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1.
Summary Embryogenic cell suspension cultures and somatic embryos of five genotypes of beech, were obtained from aged cultures derived from immature zygotic embryos cultured on solid medium containing both 2, 4-dichlorophenoxyacetic acid and N6-benzyladenine. The origin of somatic embryos was traced from single cells. Embryos remained arrested at the globular stage on liquid media, further development was achieved after plating embryogenic aggregates on Murashige and Skoog's medium with half strength major salts supplemented with glutamine and low levels of growth regulators. Cultures of different genotypes showed significant differences in maturation frequency which was not affected by the hormone treatments assayed. The frequency of conversion of embryos into plantlets was low. This frequency increased after cold storage of embryos for up to 7 months.Abbreviations BA
N6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EtOH
ethanol
- GA3
giberrellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog (1962)
- NAA
naphthalene acetic acid
- WPM
woody plant medium (Lloyd and McCown, 1980)
- Z
zeatin 相似文献
2.
Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass 总被引:1,自引:0,他引:1
Ousama M. Faizzaghmout William A. Torello 《In vitro cellular & developmental biology. Plant》1990,26(4):419-424
Summary Embryogenic callus induced from mature caryopses of perennial ryegrass (Lolium perenne L.) were placed in liquid half-strength Murashige and Skoog (MS) basal medium and supplemented with 6.0 mg/l 2,4-dichlorophenoxy
acetic acid (2,4-D), 3 g/l (w/v) casein hydrolysate (CH), and B5 vitamins, to initiate fast-growing highly embryogenic cell
suspension cultures. Newly initiated suspension cultures contained a high level of large non-embryogenic cells (NE) with relatively
few embryogenic (E) cells. Cell types were separated by discontinuous Percolls gradients or by filtering the newly initiated
cultures through 31-μm nylon mesh. The growth conditions of the E cell were optimized by testing various media components
including 2,4-D and sucrose, and subculture diluton ratio. Optimal shoot formation occurred after pretreatment of the embryogenic
cells on solidified callus maintenance medium supplemented with 60 mg/l cefotaxime for 4 weeks prior to transfer to regeneration
medium Regeneration media consisted of half-strength MS basal medium supplemented with B5 vitamins, 0.5 mg/l fluridone, and
0.5 mg/l BA. Most plants regenerated were albino with only a few green plants.
Journal Paper number MAES 2959 of the Massachusetts Agricultural Experiment Station. 相似文献
3.
A cell suspension culture was established from nodal callus ofCymbopogon martinii (Roxb.) Wats in a liquid medium containing Murashige and Skoog (1962) basal salts, vitamins, 100 mg 1–1
myo-inositol and 20 g l–1 of sucrose (MS) that was supplemented with 13.6 M 2,4-dichlorophenoxyacetic acid and 1.15 M kinetin. An initial inoculum density of 2 x 104 cells ml–1exhibited optimum cell growth. Calli were obtained 12–15 days after the suspension was plated onto semisolid medium of a similar composition. When calli were transferred to semisolid regeneration medium containing MS + 6.7 M
N
6-benzyl-adenine + 1.15 M kinetin, somatic embryogenesis and plantlet regeneration occurred after 10–25 days. There was no significant decrease in the regeneration potential of the calli even when the cultures were initiated from 47-week-old cell suspensions. Chromosome counts of cells in suspensions, calli and somatic embryos derived from cultures of different ages revealed the presence of diploids, tetraploids and octaploids. However, the 33 regenerated plants tested were all diploid, indicating that only diploid cells were capable of regeneration in vitro.Abbreviations
MS
Murashige and Skoog (1962) basal salts with vitamins (100 mg1–1
myo-inositol, 20 g1–1 sucrose)
-
2,4-D
2,4-dichlorophenoxyacetic acid
-
BA
N
6-benzyl-adenine
-
Kn
kinetin
-
MSC
MS + 13.6 M 2,4-D + 1.15 M Kn
-
MSR
MS +6.7 M BA + 1.15 M Kn 相似文献
4.
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
5.
Hypocotyl segments of 2- to 3-week-old radish (Raphanus sativus L. cv. F1 Handsome Fall) seedlings produced yellowish compact calli when cultured on Murashige and Skoog's (MS) medium supplemented with 1 mgl-1 2,4-dichlorophenoxyacetic acid (2,4-D). Upon transfer onto medium containing 6-benzyladenine and -naphthaleneacetic acid, up to 5.3% of the calli gave rise to a few somatic embryos. When subcultured for 3 to 6 months, 7% of the yellowish, compact calli produced white, compact calli which formed numerous embryos. These calli maintained their embryogenic capacity for over 18 months. When cultured on medium containing 0.1 to 3 mgl-1 2,4-D, up to 90% of longitudinally sliced somatic embryo halves produced calli with numerous secondary embryos. Embryos were transferred onto medium containing 0.1 mgl-1 2,4-D and 1 mgl-1 abscisic acid where they developed into the cotyledonary stage. Upon transfer onto half-strength MS basal medium, approximately 90% of the embryos developed into plantlets. These plantlets were successfully transplanted in potting soil and after cold treatment they were grown to maturity in a phytotron.Abbreviation 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzyladenine
- GA3
gibberellin A3
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog
- NAA
-naphthaleneacetic acid 相似文献
6.
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
7.
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
8.
Alejandro Vazquez-Tello Makoto Hidaka Takeshi Uozumi 《Plant Cell, Tissue and Organ Culture》1995,40(2):169-177
Embryogenic cell suspensions of Lavatera thuringiaca L. were established from leaf petiole and shoot regeneration was achieved when cells were plated on medium without growth regulators. We tested three methods for protoplast culture, isolated from a one-year old embryogenic cell suspension, to determine the best conditions for L. thuringiaca protoplast culture and shoot regeneration. The highest protoplast plating efficiency was obtained with the agaroseembedded method, reaching 30%, while the nursing culture method gave 5% when the protoplasts were plated over Whatman paper No. 2. However, the same nursing culture failed to produce protoplast-derived microcalluses when the protoplasts were plated on a nitrocellulose filter. The liquid thin layer method gave the lowest plating efficiency with only 0.5%. Shoot regeneration from protoplast-derived microcalluses was achieved in two steps; first, globular embryo development was favored in medium low in auxin (2,4-d and BA at 0.01 and 0.05 mg 1-1, respectively), second, the globular embryos further differentiate into shoots in medium without growth regulators or in medium containing GA3 (0.5 to 1.0 mg 1-1).Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzyladenine
- GA3
gibberellic acid
- NAA
-naphthaleneacetic acid
- IBA
indole-3-butyric acid 相似文献
9.
High-efficiency somatic embryogenesis and plant regeneration from suspension cultures of grapevine 总被引:10,自引:0,他引:10
Embryogenic suspensions of grapevine (Vitis vinifera L.) were initiated from somatic embryos of `Thompson Seedless' and `Chardonnay'. Suspension cultures consisted of proembryonic
masses (PEM) that proliferated without differentiation in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). `Chardonnay'
somatic embryos developed fully from PEMs following subculture in medium without 2,4-D; however, somatic embryo development
did not advance beyond the heart stage in `Thompson Seedless' suspension cultures. Highly synchronized development of somatic
embryos was obtained by inoculating <960-μm PEMs into liquid medium without 2,4-D. Somatic embryos were also produced in large
numbers from suspension-derived PEMs of both cultivars on semisolid medium lacking 2,4-D. Somatic embryos matured and regenerated
into plants in MS basal medium containing 3% sucrose. Using this method more than 60% of the somatic embryos regenerated plants.
More than 90% of the regenerated plants were successfully transferred to the greenhouse.
Received: 27 July 1998 / Revision received: 15 October 1998 / Accepted: 27 October 1998 相似文献
10.
Rugosa rose (Rosa rugosa) is cultivated as a garden flower and an important genetic resource for the breeding of roses (R. hybrida). This study describes culture conditions for high frequency plant regeneration from zygotic embryo explants via somatic embryogenesis in rugosa rose. Mature zygotic embryo, cotyledon, and radicle explants formed embryogenic calluses at frequencies of 38, 6.7, and 8.8% when cultured on half-strength Murashige and Skoog medium (½MS) supplemented with 2.26, 9.05, and 9.05 μM 2,4-dichlorophenoxyacetic acid, respectively. Embryogenic calluses produced numerous somatic embryos, which then developed into plantlets on ½MS without growth regulators. Regenerated plantlets were grown to whole plants in a growth chamber. 相似文献
11.
Mature zygotic embryos of balloon flower (Platycodon grandiflorum) formed embryogenic calluses at a frequency of 43% when cultured on Murashige and Skoog medium supplemented with 4.52 μM
2,4-dichlorophenoxyacetic acid (2,4-D). Cell suspension cultures were established from embryogenic calluses using MS liquid
medium with 4.52 μM 2,4-D. Following transfer to solid MS basal medium, cell suspension cultures gave rise to somatic embryos,
which then developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
12.
Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Allium cepa L.) 总被引:2,自引:0,他引:2
Somatic embryos were obtained and plants regenerated from immature embryos of onion following culture on embryogenic induction
media. Highest rates of somatic embrogenesis resulted from 0.5- to 1.5-mm immature embryos cultured on media containing 5
mg/l of picloram. Somatic embryos formed either directly on the surface of embryos or developed from compact cultures. The
production of somatic embryos was significantly affected by the addition of auxin, embryo size and cultivar. The potential
of somatic embryogenic cultures for plantlet regeneration has been maintained for over 1 year in some lines. Three types of
immature-embryo-derived cultures were characterized by histology. Some cultures were morphologically similar to immature-embryo-derived
embryogenic cultures of other monocotyledonous species. Cultures such as these have proven to be useful target tissues in
transformation studies.
Received: 16 December 1997 / Revision received: 23 February 1998 / Accepted: 13 March 1998 相似文献
13.
Somatic embryoids differentiated in suspension cultures of G. klotzschianum after 3–4 weeks of culture in a liquid medium containing glutamine (optimally, 10–15 mM). Embryogenesis occurred after a preculture of callus on a medium containing 10 mg/l of the cytokinin, 2iP. The embryoids had meristematic regions, a well formed epidermis, and formed roots and vestigial leaves. Asparagine was much less effective than glutamine in promoting embryoid differentiation. The presence of 2,4-D in the medium resulted in increased vigor of the suspension cultures and subsequently in the formation of many embryoids, but does not seem to be necessary for somatic embryogenesis in cotton.Technical Article 14646 from the Texas Agricultural Experiment Station 相似文献
14.
A procedure for the regeneration of whole cucumber plants (Cucumis sativus L. cv. Poinsett 76) by embryogenesis from cell suspension cultures is described. Embryogenic callus was initiated from the primary leaves of 14–17 day old plants. Suspension cultures of embryogenic cells were grown in liquid Murashige and Skoog basal medium containing 5 uM 2,4,5-trichlorophenoxyacetic acid and 4 uM 6-benzylaminopurine. Suspension cultures were composed of a population of cells that were densely cytoplasmic and potentially embryogenic. Differentiation of embryos was enhanced by washing the suspension culture cells with MS basal medium containing 0.5% activated charcoal and twice with MS basal medium followed by liquid shake cultures in MS basal medium. Sixty to 70 percent of the embryos prewashed with activated charcoal germinated into plantlets with normal morphology. Embryos obtained from suspension cultured cells without prewashing with activated charcoal organized into plantlets with abnormal primary leaves. Morphologically normal plantlets were obtained by excising the shoot tips and transferring them to fresh medium.Abbreviation BAP
6-benzylaminopurine
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid
- MS
Murashige and Skoog 相似文献
15.
Thirty-two barley cultivars grown in Spain, 18 of the two-row type and 14 of the six-row type, were screened for plant regeneration
from cultured immature embryos. Although there was much variation in regeneration capacity among the cultivars, plants were
obtained from all cultivars except Almunia. No statistical differences were found in the percentage of regeneration between
two- and six-row types. The influence of the auxins 2,4-dichlorophenoxyacetic acid, dicamba, and picloram on the induction
and maintenance of embryogenesis and regeneration capacity after 3–4 months in culture, were evaluated for cultivars Cobra,
Hop and Reinette. Hop had the highest rates of maintenance of embryogenic capacity and plant regeneration. The medium containing
dicamba gave the best embryogenic callus induction, maintenance and regeneration. Five regeneration media, differing in growth
regulators and micronutrient composition, as well as partial desiccation of the calli before regeneration, were tested. The
regeneration medium containing 10 μm copper sulfate gave the best results. Regeneration frequencies after 3–4 months in culture of cultivar Hop were raised from
59.5 to 93.7% in this medium. Silver nitrate and partial desiccation of the calli also enhanced plant regeneration, but the
medium containing 10 μm of silver nitrate reduced root formation.
Received: 30 October 1997 / Revision received: 3 April 1998 / Accepted: 17 April 1998 相似文献
16.
A simple and efficient protocol is described for regeneration of wild sorghum (Sorghum dimidiatum) from cell suspension cultures. Fast-growing cell suspensions were established from shoot-meristem-derived callus. Plating
of the suspension on Murashige and Skoog agar medium supplemented with 2.5 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in the formation of embryogenic calli. High-frequency (80%) somatic embryogenesis
from small cell clusters (300–400 μm) was observed when the cultures were initially maintained in liquid medium with reduced
levels of 2,4-D (0.25 mg l–1), followed by transfer to regeneration medium. Direct plating of these small clusters on regeneration medium or transfer
to liquid regeneration medium containing kinetin and 6-benzylaminopurine resulted in the development of mature somatic embryos
and plantlets. The regenerants developed to maturity and were all phenotypically and cytologically normal.
Received: 20 May 1998 / Revision received: 1 September 1998 / Accepted: 23 September 1998 相似文献
17.
Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium
containing 400 mg l−1 glutamine, 200 mg l−1 casein hydrolysate, 30 g l−1 sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l−1 gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid
MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the
same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut
water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA3 on half-strength MS medium with 0.2 g l−1 activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s−1 m−2). Plants have been successfully acclimatized in the greenhouse. 相似文献
18.
Summary Suspension cultures were initiated from somatic embryos and embryogenic callus ofDactylis glomerata L. in SH-30 liquid medium [Schenk andHildebrandt (1972) containing 30 M 3,6-dichloro-o-anisic acid (dicamba)] with or without 1.5 gl–1 casein hydrolysate. Established suspension cultures maintained in SH-30 without casein hydrolysate proliferated when cell masses underwent cell division and enlargement. These cultures contained numerous root primordia and increased in volume when the cell masses continued to grow and fragment. Embryos developed only when cell masses were plated on solidified SH-30 medium. Cultures maintained in SH-30 liquid medium with casein hydrolysate also proliferated by the growth and fragmentation of cell masses. However, these cell masses contained numerous developing embryos and possessed few or no root primordia. Embryos were either attached to cell masses by a suspensor-like structure or were free and became fully developed in the liquid medium. Newly formed embryos became callused and produced embryogenic cell masses. Embryos germinated either in liquid or on solid SH medium without dicamba. The resulting plantlets possessed green shoots and well developed roots. Plants from suspension and suspension-derived callus cultures have been established in soil and grown to maturity. 相似文献
19.
Anne Katharina Jäger Brigitte Schottländer Ulla Wagner Smitt Ulf Nyman 《Plant cell reports》1993,12(9):517-520
Cell cultures from different species of the genus Thapsia (Apiaceae) have been investigated. In one 4-yearold line of T. garganica L. spontaneous somatic embryogenesis up to the globular stage occurred in a suspension culture containing 1 mg l–12,4-dichlorophenoxyacetic acid (2,4-D). Also callus cultures of this line, previously maintained on a medium containing 1 mg l–1 2,4-D, when transferred to various media deprived of 2,4-D, produced somatic embryos that developed into plantlets. Cell culture, embryos and regenerated organs were analysed for their content of thapsigargins. The undifferentiated cell culture did not synthezise thapsigargins, but was found to produce a yet unidentified compound not present in planta. White embryos in the pre-cotyledonary stage did not synthezise thapsigargins either, but when the embryos developed to the cotyledonary stage and became green, the synthesis started. Regenerated roots and shoots also contained thapsigargins.Abbreviations BAP
Benzylaminopurine
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- EtOAc
ethyl acetate
- FDA
fluorescein diacetate
- IAA
Indole-3-acetic acid
- IBA
indole-3-butyric acid
- 2-iP
2-isopentenyladenine
- NAA
1-Napthaleneacetic acid 相似文献
20.
Miguel Jordan 《Plant Cell, Tissue and Organ Culture》1986,7(3):257-261
Cell suspension cultures of Carica candamarcensis derived from hypocotyl calli were tested concerning their in vitro embryogenic capacity to improve asexual propagation rates in this species. Somatic embryos developed in culture from cells in suspension or from microcalli. Responses were affected by nutrient media and phytohormones used. Best results were obtained by growing the cells in suspension in Nitsch and Nitsch medium containing naphthaleneacetic acid and then plating them upon the same medium containing benzyladenine, or combinations of both hormones. 相似文献