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1.
Somatic embryos were obtained from immature zygotic embryos of Cedrela fissilis Well. (Meliaceae), after a culture period of 12 months, with regular subcultures every 6–8 weeks. Callus was developed on explants in 2 months
on Murashige and Skoog (MS) medium containing 2,4 dichlorophenoxyacetic acid (2,4-D) or picloram (PIC). When the calli were
transferred to fresh medium, embryogenic tissue appeared on MS + 45 μM 2,4-D, or 22.5 μM 2,4-D + 0.4 μM 6-benzyladenine (BA),
or 20.7 μM PIC after 6 months. Sub-culture of embryogenic tissue in MS medium supplemented with 4.5 μM 2,4-D resulted in the
differentiation into somatic embryos after further 4 months. Repeated secondary somatic embryogenesis was achieved by regular
subculture on this medium. Maturation and conversion of somatic embryos into plantlets was achieved on MS medium without plant
growth regulators and the conversion frequency was approximately 12.5 %. The plantlets were successfully acclimatized in pots
with soil. Histological studies showed that somatic embryos had no detectable connection with the mother explants and that
somatic embryos in advanced stages were bipolar with shoot and root apical meristems, they contained vascular system and showed
typical characteristics of a somatic dicotyledonous embryo. 相似文献
2.
Blanca Olivia Cañedo Ortiz Martha Evelia Pérez Reyes Eugenio Pérez Molphe Balch 《In vitro cellular & developmental biology. Plant》2000,36(4):268-272
Summary Efficient plant regeneration systems via somatic embryogenesis have been developed for Acacia farnesiana and Acacia schaffneri [Leguminosae (Mimosoideae)]. The protocol used in this study consisted of placing immature, zygotic embryos of these species
in Murashige and Skoog semi-solid basal medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.65 μM kinetin to induce callus. Some parts of the callus were used for direct embryo differentiation and others for establishment
of cell suspension cultures. In the first case, somatic embryos were produced on semi-solid differentiation media without
growth regulators or with abscisic acid (ABA). The higher number of somatic embryos, 345 and 198 embryos per g callus in A. farnesiana and A. schaffneri, respectively was obtained in media without growth regulators, but adding ABA increased the percentage of embryos that reached
more advanced differentiation stages. The production of somatic embryos was achieved starting from cell suspensions only when
these suspensions were plated into the semi-solid differentiation medium. Somatic embryos germinated on medium containing
217 μM adenine sulfate with efficiencies of 69% in A. farnesiana and 47% in A. schaffneri. Some somatic embryos that developed into plantlets were acclimatized in the greenhouse, and they grew into normal plants. 相似文献
3.
Mature embryos of Acanthopanax senticosus explanted on Murashige and Skoog (MS) medium with 0.5 mg/1 2,4-D developed somatic embryos directly from swollen cotyledon and embryo axes within one to two months. When the somatic embryos were transferred to medium supplemented with 2,4-D (0.5 mg/1) or IAA (1–3 mg/1) or Zeatin (0.5 mg/1) and NAA (0.2 mg/1), additional somatic embryos developed. Most (93%) embryos germinated on the above medium without 2,4-D. Sixty-two percent of the plantlets survived in soil. Histological observations revealed that the somatic embryos originated from cell masses of epidermal and sub-epidermal origin. There was no cytological separation zone between the somatic embryos and cultured expiants. Consequently, embryos were difficult to separate from their expiant tissue. 相似文献
4.
Immature embryos of Quercus acutissima were collected weekly beginning 5 weeks post-fertilization and cultured on modified MS(Murashige and Skoog) medium containing 1,000 mg/l glutamine and 5 mM proline with different combinations of IBA(0.5–10.0 mg/l) and BA(0 or 1.0 mg/l) in light. The highest percentage of embryogenic cultures occurred on the medium containing 0.5 mg/l IBA or 1.0 mg/l BA and 0.5 mg/l IBA. Four weeks after initiation, the embryogenic cultures were transferred to MS medium without plant growth regulators and cultured for 4 weeks. The somatic embryos were then transferred to germination medium. The best germination results were achieved from WPM(Woody Plant Medium) containing 0.1 mg/l BA. Plantlets from somatic embryos were incubated on WPM supplemented with 0.2 mg/l BA for 4 weeks and plantlets with well developed shoots and roots were transplanted to perlite and peat moss(11, v/v) mixtures and placed in a culture room. After being hardened off for 8 weeks, they were transferred outdoors where they grew.Abbreviation BA
N6-benzyladenine
- IBA
indole-3-butyric acid
- GA3
gibberellic acid
- ABA
abscisic acid
- MS
Murashige & Skoog Medium
- WPM
Woody Plant medium 相似文献
5.
Isabel Santos Isabel Guimarães Roberto Salema 《Plant Cell, Tissue and Organ Culture》1994,37(1):83-86
Leaf explants of Nerium oleander L. produced masses of callus when both an auxin and a cytokinin were included in the medium. Leaves cultured on the B5 medium of Gamborg et al. supplemented with 2,4-dichlorophenoxyacetic acid (2,4-d; 9.05 M) plus benzyladenine (BA; 4.4 M) produced callus and profuse rhizogenesis was observed from callus developed from older leaves. On Murashige & Skoog medium (MS) with the same concentration of 2,4-d and BA, explants from young and mature leaves produced callus, but only that from young leaves was embryogenically competent. Globular somatic embryos were obtained when embryogenic cells were cultured on MS medium without growth regulators. Both normal and anomalous development of embryos occurred in either liquid or gelled medium. Plantlets were produced faster when mature embryos were cultured on either solid medium or placed on Sorbarod plugs soaked with this same medium but with 1% sucrose. Plantlets with three nodes were transferred to pots and acclimatized in a growth chamber and afterwards transferred to garden beds. 相似文献
6.
Somatic embryogenesis and whole plant regeneration were achieved in callus cultures derived from hypocotyl, cotyledon and leaf explants excised from seedlings of Gymnema sylvestre. Embryogenic callus was induced on Murashige and Skoog (MS) medium containing 2,4-D (0.5–5.0 M) +BA (0.5–2.0 M) and 2% (w/v) sucrose in 6–8 weeks of culture. Globular/heart stage embryos developed on induction medium. These embryos produced torpedo and cotyledon stage embryos upon sub-culturing on embryo maturation medium EM8 (medium containing MS salts, B5 vitamins, 0.5 M BA and 2% sucrose). Embryo germination and plantlet formation was achieved by sub-culturing mature embryos on fresh EM8 medium. The plantlets were acclimatized in the greenhouse. 相似文献
7.
Hamidah Musa Karim Abdul Ghani Abdul Debergh Pierre 《Plant Cell, Tissue and Organ Culture》1997,48(3):189-193
The effects of xylooligosaccharides isolated from the cell walls of Betula platyphylla var. japonica on cells and protoplasts
of Pinus radiata were examined. The addition of a semi-purified mixture of xylooligosaccharides at a concentration of 5μg.ml−1 promoted elongation of cultured cells, whereas the neutral fraction of this mixture had no effect; a similar effect was seen
in the presence of conditioned medium. The unfractionated mixture of xylooligosaccharides was also found to enhance the viability
of protoplasts prepared from cell cultures of Pinus radiata in a concentration dependent manner, highly similar to the effect
provided by addition of medium conditioned by pine cells. Such effects are considered to be due to the addition of components
that play a structural role in the cell wall of pines. It is inferred that the acidic components of the xylooligosaccharide
mixture derived from t Betula are responsible for this effect in the distant pine species. It is speculated that acidic xylooligosaccharides
operate either by replacing, or mimicking, the natural cell wall components required for growth and development of pine cultured
cells.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
8.
Friable callus was initiated from shoot apices of Gloriosa superba L. on basal MS medium supplemented with 2, 4-D (4mg L(-1)) + Kn(5 mg L(-1)) + CH(10 mg L(-1)) + CW(20%). Subculture of callus on the same medium after 4-5 weeks showed induction of large number of somatic embryos, which was confirmed with histological studies. Development of embryoids in plantlet took place when the embryogenic callus was transferred to basal MS medium supplemented with BAP (5 mg L(-1)), CH(50 mg L(-1)) +CW(20%). Roots were developed by subculturing them on to the medium containing Kn or BAP (5 mg L(-1)) and IBA (4 mg L(-1)). Plantlets were successfully transferred to pots containing mixture of soil, sand and farmyard manure (2:1:1). 相似文献
9.
High frequency plant regeneration via somatic embryogenesis has been induced from in vitro shoot-base cultures of seedlings of garden leek (Allium porrum L.). Four main steps are involved in the procedure using BDS medium: - shoot multiplication with 17.6 mM benzyladenine; - induction of nodular callus from the in vitro shoot base with 9 mM 2,4-dichlorophenoxyacetic acid; - initiation of embryogenic callus from nodular callus with 9 mM 2,4-dichlorophenoxyacetic acid +7.6 mM abscisic acid; - plant regeneration from embryogenic callus with 9.8 mM N6-(2-isopentenyl)adenine. The presence of 2,4-dichlorophenoxyacetic acid in the medium and light conditions were shown to be essential for nodular callus induction and somatic embryogenesis. Abscisic acid was not a prerequiste for somatic embryogenesis, but it significantly increased the frequency. 相似文献
10.
Plants of two cytotypes (2n=2x=20, and 2n=3x=30) of pinto peanut (Arachis pintoi Krapov. & W.C. Gregory) were regenerated through somatic embryogenesis. Embryogenic calli were induced from shoot tips or
immature leaves dissected from in vitro growing plants. In the case of the diploid peanut the best somatic embryogenesis was achieved when shoot tips were cultured
on Murashige and Skoog (MS) medium supplemented with 10 mg dm−3
Picloram (PIC) and 0.1 mg dm−3 6-benzylaminopurine (BAP) or when explants from immature leaves were cultured on MS + 10 mg dm−3 PIC + 0.01 mg dm−3 BAP. In the case of triploid peanut the highest number of somatic embryos was obtained when shoot tips were cultured on MS
+ 10 mg dm−3 PIC + 0.01 mg dm−3 BAP or when immature leaves were cultured on MS + 20 mg dm−3 PIC + 0.01 mg dm−3 BAP. Somatic embryos were converted into plants by culture on MS + 0.01 mg dm−3 naphthaleneacetic acid + 0.01 mg dm−3 BAP. Plants were successfully transferred to pots in greenhouse. 相似文献
11.
Maria Ludovina S. Guimarães Gil S. Cruz João M. Montezuma-De-Carvalho 《Plant Cell, Tissue and Organ Culture》1988,15(2):161-167
Callus cultures with globular proembryogenic structures were induced from zygotic embryos and hypocotyl segments of Cyphomandra betacea on MS medium supplemented with 2,4-D. Proembryogenic structures produced somatic embryos and plantlets on regulator-free basal medium. Pieces of embryogenic callus subcultured on medium with the same original composition gave rise to new globular structures and the potential for plantlet regeneration has been maintained for over a year. The histological examination of these proembryogenic structures suggested that somatic embryos arise from single cells. Regenerated plants are phenotypically normal, having diploid chromosome numbers (2n = 24). 相似文献
12.
Friable callus was obtained from styles and flower pedicels of Lilium longiflorum Snow Queen and the Oriental lily hybrid Star Gazer on Murashige and Skoog (MS) media containing either 2 μm dicamba or 2 μm picloram. Cell suspension cultures were established by suspending the callus of L. longiflorum Snow Queen in liquid medium containing 2 μm dicamba. Through a purification process, a fine fast-growing cell suspension was obtained. This suspension was composed of
a homogenous population of small dense cells, which tended to organise into embryo like structures (ELS). In liquid culture
with the auxin dicamba, the ELS underwent continuous callus formation. When transferred to solidified hormone-free MS medium,
the ELS germinated, forming complete plantlets. Histological investigation showed that in the ELS both shoot and root meristems
were distinctly evident. It was concluded that the ELS obtained were in fact somatic embryos.
Received: 4 April 1997 / Revision received: 13 May 1997 / Accepted: 15 June 1997 相似文献
13.
A protocol has been developed for achieving somatic embryogenesis and plant regeneration from petiole-derived callus of Heracleum candicans Wall. Callus was initiated on MS medium supplemented with 0.5 mg l–1 2,4-D and 0.5 mg l–1 BAP and subcultured on a medium containing double strength MS macrosalts, 1 mg l–12,4-D and 0.25 mg l–1 Kn. Numerous globular embryos were formed on the surface of the callus upon transfer to auxin-rich MS medium that lacked
cytokinins. The globular embryos differentiated into mature embryos only when 2,4-D was removed from the medium. Mature embryo
formation was significantly influenced by the pH of the medium and the addition of AgNO3 and ABA. Eighty-five percent of the somatic embryos were converted into plantlets when transferred to a medium supplemented
with 0.01 mg l–1 BAP and 0.01 mg l–1 IBA. The regenerated plants have been established in soil and appear to be identical to the parent plants in morphology and
chromosome number.
Received: 5 November 1997 / Revision received: 9 February 1998 / Accepted: 19 February 1998 相似文献
14.
Somatic embryogenesis was induced in immature zygotic embryos of pea (Pisum sativum L.), synthetic auxins α-naphthalene acetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-3,5,6-trichloropicolinic acid (picloram, PIC) being used. Only one (line HM-6) of 46 genotypes tested exhibited good potential for somatic embryogenesis. 2,4-D was found as the best somatic embryo inductor. Three different ways of somatic embryo conversion have been described. Plantlets from individual somatic embryos were micropropagated as somaclones and subsequently rooted. A sterile morphological mutant has been found within a group of fertile plants of T0-generation. Sufficient amount of T1-seeds is available for somaclonal variation studies. 相似文献
15.
Sylvester Elikana Anami Allan Jalemba Mgutu Catherine Taracha Griet Coussens Mansour Karimi Pierre Hilson Mieke Van Lijsebettens Jesse Machuka 《Plant Cell, Tissue and Organ Culture》2010,102(3):285-295
In Latin America and sub-Saharan Africa, tropical maize (Zea mays L.) is a major crop for human consumption. To cope with the increasing population and changing environment, there is a need
for improving tropical maize germplasm. As part of a biotechnological approach, efficient in vitro regeneration of two tropical
maize inbred lines (CML216 and CML244) was established. A number of parameters were optimized, such as age of the immature
embryos, plant media and growth regulator concentration. After 6 weeks of culture, somatic embryos that had already reached
the coleoptilar stage produced shoots after light induction and developed into fertile plants after acclimation in the soil.
The callus induction frequencies and somatic embryo-derived plantlet formation were higher when cultured with the Linsmaier
and Skoog medium than those with the Chu’s N6 basal medium. Regeneration of tropical maize shoots depended on the 2,4-dichlorophenoxyacetic
acid (2,4-D) concentration at the callus initiation stage from immature embryos. The recalcitrance of the tropical maize inbred
line TL26 to in vitro regeneration was overcome in a single-cross hybrid with the CML216 and CML244 genotypes. Remarkably,
tropical maize somatic embryos were formed at the abaxial side of the scutellum facing the medium, probably from the axis
of the immature embryos, as shown by histological sections. Upon co-cultivation, agrobacteria transiently expressed their
intronless β-glucuronidase-encoding gene at the embryogenic tissue, but not with an intron-containing gene, suggesting that
virulence genes are induced in Agrobacterium, but that subsequent steps in the T-DNA transfer are inhibited. 相似文献
16.
De Sousa Paulo Cesar Alves Souza Stênio Steferson Silva E. Meira Filipe Sathler Meira Rennan De Oliveira Gomes Hugo Teixeira Silva-Cardoso Inaê Mariê De Araújo Scherwinski-Pereira Jonny Everson 《In vitro cellular & developmental biology. Plant》2020,56(5):618-633
In Vitro Cellular & Developmental Biology - Plant - An efficient protocol is reported for in vitro plant regeneration through somatic embryogenesis in Piper aduncum, a Brazilian Amazon species... 相似文献
17.
Summary A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l–1 kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l–1 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.Abbreviations MS
Murashige and Skoog (1962)
- 2,4-D
2,4 dichlorophenoxyacetic acid
- BA
6-benzyladenine 相似文献
18.
Z. Haydu I. K. Vasil 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1981,60(5):269-273
Summary Twenty families of the lobster, Homarus americanus, were reared at 10°, 15° and 20°C with and without bilateral eye-stalk-ablation. At 20°C both eye-stalk-ablated and non-eye-stalk-ablated lobsters from each family were assigned to one of two dietary treatments: 1) frozen whole brine shrimp, or 2) a diet of cheap, local constituents. The means of growth traits improved while viability decreased with increasing temperature. The ablation technique shows promise since improved performance for growth was apparent in eye-stalk ablated lobsters, but mortality was greater. Important genetic variation of all traits was found in each of the environments tested. The genetic correlations between growth traits were reinforcing, but the correlations between growth and viability traits were mainly negative. Ease and reliability of measurement should be considered when choosing growth traits in any artifical selection program. 相似文献
19.
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 40M), 0.5 M 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 M 2,4-D and 0.5 M 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.Abbreviations PGR
plant growth regulator
- BA
benzyladenine
- TDZ
thidiazuron
- 2,4-D
2,4-dichlorophenoxyaceticacid
- NAA
-naphthaleneacetic acid
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid 相似文献
20.
A protocol was developed for the induction, maturation and germination of somatic embryos from leaf tissue of jojoba [Simmondsia chinensis (Link) Schneider]. Explants were placed on their adaxial sides in Petri dishes and maintained in darkness on half-strength
Murashige and Skoog basal medium (MS/2). Combinations of 2,4-dichlorophenoxyacetic acid (1.35–4.52 μM) with 6-benzylaminopurine
(1.33–4.43μM) and 2 synthetic cytokinins, N-(2-chloro-4pyridyl)-N′-phenylurea (1.21–4.03μM) or (E)-6-[3-(trifluoromethyl)-but-2-enylamino]
purine (1.11–3.71μM) resulted in formation of embryogenic cultures and somatic embryos. After two 30-day subcultures, embryogenic
cultures were transferred onto MS/2 medium supplemented with different auxins and cytokinins. Somatic embryo maturation, germination
and plantlet formation were achieved using 1-naphthaleneacetic acid (3.75μM) or indole-3-butyric acid (3.44μM) in combination
with BA (0.44 or 1.33μM) or F3iP (0.37 or 1.11μM). Histology confirmed each stage of development.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献