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1.
Embryogenic suspension cultures of Abies alba were established using an embryogenic suspensor mass culture originating from the zygotic embryo in immature seed explants (Schuller et al. 1989). Protoplasts were isolated from the suspension material. The protoplasts were immobilized in alginate layers in order to follow the development of single protoplasts. During the first days of protoplast culture a modified Kao and Michayluk (1975) medium proved to be necessary for subsequent divisions. The formation of proembryos succeeded within 2–3 weeks when subcultured with a modified Schenk and Hildebrandt (1972) liquid medium. Light, enhanced sugar concentration, and the addition of abscisic acid led to the formation of slightly green torpedo-shaped somatic embryos after 6–8 weeks from protoplast isolation.Abbreviations ABA
abscisic acid
- BAP
N6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- ESM
Embryonal suspensor mass (Gupta and Durzan 1986)
- KM
Kao and Michayluk (1975)
- LP
(von Arnold and Eriksson 1977)
- MES
2-(N-morpholino)ethane-sulfonic acid
- NAA
1-naphthalene-acetic acid (sodium salt)
- PVP
polyvinylpyrrolidone
- SH
Schenk and Hildebrandt (1972)
- Tween 80
polyoxyethylene-sorbitan-monooleate 相似文献
2.
Shoot cultures of Glehnia littoralis F. Schmidt ex Miq. (Umbelliferae) were established by placing shoot tip explants on Linsmaier and Skoog medium with 1 M NAA and 10 M BAP. Shoots were multiplied on the basal medium supplemented with 0.3 M NAA and 3 M BAP and rooted on medium containing either 1 M IBA or 3–10 M IAA. Plantlets survived in pots without any covering. This unique characteristic of the plantlets was ascribed partly to a well-developed cuticle on the surface of the leaf and the small ratio of surface area to fresh weight of a leaf blade in comparison with those of other species whose plantlets needed coverings after potting. The regenerated plantlets were finally transferred to soil.Abbreviations IAA
potassium indole-3-acetate
- IBA
indole-3-butyric acid
- IPA
indole-3-propionic acid
- NAA
potassium 1-naphthaleneacetate
- 2,4-D
sodium 2,4-dichlorophenoxyacetate
- BAP
6-benzylaminopurine
- 2-iP
N6-(2-isopentenyl)adenine 相似文献
3.
Intergeneric somatic hybrid plants between Hamlin sweet orange [Citrus sinensis (L.) Osbeck] and Flying Dragon trifoliate orange (Poncirus trifoliata Raf.) were regenerated following protoplast fusion. Hamlin protoplasts, isolated from an habituated embryogenic suspension culture, were fused chemically with Flying Dragon protoplasts isolated from juvenile leaf tissue. The hybrid selection scheme was based on complementation of the regenerative ability of the Hamlin protoplasts with the subsequent expression of the trifoliate leaf character of Flying Dragon. Hybrid plants were regenerated via somatic embryogenesis and multiplied organogenically. Hybrid morphology was intermediate to that of the parents. Chromosome counts indicated that the hybrids were allotetraploids (2n=4x=36). Malate dehydrogenase (MDH) isozyme patterns confirmed the hybrid nature of the regenerated plants. These genetically unique somatic hybrid plants will be evaluated for citrus rootstock potential. The cell fusion, selection, and regeneration scheme developed herein should provide a general means to expand the germplasm base of cultivated Citrus by intergeneric hybridization with related sexually incompatible genera.Abbreviations MDH
malate dehydrogenase
- CTV
citrus tristeza virus
- MT
Murashige and Tucker basal medium
- BH3
protoplast culture medium, Grosser and Chandler, 1987
- PEG
polyethylene glycol
- GA3
giberellic acid
- BA
N-(phenylmethyl)-1 H-purin-6-amine
- HCl
hydrochloric acid
Florida Agricultural Experiment Station Journal Series No. 7972 相似文献
4.
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
5.
Somatic embryos and plantlets have been obtained in quite a direct way from mesophyll protoplasts isolated from androgenetic plants of two cultivars (Loras and Tower) of Brassica napus. The procedure consists of four steps. Proembryos were induced in a medium supplemented with a cytokinin and an auxin at comparatively high concentrations. They developed to mature embryos when the auxin was reduced or omitted.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- NAA
1-naphthaleneacetic acid
Dedicated to Prof. Dr. Wilhelm Halbsguth on the occasion of his 70th birthday.On leave from the Institute of Genetic, Academia Sinica, Beijing, China 相似文献
6.
Y. Wang M. R. Huang Z. M. Wei Y. R. Sun D. M. Chen Z. H. Xu L. M. Zhang N. Xu 《Plant cell reports》1995,14(7):442-445
Simon poplar (Populus simonii) protoplasts were isolated from suspension cells, with protoplast yield of 3.8×107 g–1 F. W. They were cultured in a K8P liquid medium containing 13.57M 2,4-D, 1.07M NAA and 0.93 M KT. Protoplast culture was influenced by the plating density, osmotic pressure, and the sources and amounts of nitrogen and carbon in the culture medium. Multiple shoots were produced from protoplast-derived callus after culture on MS medium containing 4.44 M BA, 2.32M KT, 2.28 M ZT, and 0.54M NAA. Shoots 2–3 cm in height were isolated from the calli and rooted on 1/2 MS medium. After transplantation into pots, the regenerated plants grew vigorously in greenhouse.Abbreviations BA
N6-benzyladenine
- NAA
1-naphthalene acetic acid
- 2,4-D
2,4-dichlorophenoxy acetic acid
- KT
Kinetin
- ZT
Zeatin
- 2ip
2-isopentenyl-adenine
- FDA
fluorescein diacetate
- MES
2-(N-morpholino) ethane sulfonic acid
- MS
Murashige and Skoog basal medium (1962)
- K8P
Kao basal medium (1977)
- CPW
Cell and Protoplast Wash solution (Power and Davey 1980) 相似文献
7.
J. B. Teixeira M. R. Söndahl T. Nakamura E. G. Kirby 《Plant Cell, Tissue and Organ Culture》1995,40(2):105-111
Primary globular callus from immature zygotic embryos and friable embryogenic tissue derived from mature zygotic embryos were used to establish suspension cultures. Callus cultures were established either on modified Y3 or MS medium containing 475–500 M 2,4-D or 250 M picloram and 0.3% (w/v) activated charcoal. Suspension cultures of both cell lines were established in modified Y3 medium containing 10 M 2,4-D. The establishment of cell suspensions from friable embryogenic tissue took only 2 months, in contrast with suspensions from primary globular callus which took 3–5 months to establish. Embryo differentiation was observed only in cell suspensions derived from the friable embryogenic tissue after plating aliquots on regeneration medium. Germinated embryos were recovered and plantlets were successfully established under greenhouse conditions.Abbreviations CET
compact embryogenic tissue
- FET
friable embryogenic tissue
- CIM
callus induction medium
- PGC
primary globular callus
- 2,3-D
2,4-dichlorphenoxyacetic acid Y3-Eeuwens' medium
- MS
Murashige & Skoog medium
- PVP-40
polyvinylpyrrolidone
- KM
Kao & Michayluk vitamins
- ABA
abscisic acid 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.
Protoplasts were isolated from leaves of glasshouse-grown plants of Centaurea cyanus and axenic shoot cultures of Senecio x hybridus. Upon culture, using modified MS-based media, protoplasts of both systems entered division to produce callus, followed by plant regeneration. Leaf protoplasts of Callistephus chinensis entered sustained division only following the preconditioning for 24h of peeled leaf tissues on agar-solidified MS-based medium. Protoplasts were also isolated from cell suspensions of C. chinensis and divided in MS-based or KM media. However, only leaf mesophyll protoplasts of Callistephus produced callus, which developed shoots.The establishment of protoplast-to-plant protocols for these ornamental species has provided a basis for broadening their gene pools through somatic hybridisation.Abbreviations BAP
6-benzylaminopurine
- NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962)
- KM
Kao and Michayluk (1975)
- g.f.wt.
gram fresh weight 相似文献
10.
Transient gene expression in electroporated protoplasts of Eucalyptus citriodora Hook 总被引:3,自引:0,他引:3
G. Manders A. V. P. dos Santos F. B. d'Utra Vaz M. R. Davey J. B. Power 《Plant Cell, Tissue and Organ Culture》1992,30(1):69-75
Protoplasts isolated from cotyledons of Eucalyptus citriodora were electroporated using a rectangular pulse, with plasmid carrying the cat gene. The levels of transient expression and protoplast viability were influenced by the voltage and pulse duration. At a field strength of 800 V cm-1 (1000 s), a protoplast viability of 57%, and 47% conversion of 14C-chloramphenicol to its acetylated forms, were obtained. Expression levels were improved by an increase in plasmid concentration (up to 60 g ml-1), and also by the addition of carrier DNA. Gene expression was further enhanced by the addition of 40% (w/v) PEG, in the presence of the carrier DNA, to the protoplasts after electroporation.Abbreviations BAP
6-benzylaminopurine
- CAT
chloramphenicol acetyltransferase
- CPW 13M
CPW salts medium with 13% (w/v) mannitol
- DC
direct current
- FDA
fluorescein diacetate
- f. wt
fresh weight
- GUS
-glucuronidase
- K
Kao (1977)
- MES
2-N-morpholinoethane sulfonic acid
- MS
Murashige & Skoog (1962)
- NAA
-naphthaleneacetic acid
- PVP-10
polyvinylpyrrolidone (Av MW 10,000)
- TLC
thin layer chromatography 相似文献
11.
K. Sankara Rao 《Plant cell reports》1988,7(7):546-549
Rapid multiplication of axillary meristems and direct shoot development occurred from nodal explants of mature Eucalyptus tereticornis Sm. with 5.3 M NAA, 1.1 M IAA and 4.4 M BA in Murashige-Skoog medium. Repeated subcultures of the second generation shoot cultures into low cytokinin-auxin containing media (0.44–0.88 M BA+0.1 M NAA) yielded axillary microshoots in large numbers. Half-strength MS liquid medium with 4.9 M IBA, 5.5 M IAA and 5.3 M NAA for four days, half-strength semi-solid hormonefree MS medium with charcoal, and MS liquid medium without charcoal and hormones, in sequence, induced rooting of shoots in the dark. This system is suitable for the mass propagation of this difficult-to-root eucalypt.Abbreviations BA
6-benzyladenine
- IAA
indole-3-acetic acid
- IBA
-indole-3-butyric acid
- 2-iP
isopentyl adenine
- Kn
kinetin
- MS
Murashige-Skoog
- NAA
-naphthalene acetic acid
- PVP
polyvinylpyrrolidone 相似文献
12.
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
13.
F. Bekkaoui M. Pilon E. Laine D. S. S. Raju W. L. Crosby D. I. Dunstan 《Plant cell reports》1988,7(7):481-484
The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm–1 having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli -glucuronidase gene (-GUS) as a marker gene revealed very high levels of -GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since -GUS-like activity increased when plasmid DNA was present during electroporation.Abbreviations CAT
chloramphenicol acetyltransferase
- -GUS
-glucuronidase
- MUG
4-methyl umbelliferyl glucuronide
- F
microfarads
NRCC No. 29150 相似文献
14.
Jagroop S. Dahiya 《Plant and Soil》1991,134(2):297-304
A broad-host-range plasmid (pEA2-21) containing a Bradyrhizobium sp (F-4) nod DABC-lacZ translation fusion was constructed and used to monitor nod gene expression in response to pigeonpea root exudate. Two nod-inducing compounds were isolated and identified. Spectral analysis using ultraviolet absorption, infrared spectra, proton nuclear magnetic resonance, and mass spectrometry showed that the two inducers were 5,4-dihydroxy-6-(3-methyl-2-butenyl)-2, 2-dimethyl pyrano-[5, 6:7, 8]-flavanone (cajaflavanone) and 2,4,5-trihydroxy-5-isopentenyl-6, 7-dimethylchromene iso-flavanone (cajanone). When pEA2-21 was introduced into Rhizobium trifolii and R. meliloti cajanone and cajaflavanone did not induce nod gene indicating that specificity of induction appears to be influenced by the host-strain genome. 相似文献
15.
Faheem Aftab Yusuf Zafar Kouser A. Malik Javed Iqbal 《Plant Cell, Tissue and Organ Culture》1996,44(1):71-78
Embryogenic callus was developed from young leaves of sugarcane (Saccharum spp.hybrid, cv. CoL-54). A good embryogenic callus response was achieved using MS basal medium containing 2.0 mol (0.5 mg l-1) picloram under dark conditions at 27±1°C. Initiation of fast growing homogeneous cell suspension cultures was achieved in MS and AA media, both supplemented with g mol (2 mg l-1) 2,4-d and 500 mg l-1 CH. Embryogenic callus was reinitiated from embryogenic cell suspension cultures using MS medium containing 30 g l-1 sucrose, 500 mg l-1 CH and 2.26 mol (0.5 mg l-1) 2,4-d after 4–6 weeks of culture under 16-h photoperiod conditions. Plant regeneration was achieved after about 4 weeks in MS medium lacking growth regulators but containing CH (500 mg l-1) and sucrose (60 g l-1). Rooting was enhanced by transferring regenerated plantlets to half strength MS basal medium.Totipotent protoplasts with an average yield of 2.0×107 to 1.0×108 ml-1 were obtained from embryogenic cell suspension cultures at log phase, i.e., 4–5 days after transfer to fresh media. The best growth response was achieved when protoplasts were cultured in a modifed KM8P medium at the density of 2.0×105 m l-1. Protoplasts were mainly embedded in 0.8% sea plaque agarose. Division efficiency of 22.2% was achieved after 20 days of culture and 0.26% of microcolonies continued growth and formed microcalluses after 30 days of culture under dark conditions. Microcalluses were proliferated in MS medium having 2,4-d (2 mg l-1) under 16-h photoperiod. Transferring these embryogenic calluses in MS medium +9.29 mol kinetin (2 mg l-1) +5.37 mol NAA (1.0 mg l-1) + activated charcoal (200 mg l-1) for 5 weeks favoured plant regeneration. Shoots and roots were further proliferated in half strength MS basal medium for 2–4 weeks. Regenerated plants were transferred to autoclaved sand for 2 weeks under 16-h photoperiod in growth room and transferred to soil in a greenhouse to raise to maturity.Abbreviations MS
salts of Murashige & Skoog (1962) basal medium
- AA
salts of Muller & Grafe (1978) basal medium
- N6
saits of Chuet al. (1975) basal medium
- 2,4-d
2,4-dichlorophenoxyacetic acid
- CH
casein hydrolysate
- KM8P
protoplast culture medium of Kao & Michayluk (1975)
- KPR
protoplast culture medium of Kao (1977)
- P9
protoplast culture medium (Chen & Shih, 1983)
- BA
Benzyladenine
- Picloram
4-amino-3,5,6-trichloropicolinic acid
- NAA
Naphthalene acetic acid 相似文献
16.
Summary
Paecilomyces
varioti produced an extracellular xylanase and B-xylosidase when cultured in a medium containing xylan and corn steep liquor. Xylose (2%, w/v) totally inhibited production of both enzymes. The enzymes were purified and both had a pH optimum of 4.0. The xylanase had a molecular weight of 20,000, an isoelectric point of 5.2 and was inactive on all substrates tested except xylan. The -xylosidase, a glycoprotein, had a molecular weight of 67,000, an isoelectric point of 4.0 and had highest activity on p-nitrophenyl--D-xyloside. The xylanase had a Km of 49.5 mg/ml for xylan and the -xylosidase had a Km of 5.4 mM for p-nitrophenyl--D-xyloside. 相似文献
17.
Cornelis H. Hokke Jos G. M. van der Ven Johannis P. Kamerling Johannes F. G. Vliegenthart 《Glycoconjugate journal》1993,10(1):82-90
Incubation of synthetic Man\1-4GlcNAc-OMe, GalNAc1-4GlcNAc-OMe, Glc1-4GlcNAc-OMe, and GlcNAc1-4GlcNac-OMe with CMP-Neu5Ac and rat liver Gal1-4GlcNAc (2-6)-sialyltransferase resulted in the formation of Neu5Ac2-6Man1-4GlcNAc-OMe, Neu5Ac2-6GalNAc1-4GlcNAc-OMe, Neu5Ac2-6Glc1-4GlcNAc-OMe and Neu5Ac2-6GlcNAc1-4GlcNAc-OMe, respectively. Under conditions which led to quantitative conversion of Gal1-4GlcNAc-OEt into Neu5Ac2-6Gal1-4GlcNAc-OEt, the aforementioned products were obtained in yields of 4%, 48%, 16% and 8%, respectively. HPLC on Partisil 10 SAX was used to isolate the various sialyltrisaccharides, and identification was carried out using 1- and 2-dimensional 500-MHz1H-NMR spectroscopy.Abbreviations 2D
2-dimensional
- CMP
cytidine 5-monophosphate
- CMP-Neu5Ac
cytidine 5-monophospho--N-acetylneuraminic acid
- COSY
correlation spectroscopy
- DQF
double quantum filtered
- HOHAHA
homonuclear Hartmann-Hahn
- MLEV
composite pulse devised by M. Levitt
- Neu5Ac
N-acetylneuraminic acid
- Neu5Ac2en
2-deoxy-2,3-didehydro-N-acetylneuraminic acid 相似文献
18.
Growth and oxygen uptake of potato callus is faster in an oxygen-enriched atmosphere (70% oxygen, v/v; oxygen-callus) than in air (20% oxygen, v/v; air-callus). Especially the non-mitochondrial, so-called residual respiration is increased in oxygen-callus. The capacities of the mitochondrial respiratory pathways (cytochrome pathway, Vcyt and alternative pathway, Valt) are also higher in this callus. In both callus types only a small part of the alternative pathway capacity is used in uninhibited respiration. The lower oxygen uptake of air-callus at normal air oxygen pressures is partially due to diffusional impedance. Measurement of the respiratory parameters of air-callus in oxygen-saturated medium leads to higher values than measurement in air-saturated medium, although these values are still lower than those of oxygen-callus.ATP-production was calculated from the oxygen-uptake data and compared with the dry weight production of the callus to give values of 10.0 and 10.8 g dry weight produced.-mol ATP-1, for air-callus and oxygen-callus respectively. As no harmful side-effects are observed, cultivation of callus under elevated oxygen pressures may be useful, when rapid callus-growth is necessary.Abbreviations AA
antimycin A; A;
- BHAM
benzohydroxamate
- DW
dry weight
- FW
fresh weight
- 8-OHQ
8-hydroxyquinolin
- RC
respiratory control
- SHAM
salicylhydroxamate 相似文献
19.
Summary Novel sugars, oligoglucosyl-inositols, which were synthesized using CGTase fromBacillus
ohbensis, stimulated the growth ofBifidobacterium. The enzyme catalyzed transglucosylation from -1,4-maltodextrin (donor) tomyo-inositol (acceptor). Of donors examined, -cyclodextrin gave superior oligoglucosyl-inositol yield of 56.6% (w/w) based on the conversion ratio of incubated inositol. Maltosyl-inositol stimulated growth ofB.
adolescentis by 194% when compared with glucose. 相似文献
20.
Conditions which induce a transmembrane electrical potential, positive with respect to the inside of membrane vesicles, result in a substantial (4–12-fold) stimulation of the activity of membrane-associated -glucan synthetases in a membrane preparation derived from the developing cotton (Gossypium hirsutum L.) fiber. Induction of electrical potentials which are negative with respect to the inside of the membrane vesicle results in little or no stimulation of -glucan synthesis. Those products whose synthesis is stimulated are mainly -1,3-glucan, but there is also a considerable increase in -1,4-glucan. No -1,4-glucan (starch) was detected in the reaction products. A transmembrane pH gradient was found to have no effect on -glucan synthesis. The results indicate that a transmembrane electrical potential can influence, either directly or indirectly, the activity of membrane-associated polysaccharide synthetases.Abbreviations UDP-glucose
uridine-5-diphosphoglucose
- PEG
polyethylene glycol
- BTP
bistrispropane (1,3-bis[tris(hydroxymethyl)methylamino]propane)
- MES
2(N-morpholino)ethanesulfonic acid
- VAL
valinomycin 相似文献