Showing papers on "Murashige and Skoog medium published in 1993"
TL;DR: In this article, conditions for plant regeneration from explants of tomato (Lycopersicon esculentum) cv. were studied for optimizing transformation procedure and the best regeneration rate was obtained from cotyledon explants from 8-10-d-old seedlings on a modified Murashige and Skoog medium (1962) with 0.5 mg dm -3 zeatin and 0. 5 mgdm-3 indolylacetic acid.
Abstract: Conditions for plant regeneration from explants of tomato (Lycopersicon esculentum) cv. UC82B were studied for optimizing transformation procedure. The best regeneration rate was obtained from cotyledon explants from 8-10-d-old seedlings on a modified Murashige and Skoog medium (1962) with 0.5 mg dm -3 zeatin and 0.5 mg dm -3 indolylacetic acid. Tomato cultivars (UC82B, Castone, F1 Ferline, Monalbo) and a Lycopersicon peruvianum «CMV sel. INRA» were studied. The cultivar UC82B and the wild Lycopersicon species showed an efficient shoot regeneration potential. Early events in the transformation of tomato cotyledons were analysed using an Agrobacterium tumefaciens strain carrying a binary vector with an nptII (pnos) gene and a reporter GUS-intron (p35S) chimeric gene
121 citations
TL;DR: High frequency somatic embryogenesis in papaya tissue cultures was achieved by culturing hypocotyl sections from ten-day-old seedlings on half-strength Murashige and Skoog salts (MS) medium containing modified MS vitamins, and embryos developed directly from embryogenic calluses on induction medium, or, more often, they differentiated from calluses subcultured on a medium devoid of growth regulators.
Abstract: High frequency somatic embryogenesis in papaya (Carica papaya L.) tissue cultures was achieved by culturing hypocotyl sections from ten-day-old seedlings on half-strength Murashige and Skoog salts (MS) medium containing modified MS vitamins, 2.3 to 112.5 μM 2,4-dichlorophenoxyacetic acid (2,4-d), 400 mg l-1 glutamine, and 6% sucrose. Four hermaphroditic Hawaiian cultivars produced embryogenic calluses after ten to 14 weeks of culture at 27°C in the dark. Efficiency in embryogenic response of genotypes differed, ‘Kapoho’ > ‘Sunset’ > ‘Sunrise’ > ‘Waimanalo’. The frequency of embryogenesis in induction medium containing 4.5 μM 2,4-d was lowest with 3% sucrose and highest with 7% sucrose. Somatic embryos developed directly from embryogenic calluses on induction medium, or, more often, they differentiated from calluses subcultured on a medium devoid of growth regulators. Between 50 and 500 embryos were produced from each 2-mm hypocotyl section after at least two months on induction medium and two months on maturation medium. Embryos subsequently developed into normal-looking plants on MS medium. Shoot cuttings from germinated embryos and micropropagated plants were rooted with 5.0 μM indole-3-butyric acid (IBA), grown in the greenhouse, and transferred to the field.
107 citations
TL;DR: Methods for improving the efficiency of plant recovery from somatic embryos of cassava (Manihot esculenta Crantz) were investigated by optimizing the maturation regime and incorporating a desiccation stage prior to inducing germination.
Abstract: Methods for improving the efficiency of plant recovery from somatic embryos of cassava (Manihot esculenta Crantz) were investigated by optimizing the maturation regime and incorporating a desiccation stage prior to inducing germination. Somatic embryos were induced from young leaf lobes of in vitro grown shoots of cassava on Murashige and Skoog medium with 2,4-dichlorophenoxy acetic acid. After 15 to 20 days of culture on induction medium, the somatic embryos were transferred to a hormone free medium supplemented with activated charcoal. In another 18 days mature somatic embryos became distinctly bipolar and easily separable as individual units and were cultured on half MS medium for further development. Subsequent desiccation of bipolar somatic embryos resulted in 92% germination and 83% complete plant regeneration. The plants were characterized by synchronized development of shoot and root axes. Of the non-desiccated somatic embryos, only 10% germinated and 2% regenerated plants. Starting from leaf lobes, transplantable plantlets were derived from primary somatic embryos within 70 to 80 days.
106 citations
TL;DR: It is suggested that the protocol developed could be useful for cloning of mature and tested trees of P. cineraria and found essential for root induction from shoots which was 63% within two weeks.
Abstract: Genotype, age of tree, nature of explant and size (length and diameter), season of explant collection, explant position on medium, plant growth regulators and certain additives (ascorbic and citric acids, adenine sulphate, L-arginine, glutamine and ammonium citrate), incubation conditions, and subculturing period greatly influenced the in vitro clonal propagation of P. cineraria. The maximum number of 10–12 shoots were induced from the nodal shoot segment from pruned thorny adult trees on Murashige and Skoog's (MS) medium containing 0.1 mgl-1 indole- 3-acetic acid (IAA)+2.5 mgl-1 benzylaminopurine (BAP)+additives. Higher temperature (31+-2°C) and mixed (fluorescent and incandescent) light of 50 μmol m-2 s-1 photon flux density for 12 h per day photoperiod favoured shoot induction and subsequent growth. Explants from thornless trees produced 6–8 shoots per explant on MS medium containing 0.1 mgl-1 IAA+5.0 mgl-1 BAP + additives. Nodal shoot segments obtained from root and stump sprouts produced multiple shoots. Root segments differentiated into multiple shoots on MS medium containing 0.5 mgl-1 indolebutyric acid (IBA)+2.5 mgl-1 BAP.
105 citations
TL;DR: It is shown that BA was required for adventitious shoot formation but its concentration in the medium was not critical, and biotechnology could be used as a means of introducing resistance to cucurbit potyviruses by inserting segments of viral DNA into existing watermelon germplasm without altering the genetic identity of elite lines
Abstract: Adventitious shoots were obtained from watermelon [ Citrullus lanatus (Thunb.) Matsun. & Nakai] cotyledons incubated on a modified Murashige and Skoog medium containing BA. Initial experiments comparing the effects of BA (0, 5, 10, or 20 μM ) and IA4 (0, 0.5, or 5 μM ) demonstrated that BA was required for adventitious shoot formation but its concentration in the medium was not critical. The addition of IAA to medium with BA increased callus production and inhibited shoot formation. However, the percentage of responding explants in the best treatment was 90% for ‘Minilee’, 64% for S86NE, and 50% for ‘Jubilee II’ when explants were prepared from 5-day-old seedlings. Explants from nongerminated embryos or seedlings germinated for 10, 15, or 20 days produced fewer shoots. The effect of several cytokinins on shoot organogenesis was then examined using the optimized protocol. The percentage of explants with shoots and the number of shoots per explant were about two to four times higher when 5 to 10 μM BA was used compared to the most effective kinetin (20 μM ) or thidiazuron (0.1 μM ) concentration. The percentage of explants with shoots and the number of shoots per explant were greater for diploid (57% and 2.2, respectively) than for triploid (22% and 0.6, respectively) or tetraploid (20% and 0.8, respectively) lines. Chemical names used: N -(phenylmethyl)-1 H -purin-6-amine (BA); 6-furfurylaminopurine (kinetin); N -phenylN' -1,2,3-thiadiazol-5-ylurea (thidiazuron); 1 H -indole3-acetic acid (IAA). Potyviruses cause serious disease problems in commercial cucurbit plantings across the United States (Adlerz et al., 1983) and in other production areas of the world. Resistance to viruses such as zucchini yellow mosaic virus (ZYMV) exists in wild germplasm (Provvidenti, 1991); however, the introduction of this germplasm into commercial watermelon cultivars would result in the loss of favorable characteristics and require years of selective breeding to obtain acceptable virus resistant cultivars. Biotechnology could be used as a means of introducing resistance to cucurbit potyviruses by inserting segments of viral DNA (e.g., sense, antisense, coat protein genes, etc.) into existing watermelon germplasm without altering the genetic identity of elite lines. This method has proven successful for obtaining transgenic plants resistant to alfalfa mosaic virus (Loesch-Fries et al., 1987; Turner et al., 1987; Van Dun et al., 1988), cucumber mosaic virus (Chee and Slightom, 1991; Cuozzo et al., 1988; Quemada et al., 1991), potato virus x and y (Hemenway et al., 1988; Hoekema et al., 1989; Kaniewski et al., 1990; for publication 30 Mar. 1992. Accepted for publication 15 Aug. 1992. lorida Agr. Expt. Sta. J. Ser. no. R-02349. This work was supported nt from the State of Florida High Technology and Industry Council Research Grants Program. Seeds were provided by Gary W. Elmstrom, lorida Research and Education Center, Institute of Food and Agriculences, Univ. of Florida, Leesburg, Fla. Use of trade names does not dorsement of the products named nor critism of similar products not d. The cost of publishing this paper was defrayed in part by the paypage charges. Under postal regulations, this paper therefore must be arked advertisement solely to indicate this fact. orate Research Assistant. To whom reprint requests should be ad-
83 citations
TL;DR: A high level of adventitious shoot regeneration was obtained from proliferating shoots in vitro for a range of Prunus spp.
82 citations
TL;DR: Segments of cotyledon and apical tip taken from 1-week-old seedlings of Daucus carota L. were cultured for 3 weeks on hormone-free Murashige and Skoog's medium containing high amounts of sucrose and transferred to the same medium with lower sucrose concentration, resulting in somatic embryos being formed 3 to 6 weeks after the transfer.
Abstract: Segments of cotyledon and apical tip taken from 1-week-old seedlings of Daucus carota L. were cultured for 3 weeks on hormone-free Murashige and Skoog's (MS) medium containing high amounts of sucrose (0.3-0.7M) and then transferred to the same medium with lower sucrose concentration (0.09M). Somatic embryos were formed from the segments 3 to 6 weeks after the transfer. On the other hand, hypocotyl segments given the same treatment did not produce somatic embryos. When the primary culture medium contained 0.61M mannitol and 0.09M sucrose, cotyledon segments and apical tip segments also produced somatic embryos when transferred to hormone-free MS medium with 0.09M sucrose. In all cases, somatic embryos were formed directly from the explants without visible callus formation.
81 citations
TL;DR: A complete protocol for micropropagation of 4-yr-old plants of the bambooDendrocalamus longispathus is described and more than 85% of the tissue culture-raised plants were successfully transferred to soil.
Abstract: A complete protocol for micropropagation of 4-yr-old plants of the bambooDendrocalamus longispathus is described. Culture initiation was strongly influenced by the nature of the explant and the season. In vitro multiplication was achieved through forced axillary branching. Single node segments from the young lateral branches produced multiple shoots on agar-solidified Murashige and Skoog (MS) medium supplemented with 12µM benzylaminopurine (BAP) and 3µM kinetin. The shoots have been multiplied for 15 passages in liquid and thereafter for over 5 passages on semisolid MS+15µM BAP+1µM indolebutyric acid (IBA)+10% coconut water at a rate of 3.2- and 2.8-fold, every 4 wk, respectively. The nature of the propagule was a critical factor for shoot multiplication and rooting. Seventy-three percent of the shoots rooted on a modified MS medium (major salts reduced to half strength) containing 1µM indoleacetic acid, 1µM IBA, and 68µM coumarin. Through a simple in vitro hardening step, more than 85% of the tissue culture-raised plants were successfully transferred to soil.
79 citations
TL;DR: Up to 70% of the plantlets that produced roots in sucrose media enriched with indolebutyric acid were successfully acclimatized when they were dipped in a benomyl (0.075% w/v) suspension before being transplanted with care being taken to prevent over-wetting of soil.
Abstract: In vitro proliferation and rooting capacity of ‘San Castrese’ and ‘Portici’ apricots (Prunus armeniaca L.) were tested on modified MS medium enriched with varying growth regulator concentrations and sucrose (58.4 mM) or sorbitol (116.8 mM) as main carbon energy sources. The interaction of proliferation and rooting media was also studied. Proliferation of both cultivars was proportional to benzyladenine (BA) concentration and enhanced with sorbitol media. However, 8.8 μM BA was often associated with hyperhydricity, particularly when shoots were grown on sucrose media. Newly proliferated shoots elongated better on sorbitol media. The positive influence of sorbitol on proliferation and shoot growth was not due to osmotic effects. Moreover, sorbitol showed a positive carryover effect in hastening rooting of ‘Portici’. By contrast, when transferred to sorbitol rooting media, the shoots of both cultivars generally showed low rooting, with short, thick roots. Up to 70% of the plantlets that produced roots in sucrose media enriched with indolebutyric acid were successfully acclimatized when they were dipped in a benomyl (0.075% w/v) suspension before being transplanted with care being taken to prevent over-wetting of soil.
69 citations
TL;DR: Root explants did not express any new morphogenetic response in all hormonal combinations tested and plantlets were established successfully ex vitro under greenhouse mist and grown to maturity.
Abstract: Explants from 13-d old pepper (Capsicum annuum, L cv Early California Wonder) seedlings were cultured in Murashige and Skoog (MS) medium supplemented with different levels of 1-naphthalene acetic acid (NAA) and 6-benzylamino purine (BAP) Multiple shoot-buds proliferated from the cut surfaces of cotyledon, shoot-tip and hypocotyl explants in one month The best NAA to BAP combinations (mg/l: mg/l) for multiple shoot-bud regeneration of the above three explant types were 01 ∶ 50, 00 ∶ 50, and 01 ∶ 100, respectively Root explants did not express any new morphogenetic response in all hormonal combinations tested Regenerated shoot-buds were excised from the explants and cultured in 1/2X or 1X MS medium supplemented with different levels of Indole-3-acetic acid (IAA) or NAA When cultured in full strength MS medium with 05 mg/l IAA or 04 mg/l NAA, 70% of the buds rooted in one month Plantlets were established successfully ex vitro under greenhouse mist and grown to maturity
69 citations
TL;DR: Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl−1 indole-3-acetic acid or 0.1 mgl+1 indoles3-butyric acid added to MS or NN media.
Abstract: Somatic embryogenesis from leaf- and petiole-derived calli of Vitis rupestris was obtained with an efficiency of 3.2% and 4.2% of plated explants, respectively on two combinations of 6-benzyladenine and 2,4-dichlorophenoxyacetic acid (1/0.1 and 1/1 mgl−1) added to MS medium. Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl−1 indole-3-acetic acid or 0.1 mgl−1 indole-3-butyric acid added to MS or NN media. Within a 4-month culture, embryo germination occurred at a frequency of 13% of explanted embryos when chilling at 4°C was provided for two weeks and a combination of 6-benzyladenine (1 mgl−1) with indole-3-butyric acid (0.1 mgl−1) was added to NN medium supplemented with casein hydrolysate (250 mgl−1). A higher frequency (51%) was obtained in a longer culture time (9 months) when only indole-3-butyric acid was present in the medium and in absence of chilling.
TL;DR: A procedure for plant regeneration from immature seed-derived calli of rugosa rose (Rosa rugosa Thunb.) via somatic embryogenesis is described, and induced calli grew into plantlets which showed sustained growth on the MS medium containing only 0.1 M sorbitol as the sole carbon source.
TL;DR: Elite trees of Commiphora wightii (Arnott) Bhandari were selected from the wild on the basis of their content of guggul, an oleoresin to micropropagated through forced axillary branching on Murashige and Skoog's medium supplemented with benzyladenine and kinetin to study the shoot proliferating potential of the initial explants in vitro.
Abstract: Elite trees of Commiphora wightii (Arnott) Bhandari were selected from the wild on the basis of their content of guggul, an oleoresin. The selected tree was micropropagated through forced axillary branching on Murashige and Skoog's (MS) medium supplemented with benzyladenine (BA) and kinetin. Highest frequency of shoot formation was achieved on MS medium supplemented with 17.8 μM BA, 18.6 μM kinetin, 100 mg l-1 glutamine, 10 mg l-1 thiamine HCL and 0.3% activated charcoal. Seasonal changes affected the shoot proliferating potential of the initial explants in vitro. Transfer of shoots to a medium containing a lower concentration of BA (1.8 μM) and kinetin (1.9 μM) before rooting markedly stimulated shoot elongation. Shoots could be rooted by treating them with both indoleacetic acid and indolebutryic acid for 24 h in darkness and transferring them to a low-salt basal medium with activated charcoal. After rooting, transfer to a half-strength White's (modified) medium was necessary for further development of the plantlet. Regenerated plantlets were successfully established in soil.
TL;DR: Histological studies revealed various stages of somatic embryogenesis characteristic of the monocot system of ginger and efficient plant regeneration was achieved when embryogenic cultures were transferred to Murashige & Skoog medium containing 8.9 μM benzyladenine.
Abstract: Embryogenic callus cultures of ginger were induced from young leaf segments taken from in vitro shoot cultures. Among the four auxins tested in Murashige & Skoog medium, dicamba at 2.7 μM was most effective in inducing and maintaining embryogenic cultures. Efficient plant regeneration was achieved when embryogenic cultures were transferred to Murashige & Skoog medium containing 8.9 μM benzyladenine. Histological studies revealed various stages of somatic embryogenesis characteristic of the monocot system. The in vitro-raised plants have been established in soil.
TL;DR: The R 2 line from a selected variant plant showed stable expression of increased plant height and earlier maturity and the organogenic nature of regeneration from the callus tissues was confirmed.
Abstract: Plant regeneration has been achieved in two common bean lines from pedicel-derived callus that was separated from the explant and maintained through successive subcultures. Callus was induced either on B 5 or MS medium containing 2% sucrose and enriched with 0.5 or 1.0 mg thidiaznron/liter alone or plus various concentrations of indoleacetic acid. The presence of 0.07 or 0.14 g ascorbic acid/liter in the maintenance media prolonged the maintenance time. Up to 40 shoot primordia were observed in 4-week-old cultures obtained from 40 to 50 mg callus tissues on shoot-inducti on medium containing 1-mg benzyladenine/liter. These shoot primordia developed two to five excisable shoots (>0.5 cm) on medium with 0.1-mg BA/liter. A histological study confirmed the organogenic nature of regeneration from the callus tissues. The R 2 line from a selected variant plant showed stable expression of increased plant height and earlier maturity. Chemical names used: ascorbic acid, N- (phenylmethyl)-1H-pnrin-6-amine (benzyl- adenine, BA), 1H-indole-3-acetic acid (IAA), N- phenyl-N'-1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ).
TL;DR: Plants resistant to 20 μg/ml hygromycin were regenerated at a frequency of 11–36% from hypocotyl explants and were shown to be transformed by Southern blotting, GUS staining and progeny analysis.
Abstract: Twelve cultivars of Brassica juncea grown in different agroclimatic regions of the world were tested for their ability to regenerate in vitro from hypocotyl explants and, accordingly, were divided into three groups. One group of cultivars regenerated on MS medium supplemented with 2,4-D, BAP and with NAA, BAP combinations; another group regenerated only on MS with 2,4-D, BAP; and the third group showed very low regeneration on both of these combinations. Inclusion of silver nitrate in the medium was essential for high frequency of regeneration. In general, Indian cultivars were more responsive than the cultivars of CIS and Australian origin. Using the media optimal for regeneration and an Agrobacterium-based binary vector carrying hpt and gus-intron genes, conditions for genetic transformation of B. juncea hypocotyl explants were optimized. Transformation frequencies, identified by GUS staining at the initial stages of growth, were lower on MS medium with 2,4-D, BAP than on MS with NAA, BAP. Plants resistant to 20 μg/ml hygromycin were regenerated at a frequency of 11-36% from hypocotyl explants and were shown to be transformed by Southern blotting, GUS staining and progeny analysis.
TL;DR: Cell suspensions of Musa diploid species were obtained by culturing embryogenic callus initiated from immature zygotic embryos in liquid medium and plant regeneration was then achieved through somatic embryogenesis.
Abstract: Cell suspensions ofMusa acuminata sspburmannicoides andMusa acuminata sspmalaccensis were obtained by culturing embryogenic callus initiated from immature zygotic embryos in liquid medium. Plant regeneration was then achieved through somatic embryogenesis. Germination of these embryos occurred in a modified MS medium containing auxin and cytokinin. Plant recovery frequencies were 20 to 36%. This method may allow a better utilization of biotechnologies in genetic improvement of theMusa diploid species, essential for banana and plantain breeding.
TL;DR: Callus originated in microsporangial wall layers and connective tissues of anthers containing uninucleate microspores on Nitsch's or Murashige and Skoog's medium supplemented with growth regulators to induce multiple shoots induced in this anther-derived callus.
Abstract: Callus originated in microsporangial wall layers and connective tissues of anthers containing uninucleate microspores on Nitsch's or Murashige and Skoog's medium supplemented with growth regulators. A higher percentage of cultures (43) produced callus on Nitsch's medium containing 10 μM indole-3-acetic acid + 1 μM 6-benzyladenine. After 13–15 weeks, green nodular structures and prominent roots developed in 25% of the cultures on Murashige and Skoog's medium + 10 μM α-naphthaleneacetic acid + 1 μM kinetin. Multiple shoots were induced in this anther-derived callus when subcultured on Murashige and Skoog's medium augmented with 4.44 μM 6-benzyladenine + 0.53 μM α-naphthaleneacetic acid along with 18.75 μM polyvinylpyrrolidone. The excised shoots formed roots after subculturing on Murashige and Skoog's medium + 4.90 μM indole-3-butyric acid + 18.75 μM polyvinylpyrrolidone, thus developing complete plantlets. Examination of callusing anthers also revealed two- to multi-celled pollen masses with intact exine.
TL;DR: Somatic embryogenesis was greatest when cotyledon expiants were established from 18-day-old immature embryos, and the best PGR combination was 10 μM 2,4-D and 0.5 μM TDZ.
Abstract: Cotyledon expiants from immature embryos of five watermelon [Citrullus lanatus (Thunb.)Matsum. & Nakai] genotypes were incubated in the dark for three weeks on a modified MS medium containing B5 vitamins, 2,4-D (10, 20 or 40μM), 0.5 μM of either BA or TDZ, and 7 g·1-1 TC agar. Somatic embryos, some with well developed cotyledons, were observed on cotyledon expiants three to four weeks after transfer to MS medium without PGRs and 16h photoperiod. The best PGR combination for somatic embryogenesis was 10 μM 2,4-D and 0.5 μM TDZ Somatic embryogenesis was greatest (30%) when cotyledon expiants were established from 18-day-old immature embryos. Somatic embryos were germinated on MS medium without PGRs. Plants were transferred to Magenta boxes containing ProMix for three weeks before being transplanted to the field where they formed fertile male and female flowers that produced normal fruit.
TL;DR: No phenotypic variations were detected among the regenerants during both their vegetative and reproductive phases, and all the regenerateants were typically diploid, as were the various tissues of the mother plants.
Abstract: Efficient bud regeneration was obtained from a clone ofGerbera hybrida Bol. L. leaf explants cultured on modified Murashige and Skoog medium supplemented with 10 µM benzyladenine and 2.5 µM naphthalenacetic acid. The morphogenic potential varied with the developmental stage of the leaves. Up to 90% of excised developing leaves formed 3 to 5 shoots per explant. Bud regeneration was not obtained on the same medium with fully expanded leaves. Addition of 0.05 µM or 0.5 µM thidiazuron to the above medium significantly promoted regeneration from mature leaves but was ineffective for similar explants of a recalcitrant clone. The two wild speciesG. viridifolia Schultz Bip. andG. piloselloides L. Cass. also displayed specific multiplication habits and regeneration abilities. Bud regeneration occurred from callus. Chromosome counts and DNA flow cytometry indicated that all the regenerants were typically diploid, as were the various tissues of the mother plants. Afterin vitro rooting and acclimatization, no phenotypic variations were detected among the regenerants during both their vegetative and reproductive phases.
TL;DR: Organogenesis of shoots of bell pepper (Capsicum annuum L.) was achieved in fourteen cultivars on Murashige and Skoog's medium (MS medium) supplemented only with 0.4% Gellan gum.
Abstract: Organogenesis of shoots of bell pepper (Capsicum annuum L.) was achieved in fourteen cultivars on Murashige and Skoog's medium (MS medium) supplemented only with 0.4% (w/v) Gellan gum (pH 5.8). Mature seeds of cv. Shinsakigake-2 were sown on filter paper that had been wetted with sterilized water and precultured for zero to five days in under 16 hr of light per day at 25 °C. Explants, consisting of the proximal part of the hypocotyl and the radicle, were excised from the seeds and formed adventitious buds around the cut surfaces of elongated hypocotyls after four weeks of culture. When explants were subcultured on MS medium, 57% of the explants that had produced adventitious buds extended shoots after an additional three weeks of culture. Shoots were rooted on MS medium after two further weeks of culture. Chromosome numbers of all 30 regenerated plants that weexamined were normal (2n=24). The morphology of the mature plants was also normal and they set normally shaped fruits with mature seeds. Regenerated whole plants were also obtained in the case of 13 other cultivars by applying this simple procedure.
TL;DR: Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall using a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid or kinetin.
Abstract: Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall. Callus was induced on a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid (2,4-d 1 mg l-1) and kinetin (KN 0.5 mg l-1) with coconut water (CW 10% v/v) or naphthalene acetic acid (NAA 5 mg l-1) and benzylaminopurine (BAP 1 mg l-1). Somatic embryos appeared after 2–3 months or 2 subculture passages when 2,4-d or NAA induced source of the callus was transferred to a MS medium containing BAP (1 mg l-1) and NAA (0.1 mg l-1). For successful plantlet formation, the somatic embryos were transferred to a medium containing 1/4 strength MS nutrient with indole-3-butyric acid (IBA 1 mg l-1). Alternatively, the somatic embryos were dipped in a concentrated solution of IBA for 5 min and placed on a hormone free medium. Complete plantlets were formed after 4 weeks and were transferred successfully to soil.
TL;DR: Five genotypes of chickpea (Cicer arietinum L.) PG1, PG5, PG12, N59 and C235 were evaluated for induction of somatic embryogenesis and somatic embryos derived from cotyledonary segments of genotype PG12 formed callus.
Abstract: Five genotypes of chickpea (Cicer arietinum L.) PG1, PG5, PG12, N59 and C235 were evaluated for induction of somatic embryogenesis. Somatic embryogenesis was induced from immature cotyledons of genotypes PG12 and C235 and immature embryo axes of genotypes PG5, PG12 and C235. Genotypes N59 and PG1 showed no response. The maximum frequency of globular embryo formation occurred in cotyledonary segments on MS medium with 3.0 mg/l 2,4,5-trichlorophenoxyacetic acid (2,4,5-T). Further embryo development was achieved only in somatic embryos derived from cotyledonary segments of genotype PG12. Globular-stage embryos derived from immature embryo axes of PG5, C235, PG12, and cotyledonary segments of C235 dedifferentiated and formed callus. The cotyledonary stage embryos of genotype PG12 germinated on half-strength MS medium supplemented with 1 mg/l zeatin. The regenerated plants were transferred to soil and grown to maturity.
TL;DR: Change in dw/fw ratio, as well as the esterified fatty acid and sucrose concentrations correlated positively with the development of tolerance to cryopreservation, and the embryogenic calli produced from these embryos exhibited normal plant regeneration on auxin-free media.
Abstract: Spring wheat (Triticum aestivum L.) zygotic embryos were successfully cryopreserved, without the addition of exogenous cryoprotectants, using only an abscisic acid (ABA) pretreatment. Optimum survival was obtained when embryos were cultured in vitro for 10 days on semisolid Murashige and Skoog (MS) nutrient medium supplemented with 0.5 mg/L (±) ABA prior to cryopreservation. The embryos resumed growth within three days when returned to MS medium devoid of ABA but containing 2mg/L 2,4-dichlorophenoxyacetic acid. The embryogenic calli produced from these embryos exhibited normal plant regeneration on auxin-free media. Changes in dw/fw ratio, as well as the esterified fatty acid and sucrose concentrations correlated positively with the development of tolerance to cryopreservation.
TL;DR: Tissue culture techniques were applied to study the regeneration and growth of bulblets from bulb scale segments of Crinum macowanii Bak.
Abstract: Tissue culture techniques were applied to study the regeneration and growth of bulblets from bulb scale segments ofCrinum macowanii Bak. (bush- or march lily)in vitro. Shoots were induced on twin scales taken from the basal plate region of flowering-size bulbs on Murashige and Skoog (MS)-medium containing 0–20 mg l−1 NAA and BA and a modified MS medium (MMS medium) containing 1.25 mg l−1 ancymidol (A-RestTM), 0.1 mg l−1 NAA and 0.1 mg l−1 kinetin (ANK). Large bulblets could only be initiated on the latter. Subsequently the bulblets of 5 mm or more in diameter were trimmed and split in half, and secondary plantlets were regenerated on MMS-medium containing ANK or MS-medium without any growth regulators which in turn grew into bulblets suitable for splitting within 12–16 weeks. A total of 700–1000 bulblets could be obtained from each initial bulb within 12 months. Anatomical studies showed that the shoots were initiated from the epidermis and hypodermis on the abaxial surface of the meristematic tissue of the basal plate of the bulb scale. This technique is useful for the multiplication and preservation of a genotype, since plantlets regenerated in this manner should be genetically uniform.
TL;DR: Petioles, leaf discs and midribs of several olive cultivars, collected from potted greenhouse plants, field-grown and in vitro shoots, were used to test their morphogenic capacity and regeneration potential was higher in petioles collected from apical nodes than from basal ones.
Abstract: Petioles, leaf discs and midribs of several olive (Olea europaea L.) cultivars, collected from potted greenhouse plants, field-grown and in vitro shoots, were used to test their morphogenic capacity. Adventitious shoots were induced only in petioles from in vitro-grown shoots of cultivars Moraiolo, Dolce Agogia and Halkidikis, grown on Olive Medium (OM) plus 18 μM zeatin within 4 to 5 weeks. Regeneration was achieved, both on Murashige and Skoog (MS) and on modified OM, only in the dark. The highest regeneration was achieved directly from the proximal part of the petioles after 2 to 3 weeks in media containing 5 to 40 μM thidiazuron, or with both 10 μM 2-isopentenyladenine +2.2 μM 6-benzyladenine with or without low auxin concentration (not more than 2.5 μM). A few adventitious shoots were also regenerated from callus when it was shifted from auxin and cytokinin media to cytokinin only medium. The regeneration potential was higher in petioles collected from apical nodes than from basal ones. The adventitious shoots were transferred to solid half-strength MS medium supplemented with 4.5 μM zeatin for further development. Several regenerated shoots were rooted and the plantlets hardened in the greenhouse. No apparent differences regarding morphological aspects were observed among the regenerated plantlets or with those obtained by stimulation of axillary buds.
TL;DR: The ability to regenerate plants from protoplasts of passionfruit is discussed in relation to the application of somatic cell techniques for the genetic improvement of this economically important tropical woody plant.
Abstract: Enzymatic digestion of newly expanded leaves of glasshouse-grown seedlings of passionfruit released protoplasts which exhibited highest division frequency (38.6%) when plated at a density of 1.5×105 ppts ml−1 in agarose-solidified droplets of KM8P medium containing the antibiotic cefotaxime (250 μg ml−1). Cefotaxime was essential for sustained cell division. Protoplast-derived calli were cultured on agarsolidified MS medium with 5.0 mg H NAA, 0.25 mg l−1 BAP and additional vitamins. These calli regenerated shoots on transfer to MS medium with 1.0 mg l−1 BAP. Regenerated shoots were rooted in half-strength MS medium with 3.0 mg l−1 IBA and 0.5 mg l−1 NAA (7 d), followed by sub-culture to MS medium lacking growth regulators. The ability to regenerate plants from protoplasts of passionfruit is discussed in relation to the application of somatic cell techniques for the genetic improvement of this economically important tropical woody plant.
TL;DR: Embryogenic callus and adventitious buds were initiated from only two anthers of the hybrid H. annuus ×H.
Abstract: A method of plant regeneration from cotyledons ofHelianthus tuberosus, Helianthus annuus ×Helianthus tuberosus and for the backcross of the interspecific hybrids onH. annuus was developed. Induction of somatic embryogenesis and plantlet regeneration from anther culture of the interspecific hybridsH. annuus ×H. tuberosus is reported. Cotyledons were cultured on Murashige and Skoog basal medium (MS) supplemented with indole-3-acetic acid (IAA) and 6-furfurylaminopurine (kinetin) or N6-benzylaminopurine (BAP). Shoot regeneration occurred on most of the media tested, but the best results were obtained on media with a high concentration of cytokinins (BAP or kinetin: 4 mg l−1) and lower concentration of auxin (IAA: 0.5−1 mg l−1). Embryogenic callus and adventitious buds were initiated from only two anthers of the hybridH. annuus ×H. tuberosus cultured on the MS medium containing BAP (0.2 mg l−1) and 1-naphtalenacetic acid (NAA: 0.1 mg l−1). Prolonged culture of these embryogenic calli and buds on the original medium with successive subculture on MS basal medium without growth regulators resulted in embryo formation and shoot differentiation. The plantlets, after rooting, were established in soil.
TL;DR: A useful system for direct shoot regeneration from primary seedling explants and immature embryos of the American safflower cultivar is developed and numbers of regenerated shoots were comparable on media containing BAP or TDZ, althoughTDZ medium was superior in reducing shoot hyperhydricity and permitting multiple harvests of regenerateated shoots from primary explants.
TL;DR: An efficient clonal multiplication system was developed for in vitro propagation of crocin — producing Gardenia jasminoides Ellis plants and it was found that this method will be useful for rapid multiplication of Gardenia for commercial exploitation.
Abstract: An efficient clonal multiplication system was developed for in vitro propagation of crocin — producing Gardenia jasminoides Ellis plants. Murashige and Skoog's (MS) medium containing 6-benzylaminopurine (BAP 1 mg l−1) and indole-3-butyric acid (IBA 1 mg l−1) resulted in multiple shoot initiation at the rate of 21 shoots per explant in 60 d of culture. Transfer of the microshoots into liquid MS medium supplemented with BAP (5 mg l−1) with two subcultures of 15 d duration in the same medium resulted in 400 ± 25 shoots per explant. Efficient rooting was achieved in MS medium supplemented with α-naphthaleneacetic acid (5 mg l−1). The in vitro raised plants were hardened in a greenhouse and transplanted to the field successfully. The method described will be useful for rapid multiplication of Gardenia for commercial exploitation.