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Showing papers on "Murashige and Skoog medium published in 2004"


Journal ArticleDOI
TL;DR: Results indicated that shoots established at 100% regardless of media type, however, shoot height, nodes per shoot, and leaf number were highest for explants established on MS medium compared to NN or B5.

144 citations


Journal ArticleDOI
TL;DR: PCR and Southern blot analyses revealed that plants derived from hairy roots retained the Ri TL-DNA, and a significant difference in the frequency of adventitious shoot formation for each hairy-root line derived from a different cultivar.
Abstract: Hypocotyl explants of Catharanthus roseus produced hairy roots when cultured on Murashige and Skoog (MS) basal medium after infection by Agrobacterium rhizogenes. Explants gave rise to adventitious shoots at a frequency of up to 80% when cultured on MS medium supplemented with 31.1 μM 6-benzyladenine and 5.4 μM α-naphthaleneacetic acid. There was a significant difference in the frequency of adventitious shoot formation for each hairy-root line derived from a different cultivar. Plants derived from hairy roots exhibited prolific rooting and had shortened internodes. Approximately half of the plants had wrinkled leaves and an abundant root mass with extensive lateral branching, but otherwise appeared morphologically normal. Plants with hairy roots that were derived from the cultivar Cooler Apricot developed flowers with petals that were white in the proximal region, whereas the wild-type flower petals are red. PCR and Southern blot analyses revealed that plants derived from hairy roots retained the Ri TL-DNA.

103 citations


Journal ArticleDOI
TL;DR: This protocol of successful plant regeneration by asymbiotic seed germination should permit rapid propagation and conservation of this medicinally important Dendrobium species.
Abstract: Shi-hu (Dendrobium spp. or Dendrobii Herba) is one of the important traditional Chinese medicines. The commercially available crude drug in the traditional medicine market is composed mainly of three species: Dendrobium tosaense, D. nobile, and D. moniliforme. An efficient method of propagation has been developed via asymbiotic germination of seeds in vitro for the medicinally important D. tosaense. Seeds from capsules of D. tosaense collected 8–14 wk after artificial pollination germinated after being cultured on full-strength or half-strength Murashige and Skoog (MS) medium devoid of plant growth regulators and with 3% sucrose. Germination of seeds varied with the medium type and seed maturity. Germinated seedlings after transfer to MS medium with 1.5% sucrose and 8% banana homogenate or potato juice or coconut water and 20 wk of incubation developed into healthy plantlets. Well-developed plantlets were transplanted to moss or moss and tree fern or tree fern as substrates in plastic trays and transferred to a greenhouse for hardening. All plants survived, attained maturity, and developed normal flower and capsule after one and a half years. This protocol of successful plant regeneration by asymbiotic seed germination should permit rapid propagation and conservation of this medicinally important Dendrobium species.

103 citations


Journal ArticleDOI
TL;DR: Using this efficient regeneration system, plantlets were regenerated from seven elite maize inbred lines and provided a solid basis for genetic transformation of maize.
Abstract: An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l(-1) 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l(-1)) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l(-1)) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l(-1) BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l(-1) indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.

101 citations


Journal ArticleDOI
TL;DR: Bamboo somatic embryos and in vitro regenerants and potted plants flowered, but no seeds were produced and the effects of sucrose and thidiazuron (TDZ) on callus proliferation were studied.
Abstract: Current research on somatic embryogenesis of bamboo uses reproductive tissue as explants. However, it was hard to obtain the explant. Shoots of a local accession (3–4 m high) were used for multiple shoot production. In order to obtain embryogenic callus, nodal and internodal tissues from in vitro plantlets were placed on Murashige and Skoog (MS) medium supplemented with 9.2 μM kinetin (KN), 13.6 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.1% (v/v) coconut milk, and 6% (w/v) sucrose. We studied the effects of sucrose and thidiazuron (TDZ) on callus proliferation. Optimal additives to the MS medium for embryogenic callus proliferation were 0.046 μM TDZ, 13.6 μM 2,4-D and 3% (w/v) sucrose. TDZ also promoted the germination of bamboo somatic embryos. The germination rate of the somatic embryos exceeded 80% on MS-based medium supplemented with 0.455μM TDZ. Naphthaleneacetic acid (NAA) reduced germination. Well-developed plantlets were successfully transferred to soil. There was no albino mutant in subsequent culture. In vitro regenerants and potted plants flowered, but no seeds were produced.

91 citations


Journal ArticleDOI
TL;DR: Leaf explants of Paphiopedilum phiIippinense hybrids directly formed adventitious shoots from wound regions within 1 month, when cultured on modified Murashige and Skoog medium free of plant growth regulator in darkness.
Abstract: Leaf explants of Paphiopedilum phiIippinense hybrids (hybrid PH59 and PH60) directly formed adventitious shoots from wound regions within 1 month, when cultured on modified Murashige and Skoog medium (1/2-strength macro- and full-strength micro-elements) free of plant growth regulator in darkness. The combinations of 2,4-dichlorophenoxyacetic acid ((2,4-D) acid (0, 4.52 and 45.25 μM) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) (0, 0.45, 4.54 and 22.71 μM) were used to test their effects on direct shoot bud formation from two types of explants (1.5-cm long intact leaf explants and 0.5-cm long leaf segment explants). In hybrid PH59, 4.54 μM TDZ increased mean numbers of shoots per explant with leaf segment explants. In hybrid PH60, 4.52 μM 2,4-D plus 0.45 μM TDZ promoted direct shoot bud formation from leaf segment explants. In addition, three treatments (4.52 μM 2,4-D, 22.71 μM TDZ, 4.52 μM 2,4-D plus 4.54 μM TDZ) gave a higher response than control on mean numbers of shoots per explant with intact leaf explants. Healthy plantlets each with one to three roots were obtained from leaf-derived shoots after transfer onto a hormone-free medium for 22 months. These plantlets were acclimatized in a greenhouse and grew well with 100% survival rate.

84 citations


Journal ArticleDOI
TL;DR: The somatic embryogenesis methodology used did not induce major genetic changes in the somatic embryos and that the primary goal of “true-to-type” propagation was assured, indicating that no changes were induced during the embryogenic process.
Abstract: Flow cytometry was used to measure the nuclear DNA content of Eucalyptus globulus Labill. somatic and zygotic embryos and leaves in order to determine if somatic embryogenesis induces DNA content and ploidy changes in this species. Mature zygotic embryos derived from open-pollination orchard families were collected from a location in the centre of Portugal. One group was kept for nuclear DNA content and ploidy analysis, and the other group was used for establishing embryogenic cultures. Mature zygotic embryos were grown on Murashige and Skoog (MS) medium supplemented with 3% (w/v) sucrose and 3 mg l−1 α-naphthaleneacetic acid (NAA) for 3 weeks and then transferred to MS medium without growth regulators. Globular somatic embryos from approximately 8-month-old embryogenic cultures were used in the assay. DNA ploidy levels and the nuclear DNA content of mature zygotic embryos, somatic embryos and leaves from the mother field tree were determined using flow cytometry combined with propidium iodide staining. Zygotic embryos had a nuclear DNA content of 1.32 pg/2C, somatic embryos had a nuclear DNA content of 1.39 pg/2C and leaves from the field tree had a nuclear DNA content of 1.40 pg/2C. The values estimated for the somatic embryos and mother plant did not differ statistically from each other (P≤0.05), but both differed from those of the zygotic embryos (P≤0.05). These results clearly indicate that no changes were induced during the embryogenic process. However, the differences found between the field plants and zygotic embryos did suggest that some aspects must be evaluated carefully, as propidium iodide fluorescence may potentially be influenced by the presence of secondary compounds (e.g. anthocyanins, tannins) in E. globulus somatic embryos and mature leaves. Therefore we believe that the somatic embryogenesis methodology used did not induce major genetic changes in the somatic embryos and that our primary goal of “true-to-type” propagation was assured.

83 citations


Journal ArticleDOI
TL;DR: A protocol is outlined for direct and indirect regeneration of a medicinally valuable Curcuma amada Roxb using rhizome and leaf sheath explants, which revealed most of the regenerated plantlets were similar to the mother plants.
Abstract: A protocol is outlined for direct and indirect regeneration of a medicinally valuable Curcuma amada Roxb. using rhizome and leaf sheath explants. Multiple shoots were obtained from rhizome explants on Murashige and Skoog (MS) medium fortified with 4.44 µM BA and 1.08 µM α-napthaleneacetic acid (NAA). For indirect regeneration, semi-friable callus obtained from leaf sheath explants on MS medium with 9.0 µM 2,4-D was used. Transfer to 8.88 µM BA and 2.7 µM NAA containing medium produced optimum shoot initiation and development. The regenerated plantlets were transferred to the field. Random amplified polymorphic DNA (RAPD) analysis of regenerated plantlets revealed 103 scorable bands from 10 primers, including nine polymorphic bands, which were absent in control. However, most of the regenerated plantlets were similar to the mother plants.

81 citations


Journal ArticleDOI
TL;DR: A protocol has been developed for in vitro plant regeneration from cotyledonary nodes of Pterocarpus marsupium Roxb and about 52% of plantlets were successfully acclimatized and established in pots.
Abstract: A protocol has been developed for in vitro plant regeneration from cotyledonary nodes of Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes derived from 20-d-old axenic seedlings grown on Murashige and Skoog (MS) medium containing 2.22–13.32 μM benzyladenine (BA) or 2.32–13.93 μM kinetin alone or in combination with 0.26 μM α-naphthaleneacetic acid (NAA). The highest frequency for shoot regeneration (85%) and maximum number of shoots per explant (9.5) were obtained on the medium supplemented with 4.44 μM BA and 0.26 μM NAA after 15 wk of culture. A proliferating shoot culture was established by repeatedly subculturing the original cotyledonary nodal explant on fresh medium after each harvest of the newly formed shoots. Nearly 30% of the shoots formed roots after being transferred to half-strength MS medium containing 9.84 μM indole-3-butyric acid (IBA) after 25 d of culture. Fifty percent of shoots were also directly rooted as microtuttings on a peat moss, soil, and compost mixture (1∶1∶1). About 52% of plantlets were successfully acclimatized and established in pots.

79 citations


Journal ArticleDOI
TL;DR: Micropropagation protocol for an elite selection of Aloe vera syn A. barbadensis through enhanced axillary branching was standardized and the plants were successfully transferred in the soil and were morphologically similar to mother plants.
Abstract: Micropropagation protocol for an elite selection of Aloe vera syn A. barbadensis through enhanced axillary branching was standardized. Murashige and Skoog medium containing 1 mg l−1 BA and 0.2 mg l-1 IBA gave highest multiplication. Citric acid at 10mg l-1 and liquid medium improved the shoot multiplication. Hundred per cent microshoots produced rooted plantlets within 15 days of culture on hormone-free agar medium. Liquid medium during rooting stage decreased the number of shoots showing rooting response. The plants were successfully transferred in the soil and were morphologically similar to mother plants.

74 citations


Journal Article
TL;DR: This method could be useful for regenerating large number of plants as well as provide a target tissue for genetic transformation studies after induction of direct embryogenesis from inflorescence segments in Indian sugarcane.
Abstract: A protocol for direct somatic embryogenesis without an intervening callus phase was developed for sugarcane (Saccharum spp. hybrids) using immature inflorescence segments. Murashige and Skoog (MS) medium supplemented with0.5 mg/l naphthaleneacetic acid, 2.5 mg/I kinetin, 100 mg/l L-glutamine and 4% sucrose showed high frequency of somatic embryo development (54.09 ′ 2.7%), with an average 7.72 ′ 0.89 plants per explant. Embryo development was seen all over the cultured explants within four weeks of culture and the embryos germinated within a week upon transfer to basal MS medium without any growth regulators and all the plants grew normally in the greenhouse. This is a report on the induction of direct embryogenesis from inflorescence segments in Indian sugarcane. This method could be useful for regenerating large number of plants as well as provide a target tissue for genetic transformation studies.

Journal ArticleDOI
Zhihua Liao1, Min Chen1, Feng Tan, Xiaofen Sun1, Kexuan Tang1 
TL;DR: In vitro propagation can be a useful tool in the conservation of this endangered medicinal species and among the three factors, sucrose was the most important for bud initiation followed by BA, and NAA had the weakest effect.
Abstract: A rapid micropropagation protocol was established for Aloe vera L. var. chinensis (Haw.) Berger, Chinese Aloe. The effects of three factors, namely BA, NAA and sucrose, on bud initiation were evaluated by L9 (34) orthogonal design. The variance analysis of the experimental results showed that the actions of the three factors were all considerable. Among the three factors, sucrose was the most important for bud initiation followed by BA, and NAA had the weakest effect. The best medium for bud initiation was semi-solid MS supplemented with 2.0 mg l−1 BA, 0.3 mg l−1 NAA, 30 g l−1 sucrose and 0.6 g l−1 PVP (pH 5.8), on which Chinese aloe could multiply 15 times in 4 weeks. Some shoots rooted spontaneously on 1/2 strength MS medium, but the rooting percentage was improved in the presence of 0.2 mg l−1 NAA. Rooted plantlets were acclimatized to greenhouse conditions. The young plantlets from tissue culture were transplanted successfully. In vitro propagation can be a useful tool in the conservation of this endangered medicinal species.

Journal ArticleDOI
TL;DR: Callus developed on NAA and Kn-supplemented medium favored induction and maturation of embryos earlier compared to that on 2,4-D and Kn, andEmbryo-derived plantlets established in field conditions displayed morphological characters identical to those of the parent plant.
Abstract: High-frequency somatic embryogenesis and plant regeneration was achieved on callus derived from leaf (petiole and lamina) and internode explants of Centella asiatica L. Growth regulators significantly influenced the frequency of somatic embryogenesis and plant regeneration. Calluses developed on Murashige and Skoog (MS) medium fortified with 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 5.37 μM α-naphthaleneacetic acid (NAA), both with 2.32 μM kinetin (Kn), were superior for somatic embryogenesis. Callus developed on NAA and Kn-supplemented medium favored induction and maturation of embryos earlier compared to that on 2,4-D and Kn. Embryogenic callus transferred from NAA and Kn-supplemented medium to suspension cultures of half-strength MS medium with NAA (2.69 μM) and Kn (1.16 μM) developed a mean of 204.3 somatic embryos per 100 mg of callus. Embryogenic callus transferred from 2,4-D and Kn subsequently to suspension cultures of half-strength MS medium with 2,4-D (0.45 μM) and Kn (1.16 μM) developed a mean of 303.1 embryos per 100 mg of callus. Eighty-eight percent of the embryos underwent maturation and conversion to plantlets upon transfer to half-strength MS semisolid medium having 0.054 μM NAA with either 0.044 μM BA or 0.046 μM Kn. Embryo-derived plantlets established in field conditions displayed morphological characters identical to those of the parent plant.

Journal ArticleDOI
TL;DR: Using cotyledon explants excised from seedlings germinated in vitro, an efficient plant regeneration system via organogenesis was established for bottle gourd (Lagenaria siceraria Standl.)
Abstract: Using cotyledon explants excised from seedlings germinated in vitro, an efficient plant regeneration system via organogenesis was established for bottle gourd (Lagenaria siceraria Standl.). Maximum shoot regeneration was obtained when the proximal parts of cotyledons from 4-day-old seedlings were cultured on MS medium with 3 mg/l BA and 0.5 mg/l AgNO3 under a 16-h photoperiod. After 3–4 weeks of culture, 21.9–80.7% of explants from the five cultivars regenerated shoots. Adventitious shoots were successfully rooted on a half-strength MS medium with 0.1 mg/l IAA for 2–3 weeks. Flow cytometric analysis revealed that most of the regenerated plants derived from culture on medium with AgNO3 were diploid.

Journal ArticleDOI
TL;DR: Observations suggest that AtMRP5 is a putative sulfonylurea receptor that is involved in K+ homeostasis and, thus, also participates in the NaCl stress response.
Abstract: Recently, a new member of the ABC transporter superfamily of Arabidopsis, AtMRP5, was identified and characterized. In the present work, we found that AtMRP5 can bind specifically to sulfonurea when it is expressed in HEK293 cells. We also present evidence for a new role of AtMRP5 in the salt stress response of Arabidopsis. We used reverse genetics to identify an Arabidopsis mutant (atmrp5-2) in which the AtMRP5 gene was disrupted by transferred DNA insertion. In root-bending assays using Murashige and Skoog medium supplemented with 100 mm NaCl, root growth of atmrp5-2 was substantially inhibited in contrast to the almost normal growth of wild-type seedlings. This hypersensitive response of the atmrp5-2 mutant was not observed during mannitol treatment. The root growth of the wild-type plant grown in Murashige and Skoog medium supplemented with the MRP inhibitor glibenclamide and NaCl was inhibited to a very similar extent as the root growth of atmrp5-2 grown in NaCl alone. The Na+-dependent reduction of root growth of the wild-type plant in the presence of glibenclamide was partially restored by diazoxide, a known K+ channel opener that reverses the inhibitory effects of sulfonylureas in animal cells. Moreover, the atmrp5-2 mutant was defective in 86Rb+ uptake. When seedlings were treated with 100 mm NaCl, atmrp5-2 seedlings accumulated less K+ and more Na+ than those of the wild type. These observations suggest that AtMRP5 is a putative sulfonylurea receptor that is involved in K+ homeostasis and, thus, also participates in the NaCl stress response.

Journal ArticleDOI
TL;DR: Growing PEG concentration was associated with a progressive reduction in water content and increased content of endogenous free proline, and both genotypes tested followed this general trend, however, cv.
Abstract: This study was conducted to examine the response of date palm (Phoenix dactylifera L., cvs. Barhee and Hillali) calli to water stress. Callus derived from shoot tip explants was inoculated in liquid Murashige and Skoog medium containing 10 mg dm−3α-naphthaleneacetic acid, 1.5 mg dm−3 2-isopentenyladenine, and 0 to 30 % (m/v) polyethylene glycol (PEG 8000) to examine the effect of water stress. After 2 weeks, callus growth, water content, and proline accumulation were assessed. Increasing water stress caused a progressive reduction in growth as expressed in callus fresh mass, relative growth rate, and index of tolerance. Both genotypes tested followed this general trend, however, cv. Barhee was more tolerant to drought stress than cv. Hillali. Increasing PEG concentration was also associated with a progressive reduction in water content and increased content of endogenous free proline.

Journal ArticleDOI
TL;DR: A new micropropagation system for Tylophora indica, an important medicinal plant in India, using root explants as starting material was developed, with MS medium supplemented with 10.72 μM BA being the most effective in inducing FEC and somatic embryogenesis.
Abstract: The purpose of this study was to develop a new micropropagation system for Tylophora indica, an important medicinal plant in India, using root explants as starting material. Root explants cultured on MS medium supplemented with 6-benzyladenine (BA) or 2-isopentyladenine (2iP) produced organogenic nodular meristemoids (NMs) within 4 weeks. NMs induced from the cut ends of root segments showed two types of organogenic response—direct shoot bud formation and somatic embryogenesis—when maintained on induction medium. In 42% of the explants, NMs developed shoot buds directly in the presence of 10.72–26.80 μM BA. On average, 18.5±0.7 shoots per gram of NM tissue were obtained after each 4-week subculture. Elongation of microshoots and root initiation were correlated with the auxin used, with the optimal response occurring in the presence of 28.54 μM indole-3-acetic acid. In 39% of the explants, NMs dedifferentiated into friable embryogenic callus (FEC) in the presence of BA or 2iP after 12 weeks of culture. Of the different treatments, MS medium supplemented with 10.72 μM BA was the most effective in inducing FEC and somatic embryogenesis: at this concentration 64% of the cultured NMs developed FEC and, on the same medium, 89% of the FEC produced globular somatic embryos (SEs). FEC biomass increased nearly five-fold with every 4-week subculture, and about 30 SEs were recovered per gram of FEC during this period. The best conversion of mature SEs to complete plantlets was obtained on basal MS medium–42%. Plantlets derived via somatic embryogenesis and shoot organogenesis were successfully hardened (88–96%) and transferred to the field.

Journal ArticleDOI
TL;DR: The increase in the levels of antioxidant enzyme activities, for example superoxide dismutase and peroxidase, indicated the presence of excess oxygen uptake and the stressed condition of the plant tissues that arose from haemoglobin supplementation, appeared to accelerate somatic embryogenesis in cotton.
Abstract: Somatic embryogenesis in cotton (Gossypium hirsutum L.) is accelerated when the plant regeneration medium is supplemented with haemoglobin (erythrogen). In cotton SVPR 2 lines, a higher frequency of embryoid formation was observed when the medium contained 400 mg/l haemoglobin. Fresh weight of the callus, rate of embryoid induction, number of embryoids formed and the percentage of plant regeneration from somatic embryos were increased. Among the two different cultivars tested, MCU 11 showed no response to the presence of haemoglobin when compared to SVPR 2, and embryogenic callus formation was completely absent in the former. Medium containing MS salts, 100 mg/l myo-inositol , 0.3 mg/l thiamine-HCL, 0.3 mg/l Picloram (PIC), 0.1 mg/l kinetin and 400 mg/l haemoglobin effected a better response with respect to embryogenic callus induction. After 8 weeks of culture, a high frequency of embryoid induction was observed on medium containing MS basal salts, 100 mg/l myo-inositol, 0.3 mg/l PIC , 0.1 mg/l isopentenyl adenine, 1.0 g/l NH4NO3 and 400 mg/l haemoglobin. Plant regeneration was observed in 75.8% of the mature somatic embryos, and whole plant regeneration was achieved within 6–7 months of culture. The regenerated plantlets were fertile and similar to in vivo-grown, seed-derived plants except that they were phenotypically smaller. A positive influence of haemoglobin was observed at concentrations up to 400 mg/l at all stages of somatic embryogenesis. The increase in the levels of antioxidant enzyme activities, for example superoxide dismutase and peroxidase, indicated the presence of excess oxygen uptake and the stressed condition of the plant tissues that arose from haemoglobin supplementation. This increased oxygen uptake and haemoglobin-mediated stress appeared to accelerate somatic embryogenesis in cotton.

Journal ArticleDOI
TL;DR: The results suggest that sucrose is a better carbon source than glucose for organogenesis of ‘Wegierka Zwykła’, even though at lower concentrations the efficiency of the sugars was comparable, and at 2–5% concentrations glucose uptake was higher than sucrose uptake.
Abstract: Leaf explants of plum ‘Wegierka Zwykla’ were cultured on modified MS medium supplemented with 7.5 µM TDZ (thidiazuron) and 0.9 µM 2,4-D (2,4-dichlorophenoxyacetic acid), with the addition of either sucrose or glucose at a concentration of 2%, 3%, 4%, 5% or 6% (w/v). Regeneration was carried out in two steps: the first in darkness, the second in photoperiod conditions after subculture onto fresh medium. The study assessed the influence of the kind of sugar used and its concentration in the medium on organogenesis efficiency, and on sugar uptake from the medium. The results suggest that sucrose is a better carbon source than glucose for organogenesis of ‘Wegierka Zwykla’, even though at lower concentrations the efficiency of the sugars was comparable. With increasing concentration of sugars, the efficiency of organogenesis decreased, a relation more evident for media with glucose. In the first (dark) step of regeneration, the explants utilised less carbohydrates from the media than in the second step, when the main increase of explant weight took place. In the second phase, at 2–5% concentrations glucose uptake was higher than sucrose uptake.

Journal ArticleDOI
TL;DR: The rooting and in vitro growth of Dendrobium nobile Lindl (Orchidaceae) were studied using different sucrose concentrations in a modified MS medium containing half the regular concentration of macronutrients at pH 5.8.
Abstract: Sucrose is a very important component in in vitro culture media, serving as a source of carbon and energy In this paper, the rooting and in vitro growth of Dendrobium nobile Lindl (Orchidaceae) were studied using different sucrose concentrations (0 g L-1; 5 g L-1; 10 g L-1; 20 g L-1; 30 g L-1 and 60 g L-1), in a modified MS medium containing half the regular concentration of macronutrients at pH 58 Greater increases in plant height (421±06 cm) and high seedling multiplication (1:4) were observed in the 60 g L-1 sucrose treatment, even without the addition of plant hormones Sucrose concentration in the culture medium did not influence in vitro plant rooting

Journal ArticleDOI
TL;DR: The effect of vesicular arbuscular mycorrhizae (VAM) association in averting the transplantation shock was tested and proved to be highly beneficial, giving a 100% survival rate after 60 d of transplantation.
Abstract: An efficient regeneration system was developed for Kigelia pinnata L., a multipurpose tree belonging to the family Bignoniaceae. The nodal segments were cultured in vitro, and the optimum concentrations of plant growth regulators for callus induction were determined. The friable organogenic calli were derived from the basal cut end of the nodal segments. The highest yield of morphogenic callus (100%) was observed when nodal segments were cul- tured on Murashige and Skoog (MS) medium supplemented with 3 µM 2,4 dichlorophenoxyacetic acid (2, 4-D). The morphogenic callus maintained high regeneration during the first four subcultures in the callus induction medium. The maximum shoots (28/culture) were regenerated at the highest frequency of 100% when 3 µM thidiazuron (N-phenyl N' 1,2,3-thidiazol-5-yl urea) (TDZ) and 0.5 µM naphthaleneacetic acid (NAA) were added to MS medium. The emergence of multiple shoots from the calli was histologically documented. The regenerated shoots showed maxi- mum rooting on ½ MS medium containing 4 µM indole-3- butyric acid (IBA). The effect of vesicular arbuscular mycorrhizae (VAM) association in averting the transplantation shock was tested and proved to be highly beneficial, giving a 100% survival rate after 60 d of transplantation. This efficient plant regeneration system provides a founda- tion for generating transgenic plants of this multipurpose tree.

Journal ArticleDOI
TL;DR: Efficient methods for plant regeneration, via both embryogenesis and organogenesis, of Smooth Cayenne pineapple, Ananas comosus (L.) Merr.
Abstract: We have developed efficient methods for plant regeneration, via both embryogenesis and organogenesis, of Smooth Cayenne pineapple, Ananas comosus (L.) Merr. Leaf bases and core (stem) sections of in vitro shoots, produced from culture of crown tip meristem, were used as explants for plant regeneration as follows: (1) Leaf base and core section explants cultured on Murashige and Skoog (MS) medium containing 41 μM 4-amino-3,5,6-trichloropicolinic acid (picloram, P) or thidiazuron (T)/P combinations produced embryogenic tissues. Different types of embryogenic tissues (friable emryogenic tissue, embryogenic cell cluster, and chunky embryogenic tissue) have been developed with varying properties in terms of growth rate and state of development. The embryogenic tissues regenerated shoots upon culture on MS medium containing 13 μM 6-benzylaminopurine (BA) and 1μM α-naphthaleneacetic acid (NAA) followed by culture on MS medium containing 4 μM BA. (2) Crown tip meristems cultured on MS medium containing 13 μM BA followed by leaf explants cultured on MS medium with 27 μM NAA and 1 μM BA produced shoots via direct organogenesis. (3) Explants cultured on MS medium containing 5 μM T and 0.5 μM indole-3-butyric acid (IBA) produced nodular globular structures, which produced shoots upon culture on MS medium containing 1 μM BA and 1 μM gibberellic acid. Shoots obtained from all of the above methods were rooted in half-strength MS medium containing 3 μM NAA and 2.5 μM IBA. Plants were transferred to the greenhouse or shipped to Costa Rica for field trials. Somatic embryo-derived plants exhibited 21 % spininess, and organogenic-derived plants exhibited 5% spininess in the field trials.

Journal ArticleDOI
TL;DR: An efficient plant regeneration and transgenic system using somatic embryogenesis in two melon cultivars, Vedrantais and Earl’s Favourite Fuyu A, is established and the transgenic rate exceeded 2.3%, which is sufficient for practical use.

Journal Article
TL;DR: Thidiazuron (TDZ) is among the most active cytokinin like substances and induces greater in vitro shoot proliferation than many other cytokinins in many plant species and shows a higher shoot formation capacity than stem nodes.
Abstract: Thidiazuron (TDZ) is among the most active cytokinin like substances and induces greater in vitro shoot proliferation than many other cytokinins in many plant species. Leaf, stem, stem node and cotyledonary node explants of 2 extensively cultivated Turkish lentil cultivars, Ali Dayi and Kayi 91, were cultured on Murashige and Skoog (MS) media supplemented with various concentrations of TDZ. The present study was conducted to develop a rapid and efficient shoot regeneration system suitable for the transformation of lentil (Lens culinaris Medik.) using TDZ. Cotyledonary nodes and stem nodes after the initial callus stage regenerated prolific adventitious shoots via organogenesis. Shoot or callus formation was not achieved from leaf or stem explants. DMSO as a solvent for TDZ was necrotic on plant tissues and therefore TDZ was dissolved in 50% ethanol to carry out the studies. Cotyledonary nodes showed a higher shoot formation capacity than stem nodes. MS medium supplemented with 0.25 mg/l TDZ produced the highest frequency of shoot formation from cotyledonary nodes in both genotypes. Regenerated shoots (10-20 mm long) rooted in MS medium containing 0.25 mg/l indole-3-butyric acid (IBA). Rooted plantlets were finally transferred to sand in pots. Abbreviations: TDZ - thidiazuron [1 Phenyl 3-(1,2,3-thiadiazol -5YL) urea], IBA - indole-3-butyric acid, MS - Murashige and Skoog, DMSO - dimethylsulphoxide

Journal ArticleDOI
TL;DR: The higher putrescine levels occurring in WPM callus were associated with a higher arginine and ornithine content, lower γ-aminobutiric acid level, and SERK homolog expression.
Abstract: In the present work, we investigate the association of SERK gene homolog expression, polyamines (PAs) and amino acids related to putrescine synthesis (arginine and ornithine) and polyamines degradation (γ-aminobutiric acid) or S-adenosylmethionine synthesis (methionine), with the embryogenic competence in cell aggregates of Ocotea catharinensis Mez. (Lauraceae). Cell aggregates were cultivated during 7 days in woody plant medium (WPM) supplemented with 20 g l−1 sucrose, 22 g l−1 sorbitol, 400 mg l−1 glutamine and 2 g l−1 phytagel, and in Murashige and Skoog medium (MS) supplemented 20 g l−1 sucrose, 3 g l−1 activated charcoal, 2 g l−1Phytagel with and without 40 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). The cell aggregates cultivated in MS plus 2,4-D and in the WPM medium showed hybridization with a SERK gene homolog both in northern and in situ hybridization experiments. Cell aggregates cultivated in an MS basal medium, without 2,4-D, did not exhibit any hybridization signal to the SERK probe used, thus they were considered potentially non-embryogenic cells. In all three media only free polyamines were detected. The higher putrescine levels occurring in WPM callus were associated with a higher arginine and ornithine content, lower γ-aminobutiric acid level, and SERK homolog expression. Putrescine was also the major polyamine in the MS medium. In the MS plus 2,4-D medium, the levels of putrescine, spermidine and spermine were similar. Spermine exhibited similar and the lowest levels in all media. Spermidine intermediary levels occurred in the WPM and MS media. In cell aggregates methionine level was lowest in the MS plus 2,4-D medium, but similar in the MS and WPM media.

Journal ArticleDOI
TL;DR: Examination of the changes of osmotic substances under salt stress showed that accumulation of proline, glycine betaine, and total soluble sugars increased with increasing salt concentrations, indicating that the response of the callus of P. euphratica to salt stress is similar to that of the whole plant.
Abstract: The present study aimed to evaluate the response to salinity of Populus euphratica, which is more salt-resistant than other poplar cultivars, at the cellular level. To this purpose, callus was induced from shoot segments of P. euphratica on Murashige and Skoog (MS) medium supplemented with 0.5 mg l−1 (2.2 μM) 6-benzyladenine (BA) and 0.5 mg l−1 (2.7 μM) 1-naphthaleneacetic acid (NAA). Callus was transferred to MS medium supplemented with 0.25 mg l−1 (1.1 μM) BA and 0.5 mg l−1 NAA. The relative growth rate of callus reached a maximum in the presence of 50 mmol l−1 NaCl and growth was inhibited with increasing NaCl concentrations. Examination of the changes of osmotic substances under salt stress showed that accumulation of proline, glycine betaine, and total soluble sugars increased with increasing salt concentrations. The results indicate that the response of the callus of P. euphratica to salt stress is similar to that of the whole plant.

Journal ArticleDOI
TL;DR: A procedure for optimum secondary embryogenesis and maturation of Morus alba somatic embryos is described and the embryogenic competence was maintained for more than a year by repeated subculture.

Journal ArticleDOI
TL;DR: In vitro propagation of Oroxylum indicum Vent was carried out using cotyledonary node explants and in vitro developed nodal segments in MS medium with 6-benzyladenine at 4-week intervals resulted in continuous mass multiplication of shoots without any evidence of decline.
Abstract: In vitro propagation of Oroxylum indicum Vent. was carried out using cotyledonary node explants. Among the different types of cytokinins used for culture establishment, 6-benzyladenine exhibited the best response with higher concentrations (8.87 µM or above) for inducing multiple shoots. Inclusion of indole-3-acetic acid (2.85 µM) into 6-benzyladenine-supplemented medium triggered a high frequency of response as well as a proliferation of shoots. The best medium for proliferation was Murashige-Skoog (MS) medium with 6-benzyladenine (8.87 µM) and indole-3-acetic acid (2.85 µM). However, incorporation of gibberellic acid (1.44 µM) was mandatory to enhance shoot elongation. Repeated subculturing of cotyledonary node and in vitro developed nodal segments in MS medium with 6-benzyladenine (4.44 µM) at 4-week intervals resulted in continuous mass multiplication of shoots without any evidence of decline. Root induction was best (91.6%) when MS strength was reduced to one-quarter and combined with α-naphthalene acetic acid (2.69 µM) and indole-3-acetic acid (5.71 µM), with a high survival rate (70–72%) of plantlets hardened in either soil rite or soil : sand : soil rite (1 : 1 : 2).

01 Jul 2004
TL;DR: In this article, the in vitro propagation of Momordica charantia was investigated on modified modified MS medium, where shoot differentiation was obtained on MS medium supplemented with BAP Root, callus were formed on IBA and 2,4-D, respectively.
Abstract: The present investigation outlines the in vitro propagation of Momordica charantia L The explants from in vitro grown seedling were cultured on modified MS medium Shoot differentiation was obtained on MS medium supplemented with BAP Root, callus were formed on IBA and 2,4-D, respectively Shoot as well as root differentiation was obtained on medium containing BAP+IBA/NAA Multiple shoots with roots were formed on MS medium without hormones (MSO) Rooting on shoot grown occurred on medium containing IBA and 40% of the plants survived successfully, when transferred to the field

01 Oct 2004
TL;DR: A high frequency and rapid regeneration protocol was developed from shoot tip and nodal explants of Mentha piperita L. on Murashige and Skoog’s medium supplemented with either 6-benzyl amino purine (BAP) or zeatin (0.25 mg/l), and plantlets showed high survival rate in the soil and in the greenhouse.
Abstract: A high frequency and rapid regeneration protocol was developed from shoot tip and nodal explants of Mentha piperita L. on Murashige and Skoog’s (MS) medium supplemented with either 6-benzyl amino purine (BAP; 1 mg/l)) or zeatin (0.25 mg/l). The highest number of shoots (49.8) was obtained on medium containing BAP. The regenerated dwarf shoots were further elongated on MS medium supplemented with gibberellic acid (GA3; 1 mg/l). In vitro shoots were then excised from shoot clumps and transferred to rooting medium containing naphthalene acetic acid (NAA; 1 mg/l). The rooted plantlets were hardened on MS basal liquid medium and subsequently in polycups containing sterile soil and vermiculite (1:1). Plantlets, thus, developed were successfully established and finally transferred to a greenhouse. The plantlets showed high survival rate (90%) in the soil.