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1.
Parameswari Namasivayam Jeremy Skepper David Hanke 《In vitro cellular & developmental biology. Plant》2008,44(4):273-281
Napin, a storage protein, has been reported to be transcribed abundantly during the pre-embryogenic stage and associated with
the induction of Brassica napus secondary embryogenesis. In this study, we studied the distribution pattern of napin in the winter oilseed rape embryogenic
tissue in comparison to that of the non-embryogenic tissue using the indirect immunofluorescence localisation coupled with
the ultrastructural immunogold labelling techniques. Immunolocalisation studies revealed that the extracellular matrix layer
outside the outer epidermal cell wall of B. napus embryogenic tissues contained napin. This is the first study to report the extracellular localisation of napin. In addition,
we have also further characterised the expression pattern of Eg1 that encodes for napin in the B. napus embryogenic tissue. 相似文献
2.
Shinjiro Ogita Hamako Sasamoto Edward C. Yeung Trevor A. Thorpe 《In vitro cellular & developmental biology. Plant》2001,37(2):268-273
Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l−1 glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic
and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic
aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation
of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing
(2400 mg l−1) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic
aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level
of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell
masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues. 相似文献
3.
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
4.
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
5.
Embryogenic and non-embryogenic calli were induced from the Centella asiatica leaf explants on Murashige and Skoog medium supplemented with kinetin and 2,4-dichlorophenophenoxyacetic acid. The extracellular
matrix (ECM) layer was seen on the surface of embryogenic cells but not on the non-embryogenic cells. The ECM formed bridges
with net-like material between the embryogenic cells. This network like structure was believed to play an important role in
plant morphogenesis and can serve as an early structural marker of embryogenic competence in Centella asiatica calli culture. 相似文献
6.
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
7.
Summary With the aim to determine a possible relationship between somatic embryogenesis and some metabolic contents in embryogenic
and non-embryogenic calluses of sugarcane (Saccharum sp. var CP-5243), the present study was carried out. Embryogenic callus has more soluble proteins, free proline, proteolytic
activity, soluble sugars, and invertase, and lower putreseine/(spermidine + spermine) than non-embryogenic tissue. Non-embryogenic
callus has a higher peroxidase and gallic acid level, lower dry matter/fresh matter ratio, and more gross fat compared with
embryogenic callus. 相似文献
8.
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
9.
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
10.
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
11.
Summary Primary embryogenic callus ofDrosera rotundifolia and long-term cultured embryogenic callus ofZea mays possess a conspicuous extracellular matrix (ECM) around and between embryogenic cells. The structural arrangement of ECM depends on the developmental stage of the embryogenic cells. Single embryoid cells were covered with, and connected by net-like material. However, surface cells of young globular embryoids were covered with a coherent layer of ECM which forms bridges with net-like material between the cells which was gradually reduced to coarse strands. When protodermis was formed on the surface of globular embryoids, the ECM disappeared completely. The ECM network was never observed on the surface of heart- and torpedo-shaped embryoids. Safranine (especially 0.1%) stabilized the structure of ECM. Digestion with pronase E and proteinase K indicated that the ECM contains proteinaceous components. Similar developmental patterns of ECM were observed in dicotyledonous and monocotyledonous examples. The ECM represents a stable morphological structure even during long-term embryogenic culture in maize.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- Dicamba
3,6-dichloro-o-anisic acid
- ECM
extracellular matrix
- KIN
kinetin
- SEM
scanning electron microscopy
- TEM
transmission electron microscopy 相似文献
12.
The nutrient uptake of an embryogenic and of a non-embryogenic cell line of birch (Betula pendula Roth.) during cell growth and embryo production was studied in suspension culture. The embryogenic and non-embryogenic cell suspensions grew differently in the same medium. The non-embryogenic cell line started to grow without any lag period after the inoculation. It rapidly hydrolyzed sucrose in the medium to glucose and fructose and consumed the glucose as carbon source. The concentration of fructose in the medium decreased only after the depletion of glucose. The embryogenic cell line also rapidly hydrolyzed the sucrose to glucose and fructose, but the monosaccharides were consumed only after the embryos started to germinate after three weeks of culture. Both monosaccharides were then taken up at the same rate. 相似文献
13.
A method for the establishment and proliferation of developmentally stable, embryogenic suspension cultures in pecan is described,
and the growth and development of cultures characterized. Suspension cultures were generated from somatic embryos derived
from zygotic embryo cotyledon explants induced on a solidified medium with naphthaleneacetic acid. Cultures were repetitively
embryogenic and proliferated in growth-regulator-free medium. The suspensions consisted of a mixture of globular stage embryo-aggregates,
freely suspended globular embryos and pre-globular stage embryo masses. Culture growth and proembryo production were evaluated
with respect to several liquid media and pH conditions. Significant differences in growth and productivity were observed between
cultures. Pre-globular stage embryo masses collected on filter paper and overlaid on solidified medium continued ontological
development and converted into plants. Thus a method has been developed for pecan suspension culture, which presents a major
improvement in embryogenic tissue culture within the Juglandaceae.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
Cuadrado Yolanda Guerra Hilario Belen Martin Ana Gallego Piedad Hita Oscar Dorado Ana Villalobos Nieves 《Plant Cell, Tissue and Organ Culture》2001,67(2):145-151
The different invertase activities in embryogenic and non-embryogenic calli induced from explants (cotyledons, petioles, hypocotyls and leaves) obtained from Medicago arborea L. subsp. arborea seedlings were evaluated. Total invertase activity was lower in the calli with the greatest embryogenic capacity. The greatest fraction of this activity corresponded to soluble invertase. Wall-bound invertase showed maximum activity during the first two months of culture and the highest activities of this type were found in non-embryogenic calli. Extracellular invertase formed the smallest fraction of the total invertase activity evaluated. Acid and alkaline invertase activities were found in all calli but differences were detected between the embryogenic and non-embryogenic calli. In the former, the activity of both types of invertase exhibited a similar type of behaviour but different from that observed in the non-embryogenic calli. The calli with the greatest embryogenic capacity had very low levels of acid invertase and very high levels of the alkaline form. Soluble invertase – both acid and alkaline – accounted for the highest fraction after the first two months of culture and was present in lower amounts in the embryogenic than in the non-embryogenic calli. Regarding bound invertase, the highest production was seen to correspond to acid invertase. The extracellular invertase evaluated corresponded to the acid form since the alkaline extracellular invertase did not show any physiologically significant activity. 相似文献
15.
16.
Sacco C. de Vries Hilbert Booij Peter Meyerink Gert Huisman H. Dayton Wilde Terry L. Thomas Ab van Kammen 《Planta》1988,176(2):196-204
Embryogenic suspension cultures of domesticated carrot (Daucus carota L.) are characterized by the presence of proembryogenic masses (PEMs) from which somatic embryos develop under conditions of low cell density in the absence of phytohormones. A culture system, referred to as starting cultures, was developed that allowed analysis of the emergence of PEMs in newly initiated hypocotyl-derived suspension cultures. Embryogenic potential, reflected by the number of FEMs present, slowly increased in starting cultures over a period of six weeks. Addition of excreted, high-molecular-weight, heat-labile cell factors from an established embryogenic culture considerably accelerated the acquisition of embryogenic potential in starting cultures. Analysis of [35S]methionine-labeled proteins excreted into the medium revealed distinct changes concomitant with the acquisition of embryogenic potential in these cultures. Analysis of the pattern of gene expression by in-vitro translation of total cellular mRNA from starting cultures with different embryogenic potential and subsequent separation of the [35S]methionine-labeled products by two-dimensional polyacrylamide gel electrophoresis demonstrated a small number of abundant in-vitro-translation products to be present in somatic embryos and in embryogenic cells but absent in nonembryogenic cells. Several other in-vitro-translation products were present in explants, non-embryogenic and embryogenic cells but were absent in somatic embryos. Hybridization of an embryoregulated complementary-DNA sequence, Dc3, to RNA extracted from starting cultures showed that the corresponding gene is expressed in somatic embryos and PEMs but not in non-embryogenic cells.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- cDNA
complementary DNA
- PAGE
polyacrylamide gel electrophoresis
- PEM
proembryogenic mass 相似文献
17.
18.
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
19.
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