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1.
Midrib sections of Mussaenda 'Queen Sirikit', 'Do?a Luz', and 'Do?a Hilaria' were cultured on Murashige and Skoog medium (MS) supplemented with 87.7 mM
sucrose, 5 g agar l−1, 0, 5, 10 or 20 μM indole-3-acetic acid (IAA) and 0, 0.5, 1, 2.5, 5, 10, 25 or 50 μM 6-benzyladenine (BA). In addition, aseptic
5 mm shoot tips from 'Do?a Luz' cultures were excised and cultured on MS basal salts, 0.6 mM myo-inositol, 1.2 μM thiamine-HCl,
87.7 mM sucrose, 7 g agar l−1, 0, 2.5, 5, 10, 20, or 40 μM BA, 0 or 1 μM α-naphthaleneacetic acid (NAA) and 0 or 217 μM adenine sulfate at pH 5.8. Calluses
began to develop after two weeks at the cut ends of midribs when cultured on a medium containing IAA. Somatic embryos first
appeared at eight weeks but only on 'Queen Sirikit' callus. After 15 weeks, the average number of somatic embryos produced
per tube decreased as the IAA concentration increased from 0 to 20 μM. BA concentrations between 5.0 and 10.0 μM resulted
in the largest number of somatic embryos per tube. After six weeks, the total, axillary and adventitious number of 'Do?a Luz'
shoots increased as the BA concentration in the culture medium increased from 0 to 20 μM. Average shoot length and fresh weight
decreased from 0 to 40 μM BA. The addition of NAA to the culture medium reduced shoot number. Adenine sulfate in the presence
of BA reduced the total number of shoots. An ideal medium for proliferating the largest number of 'Do?a Luz' shoots would
be a MS medium supplemented with 10–20 μM BA.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
2.
Tissue culture methods for improvement of cotton has lagged seriously compared to other major crops. A method for regeneration of cotton which includes a morphogenetically competent cell suspension was needed to facilitate selection of stress-resistant variants and gene manipulation. Preliminary screening of eight strains of Gossypium hirsutum L. for embryogenic potential resulted in the production of somatic embryos in all strains. Coker 312 was selected for use in the development of a model regeneration system for G. hirsutum. Calli were initiated from hypocotyl tissues of 3-day-old-seedlings. Globular embryos were present after six weeks in culture. Calli were subcultured to liquid suspension in growth regulator-free medium. After three to four weeks, suspensions were sieved to collect globular and heart stage embryos. Collected embryos developed further when plated onto semi-solid medium. To induce germination and plantlet growth, mature embryos were placed on sterile vermiculite saturated with medium. Upon development of roots and two true leaves, plantlets were potted in peat and sand, and hardened. Mature plants and progeny have been obtained with this procedure. A high percentage of infertile plants was observed among the regenerants.Abbreviations NAA
1 naphthaleneacetic acid
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- MS
Murashige and Skoog
- BA
6 benzylamino purine
- 2i P
N6-(2-isopentenyladenine 相似文献
3.
Sulekha Hazra Anuradha V. Kulkarni Satish M. Nalawade Anjan K. Banerjee Dinesh C. Agrawal Kaza V. Krishnamurthy 《In vitro cellular & developmental biology. Plant》2000,36(6):505-510
Summary A simple and efficient method for multiple shoot induction and proliferation was achieved in six Indian cotton cultivars from
the pre-existing meristems of 21-d-old in vitro-grown seedlings. Combinations of naphthalene acetic acid (0.3–10.7 μM) and 6-benzylaminopurine (BA; 2.2 or 4.4 μM) were used for induction of shoots. The shoots proliferated and were maintained on MS (Murashige and Skoog) medium supplemented
with 4.4 μM BA. Simultaneous elongation of shoots was obtained in the same medium. Optimum multiplication was observed in cv. LRK-516
(19.7±4.6), in cotyledonary nodes isolated from the adjoining node and cultured individually in 250 ml flasks. This indicates
lateral inhibition of adjoining meristems. A positive influence of culture flasks as opposed to test tubes on the proliferation
of multiple shoots was observed in all six cultivars tested. The morphogenic response varied with genotype and the nature
of explants. Rooting of elongated shoots was achieved on MS medium devoid of growth regulators. The plantlets were transferred
to the field after hardening in the greenhouse. All plants flowered and formed bolls on maturity. 相似文献
4.
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
5.
Margaret J. Hutchinson T. Senaratna J. M. Tsujita Praveen K. Saxena 《Plant Cell, Tissue and Organ Culture》1997,47(3):293-297
A simple and efficient procedure was developed for regeneration of a tetraploid cultivar ofAlstroemeria (A. pelegrina x A. psittacina) via somatic embryogenesis in liquid cultures. Embryogenic callus induced from mature zygotic embryos, cultured on MS medium
supplemented with 40 μM NAA and 20 μM kinetin, was used as inoculum for liquid cultures. Pre-culture of the callus on MS medium
supplemented with 80 μM NAA for two days was essential for cell proliferation in the liquid medium. Embryogenic cell aggregates,
obtained by sieving through a 750 μm nylon mesh, continued to proliferate in media containing 10 or 20 μM NAA and 10 or 20
μM kinetin. When transferred to a semi-solid half strength MS medium supplemented with casein hydrolysate, cell aggregates
successfully differentiated into plantlets which later grew to maturity under greenhouse conditions. 相似文献
6.
Plant regeneration via somatic embryogenesis in many cultivars of cotton (Gossypium hirsutum L.) 总被引:4,自引:0,他引:4
Summary Embryogenic callus was formed from several cultivars of cotton (Gossypium hirsutum L.) when sections of hypocotyl and cotyledon were cultured on medium supplemented with 5 mg/liter 6-(γ, γ-dimethylallyl-amino)-purine
(2iP) and 0.1 mg/liter α-naphthaleneacetic acid (NAA) for callus initiation and proliferation, and subcultured on medium supplemented
with 5 mg/liter NAA and 0.1 to 1 mg/liter 2iP for embryogenic callus induction. It seems that a high 2iP:auxin ratio is preferred
for callus initiation and proliferation, but should be exchanged with a higher NAA:cytokinin ratio before differentiation
will occur. Embryogenic calluses were recovered at a frequency of 2 to 85% depending on the cultivar used. Coker cultivars
produced embryogenic callus faster and at higher frequencies than other cultivars. Embryogenic callus produced somatic embryos
on phytohormone-free medium. This medium was used to maintain and proliferate embryogenic callus for a perid of 18 to 24 mo.
Somatic embryos were converted to plants on a lower ionic strength medium supplemented with 0.1 mg/liter gibberellic acid
(GA3) and 0.01 mg/liter NAA. Glucose was the only carbohydrate used through all phases of tissue culture and was much better than
sucrose, on which phenolic production was very high. High temperature (30° C) and low light intensity (9 μE · m−2 · s−1) were optimal conditions for callus initiation, embryogenic callus induction, and maintenance, whereas lower temperature
(25° C) and high light intensity (90 μE · m−2 s−1) were the optimal conditions for somatic embryo maturation, germination, and plantlet development. Plants could be regenerated
within 10 to 12 wk in Cokers or 7 to 8 mo. in others. 相似文献
7.
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
8.
Miroslav Griga Marie Kubaláková Eva Tejklová 《Plant Cell, Tissue and Organ Culture》1987,9(2):167-171
The paper describes a method of somatic embryo induction in callus and suspension cultures of Vicia faba L. Callus was induced from immature cotyledons (green maturity stage) of white-flowering horse bean lines cultured on L2 medium (Phillips and Collins 1979) supplemented with 1% sucrose, 0.7% agar and different concentrations of 2,4-dichlorophenoxyacetic acid. The medium with 2.5 M 2,4-Dichlorophenoxyacetic acid was found optimum for embryogenic callus induction. Somatic embryos developed after transfer of the callus to media lower or zero 2,4-Dichlorophenoxyacetic acid and increased level of sucrose (2.5%). The release of somatic embryos from the callus was more apparent after transfer to liquid medium. There were various stages of somatic embryo development, i.e. globular, heart-shaped and torpedo ones. 相似文献
9.
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
10.
A protocol was developed for regeneration of pepper (Capsicum annuum var. Ace) through somatic embryogenesis in liquid media. For embryogenic callus formation, mature zygotic embryo explants were used on basal Murashige and Skoog medium with 9.05 M 2,4-dichlorophenoxyacetic acid and 3% sucrose. Embryogenic callus was transferred to liquid basal Murashige and Skoog medium with 4.52 M 2,4-dichlorophenoxyacetic acid and 3% sucrose in order to increase the mass of the embryogenic culture. After pretreatment with potassium citrate, cells were placed into embryo initiation medium with 6 g l-1
l-proline and a decreased (10 mM) ammonium concentration. Embryos were matured in 1.89 M abscisic acid containing half-strength Murashige and Skoog medium and converted into plants bothin vivo andin vitro at up to a 97% efficiency. 相似文献
11.
Francesco Carimi Fabio De Pasquale Francesco Giulio Crescimanno 《Plant Cell, Tissue and Organ Culture》1994,37(2):209-211
Lemon [Citrus limon (L.) Burm.] styles were treated with different growth regulators for induction of somatic embryos. Styles and stigmas were dissected from flowers and cultured on a Murashige and Skoog (MS) basal medium supplemented with 4.52 M 2,4-dichlorophenoxyacetic acid and 13.3 M 6-benzyladenine. Callus was induced from the style base 2 weeks after the treatment initiation, and embryos appeared 2 months later.Abbreviations BA
6-benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- MS
Murashige & Skoog (1962)
- NAA
-naphthaleneacetic acid 相似文献
12.
A. L. Wendt dos Santos N. Steiner M. P. Guerra K. Zoglauer B. M. Moerschbacher 《Biologia Plantarum》2008,52(1):195-199
Immature and mature zygotic embryos were used as source of explants for induction of somatic embryogenesis in Araucaria angustifolia. Embryogenic cultures (EC) were only obtained from immature zygotic embryos. Basic medium, carbon source, and genotype showed
a significant influence on the formation of stage I somatic embryos (SE). When EC were submitted to maturation conditions,
SE continued their individual development until stage II, but mature embryos were not obtained. Proteins secreted by embryogenic
cultures were, to a certain degree, genotype specific and included an extracellular class IV chitinase and β-1-3-glucanase. 相似文献
13.
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
14.
In vitro somatic embryogenesis of Glehnia littoralis Fr. schm. was observed when zygotic embryos were cultured on a medium
containing 1-naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid (0.01-10 μM), with 1 μM being the optimum. Microscopic
observations revealed globular, heart-shaped and torpedo-shaped embryo formations and plantlet regeneration. These somatic
embryos seemed to be produced directly from cells of the zygotic embryos used as explants. Of seven types of media tested,
Nitsch's medium showed the highest rate of somatic embryogenesis. Somatic embryos developed into normal plantlets and were
able to be potted.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
Somatic embryogenesis in Cucurbitaceae 总被引:1,自引:0,他引:1
Strategies based on the application of biotechnologies to crop improvement programmes generally require regeneration of whole plants from cells or tissues cultivated in vitro. In Cucurbitaceae, regeneration can occur either through a caulogenic or an embryogenic developmental pathway. Reports of somatic embryogenesis have dealt with the main cultivated crops, i.e. cucumber, melon, squash and watermelon. Somatic embryogenesis and plant recovery are obtained from numerous sources including protoplasts, but the best results are observed with explants coming from seedlings, especially cotyledons and hypocotyls. The genetic constitution of mother plants also seems to play a key role in the success of embryogenesis, but few systematic studies on genotype effect have been published. Somatic embryos can exhibit developmental abnormalities, particularly when they arise from protoplast-derived cultures. Generally, data concerning embryo yield, rate of germination and plant development and characteristics of regenerated plants and their progeny, has not been provided in previous reports. The potential use of somatic embryogenesis in cucurbit breeding programmes is stressed in this review.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine
- B5
Gamborg et al. (1968)
- CH
casein hydrolysate
- CW
coconut water
- 2,4-d
2,4-dichlorophenoxy acetic acid
- GA3
gibberellin A3
- GA4
gibberellin A4
- H
Heller (1953)
- IAA
indole 3 acetic acid
- IBA
indole 3 butyric acid
- 2ip
2 isopentenyladenine
- KIN
kinetin
- MS
Murashige & Skoog (1962)
- N
Nitsch (1951)
- N6
Chu et al. (1975)
- NAA
1 naphthalene acetic acid
- TDZ
thidiazuron
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid 相似文献
16.
Summary A somatic embryogenesis protocol was developed for Eschscholzia californica Chan. (California poppy) using embryogenic cell suspensions and optimized media conditions. Rapidly-growing, finely-dispersed
embryogenic cell suspension cultures were established from embryogenic callus and maintained in B5 liquid media supplemented
with 0.5 mg 1−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid. Culture conditions were optimized by investigating the effect of basal media composition,
gyratory shaker speed, various carbon sources, different cytokinins, and AgNO3 on the efficiency of somatic embryogenesis. After 40 d in culture, the somatic embryos that formed were counted and their
overall growth expressed as pecked cell volume. The selected media consisted of either Gamborg (B5) or Murashige and Skoog
(MS) salts and vitamins supplemented with 40 g 1−1 (117 mM) sucrose, 0.05 mg 1−1 (0.22 μM) 6-benzylaminopurine, and 10 mg l−1 (58.8 μM) AgNO3. Somatic embryo production was substantially reduced at shaker speeds above 40 rpm. Glucose and snerose were the most effective
carbon sources, whereas fructose, galactose, and maltose resulted in a reduced yield and growth of somatic embryos. The development
of somatic embryos was promoted by AgNO3 at concentrations below 10 mg l−1 (58.8 μM). A semi-solid medium containing 1.5 g l−1 Gel-rite produced the highest frequency of somatic embryo conversion, and promoted the efficient growth of plantlets. Using
the reported protocol, over 500 viable somatic embryos were produced per 25 ml of embryogenic cell suspension culture. 相似文献
17.
Somatic embryogenesis and plant regeneration from suspension cultures of Acanthopanax koreanum Nakai
High-frequency somatic embryogenesis was achieved from an embryogenic cell suspension culture of Acanthopanax koreanum Nakai. Stem segments were cultured on Murashige and Skoog (MS) medium containing auxins and cytokinins. Opaque and friable
embryogenic callus formed on MS medium with 4.5 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 μm kinetin or zeatin, but was highest on medium containing 4.5 μm 2,4-D alone. Embryogenic calli were transferred to MS liquid medium containing 4.5 μm 2,4-D and maintained by subculture at 2-week intervals. Initiation of somatic embryogenesis and development up to the globular
stage from embryogenic cell clumps occurred in medium containing 0.45 μm 2,4-D, whereas maturation and germination of somatic embryos occurred in MS medium lacking 2,4-D. Cytokinin treatment suppressed
the normal growth of embryos, but stimulated secondary somatic embryogenesis from the surfaces of primary embryos. Plants
from somatic embryos were acclimatized in a greenhouse.
Received: 14 January 1997 / Revision received: 17 June 1997 / Accepted: 5 July 1997 相似文献
18.
A. Tirajoh T. S. Kyung Z. K. Punja 《In vitro cellular & developmental biology. Plant》1998,34(3):203-211
Summary Somatic embryogenesis in American ginseng (Panax quinquefolium L.) was investigated from three explant sources (root, leaf and epicotyl) with Murashige and Skoog (MS) medium containing
different growth regulators. Mature roots and leaves obtained from 3- to 5-yr-old field-grown plants, and seedling leaves
and epicotyls from plantlets grownin vitro, were evaluated. From root and epicotyl explants, callus development was optimal with 3,6-dichloro-o-anisic acid (dicamba)
(9.0 μM) and kinetin (KN) (5.0 μM) as the growth regulators. When these calluses were transferred after 3 mo. to dicamba alone (9.0 μM), somatic embryo formation was observed at an average frequency of 15.6% in root explants after an additional 3 mo., and
2% in epicotyl explants after an additional 6 mo. No plantlets were recovered because the embryos germinated to form shoots
with no roots. From leaf explants, callus growth was optimal with α-naphthaleneacetic acid (NAA) at 10.0 μM and 2,4-dichlorophenoxyacetic acid (2,4-D) at 9.0 μM. Somatic embryos developed on this medium, with the highest frequency (40%) obtained after 3 mo. from seedling-leaf explants.
Calluses on mature leaves formed somatic embryos after 7 mo. with NAA/2,4-D at an average frequency of 30%. Transfer of these
somatic embryos to 6-benzyladenine/gibberellic acid (4.4/2.9 μM) promoted shoot development but no roots were observed. Up to 100% of germination was observed within 6 wk on half-strength
MS salts containing activated charcoal (1%) and on NAA/2,4-D (5.0/4.5 μM) with charcoal (1%). On the latter medium, somatic embryos enlarged and frequently gave rise to new somatic embryos after
a brief callusing phase. The embryos germinated through a two-stage process, involving the elongation of the root followed
by the formation of a shoot. The highest recovery of ginseng plantlets from germinated embryos was 61.0%. Following transfer
to potting medium and maintenance under conditions of high humidity and low light intensity, the plantlets elongated and developed
new leaves. A high percentage (50%) of these plants have been acclimatized to soil. 相似文献
19.
Somatic embryogenesis in plantain banana 总被引:8,自引:0,他引:8
A. Grapin J. Schwendiman C. Teisson 《In vitro cellular & developmental biology. Plant》1996,32(2):66-71
Summary A cell suspension of French Sombre plantain banana (Musa spp. AAB genome) was initiated from callus obtained from young male flowers. Histological examination enabled us to describe and
follow the evolution of the suspension consisting of: embryogenic aggregates, proembryos, nodules, and isolated cells. It
demonstrated the unicellular origin of somatic embryos, either during maintenance of the suspension or after plating on a
semisolid medium. The cells from which the embryos originated had no starch but only protein reserves. Plating 1 ml of packed
cells from the suspension led to the formation of 105 embryos of which 10 to 40% could be converted into plantlets. 相似文献
20.
Carimi Francesco Tortorici Maria Concetta De Pasquale Fabio Crescimanno Francesco Giulio 《Plant Cell, Tissue and Organ Culture》1998,54(3):183-189
Somatic embryogenesis was induced and plant regeneration was obtained in 11 different genotypes of sweet orange navel group
[Citrus sinensis (L.) Osb.] from cultures of stigma/style explants and undeveloped ovules. Explants were cultured on 3 different
modifications of Murashige and Skoog medium: 500 mg l-1 malt extract; 500 mg l-1 malt extract and 4.6 μM kinetin; and 500
mg l-1 malt extract and 13.3 μM 6-benzylaminopurine. Sucrose (146 mM) was used as carbon source. Somatic embryogenesis occurred
1–3 months after culture initiation from undeveloped ovule and stigma/style cultures of all the genotypes tested. Somatic
embryos developed into plantlets with a high frequency (74%) after transfer to Murashige and Skoog medium supplemented with
146 mM sucrose and 500 mg l-1 malt extract. Plants were successfully transferred to soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献