Showing papers on "Murashige and Skoog medium published in 1981"
TL;DR: In vitro regeneration of buds, shoots, and roots from immature leaves of 3- to 5-day-old peanut (Arachis hypogaea L. cv. Colorado Manfredi) seedlings was studied under defined nutritional, hormonal, and environmental conditions.
Abstract: The in vitro regeneration of buds, shoots, and roots from immature leaves of 3- to 5-day-old peanut (Arachis hypogaea L. cv. Colorado Manfredi) seedlings was studied under defined nutritional, hormonal, and environmental conditions. The first two leaves (2–5 mm in length) removed from aseptically germinated seeds were cultured on Murashige and Skoog medium containing vitamins as in B5 medium and 0.8% agar, supplemented with 12 combinations of naphthaleneacetic acid (NAA) (0.01 to 4 mg/L) and benzyladenine (BA) (1 and 3 mg/L). Bud regeneration occurred in all hormone combinations, but the maximum number of buds was regenerated at a concentration of 1 mg/L each of NAA and BA. Although bud regeneration was maximum with 2- to 5-mm-long leaflets, some success was also obtained with leaflets 8–13 mm long. However, no buds were regenerated when fully expanded leaflets were cultured.Development of buds into shoots was readily achieved by transferring regenerated buds into fresh medium containing 0.01 mg/L NAA and 1 mg/L BA. A few roots were induced to grow when callus with buds was also transferred to medium devoid of hormones. So far, bud regeneration from immature leaves has been induced in vitro in 5 of the 10 cultivars tested
90 citations
TL;DR: Vinca rosea protoplasts and Agrobacterium tumefaciens spheroplasts harboring octopine-type Ti plasmid were mixed and treated with polyethylene glycol or polyvinyl alcohol, which facilitated the introduction of sphero Plasmid into plant protoplast.
Abstract: Vinca rosea protoplasts and Agrobacterium tumefaciens spheroplasts harboring octopine-type Ti plasmid were mixed and treated with polyethylene glycol or polyvinyl alcohol, which facilitated the introduction of spheroplasts into plant protoplasts. After the protoplasts had been kept at 40° C for 4 days, bacteria were found to be completely eliminated from the medium. Among treated protoplasts 1–2 per 1,000 formed colonies on the Murashige and Skoog medium (1962) lacking hormones. When the colonies were isolated and subcultured, they could be maintained as clones. Octopine, an amino acid specific to crown gall, was detected in half of these clones. The phenotypic features of these putative transformants were compared but did not show any coincidental tendencies in relation to color, hardness, form, growth rate, or octopine production. The significance of this system in transformation of higher plant cells is discussed.
67 citations
TL;DR: In vitro regeneration of plants from immature leaflets of 3 day-old pea seedlings was studied under defined nutritional, hormonal and environmental conditions and shoot regeneration from the primary callus occurred within 45 to 90 days of culture in most of the hormone combinations.
Abstract: In vitro regeneration of plants from immature leaflets of 3 day-old pea (Pisum sativum L. cv. Century) seedlings was studied under defined nutritional, hormonal and environmental conditions. Immature leaflets isolated from the second and third apical leaves of aseptically germinated seeds were cultured on MS medium containing vitamins as in B5 medium, 3% sucrose, 0.8% agar and supplemented with 0.1, 1, and 10 μM concentrations of naphthaleneacetic acid (NAA) and 1 and 10 μM levels of benzyladenine (BA) in various combinations. Shoot regeneration from the primary callus occurred within 45 to 90 days of culture in most of the hormone combinations. Although the number of calli producing shoots was maximal at 10 μM levels of NAA and BA, multiple shoot regeneration was predominant at a combination of 0.1 μM NAA and 10 μM BA. Indoleacetic acid (IAA) and kinetin (K), both at 10 μM, also induced shoot regeneration. No shoots were regenerated when 10 day-old leaflets were used as explants. Root production generally occurred on non-shoot regenerating calli. Roots were induced to differentiate by transferring the regenerated shoots onto half-strength B5 medium supplemented with 1 μM NAA.
55 citations
TL;DR: Cytological analyses performed on root tips of 20 randomly chosen plantlets showed that all the analysed plants contained the diploid number of chromosomes and the method described seems to be suitable for rapid propagation of globe artichoke.
55 citations
TL;DR: Cytological studies on leaf callus cells and regenerated potato plants suggest that it may be possible to utilize somatic chromosome doubling to obtain tetraploids from outstanding dihaploid breeding clones.
Abstract: Cytological studies on leaf callus cells and regenerated potato plants suggest that it may be possible to utilize somatic chromosome doubling to obtain tetraploids from outstanding dihaploid breeding clones. The ploidy levels found in callus-derived plants were diploid, tetraploid, and octaploid, but the proportion of these was dependent on the donor genotype. L1 and L3 germ layers were studied in more than 300 plants; periclinal ploidy chimerism, an undesirable feature of colchicine doubling, was not found. Leaf callus was more efficiently induced using NAA than 2, 4-D as an auxin source in the Murashige and Skoog medium. A high proportion of dividing cells in young calli were polyploid. The frequency of doubled and octaploid plants regenerated was significantly dependent on donor genotype. The extent of polyploidization was marginally higher after callus growth on a medium containing 2, 4-D than in a medium containing NAA. In some genotypes the chromosome numbers of regenerated plants were variable, being less than tetraploid (mixohypotetraploid). After tuber propagation, the original ploidy level was maintained although mixohypotetraploidy persisted. In a few somatically doubled clones, male fertility was tested and found to be satisfactory with respect to seed-setting.
49 citations
TL;DR: Root regenerarion occurred in high frequency only when the differentiated shoots were recultured on sucrose-free 1 2 strength MS medium supplemented with 1 μM indolebutyric acid (IBA), and regenerated plantlets were maintained in vitro at 26°C for over 2 years without periodic transfer and some of them were successfully grown in pots.
41 citations
TL;DR: One to five percent of Lycopersicon peruvianum (L.) Mill undergo cell division and concomitant organization to form embryogenic-like structures when cultured in Murashige and Skoog medium (1962).
Abstract: One to five percent of Lycopersicon peruvianum (L.) Mill. leaf mesophyll protoplasts undergo cell division and concomitant organization to form embryogenic-like structures when cultured in Murashige and Skoog medium (1962) containing 3% sucrose, 9% mannitol, 1.0 mg/l kinetin (K) and 1.0 mg/l naphthalene acetic acid (NAA) at pH 5.6–5.8 (medium A). These embryogenic structures, after passing through developmental stages similar to those observed in zygotic embryogeny, are capable of forming shoots on hormone-free medium A. In medium B, wherein 0.5 mg/l of 2,4-dichlorophenoxyacetic (2,4-D) replaced the hormones (K and NAA), embryogenic structures did not develop. However, callus originating in medium B retained morphogenetic capacity as was evidenced by subsequent shoot regeneration when they were transferred to medium A with K and NAA replaced by 1.0 mg/l zeatin (Z). The potential value of incorporating this regeneration trait into Lycopersicon species and cultivated lines for use in tissue culture programs is discussed.
38 citations
TL;DR: Experiments are described which improve the protocols for initiating in vitro cultures of sugarcane and allowing efficient regeneration of plants even after 30 months of callus proliferation, but no plants have yet been regenerated derived from isolated protoplasts.
Abstract: Experiments are described which improve the protocols for initiating in vitro cultures of sugarcane and allowing efficient regeneration of plants even after 30 months of callus proliferation. Procedures adopted included use of leaf base explants, CS medium with 3 mg/l 2, 4-D and 0.25 mg/l kinetin for callus initiation and growth, MS medium with 0.5 mg/l IAA and 1 mg/l BAP for shoots, MS medium with 5 mg/l NAA and 7% (wt/vol) sucrose for rooting of shoots. Casein hydrolysate (N-Z amine) significantly shortened the lag period in the growth of sugarcane suspension cultures, but did not increase the rate of growth following the lag phase. Protoplasts isolated from two types of cultures could be grown to re-establish cell cultures but no plants have yet been regenerated derived from isolated protoplasts.
37 citations
TL;DR: The most successful combination of phytohormones for the induction of shoot development in undifferentiated callus, was 2.0 mg/1 benzyladenine and 1.0mg/1 naphthaleneacetic acid, which resulted in regeneration of whole plants from these calli at high frequencies.
Abstract: Callus cultures were established on Murashige and Skoog medium from seedling hypocotyls and roots of Slylosanlhes guyanensis (Aubl.) Sw. cv. Cook and from leaves of 6-month-old) plants. Shoots developed in primary calli derived from seedling tissue with a number of benzyladenine or kinetin and naphthaleneacetic acid combinations. Shoot formation on primary leaf callus, occurred with 2.0 mg/1 benzyladenine and 2.0 or 1.0 mg/l naphthaieneacetic acid. Undifferentiated callus from all three sources was induced and maintained on medium with 2.0 mg/1 kinetin and 2.0 mg/1 2, 4-dichlorophenoxyacede acid in the dark. Shoot formation and regeneration of whole plants from these calli were achieved at high frequencies. The most successful combination of phytohormones for the induction of shoot development in undifferentiated callus, was 2.0 mg/1 benzyladenine and 1.0 mg/1 naphthaleneacetic acid. The regenerated plants showed no phenotypic abnormalities.
32 citations
TL;DR: In this article, the anthers of three genotypes of Lycopersicon esculentum, viz. cv. HS-101, HS-102 and an F1 hybrid (Montfavet 63-4xHS-101), were cultured in various defined nutritive media.
Abstract: The anthers of three genotypes ofLycopersicon esculentum, viz. cv. HS-101, cv. HS-102 and an F1 hybrid (Montfavet 63-4xHS-101) in different stages of development were cultured in various defined nutritive media. Only anthers containing microspores in the early uninucleate stage were found to respond with the culture medium in the formation of androgenic callus. The DGII medium with 2 mg l−1 NAA and 1 mg 1−1 kinetin was found to be best for callus induction but MS medium supplemented with 2 mg l−1 2,4-D and 0.1 mg 1−1 BAP favoured proliferation and growth of the callus. The androgenic microspores followed the ‘B’ type pathway of androgenesis in the formation of callus.
31 citations
TL;DR: Protoplasts were isolated from young leaves of eggplant with a solution containing 1% cellulase R-10 and 0.2% Macerozyme R- 10 and profuse green callus formed after 4 weeks on MS medium supplemented with different combinations of growth regulators.
TL;DR: Virus-free material of IITA elite clones of sweet potato is available for distribution and can be stored in tissue culture for one year without transfer to fresh medium.
Abstract: Sweet potato vein clearing disease agents have been eliminated from 20 elite clones of sweet potato through meristem tip culture. Meristem tips of 0.25 to 0.4 mm length were cultured in a modified Murashige and Skoog medium. Plantlets were obtained after two months and transferred to soil. Indexing was accomplished by approach grafting to Ipomoea setosa. About 80% of the plants obtained by meristem tip culture indexed negatively. Rapid multiplication was accomplished using single node cutting in vitro. A multiplication rate of 4.2 /month was obtained. The material can be stored in tissue culture for one year without transfer to fresh medium. Virus-free material of IITA elite clones of sweet potato is available for distribution.
TL;DR: Preliminary to somatic hybridization between chlorophyll-deficient mutants of Nicotiana rustica and N. tabacum conditions for cell culture, protoplast production and culture and induction of morphogenesis were established.
Abstract: Preliminary to somatic hybridization between chlorophyll-deficient mutants of Nicotiana rustica and N. tabacum conditions for cell culture, protoplast production and culture and induction of morphogenesis were established. Callus and suspension cultures from wild-type and chlorophyll-deficient strains of both species were established and maintained on Murashige and Skoog medium containing either 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA). The chlorophyll-deficient phenotype was evident in cultures derived from mutants of both species. Protoplasts were readily obtained from cell cultures with N. rustica outyielding N. tabacum and mutant cells of the latter substantially outyielding the wild-type cells. In Nagata and Takebe medium protoplasts regenerated cells walls and 60–80% divided and formed cell colonies. Protoplasts from cultures on 2,4-D medium were healthier but slower to divide than those from cultures grown on an NAA-supplemented medium. Protoplasts derived from 2,4-D-g...
TL;DR: Protoplasts were isolated from the mesophyll of Digitalis lanata enzymatically and cultured in a liquid regeneration medium and developed into cell clusters with a diameter of about 4–5 mm.
Abstract: Protoplasts were isolated from the mesophyll of Digitalis lanata enzymatically and cultured in a liquid regeneration medium (D2a). Protoplast division occurred at a rate of approximately 30%. Mature cell colonies were transferred onto agar medium (D2b)where they developed into cell clusters with a diameter of about 4–5 mm. After transfer onto MS medium, these calli differentiated leaves and shoots which could be rooted on MS medium containing a low hormone concentration.
TL;DR: Cell suspension cultures were initiated from pith callus of eggplant, Solanum melongena cv.
TL;DR: Callus cultures established from the basal region of in vitro-obtained shoot buds on MS medium supplemented with CH + CW + NAA regenerated shoot buds and optimum regeneration was obtained on MS + CW (5% v/v) medium.
TL;DR: Anthers of 2 tetraploid Rosa species were plated on selected basis media, supplemented with various amounts of auxins and kinetin, at different bud stages and using various light conditions, and cytological examination showed that most cells of both species had 2 n =14 chromosomes and thus were of pollen origin.
TL;DR: It appears that although GA inhibits shoot meristem initiation from potato tuber discs, it is required for shoot development once meristems are initiated.
Abstract: The formation of adventitious shoots from potato tuber discs explanted onto a modified Murashige and Skoog (MS) medium containingN
6-benzylaminopurine (BAP) (3.0 mg/l), and α-naphthaleneacetic, acid (NAA) (0.01 mg/l), was affected by gibberellic acid (GA). The presence of GA in the explant medium was required for shoot formation and 3×10−10
M GA appeared optimum. However, microscopic examination of the tissue protuberances on the surface of the tuber discs from which shoots arose revealed that GA inhibited the formation of shoot meristems. Tuber discs cultured for 6 wk on MS medium containing BAP and NAA without GA did not initiate adventitious shoots that could be determined visually, but microscopic examination of the tissue protuberances revealed the presence of numerous shoot meristems. Subsequent transfer of these tuber discs to medium with GA but without BAP or NAA resulted in the formation of shoots from 100% of the recultrued dises. Thus it appears that although GA inhibits shoot meristem initiation from potato tuber discs, it is required for shoot development once meristems are initiated.
TL;DR: This protoplast system is technically comparable to soybean for experiments concerned with genetic manipulation involving legumes and after three weeks, cell colonies could be transferred to solid MS medium containing N-Z amine and coconut water where callus developed.
Abstract: A system was developed for protoplast isolation and culture from suspension cultured cells of winged bean,Psophocarpus tetragonolobus. Cells from a three-day-old suspension were incubated in an enzyme mixture containing 6% cullulysin, 1% Macerase, 1% desalted Rhozyme, 0.4M sorbitol, and 0.1M CaCl2 at pH 5.5. Average yields of protoplasts were 6.5 × 106 per gram fresh weight of cells. Protoplasts were cultured in modified B5 medium containing 68.4 g/l glucose, 250 mg/l xylose, 0.1 mg/l 2,4-D, 0.5 mg/l BAP, 250 mg/l N-Z amine type AS, and 20 ml/l coconut water. After 24 h of culture, the protoplasts had synthesized a new wall, and in three days had begun division. The optimum plating density was 1–2 × 103 protoplasts/ml. The division frequency ranged between 40%–60% for most experiments with a high of 72% in one experiment. After three weeks, cell colonies could be transferred to solid MS medium containing N-Z amine and coconut water where callus developed. This protoplast system is technically comparable to soybean for experiments concerned with genetic manipulation involving legumes.
TL;DR: Cambial tissues from the twigs of a mature Japanese persimmon, Diospyros kaki Thunb, cv.
Abstract: Cambial tissues from the twigs of a mature Japanese persimmon, Diospyros kaki Thunb, cv. ′Tsurunoko′, were used as the starting material in this experiment. Callli were induced on various callus induction media. Root formation in the calli was observed on Murashige and Skoog′s medium containing 1ppm NAA. KIN in combination with NAA did not inhibit to the root formation at 1ppm. Root formation in the callus depended largely on the type of medium on which the callus was isolated and subcultured and the number of successive subcultures on a given medium. When the callus was isolated on Murashige and Skoog′s medium containing NAA and KIN, roots were formed in the first passage, whereas when the callus was isolated on Murashige and Skoog′s medium containing 0.1% yeast extract or 0.1% phytone, NAA, and KIN, the first root formation was not observed until the callus was subcultured on the medium several times.Roots were never formed in calli which were isolated and subcultured on Murashige and Skoog′s medium containing 2, 4-D or IBA instead of NAA, nor were either Wolter and Skoog′s medium or a modified Wolter and Skoog′s medium containing NAA and KIN suitable for root formation.Bud initials were formed in callus which was subcultured several times on Mura shige and Skoog′s medium containing NAA and KIN. KIN was effective in promoting bud initiation in combination with NAA. These initials turned green under continuous light conditions, but they did not succeed in developing themselves into plantlets on various media. Abnormal structures were initiated in callus cultured on Murashige and Skoog′s medium containing NAA and KIN, but these structures were not identifiable as either root or bud initials.
TL;DR: Leaf explants of Salpiglossis sinuata L. were cultured on MS medium containing the auxins 2,4-D, IAA, NAA, or the cytokinins 6- BAP, 2ip, or K singly or NAA + 6-BAP combinations to determine their morphogenetic responses.
TL;DR: Plants of Iberis amara (Brassicaceae) have been regenerated from in vitro cultured leaf and stem explants and 4-dichloro phenoxy acetic acid enhanced callusing but inhibited organogenesis.