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1.
Summary A regeneration system based on repetitive somatic embryogenesis was developed for peanut (Arachis hypogaea L.). Embryogenic suspension cultures were initiated using individual somatic embryos induced from immature cotyledons cultured on a modified Murashige and Skoog medium containing 40 mg/l 2,4-D for 30 days. After transfer to a modified MS liquid medium, the somatic embryos produced masses of secondary and tertiary embryos which continued to proliferate following manual separation and subculture of the embryogenic clumps. The cultures exhibited exponential growth, and have been maintained for over one year without apparent loss of embryogenic potential. Further embryo development, germination, and conversion were achieved by placing embryo clumps onto hormone-free, solid medium. The inclusion of a desiccation period during embryo development enhanced conversion four-fold. Plants have been established in soil and appear to be phenotypically normal.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- BA
6-benzylaminopurine
- MSO
Modified Murashige and Skoog basal medium
- EM
embryogenic masses 相似文献
2.
M. A. Bailey H. R. Boerma W. A. Parrott 《In vitro cellular & developmental biology. Plant》1993,29(3):102-108
Summary Proliferative somatic embryogenesis is a regeneration system suitable for mass propagation and genetic transformation of soybean
[Glycine max (L.) Merr.]. The objective of this study was to examine genotypic effects on induction and maintenance of proliferative embryogenic
cultures, and on yield, germination, and conversion of mature somatic embryos. Somatic embryos were induced from eight genotypes
by explanting 100 immature cotyledons per genotype on induction medium. Differences in frequency of induction were observed
among genotypes. However, this step was not limiting for plant regeneration because induction frequency in the least responding
genotype was sufficient to initiate and maintain proliferative embryogenic cultures. Six genotypes selected for further study
were used to initiate embryogenic cultures in liquid medium. Cultures were evaluated for propagation of globular-stage tissue
in liquid medium, yield of cotyledon-stage somatic embryos on differentiation medium, and plant recovery of cotyledon-stage
embryos. Genotypes also differed for weight and volume increase of embryogenic tissue in liquid cultures, for yield of cotyledon-stage
embryos on differentiation medium, and for plant recovery from cotyledon-stage embryos. Rigorous selection for a proliferative
culture phenotype consisting of nodular, compact, green spheres increased embryo yield over that of unselected cultures, but
did not affect the relative ranking of genotypes. In summary, the genotypes used in this study differed at each stage of plant
regeneration from proliferative embryogenic cultures, but genotypic effects were partially overcome by protocol modifications. 相似文献
3.
Summary Procedures have been developed for the initiation and long-term maintenance of embryogenic suspension cultures of pickling
cucumber (Cucumis sativus) cultivar Endeavor and for the regeneration of normal plantlets. Embryogenic calluses from petiole explants plated on Murashige
and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BA), both at 5μM, were used to initiate the embryogenic suspension cultures. Among various growth regulator combinations evaluated for initiation
and maintenance of these suspension cultures, only MS medium with 2,4-D and BA, both at 1μM, produced cultures that were yellow, friable, and still regenerable after repeated subculture (every two wk) over a 3- to
15-mo. period. The effects of various concentrations of auxin and cytokinin in the plating medium, the addition of AgNO3, and various plating procedures were also evaluated. The highest frequency of regeneration of shoots and plantlets was achieved
by plating aggregates onto filter paper overlaid on MS medium with naphthalene acetic acid (NAA)/BA at a concentration of
2:1 or 1:1μM. The addition of activated charcoal (0.5%) or AgNO3 (30μM) in the plating medium did not enhance the frequency of plantlet regeneration. The highest frequency of normal-appearing
plantlets recovered was 42 to 46% per petri dish. The procedures described in this study can be used to increase plantlet
recovery from individual embryogenic calluses of pickling cucumber. 相似文献
4.
Suspension cultures have been established from embryogenic tissues of Pinus nigra initiated from immature zygotic embryos. The growth of tissues in liquid medium has been influenced by initial tissue weight
used for the establishment of the cultures as well as by genotype. In most of the cases initial tissue weight 0.5 g was insufficient
and the cultures showed poor growth and later degeneration. Higher amount of initial tissues (1 or 2.5 g) was more efficient
for the establishment and proliferation of somatic embryos in liquid medium. The growth of suspension cultures was also cell
line dependent. Somatic embryo maturation in liquid medium was very limited and no plantlet regeneration occurred. Cotyledonary
somatic embryos developed and produced emblings when the suspension was plated on filter paper discs and cultured on solid
maturation medium. Based on our experiments we can state that the embryogenic tissues are able to grow and proliferate in
liquid medium but somatic embryo maturation and plantlet regeneration occur only on solid medium. 相似文献
5.
Embryogenic cultures were initiated from undeveloped ovules of several polyembryonic Citrus species on a basal medium supplemented with either malt extract, 2,4-D alone, or 2,4-D in combination with BA or daminozide. Primary embryos of all responsive cultivars were harvested directly from ovule cultures; secondary embryo harvests were made from Handin orange ovule cultures and long-term embryogenic callus. Differences were observed among cultivars and treatments in percentage of responsive ovules and total number of embryos produced. The most effective treatment for embryo production varied among cultivars. Embryo germination and plant establishment frequencies were determined for this plant regeneration system. Differences among cultivars with respect to regenerate survival percentage were minimal. Plant regeneration via secondary or long-term callus-derived embryos was as efficient as from primary embryos. Critical factors influencing plant production and survival were the production of normal viable embryos, balanced germination, and successful acclimatization to the external environment. 相似文献
6.
Lambé Pascal Mutambel Hity S.N. Deltour Roger Dinant Monique 《Plant Cell, Tissue and Organ Culture》1998,55(1):23-29
Three genotypes of Pearl millet were screened in vitro for induction of embryogenic callus, somatic embryogenesis and regeneration.
Shoot apices excised from in vitro germinated seedlings or immature embryos isolated from green house established plants were
used as primary explants. The frequency of embryogenic callus initiation was significantly higher in shoot apices in comparison
with immature zygotic embryos. Moreover, differences between genotypes were minimal when using shoot apices. Friable embryogenic
calli (type II) developed on the initial nodular calli after 1 to 3 months of culture. The frequency of type II callus is
related to the composition of the maintenance medium and they were more often found in ageing cultures. The transfer of embryogenic
calli onto auxin-free medium was sufficient for inducing somatic embryo development in short-term culture (3 months) while
a progressive loss in regeneration potential was observed with increasing time of subcultures. Maturation of embryogenic calli
on medium supplemented with activated charcoal, followed by germination of somatic embryos on medium supplemented with gibberellic
acid, restored regeneration in long-term cultures.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
K. Haliloglu 《Biologia Plantarum》2006,50(3):326-330
Efficient plant regeneration system from leaf base segments of wheat (Triticum aestivum L.) was developed. The factors affecting the callus formation and regeneration capacity of leaf segments of two genotypes;
Bobwhite and Pavon 76, were investigated. The highest number of somatic embryos (SE) was obtained on Murashige and Skoog medium
supplemented with 2 mg dm−3 2,4-dichlorophenoxyacetic acid + 1 mg dm−3 naphthalenacetic acid (14.7 SE per segment). Highest frequency of embryogenic callus (96 %) and somatic embryo formation
(24.3 SE per segment) were achieved in the first segments. The highest plantlet regeneration was obtained after transfer of
embryogenic calli to regeneration medium supplemented with 1 mg dm−3 kinetin (6.3 plantlets per segment). 相似文献
8.
János Pauk Zoltán Kertész Barnabás Jenes László Purnhauser Outi Manninen Seppo Pulli Zoltán Barabás Dénes Dudits 《Plant Cell, Tissue and Organ Culture》1994,38(1):1-10
We report regeneration of fertile, green plants from wheat (Triticum aestivum L. cv. Aura) protoplasts isolated from an embryogenic suspension initiated from somatic early-embryogenic callus. The present approach combines the optimization of protoplast culture conditions with screening for responsive genotypes. In addition to the dominant effect of the culture media, the increase in fresh mass and the embryogenic potential of somatic callus cultures varied considerably between the various genotypes tested. Establishment of suspension cultures with the required characters for protoplast isolation was improved by reduction of the ratio between cells and medium and by less frequent (monthly) transfer into fresh medium. A new washing solution was introduced to avoid the aggregation of protoplasts. However, the influence of the culture medium on cell division was variable in the different genotypes. We could identify cultures from cultivar Aura that showed approximately a 9% cell division frequency and morphogenic response. The protoplast-derived microcolonies formed both early and late-embryogenic callus on regeneration medium and green fertile plants were obtained through somatic embryogenesis. The reproducibility of plant regeneration from protoplast culture based on the cultivar Aura was demonstrated by several independent experiments. The maintenance of regeneration potential in Aura suspension cultures required establishment of new cultures within a 9-month period. 相似文献
9.
Direct somatic embryogenesis from axes of mature peanut embryos 总被引:2,自引:0,他引:2
A. H. McKently 《In vitro cellular & developmental biology. Plant》1991,27(4):197-200
Summary Plant regeneration via somatic embryogenesis was obtained in peanut (Arachis hypogaea L.) from axes of mature zygotic embryos. The area of greatest embryogenic activity was a 2-mm region adjacent to and encircling
the epicotyl. Somatic embryogenesis was evaluated on Murashige and Skoog media supplemented with a variety of auxin treatments.
Maximum production occurred on medium supplemented with 3 mg · liter−1 4-amino-3,5,6-trichloropicolinic acid. Explant cultures were transferred to half-strength medium supplemented with 1 mg ·
liter−1 gibberellic acid for somatic embryo germination and early plantlet growth. Plantlets, transferred to soil, were placed in
a greenhouse and grown to maturity. 相似文献
10.
Ashok Kumar Sahrawat Suresh Chand 《In vitro cellular & developmental biology. Plant》2001,37(1):55-61
Summary An efficient method was established for high-frequency embryogenic callus induction and plant regeneration from 3-,4-, 5-
and 7-d-old coleoptile segments of Indica rice (Oryza sativa L. cv. Kasturi), Compact and friable callus developed from the cut ends and also on the entire length of the coleoptile segments
cultured on Murashige and Skoog (MS) basal medium (1962) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 4.50–18.0
μM), kinetin (2.32 μM) and sucrose (3%, w/v). High frequency embryogenic callus induction and somatic embryo development was achieved when embryogenic
calluses were transferred to MS medium supplemented with 2.25 μM 2,4-D, 2.32 μM kinetin, 490 μM
L-tryptophan and 3% (w/v) sucrose. Plant regeneration was achieved by transferring clumps of embryogenic callus onto MS medium
containing 2.85 μM indole-3-acetic acid (IAA), 17.77 μM 6-benzylaminopurine (BA) and 3% (w/v) sucrose. Histological observations of embryogenic calluses revealed the presence of
somatic embryos and also plant regeneration via multiple shoot bud formation. Three, 4- and 5-d-old coleoptile segments showed
a significantly (P<0.05) higher frequency of plant regeneration and mean number of plantlets per explant in comparison to 7-d-old coleoptile
segments. The highest frequency (73.5%) of plant regeneration and mean number of plantlets (11.9±1.0) was obtained from 4-d-old
coleoptile segments. Regenerated shoots were rooted on MS basal medium containing 4.92 μM indole-3-butyric acid (IBA) and plants were successfully transferred to soil and grown to maturity. 相似文献
11.
Immature zygotic embryos at different developmental stages were used for callus induction and regeneration studies. Immature embryos excised from fruits 77, 91, 100, 114, 128, 140 and 193 days after pollination and mature embryos were cultured on modified Y3 medium containing 500 mgl–1 cysteine, 0.5% (w/v) PVP-40, 500 M 2,4-d and 0.3% (w/v) charcoal. Compact embryogenic tissue began differentiating directly from embryo explants after 2 weeks of culture. The percentage of embryos forming compact embryogenic tissue ranged from 28.6% for 91-day-old embryos to 0% for 140-day-old and older embryos. Friable embryogenic tissue was observed in callus cultures derived from 100-day-old embryos. Although both compact and friable embryogenic tissues were successfully isolated, normal embryo and plantlet development was observed only from friable embryogenic tissue.Abbreviations ABA
abscisic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- PVP
polyvinylpyrollidone 相似文献
12.
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
13.
Somatic embryo formation and germination from immature embryo-derived suspension-cultured cells of Angelica sinensis (Oliv.) Diels 总被引:3,自引:0,他引:3
Embryogenic callus was induced from immature embryos of Angelica sinensis cultured on Murashige and Skoog (MS) basal medium. Embryogenic callus growth was more rapid on MS basal medium than on B5
or White medium. Embryogenic callus was used to establish a suspension culture and somatic embryos and germinating embryos
developed during the culture. A shaking speed of 80 rpm was found to be optimal for establishing suspension cultures, while
100 rpm produced more somatic embryos and germinating embryos with an initiation cell density of 0.2 ml packed cell volume/25
ml medium. Adding 0.3% agar to the liquid medium also stimulated the formation of somatic and germinating embryos. While no
plant growth regulators were needed for culture initiation and plant regeneration, the addition of 0.5–1 mg/l 2,4-dichlorophenoxyacetic
acid was needed to maintain the embryogenic suspension culture by preventing embryo germination. Forty percent of the germinating
embryos survived after culturing on filter paper moistened with liquid half-strength MS medium containing 3% sucrose. The
plants were successfully transferred into soil.
Received: 19 March 1997 / Revision received: 21 November 1997 / Accepted: 19 January 1998 相似文献
14.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
15.
Summary This paper investigates maintenance and proliferation of somatic embryogenesis systems for Ulmus minor and U. glabra. Proliferation occurred with subculture of embryogenic calluses. The calluses were mainly formed by friable nodules composed
of meristematic cells organized into proembryogenic cell masses (PEMs) and thin-walled vacuolated parenchymatic cells. Cotyledonary
embryos, with procambial strands and differentiation of their vascular tissues as well as visible root meristems, were identifiable
after 18d of culture on a proliferation medium with 0.44 μM benzyladenine (BA). The shoot meristem was only occasionally well developed. Somatic embryo multiplication from elm embryogenic
calluses is a clearly asynchronic system, and PEMs as well as embryos at all stages of development are observed simultaneously
at the end of subculture period. Factors affecting the proliferation of elm embryogenic callus, such as culture medium, carbon
source and genotype, were studied. Basal medium (MS) or medium supplemented with 0.44 μM BA produced the highest number of somatic embryos. Somatic embryo production was higher with sucrose or glucose than with
maltose, and significant differences were also found among the four embryogenic lines tested. The use of liquid medium with
filter paper support is an essential step for the survival of isolated somatic embryos during the germination stage. The addition
of 0.22 μM BA′ to liquid MS medium was the best treatment for germination and plantlet conversion of elm somatic embryos. 相似文献
16.
A. L. Pinto-Sintra 《Plant Cell, Tissue and Organ Culture》2007,88(3):253-265
‘Touriga Nacional’ is the most important Portuguese grapevine cultivar used for Port wine, table wine and varietal wine production.
In order to obtain a reproducible plant regeneration system that allows the application of biotechnological tools to grapevine
breeding, embryogenic cultures were induced from immature flowers of three Touriga Nacional selected clones. Gynoecia and
anthers were cultured on Nitsch and Nitsch (Science 163:85–87, 1969) basal medium supplemented with four combinations of the
growth regulators 6-benzylaminopurine (BAP), 2,4-dichlorophenoxyacetic acid (2,4-D) and indole-3-acetyl-l-aspartic acid (IASP), at 28°C, in the dark. Primary callus, observed on anthers and gynoecia in all media, produced embryogenic
callus when cultured on differentiation medium, at 24°C under light. The efficiency on induction of embryogenic callus ranged
from 1.2 ± 4.7% to 7.9 ± 13.8% in anthers, and from 17.9 ± 24.9% to 25.3 ± 22.9% in gynoecia. Seven lines of embryogenic cultures were established from the three clones. Multiplication of embryogenic
calluses was successfully obtained in maintenance medium, at 26°C, in the dark. These embryogenic calluses produced somatic
embryos when subcultured on differentiation medium, under a 16 h photoperiod. Somatic embryos were isolated and cultured on
germination medium to achieve conversion which ranged from 35.3 ± 48.5% to 72.7 ± 45.6%. The plantlets obtained were cultured in medium without growth regulators. Secondary embryogenesis
was also frequently observed in the hypocotyl-root transition region of somatic embryos. Although some morphological variation
occurred between somatic embryos, the regenerated plantlets had a normal phenotype. Maintenance of embryogenic cultures has
been achieved since 2002. 相似文献
17.
High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species 总被引:1,自引:0,他引:1
P. van der Valk O. E. Scholten F. Verstappen R. C. Jansen J. J. M. Dons 《Plant Cell, Tissue and Organ Culture》1992,30(3):181-191
The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum x A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid. The embryogenic calluses of all three Allium species were similar in appearance. For all accessions tested plants could be regenerated at a high frequency from this compact callus through somatic embryogenesis, when using kinetin supplemented MS medium (regeneration medium). Addition of abscisic acid to the regeneration medium stimulated the formation of both somatic embryos and shoots for a number of varieties. Concerning shoot regeneration from callus cultures, significant differences existed between genotypes of all accessions except one.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- VDH
Van Der Have Seed company 相似文献
18.
19.
A rapid and effective system of somatic embryogenesis and organogenesis from the in vitro needles of redwood (Sequoia sempervirens (D.Don.) Endl.) had been established. The influences of plant growth regulators (PGRs) and days of seedlings in vitro on adventitious bud regeneration and somatic embryogenesis were studied. The process of somatic embryo formation was also observed. The results showed that embryogenic callus was induced and proliferated on Schenk and Hildebrandt (SH) medium with BA (0.5 mg/l), KT (0.5 mg/l) and IBA (1.0 mg/l). SH medium containing BA (0.5 mg/l), KT (0.2 mg/l) and IBA (0.2 mg/l) effectively promoted adventitious bud regeneration. The highest frequency (66.3%) of direct somatic embryogenesis was obtained in the combination of BA (0.5 mg/l) and IBA (0.5 mg/l). The optimal days of seedling in vitro for adventitious bud and somatic embryogenesis were 30 days and 30–40 days, respectively. The developments of somatic embryos were similar to that of zygotic embryogenesis. The result of histocytological studies indicated that proteins were gradually accumulated in the process of somatic embryo formation and there were two peaks of starch grains accumulation that one was in the embryogenic callus and the other was in the globular embryos. These results indicated that starch and protein were closely related with the energy supply and the molecular base of somatic embryogenesis, respectively. 相似文献
20.
John J. Finer Ann A. Reilley Roberta H. Smith 《In vitro cellular & developmental biology. Plant》1987,23(10):717-722
Summary Maintainable, highly embryogenic suspension cultures of a wild relative of cotton (Gossypium klotzschianum Anderss.) have been obtained. Callus with no apparent organization was used to establish the liquid culture. Callus growth
conditions as well as suspension medium composition were optimized. A visual selection scheme was beneficial for the maintenance
of the embryogenic suspension. These liquid cultures have been maintained for over 10 mo. with no loss in embryogenic capacity.
The somatic embryos developed after transfer of the embryogenic tissues to a hormone-free liquid medium.
Salaries and research support were provided by State and Federal funds appropriated to OSU-OARDC. This is journal article
No. 71-87. 相似文献