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1.
Mohamed Kouider Randal Hauptmann Jack M. Widholm Robert M. Skirvin Safi S. Korban 《Plant cell reports》1984,3(4):142-145
Protoplasts could be successfully isolated and cultured from callus and suspension cultures of Malus xdomestica cv. Jonathan. Protoplast-derived colonies were recovered when the osmoticum (glucose) was gradually reduced in semi-solid 8p medium or by the use of feeder plates. Formation of embryo-like structures was induced from the protoplast-derived callus on media supplemented with IAA and BA. These structures formed roots but plants failed to develop. Protoplasts could be isolated from leaves, but not from stems or petioles. The leaf protoplasts failed to divide.List of abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- ABA
abscisic acid
- IAA
indole acetic acid 相似文献
2.
Protoplasts were isolated from cotyledons and foliage leaves of cotton (Gossypium hirsutum and G. barbadense). Cotyledon protoplasts were larger and responded to culture better than leaf protoplasts. Cotyledon derived protoplasts regenerated cell walls and formed microcolonies of 2–3 cells in G. hirsutum and 5–8 cells in G. barbadense. However, the microcolonies did not grow beyond this stage. Protoplast yield and viability, cell wall regeneration and cell division were influenced by several factors, e.g., genotype, age, tissue and growth condition of donor plant, enzyme mixture and concentration, preplasmolysis period, incubation period, and culture medium.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- GA3
gibberellic acid
- p CPA
p-chlorophenoxyacetic acid
- MES
2[N-morpholino]ethanesulfonic acid 相似文献
3.
Protoplasts of sugarbeet (Beta vulgaris L.) were isolated from cell suspension cultures and cultured in modified PGo medium. Conditions required for the efficient division of the protoplasts were investigated.The optimal combination of phytohormones was found to be 1 mg/l NAA, 0.2 mg/l 2,4-D, 0.5 mg/l zeatin. Protoplast division was also considerably stimulated by the addition of 250 mg/l casein hydrolysate, 200 mg/l yeast extract, and 20% v/v conditioned culture medium to the protoplast culture medium. The highest division rate (up to 35% of the protoplasts) was achieved at a density of 4×104- 1×105 protoplasts/ml. From the colonies callus and suspension cultures were readily obtained.Abbreviations BAP
6-Benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
Indole-3-acetic acid
- Kin
6-Furfurylaminopurine (kinetin)
- NAA
-Naphtalenacetic acid
- Zea
Zeatin 相似文献
4.
Leaf protoplasts were isolated from axenic shoot cultures of four varieties of Capsicum annuum (Americano, Dulce Italiano Florida Gynat and Nigrum) and a wild species C. chinense. Protoplasts of both species, cultured in KM8P medium and using agarose bead culture, entered division with the exception of the variety Nigrum. Cell colonies formed callus in agar-solified MS medium supplemented with zeatin and for C. annuum v. Dulce Italiano shoots were regenerated when protoplast-derived calli were transferred to MS medium with 6-BAP. Excised shoots were rooted on MS medium which lacked phytohormones.Abbreviations 6-BAP
6-benzylamino purine
- MS
Murashige and Skoog (1962)
- KM8P
Kao and Michayluk (1975)
- SDS
sodium dodecyl sulphate 相似文献
5.
Protoplasts were isolated from cell cultures of G. soja and G. tabacina, respectively. The isolation procedure employed Percoll for the separation and concentration of protoplasts. The cultured protoplasts formed cells which developed into embryo-like structures. Protoplasts also were isolated from leaf tissue of soybean cv. Williams 82. Upon culture, the protoplasts regenerated cell walls and divided to form cell cultures.Abbreviations 2,4-D
2,4-Dichlorophenoxyacetic acid BA|Benzyladenine
- BA
Benzyladenine 相似文献
6.
We report regeneration of fertile plants from barley (Hordeum vulgare L. cv. Igri) protoplasts isolated from regenerable suspension cultures initiated from anther-derived embryogenic callus. Plants were routinely regenerated from these suspension cultures, which maintained their regenerative capacity for several months. It was first possible to isolate protoplasts from suspensions after three months of culture and after four months protoplasts capable of division could be isolated. Protoplasts maintained the regenerative capacity of the donor cells and formed embryogenic callus. Green plants were regenerated from protoplast-derived calli, although the proportion of albino plantlets was high. Viable regenerants were transferred to soil and fertile plants were recovered.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- 6-BAP
6-benzylaminopurine
- PP
Protoplasts 相似文献
7.
Protoplasts were isolated from mesophyll of axenic cultures of grape, Vitis rotundifolia cv. Summit and V. vinifera cv. Cabernet Sauvignon. Enzymes effective for protoplast isolation were Macerozyme R-10 (0.5% and 0.1%) and Cellulase Onozuka R-10 (1.0% and 0.5%) for V. rotundifolia and V. vinifera, respectively. Polyvinylpyrrolidone and 2-(N-morpholino)ethanesulfonic acid were essential in the isolation media. Protoplasts were purified using flotation/centrifugation. The protoplasts of V. rotundifolia cultured in Gamborg's B5 basal medium with 2.2 M 6-benzyladenine, 4.5 M 2,4-dichlorophenoxyacetic acid and 0.4% agarose gave the best plating efficiency of conditions tried in this study. Cell division occurred within 5 to 6 days and visible microcalli developed within one month. After 6 weeks in culture, microcalli transferred to liquid medium exhibited active callus growth. Protoplasts of V. vinifera cultured under these conditions had similar results.Abbreviations MES
2-(N-morpholino)ethanesulfonic acid monohydrate
- PVP
polyvinylpyrrolidone
- PDS
potassium dextran sulfate
- BA
6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate 相似文献
8.
Callus formation from protoplasts of Artemisia sphaerocephala Krasch and some factors influencing protoplast division 总被引:2,自引:0,他引:2
Round wormwood (Artemisia sphaerocephala Krasch) seeds were germinated on Murashige & Skoog (1962) medium without plant growth regulators. The hypocotyls of seedlings were sliced and cultured on M1 medium with 2,4-dichlorophenoxyacetic acid (9.05 M) to induce callus. The induced calluses were subcultured on the same medium. Ten day old calluses were used to isolate protoplasts in an enzyme solution with 0.65 M mannitol. Protoplast yield strongly depended upon the state of callus cultures. Certain amount of hemicellulase could improve protoplast isolation. Purified protoplasts were cultured in modified Kao & Michayluk (1975) medium with 0.60 M mannitol as osmoticum, suggesting that protoplasts of A. sphaerocephala need a high initial osmolarity. Protoplasts generally divided evenly and the percentage of first division could reach 10%. Kinetin exhibited a positive effect on initial cell division. Furthermore, we studied the effect of protoplast density and vitamin C on sustained growth of protoplasts. After forty days, 1 mm calluses in diameter formed.Abbreviations CH
casein hydrolysate
- 2,4-D
2,4-dichlorophenoxyacetic acid
- KM8P
Kao & Michayluk (1975) protoplast medium
- MS
Murashige & Skoog (1962) medium
- MES-2
(N-morpholino)ethanesulfonic acid 相似文献
9.
Large yields (1.85 × 107/g.f.wt.) of viable protoplasts were obtained from leaves of axenic shoot cultures of Malus Xdomestica Borkh. cv. Greensleeves. Protoplasts cultured in liquid or agarose semi-solidified KM8P medium underwent cell wall regeneration and colony formation.Protoplast-derived cell colonies developed to callus on semi-solid KM8 medium. This is the first report of callus formation from mesophyll protoplasts of apple.Abbreviations BAP
6-benzylaminopurine
- K
kinetin
- Z
zeatin
- GA3
gibberellic acid
- IBA
3-indole butyric acid
- NAA
1-naphthalene acetic acid
- IAA
3-indole acetic acid
- ABA
abscisic acid
- f.wt.
fresh weight
- MS
Murashige and Skoog (1962) 相似文献
10.
Protoplasts were isolated from leaves, shoots, cotyledons, ray florets and callus cultures of Dimorphotheca aurantiaca (syn. D. sinuata) (Cape Marigold, Star of the Veldt) and Rudbeckia hirta, R. laciniata and R. purpurea; species of ornamental value. For Dimorphotheca, plants were regenerated from protoplasts of all sources apart from the ray floret, whilst for the Rudbeckia species, although protoplast division was induced in most cases, only leaf mesophyll protoplasts of R. hirta c.v. Marmalade gave plants. The establishment of plant regeneration for these ornamental species, from protoplasts, now provides a basis for their somatic hybridisation.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- NAA
naphthaleneacetic acid
- K
kinetin
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962)
- f.wt.
fresh weight 相似文献
11.
Summary Protoplasts were prepared from a photoautotrophic (PA) cell line of Glycine max (soybean). A yield of 75 to 90% after two to three hours digestion in a mixture of 1% Cellulase R10, 0.2% Pectolyase Y23 and 2% Driselase was obtained. Cell division and colony formation occurred from approximately 18% of the plated protoplasts. The cultured protoplasts were as sensitive to the herbicide atrazine, a photosynthetic inhibitor, as the original PA cells under the same conditions. Protoplasts and cells of a heterotrophic (HT) soybean culture were not as sensitive to atrazine. The isolated protoplasts retained the PA characteristics of the parental culture in the callus and cell suspension cultures obtained from the protoplasts. The chromosome numbers in the parental cell line and in cells derived from the isolated protoplasts (both PA and HT) were found to be largely (99%) the normal diploid number of 40.Abbreviations BA
Benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- HT
Heterotrophic
- MES
2-(morpholino) ethane sulfonic acid
- NAA
Naphthaleneacetic acid
- PA
Photoautotrophic
- PCM
Protoplast culture medium 相似文献
12.
Protoplasts were isolated from seedling roots, hypocotyls, and cotyledons of four cultivars of Helianthus annuus and from leaves of axenic shoot cultures of the wild species H. praecox, H. scaberimus and H. rigidus. Optimal culture conditions were established for the respective protoplast systems, using the agarose bead method of culture. Protoplast division was induced for all the species examined. In the case of the cultivars of H. annuus, hypocotyl and cotyledon protoplast division was sustained leading to callus formation, which in turn, could be induced to produce roots and organised meristematic regions in the presence of NAA and 6-BAP.Abbreviations 6-BAP
6-benzylaminopurine
- NAA
-naphthalene acetic acid
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog 相似文献
13.
Protoplasts from Olea europaea L. have been compared in terms of their yield, viability, cell division and callus differentiation. Viable protoplasts were isolated from in vitro cultured leaves and cotyledons by an overnight incubation in an enzyme solution containing 1–1.5% driselase and 0.5M sucrose. This method allowed high yield of purified protoplasts, which floated and formed a dark green band at the meniscus, after centrifugation. Purified protoplasts were diluted to 3×104 protoplasts·ml–1 in culture medium. After cell wall regeneration, protoplasts gradually increased their volumes under appropriate conditions. The first divisions occurred during the second week in culture. Division efficiency ranged from 5.2 to 9.8% after 20 days in culture. Two weeks later visible microcolonies developed only from cotyledon protoplasts. After 6 weeks in culture, the microcalli were transferred to a solidified culture medium with 0.6% agarose, which induced active callus growth.Abbreviations OM
olive proliferation medium, Rugini 1984
- Omg
OM for the germination of olive embryos
- OMr=OM
for root induction
- OMp=OM
for protoplasts
- OMc=OM
for callus
- BN
Bourgin and Nitsch medium 1967
- IBA
indol-3-butyric acid
- NAA
naphthalene acetic acid
- 2,4-D
dichlorophenoxyacetic acid. 相似文献
14.
A protocol for regenerating plants from callus-derived protoplasts of Pelargonium x domesticum (rega l geranium cv. Melissa) has been developed. Protoplasts were isolated from leaf-derived callus tissue on MS medium supplemented with 3.0 mg/l naphthalene acetic acid, 2.0 mg/l 6- benzylaminopurine, and 3.0% sucrose. This callus yielded 2.7×105 protoplasts/gram of tissue after a 6 hr incubation in an enzyme solution consisting of 2.0% cellulysin, 0.5% macerase, and 0.5 M sucrose. Protoplasts were plated at 1×105 protoplasts/ml in a mixture (11 v/v) of KMP8/KP liquid medium layered on the same medium solidified with 0.6% agarose. Protoplast division was initiated within 2 days, and colonies of 15 to 50 cells developed 8 wk after plating. P-calli 1–2 mm3 developed 15 wk after plating, and plants regenerated from the p-calli have been transferred to the greenhouse.Abbreviations NAA
naphthaleneacetic acid
- 6-BAP
6-benzylaminopurine
- CW
Calcofluor White
- FDA
fluorescein diacetate 相似文献
15.
S. E. Maddock 《Plant cell reports》1987,6(1):23-26
Suspension cultures have been initiated from embryogenic callus of hexaploid wheat (Triticum aestivum L.). Most commonly, these suspensions are composed of callus-like clusters (up to 2 mm in diameter). Two rapidly-growing lines (MBE6 and C82d) have been obtained, which consist of smaller aggregates of cytoplasmic cells, and these have been maintained for more than 4 years. These lines show very limited morphogenetic capacity and only a single plantlet has been regenerated, from line MBE6, after 9 months in culture. Protoplasts isolated from line MBE6 are unable to divide, but protoplasts from line C82d consistently undergo sustained divisions to form callus or secondary cell suspensions.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) medium 相似文献
16.
Protoplasts were isolated from embryogenic callus ofAbies alba L. which originated from immature seeds. The protoplasts were immobilized in alginate beads in order to follow the development of single protoplasts.Surrounding culture medium was modified from Kao and Michayluk (1975). After cell wall regeneration subsequent cell divisions lead to the formation of colonies showing an early differentiation of small meristematic cells and large vacuolated suspensor-type cells.Abbreviations 6-BA
N6-benzyladenine
- IAA
3-indoleacetic acid (potassium salt)
- KM
Kao and Michayluk (1975)
- MES
2-(N-morpholino)ethanesulfonic acid
- NAA
1-naphthalene acetic acid (sodium salt)
- PVP
Polyvinylpyrrolidone
- SH
Schenk and Hildebrandt (1972)
- Tween 80
Polyoxyethylene- sorbitan - monooleate
- WPM
(woody plant medium), Lloyd and McCown (1981) 相似文献
17.
Plant regeneration of Actinidia deliciosa var. deliciosa cv. Hayword was obtained from protoplasts isolated from petiole derived long-term callus cultures. Protoplasts were cultured in liquid medium over agarose gelled medium. Regenerated green callus, plated on solid medium, could develop shoots that rooted spontaneously in hormone-less medium. The plants obtained are growing fast in soil and present a normal phenotype.Abbreviations BAP
benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- DTT
dithiotreitol
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- Kin
kinetin
- MES
2-(N-morpholino) ethanesulphonic acid
- MS
Murashige and Skoog (1962) medium
- NAA
naphthalene-1-acetic acid
- SH
Schenk and Hildebrandt (1972) medium
This Research was supported by JNICT and INIC 相似文献
18.
Summary Rapidly proliferating and highly regenerable suspension cultures of somatic embryos of Carica papaya x C. cauliflora were used for protoplast isolation. On average, protoplast yield was 1.5×106/g fresh weight of somatic embryos. Protoplasts were first cultured in liquid KM8P-S medium for 2 weeks and then plated in the same medium solidified with 1% agarose. About 1.4% of the protoplasts developed directly into somatic embryos. Protoplast-derived somatic embryos proliferated rapidly through direct embryogenesis on modified MS medium supplemented with 1 mg/1 ABA, and developed into plantlets upon transfer to MS medium devoid of plant growth regulators. The plantlets were successfully transplanted to soil.Abbreviations MS
Murashige and Skoog medium (1962)
- KM8P
Kao and Michayluk medium (1975)
- NAA
-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
abscisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- CPW
Frearson et al. medium (1973) 相似文献
19.
Protoplasts were isolated from cell cultures of oil palm (Elaeis Guineensis). The protoplasts were cultured on a nurse medium containing oil palm cells in the presence of which protoplasts formed cell walls and divided to form cell cultures.Abbreviations NAA
-naphthalene acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- TEM
thin-section electron microscopy 相似文献
20.
Conditions for successful culture of rubber tree (Hevea brasiliensis) protoplasts were investigated. Protoplasts, derived from embryogenic callus, regenerated cell walls then underwent division when embedded in alginate and cultivated on a modified Murashige and Sook medium (9 M 2,4-dichlorophenoxyacetic acid, 0.6 M glucose, 0.93 M kinetin, NH
4
+
reduced by half) in the presence of nurse cells (tobacco feeder cell layer). The presence of nurse cells was essential to maintain viability and sustain protoplast division. Several parameters which influenced the plating efficiency were analysed, such as the density of feeder cells and the duration of contact of the feeder layer.Abbreviations BSA
Bovine Serum Albumin
- CPW
Cell and Protoplast Washing medium (Frearson et al. 1973)
- 2,4-D
2,4-dichlorophenoxyacetic acid
- 3,4-D
3,4-dichlorophenoxy-acetic acid
- PDA
Fluorescein diacetate
- FW
Fresh weight
- KIN
Kinetin
- MES
[2N-morpholino] ethane sulfonic acid
- MS medium
Murashige and Skoog (1962) medium
- PE
plating efficiency
- SAB
South American Leaf Blight
- WPM]
Woody Plant Medium (Russel and Mc Cown 1986) 相似文献