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1.
Myosin associated with the male germ cells of angiosperms interacts with actin, promoting transport of the non-motile generative
and later sperm cells in the pollen tube. Myosin localizing on the sperm cell plasma membrane seems negligible in Plumbago, as reflected by the absence of: (i) anti-myosin labeling using immunoelectron microscopy, (ii) sperm motility on actin matrices,
and (iii) electrophoretic movement changes after addition of antibody. Sperm cells injected directly into actively streaming
Nitella internodal cells, however, follow actin bundles and their movement is sensitive to ATP and Mg2+. This may be based on simple charge binding since negatively charged latex beads also migrate on actin, whereas neutral or
positively-charged latex beads do not. Sperm cells are negatively charged according to capillary microelectrophoresis, whereas
killed sperm cells, which are positively charged do not migrate. The sperm cell that normally fertilizes the egg has a higher
calculated charge (8.277 × 103 esu/cm2) compared with the sperm cell that fuses with the central cell (6.120 × 103 esu/cm2).
Received: 15 December 1998 / Accepted: 21 January 1999 相似文献
2.
Summary In the characean algaNitella, depolymerization of microtubules potentiates the inhibitory effects of cytochalasins on cytoplasmic streaming. Microtubule depolymerization lowers the cytochalasin B and D concentrations required to inhibit streaming, accelerates inhibition and delays streaming recovery. Because microtubule depolymerization does not significantly alter3H-cytochalasin B uptake and release, elevated intracellular cytochalasin concentrations are not the basis for potentiation. Instead, microtubule depolymerization causes actin to become more sensitive to cytochalasin. This increased sensitivity of actin is unlikely to be due to direct stabilization of actin by microtubules, however, because very few microtubules colocalize with the subcortical actin bundles that generate streaming. Furthermore, microtubule reassembly, but not recovery of former transverse alignment, is sufficient for restoring the normal cellular responses to cytochalasin D. We hypothesize that either tubulin or microtubule-associated proteins, released when microtubules depolymerize, interact with the actin cytoskeleton and sensitize it to cytochalasin.Abbreviations APW
artificial pond water
- Cac
cytoplasraic free calcium concentration
- DMSO
dimethyl sulfoxide
- MT
microtubule-minus
- MT+
microtubule-plus. 相似文献
3.
Summary Parallel bundles of actin filaments at the cortex-endoplasm interface provide tracks for myosin-generated cytoplasmic streaming
in characean internodes. These bundles resist disassembly or structural modification when exposed to 10 μM cytochalasin D
(CD) even though this concentration of CD rapidly (within minutes) but reversibly arrests streaming. Unexpectedly, we discovered
that prolonged treatment with lower concentrations of CD could partially disassemble the subcortical actin bundles. Actin
bundles became discontinuous following one- to several-day treatment with concentrations (6 μM) that reduced but did not arrest
streaming, and the residual fragments mostly remained parallel to the chloroplast files. When microtubules were concurrently
disassembled with tubulin-specific drugs, however, low CD concentrations (2.5–3 μM) completely arrested bulk streaming, disrupted
the largely 2-dimensional actin bundle array and caused the formation of a coarse, thick-meshed actin network that extended
from the cortex to the endoplasm. Despite such massive reconstruction, drug removal enabled cells to recover continuous parallel
bundles and streaming. Recovery was possible if both or just one of the drugs were removed. In recovered cells, the streaming
pattern frequently redeveloped in new directions that did not follow the chloroplast files, and later, chloroplast files readjusted
to the new polarity established by the actin bundles. This first report on the complete and reversible disassembly of characean
actin bundles provides new insights into the mechanism of actin bundle assembly and organization and supports the idea of
indirect interactions between actin filaments and microtubules. 相似文献
4.
Summary Reorganization of the actin cytoskeleton following cell wall puncturing of characean internodal cells was studied by immunofluorescence and confocal laser scanning microscopy. Injury locally destroyed the parallel subcortical actin filament bundles and cortical actin strands that are characteristic of unwounded regions. At wounds, a delicate three-dimensional interlaced structure of actin strands, with meshes up to 5 m wide, formed by de novo assembly of isolated filaments and by the elongation of residual subcortical actin bundles and cortical actin strands. The actin meshwork persisted for up to 2 h, corresponding to the duration of intense wound wall secretion. Actin filament bundles continuous with the subcortical bundles outside the wound then regenerated, their parallel alignment probably assisted by endoplasmic flow. Cytochalasin D concentrations that arrested cytoplasmic streaming completely inhibited the formation of the actin meshwork, wound wall deposition and recovery of actin bundles. Concentrations that only reduced streaming velocity delayed meshwork formation and wound walls were thinner than in controls. The actual amount of F-actin within the meshwork, however, was clearly greater in the presence of low cytochalasin concentrations. In late stages of recovery, the actin bundles became very thick and intervening spaces became wider thereby forming a conspicuous, three-dimensional lattice that was continuous with interwebbing subcortical bundles and cortical actin around the periphery of the wound. Our experiments suggest that actin meshwork formation is a prerequisite for plasma membrane-directed transport of vesicles involved in wounding-induced exocytosis in characean internodes. Stabilization of the meshwork by subinhibitory concentrations of cytochalasin D is probably caused by actinbinding properties of the drug that either induce bundling or impede function of associated proteins.Abbreviations AFW
artificial fresh water
- BSA
bovine serum albumin
- CLSM
confocal laser scanning microscope (microscopy)
- DIC
differential interference contrast
- DMSO
dimethyl sulfoxide
- FITC
fluorescein isothiocyanate
- MBS
m-maleimidobenzoyl N-hydroxy-succinimide ester
- PBS
phosphate-buffered saline
- SCAB
subcortical actin bundle 相似文献
5.
Summary The effects of a protein phosphatase inhibitor, calyculin A (CA), on cytoplasmic streaming and cytoplasmic organization were examined in root hair cells ofLimnobium stoloniferum. CA at concentrations higher than 50 nM inhibited cytoplasmic streaming and also induced remarkable morphological changes in the cytoplasm. The transvacuolar strands, in which actin filament bundles were oriented parallel to the long axis, disappeared and spherical cytoplasmic bodies emerged in the CA-treated cells. In these spherical bodies, actin filaments were present and the spherical bodies were connected to each other by thin strands of actin filaments. Upon CA removal, transvacuolar strands, in which actin filament bundles were aligned, and cytoplasmic streaming reappeared. A nonselective inhibitor for protein kinases, K-252a, delayed the inhibitory effect of CA on cytoplasmic streaming and suppressed the CA-induced formation of the spherical bodies. From these results, it is suggested that phosphatases sensitive to CA regulate cytoplasmic streaming and are involved in the organization of the cytoplasm in root hair cells.Abbreviations APW
artificial pond water
- CA
calyculin A 相似文献
6.
Summary Native tropomyosin from rabbit skeletal muscle introduced by intracellular perfusion intoChara cells inhibited the cytoplasmic streaming irrespective of the Ca2+ concentration. To find the action site of native tropomyosin inChara, the cytoplasmic streaming was reconstituted by introducing isolated endoplasm into actin donorChara cells from which native endoplasm had been removed. The reconstituted streaming was inhibited by pretreatment of the actin donor cells with native tropomyosin but not by that of the endoplasm, suggesting that the native tropomyosin inhibited the cytoplasmic streaming by binding toChara actin bundles. Staining of the actin bundles with FITC-labeled native tropomyosin also showed that the native tropomyosin could bind to the actin bundles. Streaming reconstituted fromChara actin bundles and skeletal muscle myosin was insensitive to Ca2+, but became sensitive on application of the native tropomyosin.Abbrevations APW
artificial pond water
- ATP
adenosine 5-triphosphoric acid
- BSA
bovine serum albumin
- EDTA
ethylene diamine tetraacetic acid
- EGTA
ethyleneglycol-bis-(-aminoethylether) N,N,N,N-tetraacetic acid
- FITC
fluorescein isothiocyanate
- FITC-NTM
fluorescein isothiocyanate-labeled native tropomyosin
- NTM
native tropomyosin 相似文献
7.
Summary The arrangement and relative stability of cortical microtubules during and after wound induction in internodal cells ofNitella flexilis andNitella pseudoflabellata were examined by immunofluorescence and by microinjection of fluorescently tagged tubulin. The formation of cellulosic wall appositions (wound walls), induced by treatment with 5×10–2MCaCl2, was identicalin young, growing cells and older non-growing internodes, suggesting that the initial microtubule pattern, which differs in growing and non-growing cells, does not influence wound wall formation. Depolymerization of microtubules with oryzalin did not alter wound wall morphology and microtubules were not detected during wound wall formation. After cessation of wound wall growth, microtubules were once again found in the wound site but these were always randomly oriented, even in young cells where the surrounding microtubules were organized into transverse arrays. Microtubules were similarly randomized in chloroplast-free windows induced by laser irradiation. Analysis of microtubule organization in living cells revealed that the microtubules in wound sites are less stable than the microtubules of adjacent transversely oriented arrays. The results indicate that although wounding can alter the relative stability and spatial organization of cortical microtubules, microtubules are neither involved in vesicle transport nor the construction of cellulosic wound walls.Abbreviations AFW
artificial fresh water
- BSA
bovine serum albumin
- DMSO
dimethyl sulfoxide
- FITC
fluorescein isothiocyanate
- PBS
phosphate-buffered saline 相似文献
8.
Actin cytoskeleton in intact and wounded coenocytic green algae 总被引:5,自引:0,他引:5
J. W. La Claire II 《Planta》1989,177(1):47-57
Summary The subcellular distribution of actin was investigated in two related species of coenocytic green algae, with immunofluorescence microscopy. Either no, or fine punctate fluorescence was detected in intact cells of Ernodesmis verticillata (Kützing) Børgesen and Boergesenia forbesii (Harvey) Feldmann. A reticulate pattern of fluorescence appears throughout the cortical cytoplasm of Ernodesmis cells shortly after wounding; this silhouettes chloroplasts and small vacuoles. Slender, longitudinal bundles of actin become evident in contracting regions of the cell, superimposed over the reticulum. Thicker portions of the bundles were observed in well-contracted regions, and the highly-convoluted appearance of nearby cortical microtubules indicates contraction of the bundles in these thicker areas. Bundles are no longer evident after healing; only the reticulum remains. In Boergesenia, a wider-mesh reticulum of actin develops in the cortex of wounded cells, which widens further as contractions continue. Cells wounded in Ca2+-free medium do not contract, and although the actin reticulum is apparent, no actin bundles were ever observed in these cells. Exogenously applied cytochalasins have no effect on contractions of cut cells or extruded cytoplasm, and normal actin-bundle formation occurs in treated cells. In contrast, erythro-9-[3-(2-hydroxynonyl)]adenine (EHNA) completely inhibits longitudinal contractions in wounded cells, and few uniformly slender actin bundles develop in inhibited cells. These results indicate that wounding stimulates a Ca2+-dependent, hierarchical assembly of actin into bundles, whose assembly and functioning are inhibited by EHNA. Contraction of the bundles and concomitant wound healing are followed by cessation of motility and disassembly of the bundles. The spatial and temporal association of the bundles with regions of cytoplasmic contraction, indicates that the actin bundles are directly involved in wound-induced cytoplasmic motility in these algae.Abbreviations EHNA
erythro-9-[3-(2-hydroxynonyl)]adenine
- MT(s)
microtubule(s) 相似文献
9.
A plant 135-kDa actin-bundling protein (P-135-ABP) isolated from pollen tubes of Lilium longiflorum (Thunb.) binds stoichiometrically to F-actin filaments and bundles them in vitro (E. Yokota et al., 1998, Plant Physiol.
116: 1421–1429). To further understand the mechanism of actin-filament bundle formation by P-135-ABP, the polarity of each
F-actin filament in bundles was examined using myosin subfragment 1 (S-1). Dissociation of F-actin filaments from bundles
organized by P-135-ABP was induced by S-1. However, F-actin filaments that remained in a bundle and decorated by S-1 showed
uniform polarity. These results indicate that P-135-ABP arranges F-actin filaments into bundles with uniform polarity and
consequently plays a key role in the orientation of cytoplasmic streaming in pollen tubes.
Received: 23 February 1999 / Accepted: 22 April 1999 相似文献
10.
Noburô Kamiya 《Journal of plant research》1986,99(4):441-467
Various methods have been used to study cytoplasmic streaming in giant algal cells during the past three decades. Simple techniques
can be used with characean internodal cells to modify the cell constitution in various ways to gain insight into the mechanism
of cytoplasmic streaming. Another method involves isolatingin vitro a huge drop of uninjured endoplasm, to examine its physical and dynamic properties. The motive force responsible for streaming
has been measured by three different techniques with similar results. Subcortical fibrils consisting of bundles of F-actin
with the same polarity are indispensable for streaming. Differential treatment of the endoplasm and ectoplasm has shown that
putative characean myosin is localized in the endoplasm. Studies of the roles of ATP, Mg2+, Ca2+, H+ etc. in the streaming have been conducted by cellular perfusion, which allows removal of the tonoplast, or by techniques
permeabilizing the protoplasmic membrane. A slow version of the movement can even be artificially reproduced by combining
characean actinin situ and exogenous myosin in the presence of Mg-ATP. The findings thus far obtained support the hypothesis that cytoplasmic streaming
in characean cells is caused by an active shearing force produced by interaction of the actin filament bundles on the cortex
with myosin in the endoplasm. 相似文献
11.
We report the first successful identification of actin, an ubiquitous contractile protein, in Tetrahymena pyriformis (strain W). We employed dimethyl sulfoxide (DMSO) as a probe to induce the formation of actin bundles in the cell nucleus [1, 2] through disruption of cytoplasmic microfilament organization [3, 4]. The cells were incubated for 30 min at 22 °C in the inorganic medium of Prescott & James [5] containing 10% DMSO, and observed under a transmission electron microscope (TEM). Microfilarment bundles were formed in interphase macronuclei, and these microfilaments, approx. 6 nm in diameter, could be decorated by rabbit skeletal muscle heavy meromyosin (HMM) in the glycerinated model. In many cases, the bundles formed closely parallel to natively existing bundles of microtubules. Interestingly, these microtubules had prominent striation with 15–16 nm periodicity. SDS-polyacrylamide gel electrophoresis was designed to show the low actin content of Tetrahymena cells in comparison with that of Dictyostelium. Actin was suggested to comprise less than 1.7% of the total protein in Tetrahymena, whereas as much as 6% was actin in Dictyostelium cells. In assessing the physiological significance of the bundle formation, we further performed HMM and myosin subfragment-1 (S1)-binding studies to clarify the organization process and the polarity of the DMSO-induced nuclear actin filaments by using the tannic acid staining technique [6]. Randomly oriented short filaments appeared in the nucleus treated with 10% DMSO for 10 min. These filaments became elongated and associated with each other to form loose bundles in the following 10 min. With 30-min treatment, the filaments were organized and large bundles with single axes developed. With these well-developed bundles, the Student's t-test was performed on 172 pairs of neighboring filaments and the probability (p) of the deviation from random polarity was 0.08, suggesting that the filaments were organized in an anti-parallel manner. The results show that the DMSO induction of nuclear actin is a powerful tool to demonstrate the existence of cellular actin in vivo and to study the mechanism of microfilament organization in relation to cell physiological activities. 相似文献
12.
Gabriel M Kopecká M Yamaguchi M Svoboda A Takeo K Yoshida S Ohkusu M Sugita T Nakase T 《Protoplasma》2006,229(1):33-44
Summary. The morphology of conidiogenesis and associated changes in microtubules, actin distribution and ultrastructure were studied
in the basidiomycetous yeast Fellomyces fuzhouensis by phase-contrast, fluorescence, and electron microscopy. The interphase cell showed a central nucleus with randomly distributed
bundles of microtubules and actin, and actin patches in the cortex. The conidiogenous mother cell developed a slender projection,
or stalk, that contained cytoplasmic microtubules and actin cables stretched parallel to the longitudinal axis and actin patches
accumulated in the tip. The conidium was produced on this stalk. It contained dispersed cytoplasmic microtubules, actin cables,
and patches concentrated in the cortex. Before mitosis, the nucleus migrated through the stalk into the conidium and cytoplasmic
microtubules were replaced by a spindle. Mitosis started in the conidium, and one daughter nucleus then returned to the mother
via an eccentrically elongated spindle. The cytoplasmic microtubules reappeared after mitosis. A strong fluorescence indicating
accumulated actin appeared at the base of the conidium, where the cytoplasm cleaved eccentrically. Actin patches then moved
from the stalk together with the retracting cytoplasm to the mother and conidium. No septum was detected in the long neck
by electron microscopy, only a small amount of fine “wall material” between the conidium and mother cell. Both cells developed
a new wall layer, separating them from the empty neck. The mature conidium disconnected from the empty neck at the end-break,
which remained on the mother as a tubular outgrowth. Asexual reproduction by conidiogenesis in the long-neck yeast F. fuzhouensis has unique features distinguishing it from known asexual forms of reproduction in the budding and fission yeasts. Fellomyces fuzhouensis develops a unique long and narrow neck during conidiogenesis, through which the nucleus must migrate into the conidium for
eccentric mitosis. This is followed by eccentric cytokinesis. We found neither an actin cytokinetic ring nor a septum in the
long neck, from which cytoplasm retracted back to mother cell after cytokinesis. Both the conidium and mother were separated
from the empty neck by the development of a new lateral wall (initiated as a wall plug). The cytoskeleton is clearly involved
in all these processes.
Correspondence and reprints: Department of Biology, Faculty of Medicine, Masaryk University, Tomešova 12, 602 00 Brno, Czech
Republic. 相似文献
13.
Cytoskeletal alterations in interphase cells of the green alga Spirogyra decimina in response to heavy metals exposure: I. The effect of cadmium 总被引:1,自引:0,他引:1
Summary. The aim of the study was to elucidate the effect of cadmium ions on the arrangement of the actin and tubulin cytoskeleton,
as well as the relationships between cytoskeletal changes and growth processes in the green filamentous alga Spirogyra decimina. Batch cultures of algae were carried out under defined conditions in the presence of various cadmium concentrations. In
control cells, the cytoskeleton appeared to be a transversely oriented pattern of both microtubules and actin filaments of
various thickness in the cell cortex; colocalization of cortical microtubules and actin filaments was apparent. Microtubules
were very sensitive to the presence of cadmium ions. Depending on the cadmium concentration and the time of exposure, microtubules
disintegrated into short rod-shaped fragments or they completely disappeared. A steep increase in cell width and a decrease
in growth rate accompanied (and probably ensued) a very rapid disintegration of microtubules. Actin filaments were more stable
because they were disturbed several hours later than microtubules at any cadmium concentration used. When cadmium ions were
washed out, the actin cytoskeleton was rebuilt even in cells in which actin filaments were completely disintegrated at higher
cadmium concentrations (40 or 100 μM). The much more sensitive microtubules were regenerated after treatment with lower cadmium
concentrations (10 or 15 μM) only.
Correspondence and reprints: Centre of Phycology, Institute of Botany, Academy of Sciences of the Czech Republic, Dukelská
135, 379 82 Třeboň, Czech Republic. 相似文献
14.
Perfused cell segments dissected from the stalk or from detached cap ray chambers of Acetabularia were used as an experimental system to study the induction of cytoplasmic contractions and concurrent cytoskeletal changes in plant cells. Immunofluorescence microscopy revealed that the actin cytoskeleton quickly rearranges upon induction of contraction by forming bundles oriented circumferentially around the affected area, whereas microtubules were not detected. Contraction is blocked by cytochalasin D or N-ethylmaleimide but is unaffected by microtubule specific inhibitors. Contraction requires external Ca2+ at concentrations of 1 μM or more, but fails to occur below 0.1 μM. Higher concentrations of Ca2+ up to 10 mM have no adverse effect. Contraction is prevented in the presence of micromolar Ca2+ by either 1 mM of the calcium channel blocker LaCl3 or 10 μM of the calmodulin inhibitor fluphenazine. Calcium ionophore A 23187 (1 μM) does not perturb wound contraction per se but causes the entire cytoplasm of wounded or unwounded cells to contract slowly. These data suggest that a localized influx of calcium ions at the wound edge causes major rearrangements in the distribution of cytoskeletal actin prior to contraction in Acetabularia. An involvement of calmodulin in calcium signaling is proposed. 相似文献
15.
Organelles in the cortical cytoplasm of the siphonous green alga Bryopsis display various types of motile activities. One of them, saltatory movement along axially oriented linear tracks is typical for mitochondria and other small particles. A method is described which allows in vitro observation of such movements in thin layers of cytoplasm extruded from the alga and attached to a poly-l -lysine coated glass surface. By comparing video recordings of motile activities with the position of cytoskeletal elements visualized by immunofluorescence in the same area of a cytoplasmic exudate, it can be shown that tracks along which particles have moved in vitro are identical with microtubules (MTs). Depolymerization of MTs in the cytoplasmic exudates by MT-specific inhibitors stops particle movement, whereas depolymerization of actin filaments with cytochalasin D disrupts actin bundles but has little effect on particle motility. These data are consistent with the model of MT guided particle transport. 相似文献
16.
Jasplakinolide reversibly disrupts actin filaments in suspension-cultured tobacco BY-2 cells 总被引:3,自引:0,他引:3
Summary. Jasplakinolide is potentially a useful pharmacological tool for the study of actin organization and dynamics in living cells,
since it induces actin polymerization in vitro and, unlike phalloidin, is membrane permeative. In the present work, the effect
of jasplakinolide on the actin cytoskeleton of living suspension-cultured Nicotiana tabacum ‘Bright Yellow 2’ cells was investigated. Actin filaments in the living cells were disrupted by jasplakinolide. The effect
of jasplakionlide on the actin cytoskeleton was concentration and time dependent. When cells were treated with a moderate
concentration (150 nM) of jasplakinolide, cortical actin filaments were disrupted preferentially, whereas actin aggregated
at the perinuclear region. With concentrations higher than 400 nM and exposure times longer than 30 min, actin filaments in
the cell disappeared completely. The effect of jasplakinolide on the actin cytoskeleton was reversible even at high concentration.
Actin bundles appeared first in the perinuclear region within 5 min, and the cortical actin array was reestablished in 15 min,
suggesting that actin filaments might be organized at this region.
Received July 31, 2001 Accepted December 14, 2001 相似文献
17.
High-frequency plant regeneration via somatic embryogenesis and organogenesis and in vitro flowering of regenerated plantlets in Panax ginseng 总被引:4,自引:0,他引:4
W. Tang 《Plant cell reports》2000,19(7):727-732
The morphogenesis ability of light yellowish globular callus derived from cotyledons of mature zygotic embryos of Panax ginseng was investigated. The optimal media for somatic embryogenesis and shoot organogenesis were MS medium containing 0.5 mg l–1 2,4-dichlorophenoxyacetic acid, 0.1 mg l–1 6-benzyladenine (BA), and 500 mg l–1 lactoalbumin hydrolysate, and SH medium supplemented with 0.5 mg l–1 α-naphthaleneacetic acid, 0.1 mg l–1 BA, and 500 mg l–1casein hydrolysate. The influences of glucose, mannose, fructose, and sorbose in the media on somatic embryogenesis and shoot
organogenesis were revealed as differences in the numbers of somatic embryos and adventitious shoots per gram of morphogenic
callus. The best regeneration of somatic embryos was obtained on medium containing glucose, with a mean of 8.7 somatic embryos
per gram of callus. The best regeneration of shoots was observed on medium containing fructose, with an average of 12.2 adventitious
shoots per gram of callus. Of the somatic embryos 95% were converted into regenerated plantlets, and 100% of adventitious
shoots rooted to form regenerated plantlets. Regenerated plants were successfully established in soil. Flowering was observed
in 5.7% of the regenerated plants derived from shoot organogenesis and in 1.4% of the regenerated plants derived from somatic
embryogenesis.
Received: 1 December 1998 / Revision received: 13 September 1999 / Accepted: 20 September 1999 相似文献
18.
Efficient shoot regeneration from hairy roots of Antirrhinum majus L. transformed by the rol type MAT vector system 总被引:1,自引:0,他引:1
Eleven independent GUS-positive hairy roots were induced by co-cultivation of leaf explants of Antirrhinum majus L. with Agrobacterium tumefaciens strain GV2260 containing the rol type MAT vector pNPI702. The MAT vector pNPI702 possesses a GUS gene under the 35 S promoter and a removal element in which
the 7.6-kb DNA fragments containing the rolA, B, C and D genes and recombinase gene with a 35 S promoter are located between two directly oriented recombination site sequences. A
total of 326 adventitious shoots regenerated from 11 independent hairy root lines cultured on 1/2MS medium without plant growth
regulators at 25 °C under a 16/8 h (day/night) photoperiod after 8 weeks of stock-culture of hairy roots and 4 weeks of culture
of the green segments of hairy roots. Regenerated plants showed either a normal or dwarf morphology. GUS activity was observed
in the hairy roots and regenerated shoots. The presence of the GUS gene in the regenerated, morphologically normal plants
was confirmed by PCR analysis.
Received: 28 February 2000 / Revision received: 18 August 2000 / Accepted: 22 August 2000 相似文献
19.