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1.
Protoplasts isolated from mesophyll cells of Eruca sativa Lam., cultured on suitable medium, underwent sustained cell divisions to form calli. The plating efficiency was found to be 0.4%. The protoplast-derived calli subsequently produced plantlets through organogenesis (15.71%) and somatic embryogenesis (11.25%). Regenerated plants exhibited normal appearance. These results indicate potential to introgress desirable traits from this wild crucifer into important oilseed and cole Brassicas by protoplast fusion and hybrid recovery.Abbreviations B5
Gamborg et al., 1968
- K3
Kao and Michayluk, 1974
- MS
Murashige and Skoog, 1962
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthalene acetic acid
- GA3
Gibberellic acid 相似文献
2.
S. B. Narasimhulu P. B. Kirti S. R. Bhatt Shyam Prakash V. L. Chopra 《Plant cell reports》1994,13(11):657-660
Camelina sativa is a wild crucifer that is reported to be resistant to Alternaria blight. Polyethylene glycol mediated fusion was attempted between protoplasts from etiolated hypocotyls of Brassica carinata and mesophyll protoplasts of Camelina sativa. The mean frequency of heterokaryons was 6.8%. Three hybrid shoots were regenerated, each from a single fusionderived callus. These shoots failed to produce roots capable of withstanding transplantation. Confirmation of hybridity was obtained from the morphology of in vitro produced leaves, somatic chromosome number in leaf tips, and restriction fragment length polymorphism for a nuclear rDNA probe. Analysis for organelle constitution using RFLPs indicated that the hybrid contained chrloroplasts derived from the wild species and mitochondria from the cultivated Brassica species.Abbreviations 2,4-D
2,4-dichlorophenoxy-acetic acid
- IAA
Indole-3-acetic acid
- NAA
-Naphthaleneacetic acid
- IBA
Indole-3-butyric acid
- GA3
gibberellic acid
- BAP
6-Benzylaminopurine
- MS
Murashige and Skoog (1962) basal medium 相似文献
3.
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
4.
Callus induction and plant regeneration were studied in 15 cultivars of the facultative apomictic species Poa pratensis L. (Kentucky bluegrass).The tissue culture responses of mature seeds and immature inflorescences were compared. Murashige and Skoog's (MS) medium, supplemented with 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and maintenance. Plants could be regenerated from compact and friable callus on MS medium devoid of 2,4-D. Plants were recovered from 14 cultivars at a high frequency (up to 79% of the callus cultures) when young inflorescences were used as the explant material and from only 3 cultivars, at a low frequency (up to 3%), with seeds. Somatic embryos were observed in callus cultures of many cultivars. Fully developed germinating somatic embryos were occasionally observed. Plant regeneration appeared to take place both via somatic embryogenesis and organogenesis. Plants were generally green but albino shoots developed at a low frequency from friable callus.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog's (1962) medium
- IAA
indole-3-acetic acid
- N6
medium of Chu et al. (1975) 相似文献
5.
Electrofusion of protoplasts from two complementary nitrate reductase deficient mutants of Nicotiana plumbaginifolia has resulted in somatic hybrid lines. Mesophyll protoplasts isolated from the cofactor mutant CNX 20 and fluorescein diacetate stained protoplasts derived from a cell suspension culture of the NA 36 line, being defective in the apoenzyme, were used in the fusion experiments. In total, 594 lines were recovered which could proliferate on a selective medium with nitrate as the sole nitrogen source. This is including 141 putative hybrid lines which were obtained after transfer of 1048 heterokaryons with a micromanipulator one day after electrofusion. The hybrid character of some of the selected lines was confirmed by nitrate reductase activity measurements. Plants were grown from hybrid calli.Abbreviations NR
nitrate reductase
- FDA
fluorescein diacetate
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
benzylaminopurine
- NAA
naphthaleneacetic acid
- NED
N-1-naphtyl-ethylenediamide hydrochloride
- PEG
polyethylene glycol
- AC
alternating current
- DC
direct current 相似文献
6.
Compact, friable and embryogenic calli were initiated from immature inflorescences and young leaf bases of one week old seedlings of Paspalum scrobiculatum cultured on MS medium supplemented with 2,4-D. A stable, embryogenic suspension culture was initiated from these calli and maintained in a liquid version of the same MS medium. Embryogenic calli and somatic embryos were obtained by plating suspension culture cells onto semi-solid medium containing 2,4-D. Complete, normal plantlets developed on 2,4-D free medium at a high frequency from somatic embryos. NAA and BAP in the medium promoted plant development.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
Abscisic acid
- MS
Murashige and Skoog (1962)
- CM
Coconut milk 相似文献
7.
Y. Asano 《Plant cell reports》1989,8(3):141-143
Embryogenic callus cultures were initiated from the mature caryopses ofZoysia japonica. Plant regeneration was through precocious germination of somatic embryos. Protoplasts were isolated from the callus and cultured in a medium solidified with agarose. Numerous calli were recovered after transferring protocolonies onto an agar medium.Abbreviations MS
Murashige & Skoog
- CH
Casein hydrolysate
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MES
2-(N-Morpholino)ethanesulfoic acid 相似文献
8.
Surface sterilized seeds and mesocotyls from sterile seedlings from Panicum bisulcatum Thumb., as well as basal parts of leaves and mesocotyls from sterile seedlings, and seeds from Panicum milioides Nees ex. Trin were used as explants to induce callus on a Murashige and Skoog medium supplemented with 2.5 to 10 mg/l of 2,4-D. Subculturing of the white callus from P. milioides and of the brown callus from P. bisulcatum on a medium containing 0.1 mg/l 2,4-D and 10 g/l sucrose led in both species to the appearance of green structures from which plants could be regenerated. Plants were regenerated by an organogenetic process in P. milioides, while P. bisulcatum plants were regenerated both via organogenesis and somatic embryogenesis. 1032 and 94 plants, from P. bisulcatum and P. milioides, respectively, were transferred into soil, and about 90% of them were grown to maturity and set seeds.Abbreviations MS
Murashige and Skoog medium (15)
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indoleacetic acid 相似文献
9.
Protoplasts from embryogenic calli of Citrus reticulata Blanco cv. Ponkan and Citropsis gabunensis (Engl.) Swing. & M. Kell (Cabon Cherry Orange), were isolated and fused using electric current. Maximum fusion frequency was obtained with AC at 75 kV/cm (1.0 MHz) for 15 s, followed by DC square-wave pulses at 1.25 kV/cm for 40 s. Fusion-treated protoplasts were cultured on MT medium containing no growth regulators, solidified with 0.6% Bacto Difco agar. Protoplast-derived calli were proliferated on MT medium containing 1 mg/l zeatin and 0.9% agar. A total of 31 lines of somatic hybrid calli were obtained by screening on the basis of chromosome count and isozyme analysis. The somatic hybrids were tetraploid (2n=36). Plants were regenerated from the calli via somatic embryogenesis. The somatic hybrid plants exhibited morphological characteristics intermediate to the parental plants.Abbreviations 2,4-D
2, 4-dichlorophenoxyacetic acid
- BA
6-benzylaminopurine
- MT
Murashige and Tucker (1969)
- PEG
polyethylene glycol
- AC
alternating current
- DC
direct current 相似文献
10.
Plantlets were obtained from leaf explants of a Labiatae tree — Leucosceptrum canum Sm. using plant tissue culture techniques. Two types of calli proliferated from the leaf explants when grown on different media, one of which was amenable to somatic embryogenesis. Differentiation of the embryoids started from the fourth passage of culture and continued up to the seventh passage. The number of embryoids decreased with the age of the callus. The capacity of such embryoids to form entire plantlets was studied using different nutrient mileux. Embryoids formed plantlets on Murashige and Skoog's (MS) medium fortified with benzylaminopurine plus indolebutyric acid. Organogenesis was observed in shoot-buds derived from explants of in vitro regenerated plantlets on MS basal medium supplemented with benzylaminopurine. Culture regenerated plantlets were transferred to MS medium without sucrose and growth hormones; finally transferred to pots containing sterile vermiculite where they are growing.Abbreviations MS
Murashige and Skoog's medium
- 2,4-D
2,4-dichlorophenoxy acetic acid
- NAA
naphthaleneacetic acid
- IAA
indoleacetic acid
- IBA
indolebutyric acid
- Kn
kinetin
- BAP
benzylaminopurine
- CW
coconut water 相似文献
11.
Gurvinder S. Cheema 《Plant cell reports》1989,8(2):124-127
Somatic embryogenesis and plantlet formation were obtained from callus and cell suspension cultures of 40-year- old Himalayan Poplar (Populus ciliata Wall ex Royle). Callus and cell suspensions were obtained by transfer of inoculum of semiorganized leaf cultures, which were maintained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), to MS with 2,4-dichlorophenoxyacetic acid (2,4-D). Reduction of 2,4-D concentration during subsequent subculture of cell suspensions resulted in the formation of embryoids. These embryoids developed further only after being transferred to agar-based MS medium supplemented with BAP and naphthalene acetic acid. Loss of embryogenic potential was observed in cell suspensions after 6 subcultures. However, callus cultures retained the embryogenic potential even after repeated subcultures for more than a year. Plantlets could be successfully hardened and grown in natural outdoor conditions.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthalene acetic acid
- MS
Murashige and Skoog (1962) medium 相似文献
12.
A method for the induction of somatic embryogenesis in callus cultures, using explants from mature leaves of Vicia narbonensis L., is described. Callus developed on a solid medium (Murashige and Skoog 1962), which was supplemented with low concentrations of picloram and benzylaminopurine. Subsequent culture was carried out in different liquid media (culture length four months). The gradual reduction of auxin and cytokinin concentrations, and the addition of glutamine and pyridoxal·HCl were favourable. Somatic embryos appeared on solid media without phytohormones.Abbreviations MS
Murashige and Skoog (1962)
- 2,4 D
dichlorophenoxy acetic acid
- KIN
6-furfurylaminopurine
- BAP
6-benzylaminopurine
- picloram
4-amino-3,5,6-trichloropicolinic acid
- NAA
1-naphthalene acetic acid
- p-CPA
parachlorophenoxy acetic acid
-
M1 - M7
media numbers (details in materials and methods) 相似文献
13.
S. L. Kothari D. C. Monte J. M. Widholm 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1986,72(4):494-502
Summary Protoplasts from different Daucus carota L. cell strains carrying resistance to glyphosate, 5-methyltryptophan, sodium selenate or selenocystine were fused in three combinations using dextran. Clones were selected for both of the resistances carried by the individual parental strains in medium with both inhibitors. No doubly resistant colonies formed from unfused controls or from protoplasts from each individual parental strain alone. Suspension cultures from the selected clones contained predominantly the additive chromosome numbers of the parental strains. Apparently the four resistances used are expressed dominantly in fusion hybrids. Analysis of mitochondrial DNA showed that recombination occurred in one fusion combination since the mitochondrial DNA in the hybrid cells was different from that of either parent as shown by restriction endonuclease fragment analysis. Mitochondrial DNA in the other two somatic hybrid combinations was parental. Thus, a dominant, nuclear resistance marker system has been developed to select efficiently for somatic hybrids in which mitochondrial DNA recombination can be studied.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- FW
fresh weight
- Glp
Glyphosate
- mt
mitochondrial
- MX
Murashige and Skoog (1962) medium containing 0.4 mg/l 2,4-D
- 5MT
5-methyltryptophan
- MXG
MX containing (5% Glucose)
- SC
selenocystine
- SS
sodium selenate 相似文献
14.
Protoplasts were isolated from cotyledon, hypocotyl and mesophyll cells of Corchorus capsularis L., a major fibre crop, by one step enzyme digestion method. They were further cultured successfully on modified KM-8p (Kao and Michayluk, 1975) medium to form microcalli. The required cultural conditions could be used to achieve 34% to 78% plating efficiency, depending upon the source of protoplasts. Hypocotyl protoplasts gave the highest plating efficiency. On transfer to regeneration medium, somatic embryos developed at high frequency. The present success is a significant step forward in the development of meaningful plant cell culture methods for application in jute.Abbreviations B5
Gamborg et al., 1968
- MS
Murashige and Skoog, 1962
- SH
Schenk and Hildebrandt, 1972
- Ad-SO4
Adenine sulphate
- BAP
6-benzylaminopurine
- NAA
1-naphthalene acetic acid
- GA3
Gibberellic acid
- 2,4-D
2,4-dichlorophenoxy acetic acid
- Kn
Kinetin
- IBA
Indole-3-butyric acid 相似文献
15.
Protoplasts were released from embryogenic suspension culture of Paspalum scrobiculatum and cultured in either liquid or semisolid KM medium supplemented with 2,4-D in the dark at 24°C with or without a feeder layer. Cell wall formation was observed in 75% of the plated protoplasts. Microcolonies developed after 10 d of culture, which in turn formed callus upon transfer to M-2 medium (Nayak and Sen, 1989). The highest plating effeciency (ca 7%) was obtained in thin-layer liquid culture. The macrocalli formed somatic embryos which regenerated to plantlets. The plantlets were grown to flowering plants upon transfer to soil.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
Fluoresceine diacetate
- MES
2-(N-Morpholino) ethanesulfonic acid
- MS
Murashige & Skoog medium (1962) 相似文献
16.
Five varieties of durum wheat: Appulo, Ofanto, Latino, Creso, and Castello (Triticum durum Desf.) adapted to the semi-arid mediterranean environment have been tested for their in vitro response. Compact, embryogenic, highly regenerable calli originated from primary callus derived through proliferation of the scutellum of immature embryos explanted in the presence of 2,4 dichlorophenoxyacetic acid. Selective subculture of the white, compact, embryogenic sectors led to the establishment of long-term cultures. Regeneration occurred on hormone-free medium either via germination of somatic embryos, or via multiple-shoot formation probably due to precocious germination of somatic embryos. The three varieties, Ofanto, Creso and Appulo, were the best responding genotypes. Callus fragmentation and two subsequent transfers onto fresh medium at 7-day intervals yielded a frequency of plant regeneration of some 25–40 plantlets per gram fresh weight callus in 21 days on Murashige and Skoog's hormone-free medium. Plantlets could be efficiently established in soil, thus confirming the possibility of biotechnological approaches with varieties of this crop species.Abbreviations E
embryogenic
- NE
non embryogenic
- MS
Murashige and Skoog's (1962) medium
- 2,4-D
2,4 dichlorophenoxyacetic acid
- DAA
days after anthesis
- FWT
fresh weight tissue 相似文献
17.
A new approach to direct somatic embryogenesis in Medicago 总被引:2,自引:0,他引:2
A highly efficient system for direct somatic embryogenesis is described. Leaf sections originating from young trifoliate leaves of Medicago falcata line 47/1–5 and Medicago sativa line No2/9R, directly produced embryos after cultivation in liquid B5IV induction medium. In comparison with indirect somatic embryogenesis the system omits the callus stage and thus allows shortening of the process of somatic embryogenesis in alfalfa by 35–40 days. It permits the avoidance of secondary changes occurring during the process of dedifferentiation. A modified B5/3H medium containing Polyethylene Glycol 6000 promoted embryo development from globular up to torpedo stage. It was clearly shown that 2.5% Polyethylene Glycol stimulated this process for both H. falcata 47/1–5 and M. sativa No 2/9R. Maturation of torpedo stage embryos was carried out on solidified or liquid abscisic acidcontaining medium. A 30M abscisic acid concentration was optimal in allowing one embryo to yield one plant. Somatic embryo conversion to plants and plant regeneration was performed on Murashige and Skoog medium. Regenerated plants showed a normal morphology.Abbreviations ABA
Abscisic acid
- B5
Medium of Gamborg et al.(1968)
- COT
Cotyledone stage embryos
- 2,4-D
2,4-dichlorphenoxyacetic acid
- FW
Fresh weight
- GA3
Gibberellin A3
- MS
Medium of Murashige and Skoog (1962)
- PEG
Polyethylene Glycol
- POLY
Polyembryos 相似文献
18.
A novel method for increasing the frequency of somatic embryogenesis in wheat tissue culture by NaCl and KCl supplementation 总被引:2,自引:0,他引:2
The effect of NaCl, KCl and LiCl on the growth and morphogeneis of tissue cultures originating from immature embryos of four wheat (Triticum aestivum L.) and one triticale (Triticosecale)varieties was investigated. The morphogenetic pathway to plant regeneration in Chinese Spring wheat was determined as incomplete somatic embryogenesis because the differentiation and subsequent germination of the shoot apices happened in the early phase of embryo development. Culture medium supplemented by NaCl suppressed the differentiation of shoot apices resulting in the development of more typical somatic embryoids. Forty mM concentrations of both NaCl or KCl increased the formation of somatic embryos in Chinese Spring. Arthur and GK Kincso wheat varieties while Lasko triticale regenerated well without the addition. The salts inhibited plantlet formation from somatic embryoids so the salts supplement should be omitted. Forty mM LiCl inhibited growth while 10mM LiCl had no effect on growth or embryogenesis.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) medium 相似文献
19.
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
20.
Protoplasts isolated from young, unexpanded leaves of the wild lettuce species, Lactuca saligna, divided to give colonies, when plated at low density in KP8 medium solidified with 1% (w/v) agarose. Sustained colony development was dependent upon the incorporation of the bead culture approach. Plant regeneration, via organogenesis, followed the transfer of colonies firstly to K8 medium and then to M/S medium supplemented with IAA and 6-BAP, both solidified with 1% (w/v) agarose. Excised shoots were rooted on agar-solidified M/S medium lacking phytohormones. The potential of this species as a source of race-non-specific mildew resistance for transfer into lettuce (L. sativa) via somatic hybridisation is discussed.Abbreviations 6-BAP
6-benzylamine purine
- IAA
indoleacetic acid
- M/S
Murashige and Skoog (1962)
- fwt
fresh weight 相似文献