Showing papers on "Murashige and Skoog medium published in 2000"
TL;DR: The nature of the explant, seedling age, medium type, plant growth regulators, complex extracts and antioxidants markedly influenced in vitro propagation of Gymnema sylvestre and the plantlets were hardened and successfully established in natural soil.
Abstract: The nature of the explant, seedling age, medium type, plant growth regulators, complex extracts (casein hydrolysate, coconut milk, malt extract and yeast extract) and antioxidants (activated charcoal, ascorbic acid, citric acid and polyvinylpyrrolidone) markedly influenced in vitro propagation of Gymnema sylvestre. A maximum number of shoots (57.2) were induced from 30 day old seedling axillary node explants on Murashige and Skoog (MS) medium containing 6-benzyladenine (1 mg l−1), kinetin (0.5 mg l−1), 1-napthalene acetic acid (0.1 mg l−1), malt extract (100 mg l−1) and citric acid (100 mg l−1). High frequency of rooting was obtained in axillary node explant derived shoots (50%) on half strength MS medium supplemented with IBA (3 mg l−1). The plantlets, thus developed, were hardened and successfully established in natural soil.
231 citations
TL;DR: Tissue culture responses of young inflorescences of a hybrid bermudagrass cultivar `Tifgreen' (Cynodon dactylon×Cynodocus transvaalensis) and a common bermUDagrasses cultivar`Savannah' (cynodondactylon) were investigated and formed a compact, nodular embryogenic structure which was highly regenerable and morphologically normal.
Abstract: In order to optimize tissue culture conditions for bermudagrass, an important warm-season turfgrass species, tissue culture responses of young inflorescences of a hybrid bermudagrass cultivar `Tifgreen' (Cynodon dactylon×Cynodon transvaalensis) and a common bermudagrass cultivar `Savannah' (Cynodon dactylon) were investigated. When cultured on Murashige and Skoog medium with 4.52 to 13.57 μM (1–3 mg l-1) 2,4-D, young inflorescence segments yielded non-embryogenic calli which were unorganized and had loosely associated, long tubular cells on the surface. However, inclusion of 6-benzyladenine (BA) in callus induction medium at a level of 0.044 μM (0.01 mg l-1) induced formation of a compact, nodular embryogenic structure on approximately 20% of the calli. Calli with such a compact embryogenic structure were highly regenerable. When young inflorescences smaller than 0.75 cm were cultured, the embryogenic structure yielded green plantlets with regeneration rates of 79.5% and 83.3%, respectively for the two cultivars. All 96 plants regenerated from calli induced in the BA-containing medium were green and morphologically normal. The embryogenic nature of the compact structure was confirmed by scanning electron microscopy.
119 citations
TL;DR: An efficient method has been developed for regeneration of complete plants via somatic embryogenesis in Corydalis yanhusuo (Fumariaceae), an important medicinal plant, using tuber-derived callus using somatic embryos to establish somatic embryo-derived plants.
118 citations
TL;DR: After 12 months of cultivation in the field, the micropropagated plants were stable and showed uniform morphological and growth characteristic and the concentration of the root specific compound, 2-hydroxy 4-methoxy benzaldehyde per plant was 2–3 fold higher in micropropAGated plants though on unit dry root biomass basis it remained the same between two sources of plants.
Abstract: Caulogenic responses of various explant types from 12-month-old plants of Hemidesmus indicus were tested Second and third visible nodes (05 cm) from the apex and root segments (05 cm) were the most and least regenerative respectively, with the formation of 937 and 26 shoots in 4 weeks on half strength MS medium supplemented with 222 μM BA and 107 μM NAA and 444 μM BA and 269 μM NAA respectively Caulogenic ability of the nodes decreased with increasing maturity Shoot buds initiated upon the young nodes on day 10 developed into 72 cm long shoots within 4 weeks thereby making a shoot elongation phase unnecessary Nodal explants of the in vitro raised shoots subcultured in the same medium produced 932 shoots of 71 cm length in 3–4 weeks, similar to those of the mature plant-derived nodes Multiplication through subculture of the nodes up to 25 passages of 4 weeks each was achieved without decline Shoot cultures were rooted in quarter salt strength MS medium containing 98 μM IBA and the rooted plants were hardened for establishment in pots at 96% rate Four months after establishment, the micropropagated plants were stable and showed uniform morphological and growth characteristic After 12 months of cultivation in the field, on an average micropropagated plant consisted of 4–5 shoots, 5–8 branches per shoot and increased root biomass (135 g) compared to the poor growth (single shoot and 2–3 branches) and root production (46 g) values obtained with plants raised from conventional rooted stem cuttings The concentration of the root specific compound, 2-hydroxy 4-methoxy benzaldehyde per plant was 2–3 fold higher in micropropagated plants though on unit dry root biomass (012% per g dry wt) basis it remained the same between two sources of plants
116 citations
TL;DR: Although bud break was dependent on BA supply, the synergistic combination of 22.2 μM BA and 2.68 μM NAA induced the optimum frequency of shoot formation as well as shoot number of Centella asiatica plants.
Abstract: A protocol is described for rapid and large-scale in vitro clonal propagation of the valuable medicinal herb Centella asiatica (L) by enhanced axillary bud proliferation in nodal segments isolated from mature plants Although bud break was dependent on BA supply, the synergistic combination of 222 μM BA and 268 μM NAA induced the optimum frequency (91%) of shoot formation as well as shoot number (4 to 5 shoots per node) Subculturing of nodal segments harvested from the in vitro derived axenic shoots on the multiplication medium enabled continuous production of healthy shoots with similar frequency MS medium supplemented with 67 μM BA and 288 μM IAA was found most suitable for shoot elongation Rooting was highest (90%) on full-strength MS medium containing 246 μM IBA Micropropagated plants established in garden soil were uniform and identical to the donor plant with respect to growth characteristics This micropropagation procedure could be useful for raising a stock of genetically homogenous plant material for field cultivation
112 citations
TL;DR: This study aimed to use somaclonal variation to select drought tolerant plants of Tagetes with significant tolerance compared with other regenerated and control plants and revealed lower water potential, greater accumulated biomass and a higher relative growth rate.
100 citations
TL;DR: The procedure reported here allows the micropropagation of H. perforatum in five months of culture and the proliferation of cell masses which could be used for studies on organic compounds of pharmaceutical interest.
Abstract: Use of Hypericum perforatum L. has increased in the past few years due to the antidepressant and antiviral activities found in extracts of this plant. As a result of its potential as a pharmaceutical, a new system was developed for in vitro culture of this species. Leaf explants were inoculated onto MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 0.45 or 4.5 μM) and 6-benzyladenine (BA, 0.44 or 4.4 μM) or kinetin (0.46 or 4.6 μM). Explants were cultivated under dark or light conditions to induce callus formation. Callus initiation was observed in all media evaluated and the highest cell proliferation was obtained from explants cultivated in the presence of 4.4 μM BA and 4.5 μM 2,4-D in the dark. Shoot induction was obtained from callus induced on 4.6 μM kinetin and 0.45 μM 2,4-D 6 weeks after transferring the callus to a MS medium supplemented with 4.4 μM BA. Roots were induced from shoots on full and half-strength MS media with or without indolebutyric acid (IBA, 4.9 μM) and the highest rooting frequencies were obtained on half-strength MS medium, regardless of the presence of IBA. Regenerated plants were easily acclimated in greenhouse conditions. The procedure reported here allows the micropropagation of H. perforatum in five months of culture and the proliferation of cell masses which could be used for studies on organic compounds of pharmaceutical interest.
98 citations
TL;DR: Direct regeneration of shoot buds was observed in MS basal medium supplemented with various concentrations of either benzyladenine (BA) or thidiazouron (TDZ) depending on the explant.
Abstract: In vitro studies were initiated with Withania somnifera (L.) Dun. for rapid micropropagation of selected chemotypes using nodes, internodes, hypocotyls and embryo explants. Direct regeneration of shoot buds was observed in MS basal medium supplemented with various concentrations of either benzyladenine (BA) or thidiazouron (TDZ) depending on the explant. Nodal explants formed multiple shoots both from pre-existing and de novo buds on Murashige and Skoog's medium (MS) containing 0.1–5.0 mg l−1 BA and a ring of de novo shoot buds on MS medium containing 0.2 and 0.3 mg l−1 TDZ. Internodal explants formed shoot buds on MS with 1.0 and 5.0 mg l−1 BA while the hypocotyl explants gave rise to multiple shoots only on MS with 0.5 mg l−1 BA. Isolated embryos gave rise to many shoot buds on MS with 0.2 and 0.3 mg l−1 TDZ. The shoot buds elongated and rooted either on MS medium with 0.01 mg l−1 BA or on half strength MS medium lacking growth regulators, which depended upon the growth regulator used in the shoot bud induction medium. Except for the embryo-derived plantlets, all other plantlets could be acclimatized with 100% success.
93 citations
TL;DR: During the first months of culture, no significant differences in the sucrose content were found between calli that produced embryos and those that did not, but during the development of somatic embryogenesis very low starch levels in the callus were found.
89 citations
TL;DR: Shoot tips from in vitro-grown, cold-hardened stock plants of white poplar were successfully cryopreserved at –196 °C by one-step vitrification, and almost 60% rooted shoots were obtained from cryop Reservation.
Abstract: Shoot tips from in vitro-grown, cold-hardened stock plants of white poplar (Populus alba L.) were successfully cryopreserved at –196 °C by one-step vitrification. After preculturing at 5 °C for 2 days on hormone-free MS medium containing different sucrose concentrations, and loading for 20 min with 2 m glycerol and 0.4 m sucrose, shoot tips were treated with the PVS2 vitrification solution and plunged directly into liquid nitrogen. Best survival rate (90%) was obtained when shoot tips were precultured on 0.09 m sucrose, hormone-free MS medium, vitrified by exposure to PVS2 solution for 60 min at 0 °C and, following cryopreservation, rewarmed at 40 °C and washed in 1.2 m sucrose solution for 20 min. Regrowth was improved by plating shoot tips on a gelled MS medium containing 1.5 μm N6-benzyladenine plus 0.5 μm gibberellic acid, while shoot rooting was achieved on MS medium containing 3 μm indole-3-butyric acid. Following this procedure, almost 60% rooted shoots were obtained from cryopreserved shoot tips.
87 citations
TL;DR: Multiple shoots were induced from nodal segments of mature trees of Pistacia vera L. on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BA) and stable regenerated plants were established in the greenhouse.
Abstract: Multiple shoots were induced from nodal segments of mature trees of Pistacia vera L. on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BA). Maximum shoot production was obtained from shoot tips taken from in vitro proliferated shoots when cultured on solidified MS medium containing 8.8 μ MB A. The multiplication rate was 20 microshoots per explant on the 30th day. Rooting of microshoots was achieved in MS medium supplemented with indole butyric acid (IBA). Rooted plantlets reassumed independent growth after a short period of acclimatisation. Stable regenerated plants were established in the greenhouse.
TL;DR: Of the six Morus species studied, the best shoot and root development was observed in M. alba, and one-step regeneration, i.e. both shoot andRoot formation, was recorded inM.
Abstract: Axillary buds obtained from in vitro shoot cultures of six mulberries (Morus alba L., M. australis Poir., M. bombycis Koidz., M. cathyana Hemsl., M. latifolia Poir., and M. nigra L.) were encapsulated in calcium alginate hydrogel containing Murashige and Skoog (1962) nutrients (MS) and 4.4 μM benzyladenine (BA). Morphogenic response of encapsulated buds to various planting media such as MS medium + 4.4 μM BA, MS basal medium, soilrite mix + half-strength MS medium, garden soil + half-strength MS medium, soilrite mix + tap water and garden soil + tap water was evaluated. Encapsulated buds of M. alba, M. bombycis, M. latifolia and M. nigra exhibited shoot development in each of the six media tested whereas that of M. australis and M. cathyana responded only to the first four media. Analysis of variance revealed that the planting medium exhibited the greatest influence on shoot development. Of the six planting media evaluated, shoot development was highest in MS medium containing 4.4 μM BA and lowest in garden soil moistened with water. Of the six Morus species studied, one-step regeneration, i.e. both shoot and root formation, was recorded in M. alba, M. bombycis and M. latifolia. Rooted shoots were retrieved from encapsulated buds of these species on all planting media tested except the one that contained BA. Root development was significantly affected by the planting medium and the plant species with planting medium contributing the maximum amount (82%) of the total variation observed. Of the five planting media tested, the percentage of root development was highest in MS basal medium. Of the six Morus species studied, the best shoot and root development was observed in M. alba. Encapsulated buds of M. bombycis, M. latifolia and M. nigra stored for 90 days and those of M. alba, M. australis and M. cathyana for 60 days at 4 °C still regenerated shoots. Plants regenerated from the encapsulated buds were hardened off and transferred to soil.
TL;DR: The development of stem callus mediated plant regeneration system for Withania somnifera is described and maximum shoot regeneration was achieved in four weeks when callus was cultured on MS medium fortified with 4.44 μM BA and 0.57 μM IAA.
Abstract: The development of stem callus mediated plant regeneration system for Withania somnifera is described. Maximum callus proliferation was obtained on Murashige and Skoog medium supplemented with 2.26 μM 2,4-D. Three-week-old, white, friable callus was used for shoot regeneration. The maximum shoot regeneration (6.2 ± 0.34 shoots/explant) was achieved in four weeks when callus was cultured on MS medium fortified with 4.44 μM BA and 0.57 μM IAA. Regenerated shoots were excised and multiplied (8.4 ± 0.43 shoots/explant) on MS medium supplemented with 4.44 μM of BA. Multiple shoots were divided into single shoots and were rooted (5.1 ± 0.49 rootlets/shoot) on half strength MS medium supplemented with 9.84 μM of IBA. After a hardening phase of 3 weeks the plantlets were transferred to the field.
TL;DR: An efficient method of micropropagation based on an increased percentage survival of explants and reduced phenol-induced browning in wild strawberry has been developed and 6-benzyladenine was found more effective than kinetin and N6-(γ,γ dimethylallyamino) purine.
Abstract: An efficient method of micropropagation based on an increased percentage survival of explants and reduced phenol-induced browning in wild strawberry has been developed. Serial transfer of nodal explants was carried out at 24-, 48- and 96-h intervals. Nodal segments cultured on Murashige and Skoog medium supplemented with 6-benzyladenine (4.0 μM) and α-naphthalene acetic acid (0.1 μM) gave the best (94.4%) explant establishment and shoot number (22.3) per explant. Of the cytokinins tested, 6-benzyladenine was found more effective than kinetin and N6-(γ,γ dimethylallyamino) purine. Excised shoots rooted on half-strength agar-gelled medium with 1.0 μM α-naphthalene acetic acid. Rooted shoots with fully expanded leaves acclimatized successfully and about 70% of plantlets survived ex vitro.
TL;DR: Subculture of the chlorotic plantlets into MSM showed that the visual symptoms of mineral deficiency disappeared in 2–4 wk, and a comparison of the mineral analysis of plantlets in both media showed a fairly large increase in Ca, Cu, Fe, Mg and S and decrease in levels of B and Cl in plantlets cultivated in MSM.
Abstract: Mineral deficiency symptoms were observed in leaves of yellow passionfruit plantlets grown in MS medium (Murashige and Skoog, 1962) with 1.0 mg l−1 (3.0 μM) gibberellic acid. Initially, leaves showed interveinal chlorosis, followed by bleaching of the leaves and retarded growth. Leaf mineral analysis was done and compared to mineral requirements suggested for passionfruit in the literature. Several modifications were made to the inorganic composition of MS medium, according to mineral deficiencies, mainly of Fe and Ca, and possible toxicity of Cl. The concentration of the elements in the new medium (MSM) was based on the mineral composition of leaves of healthy plants. The chemical equilibrium was checked using the software Geochem (Sposito and Mattigod, 1980) and final adjustments were made to ensure good availability of nutrients. To test the efficiency of the modified medium nodal segments were cultured in both MS and MSM supplemented with 3.0 mg l−1 (13.3 μM) 6-benzyladenine. After three subcultures mineral analysis of the leaves was done. Severe mineral deficiency was observed on the leaves of plantlets cultured in MS, while plantlets cultivated in MSM had green leaves. A comparison of the mineral analysis of plantlets in both media showed a fairly large increase in Ca, Cu, Fe, Mg and S and decrease in levels of B and Cl in plantlets cultivated in MSM. A slight increase or decrease in other elements was also observed. Subculture of the chlorotic plantlets into MSM showed that the visual symptoms of mineral deficiency disappeared in 2–4 wk.
TL;DR: Micropropagation of the endangered terrestrial orchid, Geodorum densiflorum (Lam.) Schltr.
Abstract: Micropropagation of the endangered terrestrial orchid, Geodorum densiflorum (Lam.) Schltr. was achieved through rhizome section culture from in vitro seed- derived rhizomes. Rhizome sections were cultured on Murashige and Skoog (MS) and Knudson C(KC) media supplemented with various growth regulators and 0.1% activated charcoal. The rhizome sections responded on MS medium. Naphthaleneacetic acid (NAA) at 2.0 μM stimulated rhizome growth. However, benzyladenine (BA) at 5.0 μM induced multiple shoots within four weeks of culture and inhibited rhizome growth. The regenerated shoots rooted on MS only or with NAA at 1.0 μM. Well-developed plantlets were transferred to community pots and then to a greenhouse where the plants have been acclimatised.
TL;DR: An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and stem explants of abnormal seedling as an explant have been developed in Gossypium hirsutum L.
Abstract: An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and stem explants of abnormal seedling as an explant have been developed in Gossypium hirsutum L. Embryogenic callus and somatic embryos have been obtained directly from the explants of cotton abnormal seedlings. Plant growth regulators influenced the induction of cotton somatic embryogenesis. The optimal medium for direct somatic embryogenesis was modified MS medium supplemented with 0.1 mg l-1 ZT and 2 g l-1 activated carbon. On this medium, an average of 28.0 and 28.1 matured somatic embryos formed from per leaf and stem explants respectively. The highest frequency of somatic embryogenesis was 100%. The somatic embryos were converted into normal plantlets when cultured on modified MS medium supplemented with 0.1 mg l-1 ZT. Upon transfer to soil, plants grew well and appeared normal. Plants could be regenerated within 60–80 days. The system of cotton somatic embryogenesis and plant regeneration described here will facilitate the application of plant tissue culture and genetic engineering on cotton genetic improvement.
TL;DR: In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds, which were germinable both in vitro and in vivo.
Abstract: Multiple shoots were efficiently regenerated from cotyledonary node and shoot tip explants of Pisum sativum within 15 days on MS medium containing B5 vitamins and supplelmented with 2.0 mgl-1 6-benzylaminopurine. The elongated shoots produced on the same medium were excised and transferred to MS medium containing half strength ammonium nitrate (8.25 gml-1) and supplemented with auxins (indole-3-butyric acid or naphthalene acetic acid) either alone or in combinations with gibberellic acid. Rooting and flowering were observed on the 7th and 15th day after their transfer to rooting medium. The flowers self-fertilised in vitro and produced mature pods within 25 days of rooting. These seeds were germinable both in vitro and in vivo. In vitro seeds sown in pots under field conditions developed into flowering plants, and subsequently produced pods with viable seeds.
TL;DR: Evaluation of explant regeneration throughout the year indicated that the incidence of browning of explants was maximum during the month of February, while dominance of the microbes in endogenously infected explants peaked in August–September.
Abstract: A protocol for the micropropagation of mature Eucalyptus tereticornis Smith has been developed using regenerated shoots from axillary bud explants. The trees were selected on the basis of their better growth rate, physical and phenotypic characteristics and freedom from disease. Regeneration was obtained in modified Murashige and Skoog (1962) medium. Evaluation of explant regeneration throughout the year indicated that the incidence of browning of explants was maximum during the month of February, while dominance of the microbes in endogenously infected explants peaked in August–September. Regeneration from primary explants was maximum during the months of March–April. Subcultures were carried out every 4 weeks. Effects of hormones and media composition on regeneration and growth were studied. Phytagel induced vitrification, while calcium chloride dihydrate reduced vitrification and induced the elongation of shoots. Best rooting was obtained with half-strength, modified MS medium supplemented with 1.0 mg/l indolebutyric acid. Plantlets were hardened in a nonsterile potting mix at high humidity and gradually exposed to the ambient environment over a period of 6 weeks, and upon transfer to field conditions the survival rate varied from 84% to 100%.
TL;DR: The results indicate that TDZ effectively stimulates sugarcane plant regeneration from embryogenic callus, and may be suitable to use in genetic transformation studies to enhance regeneration of transgenic plants.
Abstract: Efficient shoot regeneration of sugarcane (Saccharum spp. hybrid cv. CP84-1198) from embryogenic callus cultures has been obtained using thidiazuron (TDZ). Callus was placed on modified Murashige and Skoog (MS) medium containing 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D), or 9.3 μM kinetin and 22.3 μM naphthaleneacetic acid (NAA) and compared with the same MS medium supplemented with 0.5, 1.0, 2.5, 5.0 or 10.0 μMTDZ, A11 TDZ treatments resulted in faster shoot regeneration than the kinetin/NAA treatment, and more shoot production than either the 2,4-D or kinetin/NAA treatments. Maximum response, as determined by total number of shoots (26 per explant) and number of shoots greater than 1 cm (4 per explant) 4 wk after initiation, was obtained with 1.0 μM TDZ. The shoots rooted efficiently on MS medium supplemented with 19.7 μM indole-3-butyric acid (IBA). These results indicate that TDZ effectively stimulates sugarcane plant regeneration from embryogenic callus, and may be suitable to use in genetic transformation studies to enhance regeneration of transgenic plants.
TL;DR: The effect of indole-3-acetic acid or kinetin on the weight and number of microtubers formed was studied on single node cuttings of sevendifferent potato cultivars as well astransgenic lines harbouring rolB or rolC genes undercontrol of the patatin class I (B33) promoter.
Abstract: The effect of indole-3-acetic acid or kinetin on the weight and numberof microtubers formed was studied on single node cuttings of sevendifferent potato (Solanum tuberosum L.) cultivars as well astransgenic lines harbouring rolB or rolC genes undercontrol of the patatin class I (B33) promoter. Plants were cultivatedin vitro in the dark on solidified MS medium containing 1 to8% sucrose with or without phytohormones. Most of thenontransformed potato cultivars and transgenic lines responded tohormone application by an increase in tuber yield. Auxin and cytokininacted differently: IAA increased predominantly the tuber size whilekinetin increased the number of tubers. RolC transformantsdisplayed an altered response to sucrose and especially to auxin. Thedegree of phytohormone effect on tuberisation parameters depended onsucrose content of the medium and potato genotype.
TL;DR: The transformed ginseng plantlets had an actively growing root system with abundant lateral roots and the phenotypical alteration of transformed gINSeng plants might be valuable character for increasing root yield.
Abstract: Hairy roots of Panax ginseng were obtained after root disks were infected with wild-type strain Agrobacterium rhizogenes 15834. Three lines of hairy roots with different pigmentation were selected. Embryogenic callus was induced on Murashige and Skoog medium containing 1.0 mg/l 2,4-D. The frequency of embryogenic callus formation was 37.4% in a line with red pigmentation. Somatic embryo development from embryogenic callus was efficiently achieved by lowering the concentration of 2,4-D (0.5 mg/l). After the germination of somatic embryos on medium with 10 mg/l GA3, the embryos were transferred to 1/2-MS medium without GA3. The transformed ginseng plantlets had an actively growing root system with abundant lateral roots. The phenotypical alteration of transformed ginseng plants might be valuable character for increasing root yield.
TL;DR: Plant micropropagation of blueberry, blackberry and raspberry was carried out from nodal segments of adult field-grown plants, while tests carried out in both shoots and regenerated plants revealed the absence of any kind of disease.
Abstract: Summary
Plant micropropagation of blueberry (Vaccinium corymbosum L. cv. Berkeley), blackberry (Rubus sp. cv. Smoothstem) and raspberry (Rubus idaeus L. cv. Gradina) was carried out from nodal segments of adult field-grown plants. Hardwood and softwood cuttings were studied as explant sources. The cultures successfully established were softwood from all three species, and hardwood only from blueberry. Shoot-bud establishment from blueberry was achieved by culturing explants in WPM salts with MS vitamins for 15 days, and then 30 days in the same medium with 18 mM Zeatin. The best results of multiplication were obtained in the same medium with 25 mM 2iP. For blackberry, shoot-bud establishment was achieved by culturing explants in MS medium for 15 days, and then in the same medium with 4 mM BA and 0.25 mM IBA. This medium was also the best for blackberry multiplication. Raspberry explants (cvs Gradina and Willamette) were cultured in MS medium for 15 days and then transferred to MS medium supplemented with 4 μm BA and 0.25 mM IBA. After 30 days of culture, only ‘Gradina’ explants survived, from which shoot-bud establishment was obtained in a modified MS medium (Anderson's macronutrients except calcium, with Sequestrene as the iron source) with the same growth regulators. Multiplication was achieved by subculturing explants in the same medium either with 4 mM BA plus 0.25 mM IBA or with 8 mM BA plus 0.25 mM IBA. Shoots of at least 1 cm in length from all species were rooted ex vitro in a mixture of peat and Perlite (1:1, v/v) in a mist chamber, and 100% of rooting plants were acclimated. Bacterial, fungal and viral diseases were detected in stock plants, while tests carried out in both shoots and regenerated plants revealed the absence of any kind of disease.
TL;DR: Calli developed further along a route of production of protocorm-like bodies and eventually formed plantlets that could be transplanted to pots and grew well.
Abstract: Totipotent calli of a Paphiopedilum hybrid (Paphiopedilum callosum ‘Oakhi’ × Paph. lawrenceanum ‘Tradition’) were induced from seed-derived protocorms on a 1/2 strength Murashige–Skoog medium plus 1–10 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1–1 mg l−1 1-phenyl-3-(1.2.3-thiadiazol-5-yl)urea (TDZ). These calli grew well when subcultured on the same medium, but proliferated more on 1/2 MS medium plus 5 mg l−1 2,4-D and 1 mg l−1 TDZ. Calli developed further along a route of production of protocorm-like bodies and eventually formed plantlets that could be transplanted to pots and grew well.
TL;DR: An anatomical study confirmed shoot regeneration via direct organogenesis and micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium.
Abstract: The establishment of a micropropagation protocol for Pothomorphe umbellata was carried out using leaf segments cultured on 1/4 strength Murashige and Skoog medium supplemented with 0.5 mg l-1 6-benzyladenine, 0.1 mg l-1 gibberelic acid added with 10 g l-1 sucrose. Rooting was achieved using MS medium devoid of growth regulators. An anatomical study confirmed shoot regeneration via direct organogenesis.
TL;DR: Callus cultures were established from excised embryos of Nothapodytes foetida on Murashige and Skoog medium supplemented with picloram and 9-methoxy-camptothecine as determined by TLC, UV, HPLC, electron spray mass spectral analysis.
Abstract: Callus cultures were established from excised embryos of Nothapodytes foetida on Murashige and Skoog medium supplemented with picloram (2 mg l−1) and 3% (w/v) sucrose. The embryos developed into callus after 4 weeks of incubation at 25 ± 2 °C in dark. The cultures produced camptothecine and 9-methoxy-camptothecine as determined by TLC, UV, HPLC, electron spray mass spectral analysis.
TL;DR: A continuously growing callus was obtained from immature endosperm of Morus alba L Cv S-36 cultured on Murashige and Skoog medium containing 5 μm 2,4-dichlorophenoxyacetic acid and each of the ten plants examined cytologically was triploid.
Abstract: A continuously growing callus was obtained from immature endosperm of Morus alba L Cv S-36 cultured on Murashige and Skoog medium containing 5 μm 2,4-dichlorophenoxyacetic acid. Shoot buds were produced when the callus was subcultured on a medium containing a cytokinin or a cytokinin and 1-naphthaleneacetic acid (NAA). The maximum number of shoots was formed on the medium containing thidiazuron (1 μM), or benzylaminopurine (5 μM) and NAA (1 μM). Shoots were multiplied by forced axillary branching and rooted in vitro. Endosperm-derived plants were established in soil. Each of the ten plants examined cytologically was triploid (3 n=42).
TL;DR: A continuous system of mass propagation of bulblets was achieved through in vitro scale formation (secondary explants) on MS medium supplemented with 23 μM kinetin and 0.5 μM NAA, as well as scale proliferation on MS basal liquid stationary medium.
Abstract: A simple, rapid and cost-effective in vitro scheme has been proposed for mass propagating two cultivars of Asiatic lily hybrids. An average of seven bulblets was formed after 17 d when 1×1 cm2 bulb scale segments (explants) were cultured on Murashige and Skoog (MS) medium with 3% sucrose and 0.5 μM α-naphthaleneacetic acid (NAA). On MS medium containing 0.5 μM NAA and 6 or 9% sucrose, depending on the cultivar, large numbers of bulblets of increased size (3.5–5.0 cm in circumference) were formed under a 16/8 h photoperiod. A continuous system of mass propagation of bulblets was achieved through in vitro scale formation (secondary explants) on MS medium supplemented with 23 μM kinetin and 0.5 μM NAA, as well as scale proliferation on MS basal liquid stationary medium. Upon transplantation all bulblets sprouted, of which 40% flowered in the first season. Under ideal conditions, ca. 9.68×105 bulblets can be produced from a single scale segment in 1 yr by following the systematic propagation steps proposed here.
TL;DR: The HDL clones closely resemble the parent in having higher content of citronellol than geraniol while the LDL clones contain almost equal contents of citronesol and Geraniol in their essential oils as revealed by gas chromatography analysis.
TL;DR: In vitro morphogenesis via organogenesis was achieved from callus cultures derived from hypocotyl explants of Acacia sinuata on MS (Murashige and Skoog, 1962) medium and Rooted plantlets were hardened and successfully established in the soil.
Abstract: In vitro morphogenesis via organogenesis was achieved from callus cultures derived from hypocotyl explants of Acacia sinuata on MS (Murashige and Skoog, 1962) medium. Calli were induced from hypocotyl explants excised from 7-day-old seedlings on MS medium containing 3% sucrose, 0.8% agar, 6.78 μM 2,4-dichlorophenoxyacetic acid and 2.22 μM 6-benzylaminopurine. Regeneration of adventitious buds from callus was achieved when they were cultured on MS medium supplemented with 10% coconut water, 13.2 μM 6-benzylaminopurine and 3.42 μM indoleacetic acid. Addition of gibberellic acid (1.73 μM) favored shoot elongation. Regenerated shoots produced prominent roots when transferred to half strength MS medium supplemented with 7.36 μM indolebutyric acid. Rooted plantlets, thus developed were hardened and successfully established in the soil. This protocol yielded an average of 20 plants per hypocotyl explant over a period of 4 months.