Showing papers on "Murashige and Skoog medium published in 1987"
TL;DR: Basal leaf segments of 3 to 4 week old maize seedlings plated on SH medium with 30 μM dicamba produced embryogenic callus and/or somatic embryos that germinated and the resulting seedlings could be established in culture tubes.
Abstract: Basal leaf segments of 3 to 4 week old maize (Zea mays L.) seedlings plated on SH medium with 30 μM dicamba produced embryogenic callus and/or somatic embryos. Histological evidence showed that some of the embryos arose directly from the explant. When leaf segments with embryos were transferred to MS medium with 1.0 μM NAA, 1.0 μM IAA, 2.0 μM 2iP, and 60 g/l sucrose, the embryos germinated and the resulting seedlings could be established in culture tubes. These responses were obtained from three inbred lines, CHI31, S615, and S7.
593 citations
TL;DR: Histology of embryogenic callus demonstrated the presence of lipid-like substances and abundant starch in Vitis longii embryos, which possessed typical epidermal, cortical, and vascular tissue and Somatic embryos were attached to callus by narrow to wide suspensor-like structures.
Abstract: Anthers and ovaries of Vitis longii ‘Microsperma’ produced embryogenic callus when cultured on solidified Murashige and Skoog medium with 5μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1μM benzyladenine (BA). The initial callus was short-lived. However, long-term embryogenesis from callus was maintained through serial transfers by careful selection of clustered embryos with subtending callus. Alternatively, long term culture maintenance was through secondary embryogenesis which occurred directly from previously formed embryos on medium lacking growth regulators. Somatic embryos were white, exhibited frequent pluricotyly and tended to be larger than zygotic embryos. Histology of embryogenic callus demonstrated the presence of lipid-like substances and abundant starch. Somatic embryos were attached to callus by narrow to wide suspensor-like structures and possessed typical epidermal, cortical, and vascular tissue. Embryo cells contained abundant lipid-like accumulations but no starch. Embryos germinated when placed on medium containing 1μM BA and produced plants of normal appearance.
113 citations
TL;DR: The ability of somatic embryos to germinate was closely correlated with embryo normality, and was influenced little by the hormone content of germination media.
Abstract: Immature soybean (Glycine max (L.) Merr) embryos, or cotyledons isolated from them, were cultured on modified MS medium containing B5 vitamins and NAA (50 μM) to induce somatic embryogenesis. The effects of media variables, dissection treatments and light conditions were investigated in this system. The efficiency of embryogenesis increased as sugar concentration decreased from 12 to 1.5%; sucrose and glucose were similarly effective as carbon sources. In an examination of the effects of medium pH, values between pH 5.0 and 7.0 gave similar embryogenesis efficiencies, but the frequency of normal embryos was greater in media with low pH values. In buffered medium (10 mM MES), a pH of 5.0 was inhibitory to embryogenesis, and most normal embryos were produced at pH 5.5. Under various dissection treatments, embryogenesis efficiency and root and callus production were increased by tissue damage. Somatic embryogenesis was observed both in darkness and in light, although embryo development was impaired under high light (80 μE m-2 s-1) conditions. The ability of somatic embryos to germinate was closely correlated with embryo normality, and was influenced little by the hormone content of germination media. Of various media tested for their ability to support the growth of germinated embryos, a medium based on hydroponic nutrient salts, supplemented with yeast extract, and gelled with Difco-Bacto agar gave the best plantlet growth.
92 citations
TL;DR: Factors influencing in vitro multiplication and rooting of 8 peach scion cultivars and one rootstock, as well as acclimatization and genetic stability of these cultivars, are described.
Abstract: Success at propagating peach (Prunus persica (L.) Batsch) scion cultivars in vitro has been limited. This study describes factors influencing in vitro multiplication and rooting of 8 peach scion cultivars and one rootstock, as well as acclimatization and genetic stability of these cultivars. Shoot multiplication was best when 8.8 μM 6-benzylamino purine (BA) was added to the shoot proliferation medium. Maximum rooting occurred when shoots were placed on 1/2-strength Murashige and Skoog (MS) medium, stored in the dark at 4°C for 35 to 40 days and then incubated on rooting medium in the dark at 26°C for 14 days. All cultivars exposed to 1/2-strength MS medium supplemented with 28.5μM of either indoleacetic acid (IAA), indolebutyric acid (IBA) or α-napthaleneacetic acid (NAA) rooted best on NAA medium. A 5-fold reduction in NAA concentration to 5.8 μM, the use of IAA plus phenolic rooting cofactors, and length of time shoots were in vitro prior to rooting each increased the percentage of rooting for most cultivars. No plant loss occurred during acclimatization. Cytogenetic analysis of micropropagated plants indicated that all plants were diploid, 2n=2x=16. Examination of the performance of in vitro propagated plants under field conditions is now in progress.
84 citations
TL;DR: A reproducible method for regeneration of plants from primary leaf tissue of 27 varieties of soybean (Glycine max), encompassing maturity groups 00 to VIII, has been developed and Stimulatory effects were attained if pyroglutamic acid was added directly to the medium or if it was formed in situ as a result of chemical transformation of glutamine during autoclaving.
Abstract: A reproducible method for regeneration of plants from primary leaf tissue of 27 varieties of soybean (Glycine max), encompassing maturity groups 00 to VIII, has been developed. Progeny from seeds recovered from regenerated plants appear normal. Best regeneration was from leaf explants (2.1–4.0 mm) obtained from 5 day old seedlings. While 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) was demonstrated to be essential for regeneration, addition of benzyladenine (BA) was found to enhance regeneration. Of the 6 other auxins tested, only picloram induced any regenerative response. Using identical volumes of medium and other conditions, regeneration could be obtained in 95 × 25 mm glass culture tubes but not in 60 × 15 mm Petri dishes. The regeneration of soybeans from primary leaf tissue was shown to be greatly enhanced by pyroglutamic acid (5-oxoproline). Stimulatory effects were attained if pyroglutamic acid was added directly to the medium or if it was formed in situ as a result of chemical transformation of glutamine during autoclaving. The “active” component produced by autoclaving glutamine was not a conjugate of glutamine with inorganic salts or another organic component of the medium. Filter-sterilized glutamine was shown to be inhibitory to regeneration. Murashige and Skoog (MS) and Schenk and Hildebrandt (SH) basal media were compared to Gamborg B5 medium. All contained 0.1 mg/l 2,4,5-T, 40 mg/l adenine sulfate and 10 mM pyroglutamic acid. No regeneration occurred when MS medium was used. Growth and appearance of callus growing on SH and B5 media with the additives were similar. The incidence of regeneration among cultures growing on SH medium was only one third compared to cultures grown on B5 medium.
75 citations
TL;DR: Shoot tips of Seckel pear and Almey crabapple were cultured on liquid MS medium containing 8.8 μM BA and changes in shoot proliferation and growth and in nutrient and carbohydrate composition of the medium were determined.
Abstract: Shoot tips of Seckel pear and Almey crabapple were cultured on liquid MS medium containing 8.8 μM BA. Changes in shoot proliferation and growth and in nutrient and carbohydrate composition of the medium were determined during a 9 week culture period. Whereas shoot proliferation in crabapple increased linearly during the culture period, it levelled off after week 4 in pear. Explant dry weight in both genera showed a linear increase over time. Culture medium pH decreased in the initial weeks and increased thereafter. There was a rapid decline in medium P and Fe concentration with both genera and of Zn in the medium of crabapple. In no instance did the rate of depletion of any nutrient from the medium of pear cultures exceed that measured in the crabapple medium. The decline in sucrose concentration in the medium was similar for both genera and was accompanied by an increase in the level of glucose and fructose. At the end of the culture period slightly over half of the initial carbohydrate level remained in the medium.
70 citations
TL;DR: Somatic embryo (embryoid) formation from immature-embryo-derived calli was quantified in replicated experiments involving 10Triticum aestivum L. genotypes and there were highly significant differences among genotypes, media, and individual plants from which explants were taken.
Abstract: Somatic embryo (embryoid) formation from immature-embryo-derived calli was quantified in replicated experiments involving 10Triticum aestivum L. genotypes. Several published media formulations, which had previously been optimized for wheat tissue culture, were tested for each genotype. Embryos from each plant were randomly assigned to each medium. Percentage precocious germination of immature embryos and mean percentage scutellar callus per explant were recorded. Embryoids per callus were determined by microscopic examination at 28 and 56 days. There were highly significant differences among genotypes, media, and individual plants from which explants were taken. A medium based on double the Murashige and Skoog (MS) inorganic salt concentration was significantly better than other media. Inclusion of all MS vitamins appeared essential for optimal response. Two genotypes were tested in a second experiment where both 3,6-dichloro-o-anisic acid (9.05 μM) and 6-furfurylaminopurine (0.46 μM) were substituted for 2,4-dichlorophenoxyacetic acid (4.52 μM) in either double or normal MS medium. This substitution significantly increased embryoid formation at 28 days. Additions of either 6-furfurylaminopurine or coconut water increased precocious germination of both embryo explants and embryoids.
68 citations
TL;DR: Callus and plant regenertion were induced from shoot portions of mature embryos (dry seeds) of five high tannin Sorghum bicolor (L.) Moench cultivars to induce regeneration frequencies of 11–48%.
Abstract: Callus and plant regenertion were induced from shoot portions of mature embryos (dry seeds) of five high tannin Sorghum bicolor (L.) Moench cultivars. The explants were cultured on Murashige and Skoog medium with altered concentrations of 5 salts, supplemented with 150 mg/L L-asparagine, 5mg/L 2,4-Dichlorophenoxyacetic acid and 0.05mg/L kinetin. Calli which were yellow and globular were formed with 70–90% frequencies. The subculture medium which gave best results was MS with 2mg/L 2,4-Dichlorophenoxyacetic acid and 0.5mg/L kinetin. Plants were regenerated on MS medium supplemented with 150mg/L L-asparagine and 0.2mg/L kinetin with regeneration frequencies of 11–48%.
53 citations
TL;DR: It is demonstrated that shoot tip explants from mature trees are capable of forming multiple shoots and addition of activated charcoal showed beneficial effects on rooting percentages and plantlet growth.
43 citations
TL;DR: Rapid propagation of Nephrolepis cordifolia was achieved by means of runner tip culture in vitro by using green globular bodies for the multiplication stage, and 160 000 plantlets could be obtained from one runner tip in 6 months.
42 citations
TL;DR: A successful technique for the initiation and proliferation of shoots from epicotyl tissue of soybean, Glycine max, has been developed and is described, and some variation in numbers of shoots obtained was observed.
Abstract: A successful technique for the initiation and proliferation of shoots from epicotyl tissue of soybean, Glycine max, has been developed and is described. Fertile plants were recovered. Seeds were germinated on Murashige and Skoog medium containing 5 μM benzyladenine. Explanted epicotyl sections were induced to form callus and shoots on Schenk and Hildebrandt medium containing 5.2 mM monobasic ammonium phosphate, 74 μM 3-aminopyridine, and 20 μM kinetin for five weeks. Shoot proliferation was maintained on N6 medium containing 1.75 mM ammonium sulfate, 2.1 nM picloram, and 0.1 μM benzyladenine. Shoots rooted on Gamborg's B5 medium without growth regulators. Shoot-forming cultures were maintained for 60 months. Although all varieties tested produced shoots, some variation in numbers of shoots obtained was observed.
TL;DR: Somatic embryogenesis in pejibaye or peach palm was induced from callus derived from in vitro cultured shoot tips of young field-grown plants using a modified Murashige and Skoog medium, in which somatic embryos, as well as shoot primordia, and finally complete plantlets were formed.
Abstract: Somatic embryogenesis in pejibaye or peach palm (Bactris gasipaes H.B.K.) was induced from callus derived from in vitro cultured shoot tips of young field-grown plants using a modified Murashige and Skoog medium supplemented with 5 mg L−1 of N6-benzyladenine (BA) and 0.06 mg L−1 of picloram for three months in the dark; this was followed by an additional three months with the same medium and incubation conditions, but using 0.03 mg L−1 of picloram. The cultures were then transferred to light on a medium without hormones. This led to the formation of morphogenic callus, in which somatic embryos, as well as shoot primordia, and finally complete plantlets were formed. These plantlets continued to grow on transfer to the greenhouse.
TL;DR: Microtubers produced from the cultivar Superior were shown to contain the same characteristic group of proteins as field-grown tubers, and patatin mRNA is readily detectable in developing microtubers 15 d after transfer of the cuttings to inductive medium.
Abstract: Nodal cuttings from micropropagated potato plantlets give rise to microtubers when placed on Murashige and Skoog medium containing 6% sucrose and 2.5 mg/liter kinetin and incubated in the dark at 19 degrees C. Microtubers produced from the cultivar Superior were shown to contain the same characteristic group of proteins as field-grown tubers. As with field-grown tubers, the 40,000-dalton major tuber glycoprotein, patatin, accumulated to high levels in microtubers, reaching 3.7 +/- 0.2 mg/g fresh weight after 90 d. Also in agreement with field-grown plants, stems and leaves of micropropagated plantlets did not contain detectable levels of patatin, but small amounts of an electrophoretically distinct form accumulated transiently in roots. Patatin mRNA is readily detectable in developing microtubers 15 d after transfer of the cuttings to inductive medium. Patatin mRNA was also present in roots, but as with field-grown plants, was 50- to 100-fold less abundant and could be distinguished from that in tubers by primer extension. Microtuber development and patatin accumulation were inhibited by gibberellic acid.
TL;DR: A regulatory role for endogenous ABA content, possibly mediated by emergence-induced water stress, in the control of aerial leaf development in Hippuris is suggested.
TL;DR: Rooting of shoots in vitro was achieved on MS medium (× 12 macronutrients) containing 0.5–1.0 mg/l indolebutyric acid (IBA) and plantlets produced in vitro were established readily in soil compost.
TL;DR: Embryogenic and organogenic calli of B73 were maintained for more than two and one half years without losing regenerability, and no morphological variants were observed in the R1 progeny.
Abstract: Calli were induced from mature embryos of maize (Zea mays L.) inbred lines A632, B73 and Mol7 on MS medium supplemented with 1-2 mg/1 2,4-dichlorophenoxyacetic acid. Callus induction frequency ranged from 23-100%, with Mol7 having the highest frequency. Plants were regenerated from 4-5% of the B73 and Mol7 explants. Embryogenic and organogenic calli of B73 were maintained for more than two and one half years without losing regenerability. Of 95 regenerated plants, only one R0 plant with abnormal pollen was detected, and no morphological variants were observed in the R1 progeny.
TL;DR: Results indicate successful formation of cybrids between D. capillifolius and D. carota by protoplast fusion, and novel restriction fragments, not present in mtDNA digests from both parents, were observed in mtDNAs of regenerated plants.
Abstract: Protoplasts of Daucus capillifolius isolated from a suspension culture (chromosome number above 60) were X-irradiated over lethal dose (60 krad) just prior to fusion. Protoplasts from D. carota cell line (chromosome number 17) were treated with 15 mM iodoacetamide and fused with the X-irradiated protoplasts. Putative cybrid plants were regenerated on Murashige and Skoog medium (MS) lacking 2,4-D. The regenerated plants possessed chromosome numbers of 17 (2n−1) or 34 (4n−2) and an identical leaf morphology to D. carota. Their mitochondrial DNAs (mtDNAs) were analysed with restriction endonucleases. Novel restriction fragments, not present in mtDNA digests from both parents, were observed in mtDNAs of regenerated plants. These results indicate successful formation of cybrids between D. capillifolius and D. carota by protoplast fusion.
TL;DR: Leaf explants of Phyllostachys viridis (Young) McClure were cultured on Murashige and Skoog medium supplemented with 9×10-6 M 2,4-dichlorophenoxyacetic acid and developed within two weeks and were later successfully transferred to the field.
Abstract: Leaf explants of Phyllostachys viridis (Young) McClure were cultured on Murashige and Skoog medium supplemented with 9×10-6 M 2,4-dichlorophenoxyacetic acid. Numerous embryoids were observed. On transfer to Murashige and Skoog medium lacking hormones, plantlets developed within two weeks and were later successfully transferred to the field.
TL;DR: Out of 2C regenerated plantlets, 11 were diploids (2n = 14) arid 9 aneusomatic (chromosomal mosaics) with chromosome numbers ranging from 12 to 16, showing de novo origin of the shoots via organogenesis.
Abstract: Plant regeneration was achieved from immature embryo-derived, calli of Pisum sativum. Embryo axes were separated from cotyledons and cultured on different media containing BAP and NAA until plantlet regeneration. Rooting of the plantlets was obtained on MS medium supplemented with 2 mg/1 IBA. Frequency of regeneration was shown to be under the influence of the genotype. Histological preparations showed de novo origin of the shoots via organogenesis. Out of 2C regenerated plantlets, 11 were diploids (2n = 14) arid 9 aneusomatic (chromosomal mosaics) with chromosome numbers ranging from 12 to 16.
TL;DR: Cultures were established from the shoot apices (1–2 mm) initiated on a medium composed from Knop's macroelements and Murashige and Skoog's microelements in the presence of 0.5 mg l−1 benzyladenine and survival of potted plants was about 90%.
TL;DR: Rooting was induced in cultures with multiple shoots by subculturing explants on a half-strength MS medium supplemented with 2 mgl-1 indole-3-acetic acid (IAA).
Abstract: Explants of shoot tips, internodal stem sections, and leaf segments of Lisianthus, Eustoma grandiflorum (Griseb.) Schinners, ‘Dwarf Purple’ were cultured in vitro on modified Murashige and Skoog (MS) media. Explants of shoot tips and internodal stem sections developed into multiple shoots, whereas, leaf segments turned chlorotic on a medium supplemented with 3 mgl-1 benzyladenine (BA) and 0.2 mgl-1 naphthalene acetic acid (NAA). Shoot proliferation was obtained on shoot tips and leaf segments with 3 mgl-1 BA, but internodal stem sections became necrotic and died on this medium. Rooting was induced in cultures with multiple shoots by subculturing explants on a half-strength MS medium supplemented with 2 mgl-1 indole-3-acetic acid (IAA). Rooted plantlets were successfully transferred to soil.
TL;DR: Plants of Solanum melongena were propagated under in vitro conditions by subculture of terminal and lateral cuttings on MS medium +20 gl-1 sucrose + Morel and Wetmore vitamins at 1/8 strength and 7gl-1 agar to identify embryos-like structures and some of embryo-like structure developed into plantlets.
Abstract: Plants of Solanum melongena were propagated under in vitro conditions (27°C, 12h/day illumination at 62 μEm-2s-1, 60% humidity) by subculture of terminal and lateral cuttings on MS medium +20 gl-1 sucrose + Morel and Wetmore vitamins at 1/8 strength and 7 gl-1 agar. Lamina, petioles and stems of 3-week-old cuttings were used as sources of protoplasts. The best mean yield of protoplasts was obtained from the lamina with 9,030×103 protoplasts per gram of tissue. Petioles and stems yielded respectively 3,144×103 and 1,220.4×103 protoplasts per gram of tissue. first division of petiole and stem protoplasts occurred within 48 h, while lamina protoplasts underwent division after 3–4 days of culture in KM8p medium +2,4-D(0.2 gl-1) + zeatin (0.5 mgl-1) + NAA (1 mgl-1) and 0.35M glucose as osmoticum. The highest percentage of dividing cells was obtained from petiole material, estimated at 33.4% after 7 days, compared to 23.8% and 19.4% respectively for stem and lamina protoplasts. When BAP replaced zeatin in KM8p, the division percentage of lamina protoplasts was reduced to 10–15%. When transferred to regeneration medium, all calli derived from KM8p + zeatin formed deep-green spots identified as embryo-like structures, while only few calli from KM8p + BAP underwent shoot organogenesis without formation of green spots. Some of embryo-like structure developed into plantlets with a frequency of 1–2 plantlets per callus especially on MS medium + zeatin (4 mgl-1) + IAA (0.2 mgl-1). Maintaining protoplast-derived calli on MS + BAP (0.5 mgl-1) + NAA (0.5 mgl-1) for more than 3 weeks resulted in a decrease and loss of cell totipotency.
TL;DR: Results indicate that the type of basal medium, viz.
Abstract: Time-course experiments have been carried out to investigate the relationship between growth and alkaloid accumulation in A. altissima cell suspension cultures. Results indicate that the type of basal medium, viz. Murashige and Skoog, Linsmaier and Skoog, Schenk and Hildebrandt or Gamborg's B5, has a significant influence on both growth and alkaloid production.
TL;DR: Formation and growth of successive leaves yielded small, elongated, adventitious shoots with constricted bases, and transferral to a basal MS medium with 1 mg/l 1-naphthaleneacetic acid (NAA) led to the formation of rooted plantlets.
Abstract: Isolated tips (approx. 2 mm long) from aseptic, multiplying shoot cultures of the triploid dessert banana clone ‘Highgate’ were tested for their morphogenetic responsiveness to hormone treatments on semisolid media. Medium containing Murashige and Skoog (1962) salts, p-chlorophenoxyacetic acid, and kinetin produced a compact calloid mass. Protuberances disclosed by SEM as rounded, button-shaped, and pointed outgrowths resembling fasciated shoots were formed in profusion. Sections showed many meristematic regions, some associated with distinct leaf primordia. Formation and growth of successive leaves yielded small, elongated, adventitious shoots with constricted bases. Transferral to a basal MS medium with 1 mg/l 1-naphthaleneacetic acid (NAA) led to the formation of rooted plantlets.
TL;DR: Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and SkOog (1/2 MS) media containing high levels of growth regulators, confirming the embryogenic pathway of differentiation.
Abstract: Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes ≤8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity.
TL;DR: Protoplasts isolated from hypocotyls of three-day-old seedlings of Brassica carinata (Braun) cv R-2128 were cultured in a modified Nitsch and Nitsch liquid medium and developing microcalli were transferred onto MS medium which produced plantlets.
Abstract: Protoplasts isolated from hypocotyls of three-day-old seedlings of Brassica carinata (Braun) cv R-2128 were cultured in a modified Nitsch and Nitsch liquid medium containing 13% sucrose, 04% Ficoll, 025 mg/l BA, 05 mg/l NAA and 05 mg/l 2,4-D The density of medium caused the protoplasts and the developing microcalli to float on the surface of the liquid medium whereas all debris and lysed cells sank to the bottom of the culture plate After 4–6 weeks developing microcalli were approximately 05 mm in diameter and were transferred onto MS medium containing 3% sucrose, 04% agarose, 200 mg/l casein hydrolysate, 5 mg/l BA and 05 mg/l NAA, pH 57 Approximately 20% of the calli transferred to this medium produced plantlets
TL;DR: Highly morphogenic callus cultures were isolated from stamens of a wild peanut species, Arachis paraguariensis, and grafting to stems of rooted seedlings was used to obtain plants from regenerated shoots.
Abstract: Highly morphogenic callus cultures were isolated from stamens of a wild peanut species, Arachis paraguariensis. These cultures were initiated on modified N6 medium containing 0.2 mg1l-1 4amino-3,5,6-trichloro-picolinic acid (picloram) and 0.5 mg l-1 6-benzylaminopurine (BAP) and were maintained on modified N6 medium with 0.008 mg l-1 picloram and 0.25 mg l-1 BAP. Buds formed on the calli growing on the maintenance medium developed into shoots when they were transferred to a MS salts based medium with no hormones. The cultures could also be maintained as a suspension culture in N6 liquid medium. When cell clumps larger tham 840 μm were collected from the suspension culture and transferred to MS medium without hormones, they formed shoots in liquid culture. Root formation rarely occurred in agar or liquid cultures. Therefore, grafting to stems of rooted seedlings was used to obtain plants from regenerated shoots. Eight out of 50 field grown plants produced viable seed.
TL;DR: The plantlets formed were tetraploid in nature by cytological observations of the root tips and exhibited 80–90% success in establishment under glass house and field conditions.
Abstract: A tissue culture procedure was developed for the establishment and propagation of a colchi-autotetraploid of Rauvolfia serpentina for possible commercial exploitation. Multiplication of autotetraploid shoots was obtained either through axillary bud elongation on Murashige and Skoog [1] medium (MS) containing 2.65 μM (0.5 mgl−1) α-naphthaleneacetic acid and 0.33 μM (0.05 mgl−1) kinetin, or via multiple shoot formation on MS medium supplemented with 4.44 μM (1.0 mgl−1) 6-benzylaminopurine and 0.53 μM (0.1 mgl−1) α-naphthaleneacetic acid. Rooting could be induced by transferring the shoots to MS medium containing 7.95 μM (1.5 mgl−1) α-naphthaleneacetic acid alone. The plantlets, thus formed, were tetraploid in nature by cytological observations of the root tips. They exhibited 80–90% success in establishment under glass house and field conditions.
TL;DR: Callus cultures established on modified Murashige and Skoog medium from hypocotyl, leaf and stem tissues of the tree legume Albizia lebbek (L.) Benth showed no morphological abnormalities and cytological investigations revealed no change in chromosomal complement.
TL;DR: Genotypic differences among carinata accessions for regeneration were observed and compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l).
Abstract: Protocols of plant regeneration have been developed for Brassica carinata for creating somaclonal variation for plant type and adaptability, so that this species can fit into cropping systems in Indian agriculture. The response of cotyledonary and stem explants was assessed for callus induction and shoot regeneration on MS and B 5 basal media containing different combinations of auxin and cytokinin concentrations. MS medium supplemented with BA and NAA favoured callus induction. Supplementing MS with combinations of BA and IAA, as also with BA alone, regenerated shoots from the ex pi ants with a high frequency. The frequency of shoot regeneration and the mean number of shoots per explant were higher in cotyledons than in stem explants on identical growth regulator combinations. On B 5 medium, supplemented with BA (2 mg/l) and IBA (0.4 mg/l), compact callus was produced which regenerated shoots on transfer to medium containing BA (0.8 mg/l). Genotypic differences among carinata accessions for regeneration were also observed.