgrass: Influence of 2,4-d and picloram in combination with benzyladenine on callus initiation and regeneration

Abstract: The objective of this study was to determine the types and concentrations of auxins and cytokinins that would result in effective culture initiation and plant regeneration of switchgrass (Panicum virgatum L.). Whole caryopses and young seedling explants were cultured on MS medium containing different concentrations of picloram or 2,4-dichlorophenoxyacetic acid (2,4-d) in combination with benzyladenine. Best results from mature caryopses were obtained with 11.3–45.0 μM 2,4-d in combination with 15.0 or 45.0 μM benzyladenine (BA). More than 300 plants were obtained per embryogenic callus from some treatments after the second transfer to regeneration medium (90 days after initiation of the cultures). Regeneration was obtained from young seedling explants with both auxins. However, picloram was more effective over a wider range of BA concentrations than 2,4-d. Protocols developed during this study were used to regenerate hundreds of plants which could easily be established in the field.

Improved shoot-tip micropropagation of Pistacia vera L. and the beneficial effects of methyl jasmonate

Abstract: Micropropagation of Pistacia vera 'Mateur' was improved by adding MeJA to the multiplication and rooting media. Shoot-tip cultures established from grafted trees were maintained on a modified Murashige and Skoog medium containing 5 µ M BA and 0.05 µM IBA. Adding 0.3, 1, or 3.2 µM MeJA improved shoot multiplication rates 2.5, 3.0, and 2.3, respectively. There was a significant interaction between the effects of auxin and temperature on the percentage of shoots forming roots. At 25C, the percentage of shoots forming roots was higher in the presence of NAA than IAA or IBA, whereas, at 28C, there was no difference among the auxins. Adding MeJA to the best auxin treatments-31.6 µ M NAA at 25C and 31.6 µM IAA at 28C-increased the percentage of shoots forming roots and number of roots per shoot but decreased root length. More than 80% of the shoots rooted at 25C when 1 µM MeJA was added to the root induction medium, which contained 31.6 µM NAA, and the root elongation medium, without auxin. The large number of short roots induced by MeJA facilitated plantlet transfer to soil and acclimation. Chemical names used: methyl jasmonate (MeJA); N 6 -benzyladenine (BA), indole-3-butyric acid (IBA), α- naphthaleneacetic acid (NAA), indole3-acetic acid (IAA).

In vitro propagation ofCereus peruvianus mill. (cactaceae)

Abstract: Cereus peruvianus seedlings were used as a source of stem explants to determine the effective conditions for inducing and maintaining callus tissues in a state of rapid growth, as well as to obtain plants regenerated from callus cultures. Factorial combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin in MS medium were tested, and we concluded that the 18.1µM 2,4-D and 18.6 or 27.9µM kinetin combinations were suitable for callus induction. The cactus shoots were produced from the friable callus; root elongation occurred within 2 wk in medium without 2,4-D and with 18.6µM kinetin. This method can be used to rapidly produce manyC. peruvianus plants.

Role of liquid versus agar-gelled media in mass propagation and ex vitro survival in bananas

Abstract: Shoot cultures from meristem explants of three cultivars of banana, ‘Cavendish’, ‘Bluggoe’ and ‘Silk’ were established on Murashige and Skoog medium with 8.9 μM benzyladenine and 0.98 μM indolebutyric acid. Different types of media, such as agar-gelled, agitated liquid and static liquid, were assessed for their ability to support shoot multiplication and ex vitro survival. Liquid media were found better for shoot multiplication whereas agar-gelled medium supported maximum ex vitro survival. For maximum plantlet production, growing in static liquid medium followed by a brief culture on agar-gelled medium has been suggested.

Plant regeneration from mesophyll protoplasts of lisianthus (Eustoma grandiflorum) by adding activated charcoal into protoplast culture medium

Abstract: Plant regeneration from isolated protoplasts of 8 cultivars of lisianthus, Eustoma grandiflorum (Griseb.) Schinners, has been established by using activated charcoal. Protoplasts were isolated from lisianthus leaves grown in vitro and started to divide within 3–4 days of culture, but successful colony formation was only achieved by adding gellan gum blocks containing 1% (w/v) activated charcoal immediately after culture. Colonies consisting of as many as 50–100 cells formed after 30 days of culture and were transferred to fresh medium for callus proliferation and shoot regeneration, respectively. These shoots rooted on MS medium containing 0.5 mg l−1 indolebutyric acid(IBA) and the plantlets were finally transplanted to pots. Morphological characteristics, growth habit and pollen fertility of protoplast-derived plants of one cultivar were not different from those of seed-grown plants as control.

Agrobacterium -mediated Transformation of Torenia (Torenia fournieri)

Abstract: A method for Agrobacterium -mediated transformation of torenia (Scrophulariaceae) is reported. Leaf segments of torenia plants grown in vitro were infected with the Agrobacterium strain LBA4404 harboring a binary vector containing the neomycin phosphotransferase IIgene, P -glucuronidase (GUS) gene lvith an intron and hygromycin phosphotransferase gene. The infected explants were cultured on Murashige and Skoog medium supplemented with 1 mg/l benzyladenine, 100 mgfl carbenicillin and 300 mg/l kanamycin (selection medium) for regeneration. Twelve shoots out of 67 shoots which regenerated from green compact calli on the selection medium were GLTS-positive. Polymerase chain reaction analysis confirmed the presence of the GUS gene in the GUS-positive plants. Mendelian inheritance of the GUS activity in the progenies of the transformed plants was demonstrated.

Factors affecting in vitro shoot proliferation and ex vitro establishment of mangosteen

Abstract: Shoot proliferation has been achieved in Garcinia mangostana L using seed explants Maximum mean number of shoots per explant (168) was obtained from cultures on Murashige and Skoog medium supplemented with 40 mM 6- benzyladenine, and 25 mM α-naphthaleneacetic acid and kept at 30 °C under an 8 hour photoperiod Cultures on the same medium but supplemented with 2 g l-1 activated charcoal produced fewer shoots However, growth of these shoots was more organized and 75% rooting was obtained Woody Plant Medium was not a suitable medium for shoot proliferation Ex vitro establishment was best obtained on planting medium consisting of sand, soil and organic material (3:2:1)

Rapid in vitro multiplication of jujube through mature stem explants.

Abstract: Stem explants obtained from a mature tree of Ziziphus mauritiana Lamk were grown on modified Murashige and Skoog medium containing 3800 mg l-1 potassium nitrate, 2475 mg l-1 ammonium nitrate, 11 μM benzyladenine and 0.5 μM indole-3-acetic acid. During successive subcultures 15–20 shoots per inoculum were produced. Rooting was induced by pretreatment with 50 μM indolebutyric acid or 1-naphthaleneacetic acid for 24 h followed by transfer to auxin-free White's medium. Plantlets grew well in a soil and vermiculite mixture.

Vegetative propagation of adult Eucalyptus grandis X urophylla and comparison of growth between micropropagated plantlets and rooted cuttings.

Abstract: Methods for the production of micropropagated plantlets and rooted cuttings were developed and used to vegetatively multiply adult Eucalyptus grandis X urophylla. Rooting success was less than 5% when cuttings excised from twigs of 3-year-old trees were used. The rooted cuttings were grown in the greenhouse as explant- or cutting-donors and maintained at a height of 30 to 100 cm by trimming back periodically. Good rooting success (95%) of cuttings was obtained for epicormic shoots produced from donor plants after trimming 5 times. Explants of both apical and axillary buds taken from the donor plants produced multiple shoots when cultured in vitro. In vitro multiple shoot production was optimal on MS medium containing 0.1 mg/l BA and 0.01 mg/l NAA averaging 13.7 shoots per explant in a 40-day culture period. Shoot elongation was accelerated on a modified MS medium containing half strength potassium nitrate and sucrose. Elongated shoots excised at approximately 1.5 cm in length were successfully rooted on media with NAA or IBA concentrations ranging from 0.1 to 10 mg/l. Root formation was optimal on medium consisting of full strength MS basal macro elements and vitamins, half strength micro elements, 1% sucrose and supplemented with 0.3 mg/l IBA. In the field test, no significant differences were found in tree height and DBH between micropropagated plantlets and rooted cuttings at 1 and 3 years old, with the exception at 2 years old. A considerable difference arose between the 2 types of vegetative propagules in physiological response to flowering, caused by dissimilar degrees of rejuvenation.