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1.
Embryogenic calli were initiated from embryonic explants of Pinus roxburghii using female gametophytes containing immature pre-cotyledonary embryos. Zygotic embryos were collected at different developmental
stages and cultured on various media. Initiation of embryogenic calli was achieved in pre-cotyledonary zygotic embryos of
a 0.1-mm to 1.2-mm embryonal head on Douglus fir cotyledon revised medium (DCR) medium supplemented with 2,4-D or NAA and
BA. Embryogenic callus development was initiated from the suspensor region of immature embryos. The method of immature embryo
culture was significant as rapid embryogenic callus development occurred in megagametophytes where the suspensor was stretched
onto the medium from the cut micropylar end. Sixty embryogenic lines were established from 2500 explants cultured during one
season. A pro-embryo with six to eight meristematic cells and suspensor of six to ten long, vacuolated cells dominated the
early phase of the callus development. Cleavage polyembryony occurred in proliferating callus, constituting a method of multiplication
of these somatic embryos. Somatic embryos developed to stage-I and stage-II embryos on DCR medium supplemented with 5 μM 2,4-D or 10 μM NAA.
Received: 30 June 1999 / Revision received: 15 November 1999 / Accepted: 3 December 1999 相似文献
2.
An efficient method of repetitive somatic embryogenesis and plant regeneration of two carnation cultivars (Sagres and Impulse)
was established using a two-step protocol. In the first step, embryogenic tissue were induced from petal explants on MS culture
medium containing 9% sucrose (w/v), 9 μM 2,4-D and 0.8 μM BA. In the second step, embryogenic tissue transferred onto the
MS medium containing 3% sucrose supplemented with different concentrations of picloram (0.8, 2, 4, 8 and 16 μM) to produce
primary somatic embryos. Precotyledonary clumps and cotyledonary somatic embryos were then isolated and subcultured onto the
same media as the second step where they formed secondary somatic embryos in repetitive cycles. Cotyledony somatic embryos
were converted into plantlets when they transferred onto the growth regulator-free half-strength MS medium followed by being
acclimated to the greenhouse conditions. 相似文献
3.
Mikihisa Umehara Shinjiro Ogita Hamako Sasamoto Hiroyuki Koshino Takemichi Nakamura Tadao Asami Shigeo Yoshida Hiroshi Kamada 《In vitro cellular & developmental biology. Plant》2007,43(3):203-208
In Japanese larch (Larix leptolepis Gordon), a well-developed suspensor forms during somatic embryogenesis. The suspensor is the essential tissue for development
of the embryo proper. In high-cell-density culture, the embryogenic cells proliferate, but no somatic embryos form because
suspensor development is suppressed. Previously, we identified vanillyl benzyl ether (VBE) as a novel factor suppressing suspensor
development from the high-cell-density conditioned medium (HCM), but the inhibitory effect of VBE was weaker than that of
HCM added. Therefore, this study attempted to identify another inhibitory factor in the culture medium. Induction of somatic
embryos was performed in a medium containing both VBE and a fraction of each chromatogram extracted from the culture medium.
Results of the bioassay showed that a fraction had strong inhibitory activity with VBE, but weak activity without it. By physicochemical
analyses of the fraction, 4-[(phenylmethoxy)methyl]phenol was identified as an inhibitory factor of larch somatic embryogenesis. 相似文献
4.
Małgorzata Malik 《Plant Cell, Tissue and Organ Culture》2008,94(3):337-345
Alternative procedures for the production of Narcissus L. somatic embryos were investigated. Somatic embryogenesis was initiated on ovary explants isolated from cv. Carlton bulbs,
chilled for 12 weeks at 5°C. The explants were cultured on MS media with 3% sucrose and growth regulators: Picloram or 2,4-D
(10 or 25 μM) and BA (1 or 5 μM) for 12 weeks in the culture systems: continuous cultivation on solid media, continuous cultivation
in liquid media and sequential cultivation using cycles in liquid and solid media. Two types of somatic embryogenesis, indirect
and direct, were observed. The developmental pathway depended on the period of exposure to liquid media. Somatic embryos were
formed via embryogenic nodular callus on solid media. 2,4-D and BA stimulated the process. The 4-week and 8-week liquid medium
treatments resulted in the development of somatic embryos directly from the ovary explant tissue. The highest number of somatic
embryos was noted under the influence of 25 μM 2,4-D and 5 μM BA in explants cultivated for 8 weeks in liquid medium and then,
for 4 weeks, on solid medium. The effects of inoculum density on biomass increase and the formation of somatic embryos in
cultures obtained on a medium with 25 μM 2,4-D and 5 μM BA were also checked. The highest biomass increase was observed after
subculturing in liquid medium containing 0.5 μM NAA and 5 μM BA when the density of inoculum was 0.5 g/25 ml of the medium.
The highest number of somatic embryos was noted when the density of inoculum was 1.5 g/25 ml. 相似文献
5.
D. A. Steinmacher G. C. Cangahuala-Inocente C. R. Clement M. P. Guerra 《In vitro cellular & developmental biology. Plant》2007,43(2):124-132
The factors affecting the induction and development of somatic embryos and plantlet acclimatization of peach palm (Bactris gasipaes Kunth) were evaluated to establish an efficient regenerative protocol based on somatic embryogenesis. Mature zygotic embryos
were cultured in Murashige and Skoog (MS) medium supplemented with 0–40 μM of picloram (4-amino-3,5,6-trichloropicolinic acid)
and 0 or 5 μM of 2-isopentyladenine (6-dimethylaminopurine) (2-iP). After 5 mo. in culture embryogenic callus arose from primary
calli. Picloram (10 μM) was effective in inducing embryogenic calli in 9.8% of the explants. The use of 1 μM of AgNO3 enhanced embryogenic competence. Embryogenic calli showed an organized structure, a globular aspect, and were white to yellowish
in color. Histological analyses showed that cell proliferation arose from subepidermal cells adjacent to vascular bundles,
resulting in primary callus formed by a meristematic zone from which somatic embryos arose. Protein profile analyses revealed
two high molecular mass bands in these embryogenic calli, but not in other tissues. Embryogenic calli were transferred to
a culture medium containing 40 μM of 2,4-dichlorophenoxyacetic acid, 10 μM of 2-iP, plus 1 g l−1 of glutamine, hydrolyzed 0.5 g l−1 casein, and activated 1.5 g l−1 of charcoal. Morphogenetic responses achieved in this medium were the development of somatic embryos, rooting, and loss of
embryogenic capacity. Somatic embryos were converted to plantlets on MS medium plus 24.6 μM of 2-iP and 0.44 μM of naphthalene
acetic acid. Plantlets were maintained in MS medium with activated charcoal (1.5 g l−1) until they were 6 cm tall, and then acclimatized. After 16 wk, 84.2 ± 6.4% survival was observed.
M. P. Guerra and C. R. Clement are Fellows of the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasília,
DF. 相似文献
6.
Maurecilne Lemes da Silva Daniela Lopes Paim Pinto Miguel Pedro Guerra Eny Iochevet Segal Floh Cláudio Horst Bruckner Wagner Campos Otoni 《Plant Cell, Tissue and Organ Culture》2009,99(1):47-54
The objective of the present work was to induce somatic embryogenesis from zygotic embryos of Passiflora cincinnata Masters. Zygotic embryos formed calli on media with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and
4.5 μM benzyladenine (BA) after 30 days of in vitro culture. A concentration of 18.1 μM 2,4-D resulted in the largest number
of somatic embryos. Embryogenic calli were yellowish and friable, forming whitish proembryogenic masses. Morphologically,
embryogenic cells were small and had large nuclei and dense cytoplasm, whereas non-embryogenic cells were elongated, with
small nuclei and less dense cytoplasm. Calli cultured under white light on basal Murashige and Skoog’s medium with activated
charcoal produced embryos in all developmental stages. There were differences among the treatments, with some leading to the
production of calli with embryos and some only to callus formation. Some abnormalities were associated with somatic embryos,
including fused axes, fused cotyledons and polycotyledonary embryos. Production of secondary somatic embryos occurred in the
first cycle of primary embryo development. Secondary embryos differentiated from the surface of the protodermal layer of primary
embryos with intense cell proliferation, successive mitotic divisions in the initial phase of embryoid development, and a
vascular system formed with no connection to the parental tissue. This secondary embryogenic system of P. cincinnata is characterized by intense proliferation and maintenance of embryogenic competence after successive subcultures. This reproducible
protocol opens new prospects for massive propagation and is an alternative to the current organogenesis-based transformation
protocol. 相似文献
7.
Katharina Hristoforoglu Josef Schmidt Harald Bolhar-Nordenkampf 《Plant Cell, Tissue and Organ Culture》1995,40(3):277-284
Mature zygotic embryos of Abies alba mull were placed on a modified MCM medium (basal medium-BM) with 2.2 M benzyladenine and 2.3 M kinetin to induce embryogenic suspensor masses (ESM). These ESM proliferated on induction medium supplemented with 0.2 M 2,4-dichlorophenoxyacetic acid. From 61 ESM lines induced, 36 are still in culture after 2 years, of which 18 show embryogenic potential indicated by spontaneous formation of globular somatic embryos on the proliferation medium supplemented with 500–1000 mg l-1 casein hydrolysate and 500 mg l-1
l-glutamine. ESMs from cell line 2/56 were conditioned 1 week on BM with 58 mM sucrose and 10 g l-1 activated charcoal for maturation of somatic embryos. Maturation was achieved on BM containing 20 M (±)cis-trans-abscisic acid in combination with 111 mM maltose. Organic nitrogen supplements improved the proliferation rate of cell line 2/56 as well as the maturation and vitality of the somatic embryos. Partial drying was necessary for subsequent root development. Plantlets with a root, primary needles and a terminal bud developed on BM when a combination of 30 mM sucrose and 50 mM maltose was provided as carbon source.Abbreviations BM
basal medium
- BA
benzyladenine
- ESM
embryogenic suspensor mass
- 2,4-d
2,4-dichlorophenoxyacetic acid
- CH
casein hydrolysate
-
l-gln
l-glutamine
- ABA
(±) cis-trans-abscisic acid 相似文献
8.
Wang Xiao Xue-Lin Huang Xia Huang Ya-Ping Chen Xue-Mei Dai Jie-Tang Zhao 《Plant Cell, Tissue and Organ Culture》2007,90(2):191-200
A protocol for plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis was developed. Viable protoplasts were isolated from embryogenic cell suspensions
at a yield of 1.2 × 107 protoplasts/ml packed cell volume (PCV). Liquid and feeder layer culture systems with medium-A and medium-B were used for
protoplast culture. In liquid culture system, medium-B was more efficient for inducing cell division (17.5% at 14 days) and
colony formation (6.7% at 28 days) than medium-A. However, all protoplast-derived cell colonies (PDCC) obtained from liquid
culture system could not develop further. In feeder layer culture system, there was no significant difference between medium-A
and medium-B on cell division and colony formation of the cultured protoplasts, and the cell division frequency at 14 days
and colony formation frequency at 28 days were 24.5% and 11.2%, respectively, in medium-B. Comparative study on the effects
of BAP (2.2 μM, 4.4 μM, 8.8 μM), zeatin (0.4 μM, 0.8 μM, 1.2 μM) and TDZ (0.2 μM, 0.4 μM, 0.6 μM) on embryo formation of PDCC
from feeder-layer culture indicated that TDZ was best. TDZ at 0.4 μM induced 7906 mature embryos per ml PCV PDCC, which was
4-fold the frequency as with BAP at 4.4 μM, 7.5-fold as with zeatin at 0.8 μM and 150-fold as control medium (no mentioned
cytokinins) after 45 days on M3 medium. About 44% of the mature embryos were converted into plantlets with poor root system
after subculture on M4 medium. Root further development of regenerated plantlets was promoted by addition of activated charcoal
(AC) to MS basal medium. 相似文献
9.
Susana A. Avilés-Viñas Carlos A. Lecona-Guzmán Adriana Canto-Flick Stephanie López-Erosa Nancy Santana-Buzzy 《Plant biotechnology reports》2013,7(3):277-286
Somatic embryo-like structures were produced from the hypocotyls of aseptic plants of Capsicum chinense. Different concentrations of 2,4-dichlorophenoxyacetic acid (0, 4.5, 9.05 μM), several exposure times of the explant to this auxin (15, 30, 45, 60 days) and the development of somatic embryos cultured in a solid and/or liquid medium were evaluated. As a result, a novel system of regeneration via direct somatic embryogenesis in liquid medium was established, with an efficiency of 1.77 × 104 somatic embryos per liter of medium. Critical stages of embryogenesis, including cellular acquisition of morphogenetic competence, suspensor formation, and development and maturation of somatic embryos, were identified by histological analysis and scanning electron microscopy. Our results show a promising new outlook on the in vitro regeneration of this species. Contrary to what has been reported to date for the Capsicum genus, it is a species of plants with higher embryogenic potential in vitro. 相似文献
10.
We elucidated the relationship between cell proliferation and somatic embryogenesis in the culture of carrot cotyledons. Fresh
weights of the cotyledon expiants were determined every five days while being cultured on a medium containing 2,4-D. Callus
production increased exponentially from Day 20 to Day 25, showing a two-fold rate of proliferation. To examine the embryogenic
potential of the callus, we pre-cultured cotyledon explants on an MS medium with 2,4-D, then transferred them to an MS basal
medium at five-day intervals. Somatic embryos formed most frequently when the cotyledons were pre-cultured for 20 days on
an MS medium that contained 5 μ2,4-D. The frequency of somatic embryo formation was 81%, while that of normal embryos with
two cotyledons was 51% among those formed on a hormone-free medium. We used FACScan analysis to relate the embryogenic potential
of the callus to the S phase in the cell cycle of cultured cells. The S phase was high after 25 days of culture on the medium
with 5 μM 2,4-D. In contrast, the frequency of normal embryogenesis was higher at Day 20 of the pre-culture period. Culturing
embryogenic calli on a medium with 5 μM 2,4-D was most favorable for producing somatic embryos with two cotyledons. We verified
that active somatic embryogenesis was apparently related to cell division activity; somatic embryos induced from actively
dividing cells were apt to accompany cotyledonary abnormality. 相似文献
11.
A method for secondary somatic embryogenesis was developed on embryos derived from embryogenic callus formed on Hepatica nobilis seedlings. Somatic embryogenesis (SE) was induced on seedlings (on the hypocotyl and epicotyl parts) grown on the Murashige and Skoog (1962) medium (MS) supplemented with 1 µM naphthaleneacetic acid (NAA), and/or 0.1 µM 6-benzyladenine (BA) and on medium without plant growth regulators (PGR). The best response of embryogenic callus formation was observed on the medium containing 1 µM NAA alone or with 0.1 µM BA. Individual somatic embryos, formed on embryogenic callus on the medium without PGR (MS0), at heart, torpedo and cotyledonary stage, were transferred to the media where secondary somatic embryo formation and development into plantlets occurred. Although the most efficient repetitive cycles of secondary SE were recorded for all stages of somatic embryos (heart, torpedo, cotyledonary) on the MS0 medium (77.8–87.4 %), secondary somatic embryos were also obtained on all media supplemented with cytokinins. The best rate of somatic embryos germination was achieved on MS media with 0.2 µM NAA and 2 µM BA, and 0.1 µM NAA and 1 µM BA (48.8–52.0 %) when more mature embryos (cotyledonary stage) were used. Plantlets grown from somatic embryos were successfully acclimatized to greenhouse conditions. 相似文献
12.
Zygotic embryos at different developmental stages were tested for their potential in the initiation of embryogenic suspensor
mass (ESM) lines using immature seeds of Pinus rigida × P. taeda. The highest frequency (1.1%) of ESM was obtained with explants from cones collected on July 1. All excised embryos of the
July 1 collection were at the early proembryo stage. Two different culture media were compared. Forty-eight ESM lines were
initiated on Pinus taeda basal medium (P6) (0.97%) with 13.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.4 μM benzyladenine (BA). However, only
four ESM were obtained on a modified Murashige and Skoog medium (MSG; 0.55%). Most of the ESM arose from the seeds that were
at the stages ranging from late cleavage polyembryony to the early staged proembryo. Out of 52 lines (0.46%) that were produced
from 11,388 explants, only two viable lines (0.018%) (PRT11 and PRT28) survived. As for somatic embryo maturation, the highest
number (224/g−1 FW) of matured cotyledonary somatic embryos (line PRT 28) was obtained on a medium containing 100 μM abscisic acid (ABA),
0.2 M maltose, and 1.2% gellan gum. For germination of the somatic embryos, the cotyledonary somatic embryos derived from
maturation medium were transferred on half-strength Litvay medium (LM) plus 0.4% gellan gum. The germination rates were high
(71.4–96.3%) regardless of the concentrations of either ABA or gellan gum in the maturation medium. Approximately 500 somatic
plants were recovered from the germination medium and transferred to the green house; finally most of them were transplanted
successfully to the experimental field. 相似文献
13.
Xingyu Yang Jinfeng Lü Jaime A. Teixeira da Silva Guohua Ma 《Plant Cell, Tissue and Organ Culture》2012,109(2):213-221
Primulina tabacum is a rare and endangered species that is endemic to China. Establishing an efficient regeneration system is necessary for
its conservation and reintroduction. In this study, when leaf explants collected from plants grown in four ecotypes in China
are incubated on Murashige and Skoog (MS) medium containing 5.0 μM thidiazuron (TDZ) for 30 days, then transferred to medium
containing 5.0 μM 6-benzyladenine (BA), adventitious shoots are then observed. Conversely, when leaf explants are incubated
on medium containing 5.0 μM BA for 30 days, then transferred to medium containing 5.0 μM TDZ, somatic embryogenesis is induced.
This indicates that somatic embryogenesis and shoot organogenesis could be switched simply by changing the order of two cytokinins
supplemented in the culture medium. Histological investigation has revealed that embryogenic cells are induced within 30 days
following incubation of explants in medium containing TDZ. Only if embryogenic cells were induced, TDZ could enhance somatic
embryogenesis and BA could stimulate shoot organogenesis. When comparing explants from different ecotypes, leaf explants
from Zixiadong in Hunan Province could induce low numbers (1–2) of either somatic embryos or adventitious shoots on medium
containing either 5.0 μM TDZ or 5.0 μM BA, respectively. Whereas, leaf explants from plants collected from the other three
ecological habitats could induce 50–70 somatic embryos/adventitious shoots per explant. Moreover, somatic embryos could induce
secondary somatic embryogenesis and adventitious shoots on different media. All regenerated shoots developed adventitious
roots when these are transferred to rooting medium, and over 95% of plantlets have survived following acclimatization and
transfer to a potting mixture (1:1, sand:vermiculite). 相似文献
14.
Cerda Francisca Aquea Felipe Gebauer Marlene Medina Consuelo Arce-Johnson Patricio 《Plant Cell, Tissue and Organ Culture》2002,70(3):251-257
Embryogenic cultures from immature zygotic embryos of Pinus radiata seeds were established on semisolid proliferation medium with 2,4-D and BAP. Growing embryogenic masses containing embryonal cells and suspensor cells were subcultured on this media every 2 weeks. After 10 weeks, embryogenic masses (1.5 cm diameter) were transferred to a maturation medium containing ABA. Fully developed somatic embryos were obtained in this medium after 12 weeks. Embryogenic masses were genetically transformed using Agrobacterium tumefaciens. The pBI121 vector containing -glucuronidase (uidA) and the neomycin phosphotransferase (nptll) genes was introduced into this tissue. After co-cultivation with Agrobacterium, the embryogenic tissues were transferred to a selection media containing geneticin and carbenicillin. After 1 month of selection, histochemical assays showed extensive GUS positive activity zones in the transformed embryogenic tissues. Under light microscope, blue crystals were seen inside the embryogenic and suspensor cells, and also completely blue somatic embryos were obtained. The uidA gene was also detected by PCR analysis in genomic DNA isolated from transformed embryogenic tissues. These results indicate stable transformation of P. radiata somatic embryogenic tissues using Agrobacterium-mediated transformation. 相似文献
15.
S. Zdravković-Korać J. Milojević Lj. Tubić D. Ćalić-Dragosavac N. Mitić B. Vinterhalter 《Plant Cell, Tissue and Organ Culture》2010,101(2):237-244
A protocol has been developed for somatic embryogenesis and subsequent plant regeneration in Allium schoenoprasum L. Calli were induced from root sections isolated from axenic seedlings and cultivated on media containing either Murashige
and Skoog’s (MS) or Dunstan and Short’s mineral solution supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) in
combination with 6-benzylaminopurine (BA), 6-furfurylaminopurine (Kin) or thidiazuron (TDZ) at 1, 5 or 10 μM. The highest
frequencies of callus induction were achieved on media with 5 μM 2,4-D in combination with 5 μM TDZ or 10 μM BA (78.9% and
78.4%, respectively). Calli were then transferred to 1 μM 2,4-D, where compact yellow callus turned to segmented yellowish
callus with transparent globular somatic embryos at the surface. Calli that were previously grown on media with 5 μM 2,4-D
in combination with 10 μM BA or 10 μM TDZ showed the highest frequencies of embryogenic callus formation (45% and 42%) as
well as mean number of somatic embryos per regenerating callus. The choice of mineral solution formulation did not significantly
affect callus induction or embryogenic callus formation. The embryos could complete development into whole plants on plant
growth regulator (PGR)-free medium, but inclusion of Kin (0.5, 2.5 and 5 μM) in this phase improved somatic embryo development
and multiplication. Subsequently transferred to 1/2 MS PGR-free medium, all embryos rooted and the survival rate of the plants
in a greenhouse was 96%. 相似文献
16.
Somatic embryogenesis in wild cherry (Prunus avium) 总被引:3,自引:0,他引:3
Garin Elisabeth Grenier Emmanuel Grenier-De March Ghislaine 《Plant Cell, Tissue and Organ Culture》1997,48(2):83-91
Indirect somatic embryogenesis was obtained inPrunus avium L. from either somatic or zygotic embryos. An embryogenic line
was established by reinduction of embryogenic calluses from somatic embryos. The line was maintained for more than 3 years
through 6 generations of embryogenic cultures. In the last 2 generations, more than 50% of the explants were embryogenic.
Embryos at different stages of development were produced. Among cotyledonary-stage embryos, 50% had two cotyledons and a distinct
hypocotyl, 43% had one or more than 2 cotyledons and 7% had fused cotyledons. Most of the embryos were translucent and conversion
into plantlets was very rare. Secondary embryos could be observed to occur with low frequency from cultured somatic embryos
and from embryos emerging from calluses. Indirect somatic embryogenesis was also induced from immature zygotic embryos. From
one donor tree, 51% of the explants were embryogenic when cultured on a medium containing 0.9 μM kinetin, 0.9 μM BA and 0.5
μM NAA.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
17.
S. Paul A. Dam A. Bhattacharyya T. K. Bandyopadhyay 《Plant Cell, Tissue and Organ Culture》2011,105(2):271-283
A reproducible protocol for direct and indirect somatic embryogenesis was established in a small aromatic tree, Murraya koenigii. Embryogenic callus was obtained from 90% zygotic embryonic axis (ZE) and 70% cotyledon (COT) explants in Murashige and Skoog
(MS) basal medium supplemented with 8.88 μM 6-benzyladenine (BA) and 2.675 μM α-naphthaleneacetic acid (NAA). Globular somatic
embryos were induced and further matured from such embryogenic callus by subsequent culture on the same basal media containing
thidiazuron (TDZ) (2.27–9.08 μM). The highest frequency of somatic embryos (14.58 ± 0.42) was recovered from ZE-derived callus
after 6 weeks. The age and type of explant and concentration of TDZ played an important role in the development of somatic
embryos. Explants excised from 60-day-old seed differentiated from 96.67% of ZE explants and 86.67% from COT explants when
cultured on MS basal medium supplemented with 4.54 and 9.08 μM TDZ, respectively, after 4 weeks. The best result obtained
for the average frequency of somatic embryos (11.28 ± 0.32) was from ZE explants, which was significantly higher than COT
explants (7.34 ± 0.97). Most of the somatic embryos (above 95%), irrespective of their origin, germinated after 4 weeks in
1/2 MS basal media containing 2.32 μM kinetin (KN) and 1.07 μM NAA. Well-rooted plantlets were successfully acclimatized.
Histological analysis and scanning electron micrographs confirmed the initiation, development, and germination of somatic
embryos from both explants. 相似文献
18.
Konstantin V. Kiselev Anna V. Turlenko Yuri N. Zhuravlev 《Plant Cell, Tissue and Organ Culture》2009,99(2):141-149
A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium
(MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-d) after 4 weeks of culture in darkness. A higher response (66%) of embryogenic callus was induced on 0.45 μM 2,4-d. Higher numbers of globular- (31), heart- (17), torpedo- (12), and cotyledon-stage (8) embryos per explant were obtained
by culturing embryogenic callus on MS with 3% sucrose, 0.24% Phytagel™, and devoid of growth regulators after 8 weeks culture
in darkness. Continuous sub-culturing of embryogenic callus on medium containing 2,4-d yielded only compact callus. Desiccation of embryos for 3 days in darkness at 25 ± 2°C followed by cold storage at 4°C in
darkness for 8 weeks favored embryo germination and development of plantlets. Cotyledon-stage embryos subjected to desiccation
and chilling treatment cultured on MS with 3% sucrose, 0.24 Phytagel™, 0.44 μM 6-benzylaminopurine (BA), and 0.29 μM gibberellic
acid germinated at a higher frequency (61%) than with 0.44 μM BA alone and control cultures. Germinated plantlets developed
a shoot and root, were acclimatized successfully, and maintained in a growth room for plantlet development. 相似文献
19.
Zanderluce Gomes Luis Jonny Everson Scherwinski-Pereira 《Plant Cell, Tissue and Organ Culture》2014,118(3):485-496
An improved protocol for plant regeneration via somatic embryogenesis was developed using mature macaw palm (Acrocomia aculeata) zygotic embryos as initial explant. For induction of the embryogenic callus (EC), two basic media (BM) were tested consisting of Murashige and Skoog and Eeuwens (Y3) salts with 30 g L?1 sucrose, 0.5 g L?1 glutamine and 2.5 g L?1 Phytagel. The 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6-trichloro-picolinic acid (picloram) and 2,4-dichlorophenoxyacetic acid (2,4-D) auxins were added to the culture media at concentrations of 0, 1.5 or 3.0 mg L?1. After 240 days, the embryogenic calli were transferred to the respective BM media with auxin concentrations reduced to 0.5 or 1.0 mg L?1 in order to differentiate the somatic embryos (SEs). Plant regeneration was performed on the BM media without growth regulators. Embryogenic calli were observed after 180 days of culture and in all treatments with auxin. The Y3 medium showed the best EC formation results (60.8 %). These calli showed yellowish coloration, compact consistency and nodular aspect. After 60 days in differentiation medium, SEs were verified in different stages of development. Histological analysis showed that the SEs were formed from a nodular EC. The SEs generally presented unicellular origin with suspensor formation, and at the end of development, bipolar embryos were observed. The plant regeneration frequency reached levels up to 31.9 % when using induction medium consisting of Y3 associated to 1.5 mg L?1 of 2,4-D and the subsequent auxin reduction to 0.5 mg L?1 in the differentiation stage. Regenerated plants showed normal development, with root and aerial part growth. 相似文献
20.
Immature embryos of Cytisus laburnum L. were cultivated in vitro and four culture media, different techniques of substrate preparation, sucrose concentration and the effect of suspensor
removal were tested. The best results were obtained with N6 medium supplemented with 2 mg dm−3 glycine and set up using a double-layer culture system, in which the top layer had a higher osmotic potential than the bottom
one. These conditions allowed normal embryogenic development in up to 45 % of early globular embryos, that were able to develop
until a complete maturity. Osmotic potential and mineral nutrients of the medium demonstrated to be crucial for the successful
culture and their effects were dependent on embryo age at the time of excision. The presence of an intact suspensor showed
to be beneficial only for early globular embryos while older developmental stage embryos were not significantly affected.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献