Showing papers on "Murashige and Skoog medium published in 1988"
TL;DR: In leaf discs and younger and older zygotic embryos, only callus and root formation was observed, and somatic embryogenesis, approx.
Abstract: Leaf discs from olive (Olea europaea L.) grown in vitro and immature zygotic embryos collected at 50, 75, 90 and 105 days after full bloom were tested for their somatic embryogenic capacity. The embryos were grown in half-strength MS medium and half-strength OM medium with BAP combinated with either 2,4-D or NAA. Incubation was either in an initial dark period followed by 16h daylight or in 16h daylight throughout. Somatic embryogenesis, approx. 40%, mostly directly from the embryos, was observed only in 75-day-old embryos in medium containing low cytokinin and auxin concentrations. Differentiation was inhibited by 2,4-D whereas NAA did not. In leaf discs and younger and older zygotic embryos, only callus and root formation was observed. Somatic embryos were germinated and then potted-up to soil.
71 citations
TL;DR: In this paper, the authors induced ginger with or without leaf primordia to form shoots on three-quarter strength Murashige-Skoog's (MS) medium containing sucrose 6%, coconut milk (CM) 20%, ascorbic acid (AA) 100 mg l−1, glutamine (GL) 400 mg l − 1, activated charcoal (AC) 250 mg l-1, 6-benzylaminopurine (BAP) 0.5 mg l+1, indolebutyric acid (IBA) 0
Abstract: SummaryMeristems of ginger with or without leaf primordia were induced to form shoots on three-quarter strength Murashige-Skoog’s (MS) medium containing sucrose 6%, coconut milk (CM) 20%, ascorbic acid (AA) 100 mg l−1, glutamine (GL) 400 mg l−1, activated charcoal (AC) 250 mg l−1, 6-benzylaminopurine (BAP) 0.5 mg l−1, indolebutyric acid (IBA) 0.4 mg l−1 and agar 0.8%. Meristem-derived shoots exhibited consistent multiplication on three-quarter strength MS medium containing sucrose (3%), AA (100 mg l−1), AC (100 mg l−1), BAP (4–5 mg l−1) and agar (0.8%). Liquid media (agitated or static) were less effective than a solid (agar-gelled) medium for micropropagation. Kinetin and naphthalene acetic acid (NAA) incorporated at various levels (0.01–0.8 mg l−1) with or without added BAP and IBA neither improved plantlet formation nor enhanced shoot multiplication. The in vitro plants were successfully established in vivo and the rhizome yield was comparable with that of plants grown by conventional methods.
68 citations
TL;DR: Shoots obtained from the proliferation stage were rooted on half-strength MS medium containing indole-3-butyric acid and α-naphthaleneacetic acid and Regenerated plantlets were successfully established in soil under field conditions.
68 citations
TL;DR: Various explant sources of Populus nigra × P. maximowiczii were used to examine the effects of growth hormones on morphogenesis in vitro and initial experiments indicated that punctured leaves were superior to non-punctured ones for both callus growth and formation of shoots and roots on MS medium containing various types and concentrations of growth hormone.
Abstract: Various explant sources of Populus nigra × P. maximowiczii were used to examine the effects of growth hormones on morphogenesis in vitro. Initial experiments indicated that punctured leaves were superior to non-punctured ones for both callus growth and formation of shoots and roots on MS medium containing various types and concentrations of growth hormones. After 6 weeks in culture, an average of 178 shoots, 129 roots and 3.1 g fresh weight of callus were directly produced from the abaxial side of each punctured leaf. The best combinations of growth hormones for shoot, root and callus proliferation were 0.88 μM BA plus 0.05 μM 2,4-D, 0.44 μM BA plus 2.69 μM NAA and 0.44 μM BA plus 2.26 μM 2,4-D, respectively. Embryoids were also formed on callus derived from punctured leaves. The number of embryoids varied from 0 to 30 per punctured leaf. Adventitious shoots also developed simultaneously with the embryos. Embryoids were removed with a scalpel at the early developmental stages and placed on MS medium lacking growth regulators. Regenerated plantlets were transferred to pots containing vermiculite for normal growth in the greenhouse.
67 citations
TL;DR: All cultivars gave the same response in tests of root formation, but shoot regeneration from callus culture of cucumber cotyledons was dependent on genotype with cultivar Manchoonchoungjang exhibiting the best shoot differentiation capability among the genotypes examined.
Abstract: The growth and differentiation of callus tissues derived from cotyledons of ten cultivars ofCucumis sativus L. were investigated. Cotyledonary explants from all ten cultivars formed callus tissue on Murashige and Skoog (MS) medium supplemented with 0.5 μM 2,4-dichlorophenoxyacetic acid and 5 μM 6-benzylaminopurine. Fresh weight of the callus tissues averaged 1 to 8 g per flask after five weeks of culture. Shoot development was achieved in three cultivars, Hukchinju, Manchoonchoungjang and Seoul, on MS medium supplemented with 0.5 μM α-naphthaleneacetic acid and 5 μM 6-benzylaminopurine. Reducing the 6-benzylaminopurine concentration to 0.01 μM resulted in root formation on callus tissues and on shoots transferred to this medium. All cultivars gave the same response in tests of root formation, but shoot regeneration from callus culture of cucumber cotyledons was dependent on genotype with cultivar Manchoonchoungjang exhibiting the best shoot differentiation capability among the genotypes examined. Examination of mitotic metaphase from the regenerants revealed that all were tetraploid.
64 citations
TL;DR: Conditions were established for efficient plant regeneration from cotyledon explant calli in different cultivars of Brassica juncea, B. campestris and B. carinata on Murashige & Skoog's (MS) medium supplemented with various combinations of cytokinins and auxins, but regeneration frequency varied with genotype and the different growth hormone combinations in media.
Abstract: Conditions were established for efficient plant regeneration from cotyledon explant calli in different cultivars ofBrassica juncea, B. campestris andB. carinata on Murashige & Skoog's (MS) medium supplemented with various combinations of cytokinins and auxins. Regeneration frequency, however, varied with genotype and the different growth hormone combinations in media. Almost in all species, MS medium with zeatin (1.0 mg 1-1) and IAA (0.1 mg l-1) was found to be best for shoot organogenesis followed by the ones containing high kinetin (2.0 mg l-1) and low IAA (0.02 or 0.2 mg l-1) concentrations. On these media, the cotyledonary explants invariably underwent callusing followed by multiple shoot formation, which could be separated and subcultured for further propagation. Number of shoots per cotyledon explant cultured varied from 0 to as many as 50. InB. juncea andB. campestris, the regeneration frequency declined sharply in the absence of auxin in medium. BAP in combination with NAA yielded no or a reduced number of shoots. Shoot organogenesis also declined with the reduction in photoperiod from continuous light to 16 hours. Shoots were easily rooted during prolonged incubation on the same medium and whole plants were transferred to pots in the greenhouse and grown to maturity.
62 citations
TL;DR: Of the explants tested, the best morphogenetic responses were obtained from nodes and the poorest from leaves, and Root formation from explants was achieved only in media with NAA or IAA.
Abstract: The influence of culture medium and explant on callus and shoot formation of lentil (Lens culinaris Medik.) has been studied. Three different explants (shoot-tip, first node and first pair of leaves) from three Spanish lentil cultivars were cultivated on two basal media: Murashige and Skoog medium (MS) and medium with mineral salts of MS medium plus vitamins of Gamborg's B5 medium (MSB), supplemented with growth regulators. Media with 2,4-D induced the formation of calli in all explants, but no organ regeneration was obtained from these calli. Multiple shoot formation was obtained from 33% to 92% of the explants in media supplemented with 2.25 mg l−1 of BA and 0.186 mg l−1 NAA+2.25 mg l−1 BA; in the other media one to two shoots per explant were formed in 10 to 98% of the explants. Root formation from explants was achieved only in media with NAA or IAA. Of the explants tested, the best morphogenetic responses were obtained from nodes and the poorest from leaves.
62 citations
TL;DR: Higher levels of sucrose caused browning of green calli and also inhibited differentiation into embryos and shoot buds and by selective sub-culturing of 0.1 cm3 pieces of embryogenic calli on MS+10−5M BAP, 46% of the cultures produced somatic embryos and the latter germinated into plantlets on Knop's medium.
Abstract: Hypocotyl explants of 1 and 10 mm lengths were excised from 12-day-old in vitro-grown seedlings of Albizia richardiana The larger pieces, after 40 days of culture, developed shoots along with green calli on B5 + BAP (10(-7)-10(-5)M), while the smaller segments produced only green calli on B5+BAP (10(-7)-10(-4)M) medium Some of the green calli turned morphogenic and started producing somatic embryos with the 2nd sub-culture and shoots from 7th sub-culture onwards Calli retained the morphogenic potential even after repeated sub-culturing for over two years The number of embryos in an embryogenic culture varied from 2 to 20 per callus mass of 5-65 cm(3) Sucrose at the 2% level in MS medium was optimal for embryogenesis while 4% was optimal for shoot bud differentiation Higher levels of sucrose (6-10%) caused browning of green calli and also inhibited differentiation into embryos and shoot buds By selective sub-culturing of 01 cm(3) pieces of embryogenic calli on MS+10(-5)M BAP, 46% of the cultures produced somatic embryos The latter germinated into plantlets on Knop's medium
58 citations
TL;DR: A procedure has been developed for the clonal propagation of Picrorhiza kurroa Royle ex Benth through shoot tip culture and addition of indole-3-acetic acid to the kinetin containing medium showed marked improvement in the growth of regenerated shoots.
Abstract: A procedure has been developed for the clonal propagation of Picrorhiza kurroa Royle ex Benth. through shoot tip culture. Murashige and Skoog's medium (1962) supplemented with kinetin (3.0 to 5.0 mg/l) supported rapid proliferation of multiple shoots from the explants. Addition of indole-3-acetic acid (1.0 mg/l) to the kinetin containing medium showed marked improvement in the growth of regenerated shoots. However, presence of IAA in the medium did not alter the frequency of shoot multiplication. Rooting was readily achieved upon transferring shoots onto MS medium containing ∝-naphthaleneacetic acid (1.0 mg/l). Plantlets were successfully transferred to soil.
55 citations
TL;DR: Hypocotyl-derived callus cultures of Brassica campestris L. pekinensis cv.
Abstract: Hypocotyl-derived callus cultures of Brassica campestris L. ssp. pekinensis cv. Kim-jung (Chinese cabbage) were grown on Murashige and Skoog medium containing no additional salt, NaCl or Na2SO4. Na2SO4 was more than twice as inhibitory in comparison to the same concentration of NaCl when growth and fresh:dry weight ratios of established callus were measured. Levels of protein, starch, sucrose and α-amino nitrogen were not significantly altered in salt-grown callus. Concentrations of reducing sugars and chlorophyll were 2–3 times greater in callus grown on either salt. Proline concentration increased 15–20 fold on the highest levels of salt. Final concentrations (reached in 20–24 days) were closely correlated to the initial Na+ concentration of the medium, regardless of salt type. The osmotic potential in callus transferred to NaCl or Na2SO4 reached a maximum negative value after 16 days. For both salts, subsequent increases were correlated to increases in fresh:dry weight and growth. On both salts, turgor remained relatively constant (0. 6–0.75 MPa). Changes in Na+, K+, Mg2+ and Ca2+ content were correlated to initial Na+ concentration in the medium, not salt type. Accumulation of Na+ was accompanied by loss of K+ and Mg2+. Six to seven times less sulfate was measured in callus grown on Na2SO4 than chloride in callus grown on similar concentrations of NaCl.
51 citations
TL;DR: Following rooting, protoplast-derived plants of pear were transferred to the glasshouse where they completed acclimatization and exhibited shoot regeneration on half-strength Murashige and Skoog medium.
Abstract: Leaf protoplasts of axenic shoot cultures of Pyrus communis L. cv. Williams' Bon Chretien (syn. Bartlett) underwent cell wall regeneration and division to give multicellular colonies in a modified Murashige and Skoog medium which lacked ammonium ions, but supplemented with 1-naphthaleneacetic acid (NAA), 4-indole-3yl-acetic acid, 6-benzylaminopurine (BAP) and casein hydrolysate. Protoplast-derived colonies gave callus on Murashige and Skoog salts medium with NAA and BAP and exhibited shoot regeneration on half-strength Murashige and Skoog medium supplemented with 0.2 mg 1−1 4-indole-3yl-butyric acid, 2.0 mg 1−1 BAP, 0.2 mg 1−1 gibberellic acid, 50 mg 1−1 casein hydrolysate and 10 mg 1−1 Ca-pantothenate. Following rooting, protoplast-derived plants of pear were transferred to the glasshouse where they completed acclimatization.
TL;DR: Alkaloid contents of clonally-propagated plants showed less variation than those of the normally grown plants and multiple shoot forming ability does not decrease at least for 6 generations.
Abstract: Shoot tips and axillary buds of ACONITUM CARMICHAELI Debx cultured in Murashige and Skoog medium supplemented with 5 ppm BAP at 25 +/- 1 degrees C under continuous light for 6 weeks formed multiple shoots with an average of 7 shoots per culture Multiple shoot forming ability does not decrease at least for 6 generations Subsequent culture of these shoots on MS medium supplemented with 05 ppm IAA at 20 +/- 1 degrees C under continuous light for 6 weeks resulted in root formation The plantlets were transplanted successfully in the soil Alkaloid contents of clonally-propagated plants showed less variation than those of the normally grown plants
TL;DR: Shoot tip cultures from 2- to 3-d-old seedlings of sorghum bicolor (L.) Moench cv.
Abstract: Shoot tip cultures from 2- to 3-d-old seedlings ofSorghum bicolor (L) Moench cv IS3620C develop highly embryogenic callus from which plants can be regenerated when transferred to plant growth regulator-free medium Isolated shoot tips were cultured on Murashige and Skoog medium supplemented with 25 mg/liter 2,4-dichloro-phenoxyacetic acid and 005 mg/liter kinetin Purple pigmentation characteristic of sorghum cultures on growth regulator-free medium is virtually eliminated with the shoot tip culture Embryogenic callus is white and hard with an undulating appearance but can be separated into multiple bipolar structures by application of gentle pressure The well-developed embryos have a cup-shaped scutellum These germinate like zygotic embryos and develop root-shoot axis Lack of vascular connections to the parent tissue and the synchronous development of the plumule and radicle suggest that these embryos may be of unicellular origin In contrast, when the entire seedling serves as the explant, all meristematic centers in the shoot, including the coleoptile sheath close to the apical meristem respond to plant growth regulators in the medium by callus formation Upon subsequent reculture onto growth regulator-free medium several modes of development occur The differential response of these tissues to identical culture conditions indicate the presence of different population of cells that respond differently to exogenous plant growth regulators
TL;DR: Leaf protoplasts were isolated from axenic shoot cultures of four varieties of Capsicum annuum (Americano, Dulce Italiano Florida Gynat and Nigrum) and a wild species C. chinense and protoplasts of both species entered division with the exception of the variety Nigrums.
Abstract: Leaf protoplasts were isolated from axenic shoot cultures of four varieties of Capsicum annuum (Americano, Dulce Italiano Florida Gynat and Nigrum) and a wild species C. chinense. Protoplasts of both species, cultured in KM8P medium and using agarose bead culture, entered division with the exception of the variety Nigrum. Cell colonies formed callus in agar-solified MS medium supplemented with zeatin and for C. annuum v. Dulce Italiano shoots were regenerated when protoplast-derived calli were transferred to MS medium with 6-BAP. Excised shoots were rooted on MS medium which lacked phytohormones.
TL;DR: The response in culture with regard to compact callus induction, embryogenesis and plant regeneration was determined for different varieties and there was no consistent effect of the growth conditions of the donor plants or the 2,4-D concentration of the medium on this response.
TL;DR: Ferulic acid, a probable precursor of vanillin the dominant flavor component of vanilla extract, was topically applied to V. planifolia callus in the dark on a modified Murashige and Skoog medium at 26°C.
Abstract: Vanilla planifolia callus was maintained in the dark on a modified Murashige and Skoog medium at 26°C. Ferulic acid, a probable precursor of vanillin the dominant flavor component of vanilla extract, was topically applied to V. planifolia callus. Callus treated with ferulic acid experienced immediate fluid uptake followed by a delayed and reduced growth response. Over the 28 day period, vanillin concentration in untreated callus increased by a factor of 1.2, and vanillin concentration increased 1.7‐fold after application of a 1 mM ferulic acid solution. The addition of 10 mM ferulic acid decreased vanillin production.
TL;DR: Eggplant (Solarium melongena L.) cotyledons grown on Murashige and Skoog medium with naphthaleneacetic acid formed callus, roots, and somatic embryos.
Abstract: Eggplant (Solarium melongena L.) cotyledons grown on Murashige and Skoog medium with naphthaleneacetic acid formed callus, roots, and somatic embryos. Low levels of naphthaleneacetic acid (0.1 – 0....
TL;DR: Embryogenesis of PANAX GINSENG was induced from young flower buds via callus during a 3 months culture period and the plantlet was transferred to vermiculite.
Abstract: Embryogenesis of PANAX GINSENG was induced from young flower buds via callus during a 3 months culture period. Matured embryos could be germinated on the 1/2 Murashige and Skoog's medium supplemented with GA (3) (1.4 microM)-BAP (2.2 microM) and 1.5% sucrose. In the medium containing 1.4 microM GA (3) and 11.1 microM BAP, a multiple shoot complex having 8 shoots per segment was formed from a single shoot set. On the other hand, the addition of 11.1 microM BAP and 1.4 microM GA (3) to the medium stimulated the flower bud formation. For root formation of shoots, the MS medium supplemented with 5.4 microM NAA was the most favourable. Subsequently the plantlet was transferred to vermiculite.
TL;DR: A tissue culture procedure has been developed for the rapid multiplication of VALERIANA WALLICHII D C through shoot tip and axillary bud explants, which has implications in the early introduction of an elite population as well as its improvement.
Abstract: A tissue culture procedure has been developed for the rapid multiplication of VALERIANA WALLICHII D C. through shoot tip and axillary bud explants. MS medium containing Kn or BAP (5.0 mg/l (-1)) in combination with IAA (1.0 mg/l (-1)) induced an optimal growth of shoots within 6-8 days from both apical and axillary bud explants. The roots developed on the same medium within 2-3 weeks. Hardening of IN VITRO grown plantlets in pots under glass-house conditions was dependent upon the temperature and humidity. A cold-temperate climate favoured early establishment. Following the given procedure, a large number of plants have been established under field conditions at two locations. The method has implications in the early introduction of an elite population as well as its improvement.
TL;DR: The results suggest that cytokinins and ethylene both play roles in natural lateral bud initiation and that the cytokinin function involves two stages of the process.
Abstract: Aechmea victoriana var discolor L. B. Foster and Aechmea dactylina Bal. are commercially propagated in vitro through lateral shoot growth. A modified Murashige and Skoog medium is used which contains both BA and IAA. These growth substances were shown in the present study to synergistically stimulate the production of ethylene by the cultured plants. The stimulation of ethylene production is correlated with the outgrowth of the lateral buds. The rise in ethylene production was concluded to induce lateral shoot growth, because: (a) outgrowth of the shoots was blocked by preventing an increase in ethylene production, (b) 1-aminocyclopropane-1-carboxylic acid (ACC), the natural precursor of ethylene biosynthesis, substituted for IAA in the promotion of ethylene production and lateral bud outgrowth. Although ACC could substitute for IAA, it could not substitute for BA; therefore, cytokinins are concluded to be essential for lateral bud outgrowth in vitro in Aechmea. These results suggest that cytokinins and ethylene both play roles in natural lateral bud initiation and that the cytokinin function involves two stages of the process.
TL;DR: Embryos from young seeds of three Citrus species, ‘Washington’ navel orange, yuko and ponkan, were cultured on a Murashige and Skoog medium with 0.16 M sucrose, 0.5 g l−1 malt extract and 50 μM kinetin and maintained an embryogenic capacity after sub-culture for about 2 years.
TL;DR: A comparison of BA and N6-(Δ2-isopentenyladenine) (2iP) showed that 2iP was not effective in promoting shoot proliferation, and was found to proliferate with increasing concentrations ofBA and to produce callus and poorer quality shoots in the presence of NAA and the absence of BA.
Abstract: A micropropagation system was developed to facilitate the release and subsequent testing of unique pink- or white-flowered selections of Yucca glauca. Shoot tip explants from mature plants were cultured on Murashige and Skoog medium supplemented with factorial combinations of α-naphthaleneacetic acid (NAA) (0.0 to 3.2 μM) and 6-benzylaminopurine (BA) (0.0 to 45 μM). Shoots were found to proliferate with increasing concentrations of BA and to produce callus and poorer quality shoots in the presence of NAA and the absence of BA. The response to BA and NAA was similar among 3 genotypes examined. A comparison of BA and N6-(Δ2-isopentenyladenine) (2iP) showed that 2iP was not effective in promoting shoot proliferation. Shoot tips rooted in the absence of growth regulators or in the presence of low concentrations of indole-3-butyric acid (IBA). Plantlets were successfully acclimated to greenhouse conditions.
TL;DR: Phragmites australis tissue cultures established long-term regenerable cultures and indicated both the presence of embryogenic-type cells and the bipolar development of regenerated plants.
Abstract: Phragmites australis tissue cultures were initiated from mature seeds on MS medium supplemented with 1 mgl-1 each of 2,4-D and IAA. Cultures displayed typical embryogenic callus that was compact and bright yellow. Selection for embryogenic callus established long-term regenerable cultures. Removal of auxin from the basal medium allowed numerous complete plants to be recovered from the cultures. Histological study indicated both the presence of embryogenic-type cells and the bipolar development of regenerated plants.
TL;DR: Axillary shoot induction and plant regeneration were obtained in Plantago ovata and the regenerated plants were similar to the control plants in karyotypic and phenotypic details.
Abstract: Axillary shoot induction and plant regeneration were obtained in Plantago ovata. The optimum medium for inducing axillary shoots was Murashige & Skoog (MS) medium [5] supplemented with 4.6 μM kinetin and 0.05 μM NAA. Rooting of shoots was best on half-strength MS medium containing 5.0 μM IBA and 0.05 μM kinetin. The regenerated plants were similar to the control plants in karyotypic and phenotypic details.
Patent•
23 May 1988
TL;DR: In this article, 2,4-dichlorophenoxyacetic acid is the preferred auxin for the development of the shoot tip formation of a plantlet in soil or potting medium.
Abstract: Seeds of Glycine max, Glycine soja and hybrids thereof are produced by propagating embryogenic tissue containing multiple immature somatic embryos directly or serially by culturing particular cotyledonary tissue or derivation product thereof on media containing auxin selected from the group consisting of chloro-substituted phenoxyacetic acid, methyl derivative thereof, dicamba or picloram, incubating individual immature somatic embryos or embryogenic tissue containing such in a maturation stage to produce mature somatic embryos, germinating the mature somatic embryos to cause shoot tip formation, cultivating the germinated embryos to provide plantlets with 2 to 5 nodes and roots, cultivating the plantlets to provide whole fertile plants which flower and bear seeds, and recovering the seeds. Culturing to provide immature somatic embryos is carried out utilizing auxin in a concentration within the range of 2.5 to 1000 ppm effective to foster normal development of immature somatic embryos. 2,4-dichlorophenoxyacetic acid is the preferred auxin. Maturation is carried out in B5 or MS medium containing indolebutyric acid and abscissic acid or activated charcoal and sucrose, and germination is carried out in B5 or MS medium containing indolebutyric acid and gibberellic acid or MS medium containing filter sterilized L-proline and sucrose. Plantlet formation is carried out in B5 or MS medium without growth regulators. Plant and seed formation is carried out in soil or potting medium.
TL;DR: Callus culture of saffron was investigated to promote its efficiency by the use of phytohormones combined with a suitable range of temperature for culture to develop an efficient regeneration system for saffrons breeding.
Abstract: In order to develop an efficient regeneration system for saffron breeding, callus culture of saffron was investigated to promote its efficiency by the use of phytohormones combined with a suitable range of temperature for culture. Callus induction was carried out on MS medium (MURASHIGE and SKOOG, 1962) with 2, 4-D(D) and Zeatin(Z), after sterilization of the bulbs of commercial origin (Sakata Seed Corp.) by standard methods. The callus was subcultured three times to obtain a complete dedifferentiation. In one method, the callus obtained after the 3-rd transfer was cultured by gyrating it after transfer to MS liquid medium supplemented with 1 mg/l, 2, 4-D. After 7∼10 days of culture, the callus crumbled and developed into a small spherical organ-like nodule. When this nodule was cultured continuously, it was able to grow, but after transfer onto MS medium supplemented with 0.1mg/l 1-naphthaleneacetic acid (N 0.1) and 1.0 mg/l 6-benzylaminopurine (BA 1) or MS (N 1, BA 3), the nodule enlarged irrespective of the conditions of the culture (liquid or solid). Shoots regenerated on thc surface of the enlarged tissues after 3 months of culture on solid medium with similar composition at 25°C. These shoots were transplanted and cultured under aseptic conditions. On the other hand, the callus was cultured on the MS (N 0.1, BA 1) medium at 15°C and 20°C, it became an organogenetic tissue with a smooth surface after 3 months. Then this tissue turned green and regenerated shoots or rooted partially.
TL;DR: Maximum productivity was achieved when the cell line K 3 OHD was used with an initial cell density of about 20% and all of the digitoxin added was biotransformed within 12 days of incubation yielding the main product deacetyllanatoside C (88%) together with purpureaglycoside A (12%) both of which accumulated in the cells.
Abstract: Suspension-cultured DIGITALIS LANATA cells, known to form beta-methyldigoxin from beta-methyldigitoxin without any by-products, were not able to 12beta-hydroxylate digitoxin efficiently when cultivated in the cell culture medium devised by Murashige and Skoog. Most of the substrate added was merely glucosylated at its 16'-O-position leading to purpureaglycoside A as the main biotransformation product after 9 days of incubation. An 8% glucose solution (pH 5.5) turned out to be a suitable production medium for an efficient 12beta-hydroxylation of digitoxin. A two-stage procedure was developed in which DIGITALIS cells were grown in a modified Murashige and Skoog medium for 10 days and then transferred into 8% glucose medium. With regard to an effective 12beta-hydroxylation of digitoxin, maximum productivity was achieved when the cell line K 3 OHD was used with an initial cell density of about 20%. The substrate was added in one batch (190 mg digitoxin per flask, i.e. 0.5 gl (-1)) 3 days after transfer of cells to production medium. Under these conditions all of the digitoxin added was biotransformed within 12 days of incubation yielding the main product deacetyllanatoside C (88%) together with purpureaglycoside A (12%) both of which accumulated in the cells.
TL;DR: Plantlets obtained by organogenesis from cultured anthers of Populusdeltoides (Bartr) were found to be haploid and two plants developed male terminal inflorescences, but died shortly thereafter.
Abstract: Plantlets were obtained by organogenesis from cultured anthers of Populusdeltoides (Bartr.). Anthers formed callus in the dark on modified Murashige and Skoog medium supplemented with 9.0 μM 2,4-dichlorophenoxyacetic acid and 4.7 μM kinetin. Anther calli were differentiated into shoots by sequential transfer in the light onto Murashige and Skoog medium containing 4.4 μM benzylamino purine and 1.1 μM naphthaleneacetic acid for 4 weeks, followed by several transfers to woody plant medium with 2.2 μM benzylamino purine and 1.1 μM naphthaleneacetic acid. The shoots that formed were rooted by excising and transferring to woody plant medium supplemented with 1.0 μM indole-3-butyric acid. A few of these plants were found to be haploid. Two plants developed male terminal inflorescences, but died shortly thereafter.
TL;DR: In vitro regeneration of plantlets was achieved from callus of shoot tips and shoot segments of over 50-year-old ‘elite’ trees on Murashige & Skoog's medium containing naphthaleneacetic acid (NAA) and benzylaminopurine (BAP).
Abstract: A successful procedure was established for in vitro mass multiplication of Indian rosewood (Dalbergia latifolia Roxb.). In vitro regeneration of plantlets was achieved from callus of shoot tips and shoot segments of over 50-year-old ‘elite’ trees on Murashige & Skoog's medium containing naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). For rooting, regenerated shoots from the calli were excised and first treated with White's liquid medium or half-strength Murashige & Skoog's medium, supplemented with indole-3-acetic acid, indole-3-butyric acid and naphthaleneacetic acid for 48 h to 72 h. Following this treatment, plantlets were transferred to hormone-free half-strength MS medium. Rooted plantlets were then transferred to pots and grown in the greenhouse.
TL;DR: Procedures were developed for micropropagation of Alnus cordata through in vitro axillary shoot multiplication of axillary bud explants cultured in Murashige & Skoog (MS) medium to minimize excessive callus growth, abnormally thick and brittle leaves, inhibition of shoot elongation, and senescence.
Abstract: Procedures were developed for micropropagation of Alnus cordata through in vitro axillary shoot multiplication of axillary bud explants cultured in Murashige & Skoog (MS) medium. Establishment of cultures from plants grown in the field was very difficult due to bacterial contamination and phenolic oxidation in explants causing severe browning. Explants were first cultured on an MS medium containing 4.4 μM 6-benzyladenine and 87.6 mM sucrose (initiation medium) for 7 days and then transferred to an MS medium containing 1.1 μM 6-benzyladenine and 333 mM glucose (multiplication medium) for a further 20–25 days. It was necessary to transfer cultures from initiation medium to multiplication medium after 7 days to minimize excessive callus growth, abnormally thick and brittle leaves, inhibition of shoot elongation, and senescence. Shoot multiplication comparable to the above method was achieved by culture of axillary bud explants in MS medium supplemented with 1.1–4.4 μM 6-benzyladenine and 333 mM glucose 4–5 weeks after culture. Shoots rooted in MS medium (1/2 x macro-nutrients) supplemented with 1.2–4.9 μM indolebutyric acid. Also, 98% rooting was achieved when cultures were treated with 625 mgl-1 indolebutyric acid for 24 h at the end of the shoot production stage and rooted in vivo as mini-cuttings. Plantlets established well in soil.