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1.
Comparison of somatic embryogenesis and embryo conversion in commercial soybean cultivars 总被引:4,自引:0,他引:4
Six commercially important soybean cultivars and one control cultivar were compared for differences in induction-efficiency of somatic embryogenesis, primary embryo yield, and embryo conversion. Cotyledons from immature seeds of similar developmental stage for all soybean cultivars were used for embryo induction. The experiments utilized a Latin square design to exclude the effect of differential lighting and position due to plate location in the growth chamber on the embryogenesis process. Results indicated that the efficiency of embryo induction and yield of primary somatic embryos were genotype-dependent. In contrast, no dependence on genotype was observed for the conversion of embryos to form roots and shoots. The percentage of cotyledons that gave a positive embryogenic response ranged from 26 to 89% for the soybean cultivars tested. The average number of primary globular-stage embryos per responding cotyledon after one month on induction medium ranged from 6 to 13 among the seven cultivars. Conversion frequencies for all genotypes ranged from 27 to 45%. 相似文献
2.
An acidic esterase as a biochemical marker for somatic embryogenesis in orchardgrass (Dactylis glomerata L.) suspension cultures 总被引:2,自引:0,他引:2
The isoenzyme pattern of esterases (EC 3.1.1.2) secreted into the medium of orchardgrass (Dactylis glomerata L.) embryogenic suspension cultures during defined stages of somatic embryogenesis was compared with that of non-embryogenic
suspension cultures during unorganised cell proliferation. Isoelectric focusing revealed the presence of 7–14 predominantly
acidic isoforms. Comparison with the corresponding cell-wall isoenzyme pattern showed minor, mainly quantitative differences.
The pattern of intracellular soluble esterases did not change markedly during somatic embryo development. A unique esterase
whose migration in two-dimensional gel electrophoresis corresponds to an apparent molecular mass of 36 kDa and pI=3.8 was
detected only in embryogenic cultures at very early stages of development. Since this isoform appeared long before morphological
changes had taken place, it could possibly be used as a biochemical marker for embryogenic potential in D. glomerata L. suspension cultures.
Received: 6 June 2000 / Revision received: 17 July 2000 / Accepted: 17 July 2000 相似文献
3.
4.
Maira Oropeza Ana Karina Marcano Eva De García 《In vitro cellular & developmental biology. Plant》2001,37(2):211-216
Summary Previous results have shown that some proteins secreted in the culture medium are involved with the formation of embryogenic
cells and can modify somatic embryo differentiation. Undifferentiated cell suspensions grown in the presence of 13 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and obtained from embryogenic and non-embryogenic callus were used to study these
events in sugarcane plants (cv.PR-62258). The cell suspension growth curves were determined and soluble proteins were extracted
from embryogenic and non-embryogenic callus and culture medium from cell suspensions. In embryogenic callus we detected 1.43
times more protein than in non-embryogenic callus and the electrophoretic protein patterns show specific polypeptides for
both callus types. In embryogenic callus we detected a cluster of four polypeptides in the range of 38–44 kDa and another
polypeptide of 23 kDa that were not observed in non-embryogenic callus. In nonembryogenic callus there is a 35-kDa polypeptide
that was not detected in embryogenic callus. In the case of extracellular proteins, the medium from embryogenic cell suspensions
contained four polypeptides of 41, 38, 34 and 28 kDa that were slightly detected in the medium from non-embryogenic cell cultures;
we also detected a band at 15 kDa that could not be observed in the medium from non-embryogenic cell suspensions. These results
suggest that the development of embryogenic callus and cell suspensions is related to the type and amount of intracellular
proteins in the callus cells and to the secreted proteins from these cells into the medium. 相似文献
5.
Anzidei M. Bennici A. Schiff S. Tani C. Mori B. 《Plant Cell, Tissue and Organ Culture》2000,61(1):69-79
Different NAA plus kinetin or BA combinations were tested on Francia Pernod fennel seedlings for callus induction and plant
regeneration. Callogenesis from hypocotyls was obtained in all auxin/cytokinin-containing media. The organogenic response
was observed especially in presence of NAA plus kinetin. The highest frequency of shoot regeneration was found when the auxin
and kinetin were used at a 1:1 ratio. Moreover, a prolonged culture period increased shoot formation. Somatic embryogenesis
was tested on several fennel populations. The results gave evidence of the genotypic importance. Two different protocols were
used for somatic embryo induction. Using the first protocol among the different fennel genotypes tested, only Francia Pernod
showed embryogenic capacity. In this case, from a primary non-embryogenic callus cultured for 12 months in presence of 2,4-D,
an embryogenic secondary callus was produced. When transferred to the medium without 2,4-D (agarized or liquid), this gave
embryogenic plants in high frequency. As far as the second embryogenic method is concerned, secondary embryogenic callus developed
only in the presence of 2,4-D plus kinetin in Francia Pernod genotype. Thereafter, the replacement of those growth regulators
by GA3 into the medium greatly increased the somatic embryo development, especially in `Francia Pernod', but also in `Aboca erbe'
callus, a population with a very poor embryogenic capacity. In Francia Pernod, the primary and secondary (embryogenic) calli
showed different morphological and histological responses, either when the secondary callus was induced by 2,4-D alone or
by 2,4-D plus kinetin. Ontogenetic processes leading to somatic embryo formation are described in this context.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
6.
Media from embryogenic and non-embryogenic cell suspension cultures were analysed for protein content, electrophoretic protein
patterns, glycoproteins and activity of peroxidases and β-glucosidases in order to characterize the physiological status of
the cultures. On a dry mass basis the amount of extracellular proteins per cell was greater in embryogenic suspensions than
in non-embryogenic suspensions. Non-embryogenic suspensions contained unidentified slimy compounds which were not present
inembryogenic cultures. The extracellular Concanavalin A-specific glycoproteins gave different isoelectric focussing patterns
and thus enabled embryogenic and non-embryogenic cultures to be differentiated. The extracellular peroxidase activity per
cell dry mass was far greater in embryogenic than in non-embryogenic cultures. The isoenzymes differed in number and composition
of the anionic bands. β-glucosidases were found in the same range of activity in both culture types, but the time course of
enzyme activity during cultivation was significantly different. In the embryogenic culture the activity was correlated with
dry mass increase, whereas in the non-embryogenic suspension the activity reached maximum during the linear growth phase.
Polyphenoloxidase which was recently recognized as an intracellular marker for embryogenic stages was not released into culture
media.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
Tanoh Hilaire Kouakou Pierre Waffo-Téguo Yatty Justin Kouadio Josep Valls Tristan Richard Alain Decendit Jean-Michel Mérillon 《Plant Cell, Tissue and Organ Culture》2007,90(1):25-29
Studies of phenolic compounds were performed during cell suspension cultures in relation with the induction of embryogenic
structures in two cultivars of cotton. Coker 312 produced embryogenic structures, unlike R405-2000 which was found to be a
non-embryogenic cultivar. Embryogenesis induction in Coker 312 was strongly linked to a higher content of caffeic, ferulic
and salicylic acids and to the appearance of p-coumaric acid, benzoic acid, trans-resveratrol, catechin and naringenin. 相似文献
8.
Vanildo Silveira Aline Martins de Vita Amanda Ferreira Macedo Maria Fernanda Ribeiro Dias Eny Iochevet Segal Floh Claudete Santa-Catarina 《Plant Cell, Tissue and Organ Culture》2013,114(3):351-364
Differences in competence acquisition and subsequent embryo maturation in embryogenic and non-embryogenic callus of sugarcane var. SP79-1011 were evaluated using histomorphological analysis, growth curves, numbers of somatic embryos, and polyamine contents. Embryogenic callus was formed by cells with embryogenic characteristics such as a rounded shape, prominent nuclei, a high nucleus: cytoplasm ratio, small vacuoles and organized globular structures. However, non-embryogenic callus presented dispersed, elongated and vacuolated cells with a low nucleus: cytoplasm ratio; these characteristics did not allow for the development of somatic embryos even upon exposure to a maturation stimulus. These results suggest that non-embryogenic callus does not acquire embryogenic competence during induction and that maturation treatment is not sufficient to promote somatic embryo differentiation. The use of activated charcoal (AC; 1.5 g L?1) resulted in a higher somatic embryo maturation rate in embryogenic callus but did not yield success in non-embryogenic callus. Embryogenic callus incubated with control (10 μM 2,4-dichlorophenoxyacetic acid) and maturation (1.5 g L?1 AC) treatments for 28 days showed similar patterns of total free polyamines; these results differed from the results observed with non-embryogenic callus, suggesting that embryogenic callus already exhibits a characteristic pattern of endogenous polyamine levels. At 28 days of culture with maturation treatment, embryogenic callus exhibited significantly higher levels of free Spm than embryogenic callus incubated with control treatment and non-embryogenic callus incubated with both treatments. This result suggests that Spm could be important for the acquisition of embryogenic competence and somatic embryo maturation in sugarcane var. SP79-1011. 相似文献
9.
Slowly activating vacuolar channels (SV), were examined in embryogenic and non-embryogenic cultures of winter wheat using
a patch-clamp technique. Four different types of cultures were examined: embryogenic and non-embryogenic calli from embryos,
embryogenic and non-embryogenic calli from inflorescences. In a cell-attached mode single SV channel events were recorded.
Unitary conductance of single SV channels was between 37 pS and 48 pS and did not significantly depend on the kind of the
culture, although it was a tendency that SV channels of embryogenic calli possessed lower unitary conductance than those of
non-embryogenic. 2,4-D caused significant lowering of unitary conductance from 48±6 pS in the control culture of embryogenic
embryos to 28±6 pS in vacuoles treated. The SV channel density was estimated as 0.34 μm−2. 相似文献
10.
We identified and isolated a monoclonal antibody (MAb 3G2) raised against extracellular proteins from microcluster cells of
orchard grass (Dactylis glomerata L.) embryogenic suspension culture. MAb 3G2 recognized with high specificity an antigen ionically bound within the primary
cell wall and in the culture medium of microcluster cells. Two-dimensional polyacrylamide gel analysis and blotting of proteins
on PVDF membrane showed that MAb 3G2 detected a single polypeptide of apparent molecular mass of 48 kDa and an isoelectric
point (pI) of 5.2, designated EP48. A transient expression during somatic embryogenesis was observed for EP48. Indirect immunofluorescence
showed that this protein highly accumulated in the cell walls of some single cells, microclusters and partly in proembryogenic
masses (PEMs), but not in globular embryos of the embryogenic cell line and microclusters from the non-embryogenic cell line.
Signal intensity varied between individual cells of the same population and in successive stages of somatic embryo development.
Screening of several D. glomerata L. embryogenic and non-embryogenic cell lines with MAb 3G2 indicated the presence of ECP48 in only embryogenic suspension
cultures at early stages of embryo development long before morphological changes have taken place and thus it could serve
as an early marker for embryogenic potential in D. glomerata L. suspension cultures. 相似文献
11.
12.
Maria Carolina Andrade Nascimento-Gavioli Gabriela Claudia Cangahuala-Inocente Douglas Steinmacher Joseph Francis Ree Neusa Steiner Miguel Pedro Guerra 《In vitro cellular & developmental biology. Plant》2017,53(1):33-40
Both embryogenic and non-embryogenic peach palm (Bactris gasipaes Kunth) cultures arise during somatic embryogenesis induction, and both tissue types are often observed growing side-by-side from the same explant. To better understand why this occurs, samples from each tissue type were analyzed for their endogenous concentrations of indole-3-acetic acid (IAA), abscisic acid (ABA), polyamines, and amino acids with high-performance liquid chromatography and for total phenolics with spectrophotometry. Embryogenic cultures contained significantly higher concentrations of IAA, ABA, and total amino acids, whereas non-embryogenic tissue contained more total polyamines and phenolics. The greater IAA concentrations in embryogenic cultures supported the role of that hormone as a marker of embryogenic potential. Putrescine was especially prevalent in non-embryogenic cultures; however, the decreased putrescine/spermine + spermidine ratio in embryogenic cultures added support to the conclusions of previous studies in other species that this can serve as a marker of embryogenic competence. Though embryogenic cultures contained higher total amino acids, each culture type had different concentrations of specific amino acids. 相似文献
13.
Embryogenic callus cultures of Ipomoea batatas Poir. produce fast growing non-embryogenic material which soon dominates the cultures. Our objective was to selectively enhance the proliferation of the embryogenic fraction. For this, the effect of BAP and 2,4-D concentrations on growth of embryogenic and non-embryogenic callus were studied and consequently, nutrient media for the production and indefinite maintenance of embryogenic callus without embryo formation were defined. Selective proliferation of embryogenic callus was obtained on solid media with 10 M 2,4-D and 1 M BAP and in liquid media with 5 M 2,4-D. Selective proliferation of non-embryogenic callus occurred in liquid medium with 1 M 2,4-D. In embryogenic liquid culture, embryos were produced with 0–2 M 2,4-D. Increasing 2,4-D concentration from 0 to 2 M in these cultures restricted embryo development.Abbreviations 2,4-D =
2,4-dichlorophenoxyacetic acid
- BAP =
6-benzylaminopurine 相似文献
14.
Unopened leaves, petioles and fully opened leaves from micropropagation cultures of five Vitis rotundifolia Michx. varieties were cultured on induction medium to study their embryogenic response. Among the various explants tested,
the maximum number of varieties produced embryogenic cultures from unopened leaves followed by fully opened leaves and petioles.
Based on morphological differences, two types of embryogenic cultures were identified. Friable cultures typically arose as
proembryonic masses (PEM) on induction medium, whereas somatic embryo production without an intervening PEM stage was observed
in compact cultures. Of the five varieties tested, the highest frequency of embryogenic response was observed from fully opened
leaves of ‘Supreme’ and unopened leaves and petioles of ‘Delicious’. Attempts to initiate suspension cultures from varieties
resulted in proliferation and maintenance of ‘Alachua’ and ‘Carlos’ cultures in liquid medium for 16 weeks. Embryogenic potential
of varieties was studied on cultures growing on embryo development medium. The maximum number of cotyledonary stage somatic
embryos from 0.2 g proembryonic masses were observed in ‘Carlos’ (379.3) followed by ‘Alachua’ (350.0) and ‘Delicious’ (305.0).
Cotyledonary stage somatic embryos germinated when cultured on Murashige and Skoog medium containing 1 μM Benzyladenine (BA).
Although high embryo germination rates (80–100%) were observed in the varieties tested, plant recovery from germinated somatic
embryos ranged from 6–47%. Embryogenic cultures could be maintained on X6 medium and used in genetic engineering studies. 相似文献
15.
Somatic embryogenesis and long term maintenance of embryogenic lines from fox grapes 总被引:3,自引:0,他引:3
Motoike S. Y. Skirvin R. M. Norton M. A. Otterbacher A. G. 《Plant Cell, Tissue and Organ Culture》2001,66(2):121-131
In the present paper, a method for the induction and long-term maintenance embryogenic cultures for Vitis × Labruscana `Niagara' and `Fredonia' is reported. Embryogenic cultures from these two cultivars were induced in an embryogenesis establishment
medium (CIM) from ovaries obtained from flowers 10–14 days pre-anthesis. The embryogenic lines obtained in this experiment
have been stably maintained for more than 2 years, through repeated subcultures on a long-term maintenance medium (LTMM) without
loss of embryogenic competence. Somatic embryo regeneration and maturation have been successfully achieved after 30 days of
cultivating embryogenic cultures in an embryo regeneration medium (EDMM), supplemented with charcoal and polyethylene glycol.
The somatic embryos were successfully germinated in two different media, `Fredonia' germination medium (FGM) and `Niagara'
germination medium (NGM), and converted into normal looking plants on a conversion medium (CM).
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
Efficient procedure for grapevine embryogenic suspension establishment and plant regeneration: role of conditioned medium for cell proliferation 总被引:1,自引:0,他引:1
Ben Amar A Cobanov P Boonrod K Krczal G Bouzid S Ghorbel A Reustle GM 《Plant cell reports》2007,26(9):1439-1447
An efficient system for the establishment and multiplication of highly prolific embryogenic cell cultures of grapevine (Vitis sp.) was developed. Using anther-derived pro-embryogenic masses as starting material, cell suspensions of different grapevine cultivars (Tempranillo, Cabernet-Sauvignon) and rootstocks (Kober 125 AA, Kober 5 BB, 110 Richter) were initiated in liquid medium containing NOA (1.0 mg l(-1)) and BAP (0.25 mg l(-1)) as growth regulators. Conditioned medium was recovered and utilised for establishing new, highly totipotent cell cultures. The suspensions obtained, showed embryogenic competence resulting in somatic embryo induction and subsequent plant regeneration. In this study, a simplified establishment procedure for grapevine embryogenic cell suspension allowing the fast multiplication of embryogenic material is described. Evidence for the promoting effect of the protein fraction derived from conditioned medium, on cell proliferation was found. In bioassays, addition of ss-D: -GlcY affect cell proliferation suggesting that arabinogalactan proteins are required for growth processes in grapevine cell cultures. 相似文献
17.
Catabolism of putrescine and spermidine in embryogenic and non-embryogenic callus lines of Picea abies 总被引:2,自引:0,他引:2
The oxidation of putrescine and spermidine were studied in embryogenic and nonembryogenic cell cultures of Picea abies (L.) Karst., with [1,4-14 C]-putrescine and [1,4-14 C]-spermidine as substrates. Activities of putrescine and spermidine oxidation varied at every developmental stage in both cultures. Putrescine was oxidized ca 5 times as fast both in embryogenic and non-embryogenic tissue as spermidine. Diamine and especially polyamine oxidase activity increased markedly in both tissues towards the end of the culturing. In maturing embryos and in ageing non-embryogenic cultures, enzyme activities were lower than in non-differentiated embryogenic calli. Aminoguanidine (1 m M ) inhibited di- and polyamine oxidation in non-embryogenic tissue by >60% and >30%, respectively. The pH optimum for putrescine oxidation was 8.0, but in non-embryogenic tissue spermidine was degraded even more actively at pH 5.0. [14 C]-Spermidine was catabolized to [14 C]-putrescine. Pyrroline dehydrogenase activity was observed in non-embryogenic spruce tissue cultures. 相似文献
18.
M. M. H. Kristensen J. I. Find F. Floto J. D. MØller J. V. NØrgaard P. Krogstrup 《Protoplasma》1994,182(1-2):65-70
Summary The development of somatic embryos in an embryogenic suspension culture ofPicea sitchensis was followed every day for two weeks after thawing from liquid nitrogen (LN2). Only a few cells, primarily located at the periphery of the embryonic region of the embryos, survived cryopreservation in LN2. Surviving cells were classified into two groups: embryogenic cells (EC) and non-embryogenic cells (NEC), based on their morphology and embryogenic competence. The dense cytoplasmic EC underwent organized growth and differentiation with first divisions occurring after 24 h, and embryo formation 6–8 days after thawing from LN2. No evidence of asymmetrical divisions or free-nuclear stages was found during somatic embryo formation. NEC had less dense cytoplasm with numerous small vacuoles. One to five days after thawing the NEC became progressively more vacuolated and elongated. Histological examination revealed no mitotic activity in NEC, and six days after thawing NECs were seen as single cells or unorganized cell aggregates. Two weeks after thawing the appearance of the cryopreserved cultures was comparable to that of the untreated cultures.Abbreviations EC
embryogenic cells
- ECC
embryogenic cell clusters
- FDA
fluorescein diacetate
- GMA
glycol methacrylate
- LN2
liquid nitrogen (–196°C)
- NEC
non-embryogenic cells 相似文献
19.
20.
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