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1.
X. Niu X. Li P. Veronese R. A. Bressan S. C. Weller P. M. Hasegawa 《Plant cell reports》2000,19(3):304-310
Substantial improvement in peppermint (Mentha x piperita L. var. Black Mitcham) genetic transformation has been achieved so that the frequency of transgenic plants regenerated (percent
of leaf explants that produced transformed plants) was 20-fold greater than with the original protocol. Essential modifications
were made to conditions for Agrobacterium tumefaciens co-cultivation that enhanced infection, and for selection of transformed cells and propagules during regeneration. A systematic
evaluation of co-cultivation parameters established that deletion of coconut water from the co-cultivation medium resulted
in substantially increased transient β-Glucuronidase (GUS) activity, in both the frequency of explants expressing gusA and the number of GUS foci per explant (>700 explants). Co-cultivation on a tobacco cell feeder layer also enhanced A. tumefaciens infection. Enhanced transformation efficiencies were further facilitated by increased selection pressure mediated by higher
concentrations of kanamycin in the medium during shoot induction, regeneration, and rooting: from 20 to 50 mg/l in shoot induction/regeneration
medium and from 15 to 30 mg/l in rooting medium. Raising the concentration of kanamycin in media substantially lowered the
number of "escapes" without significant reduction in plant regeneration. These modifications to the protocol yielded an average
transformation frequency of about 20% (>2000 explants) based on expression of GUS activity or the tobacco antifungal protein,
osmotin, in transgenic plants. Genetic transformation of peppermint has been enhanced to the extent that biotechnology is
a viable alternative to plant breeding and clonal selection for improvement of this crop.
Received: 7 December 1998 / Revision received: 27 April 1999 / Accepted: 14 May 1999 相似文献
2.
Effect of 1,2-benzisoxazole-3-acetic acid on adventitious shoot regeneration and in vitro rooting in apple 总被引:3,自引:0,他引:3
The effect of 1,2-benzisoxazole-3-acetic acid (BOA), compared to 1-naphthaleneacetic acid (NAA), on adventitious shoot formation
in leaf portions and compared to indolebutyric acid (IBA), on in vitro rooting in the apple (Malus domestica Borkh) cultivars McIntosh and Gala, and one rootstock, Jork 9, was investigated. BOA at 43.0 μm or 2.7 μm at NAA in combination
with 17.8 μm benzyladenine (BA), induced the highest number of explants to produce adventitious shoots in Jork 9. In Gala,
the combination of 21.5 μm BOA with 1.0 μm thidiazuron (TDZ) or with 22.0 μm BA induced the highest regeneration percentages,
58 and 54%, respectively, giving more satisfactory results than NAA (where only 42% of leaf explants exhibited shoot formation).
In McIntosh, the highest percentage of regeneration was obtained with 1.3 μm NAA and 22.0 μm BA, while 51% was the highest
response obtained with the BOA treatment. The combination of BOA with TDZ completely inhibited regeneration activity in leaf
portions of this cultivar. The shoots of all the genotypes obtained with the most morphogenetic NAA or BOA treatments were
excised, multiplied and successfully rooted and hardened. The results demonstrate that the synthetic auxin BOA is active in
inducing shoot regeneration from leaf explants of apple and that the activity of BOA in plant regeneration is genotype dependent.
When BOA was used to induce rooting in apple microcuttings, lower rooting percentages were obtained than with IBA, showing
that the effect of BOA in inducing root formation is very low and that it cannot be used routinely to replace IBA in the in
vitro rooting of microcuttings.
Received: 18 June 1998 / Revision received: 4 January 1999 / Accepted: 29 January 1999 相似文献
3.
M. G. C. Costa F. T. S. Nogueira M. L. Figueira W. C. Otoni S. H. Brommonschenkel P. R. Cecon 《Plant cell reports》2000,19(3):327-332
Cotyledon explants of tomato (Lycopersicon esculentum Mill. cvs 'Santa Clara', 'Firme' mutant, 'IPA-5' and 'IPA-6') were excised from 8- to 10-day-old in vitro-grown seedlings.
Four different shoot induction media supplemented with timentin (300 mg l–1) were screened. When cotyledon explants were cultured on MS-based medium with 1.0 mg l–1 zeatin plus 0.1 mg l–1 IAA and supplemented with timentin, higher regeneration frequencies and a greater number of elongated shoots were obtained.
It was observed that timentin caused an increase in the morphogenesis of in vitro cotyledon explants of tomato cultivars.
In two of three cultivars tested, rooting of shoots was positively influenced, both in the presence and absence of timentin
in the rooting medium, among shoots regenerated from explants derived from timentin-supplemented medium. The results confirm
those of a previous investigation on the beneficial effects of this class of antibiotics on tomato regeneration and, consequently,
its reliability for use in the transformation of this species.
Received: 1 December 1998 / Revision received: 19 May 1999 · Accepted: 3 May 1999 相似文献
4.
A simple suspension culture system of Platycerium bifurcatum was developed where sporophytes could be regenerated directly from leaf cells or indirectly through an aposporous gametophyte
stage under the same culture conditions. Single cells and aggregates of up to 100 cells developed aposporous gametophytes
which later gave rise to sporophytes. Such gametophytes started apogamy when they were mostly less than 0.7 mm in length,
bearing only rhizoids. In most cases, only one sporophyte was regenerated from one gametophyte. Aggregates of 500–1000 or
more cells, on the other hand, regenerated sporophytes directly. Intercellular interaction was considered to be the physiological
cause, and the separation of leaf cells to a certain degree drove the cells to embark on different regeneration paths. It
is suggested that the possible existence of a threshold size of cell aggregates separates the two regeneration patterns.
Received: 3 March 1997 / Revision received: 11 April 1997 / Accepted: 3 June 1997 相似文献
5.
An efficient plant regeneration procedure has been established from hypocotyl explants of the common ice plant, Mesembryanthemum crystallinum L, a halophytic leaf succulent that exhibits a stress-induced switch from C3 photosynthesis to crassulacean acid metabolism
(CAM). Somatic embryos were initiated and developed up to globular and heart stages in Murashige and Skoog (MS) media supplemented
with 3% sucrose, 0.6% bacto-agar, 80 mM NaCl, 5 μM 2,4-D and 1 μM kinetin. High frequency regeneration occurred when somatic
embryos were germinated on media that lacked 2,4-D. High cytokinin treatment suppressed normal growth of embryos and favored
abnormal embryo proliferation. Without growth regulators, regenerated plants rooted on MS medium with 100% efficiency. Mature,
regenerated plants were fertile and morphologically identical to seed-derived plants.
Received: 29 April 1999 / Revision received: 2 July 1999 · Accepted: 12 July 1999 相似文献
6.
A new reliable protocol for the induction of adventitious shoot formation and plant regeneration from apple callus has been
developed. High regeneration frequency was obtained with this method in four different genotypes (Jork9, M26, Gala and McIntosh)
and callus maintained regeneration ability for several months. The procedure consists of inducing vegetative shoot apices,
excised from in vitro shoots, for 20 days in darkness on an MS medium without glycine, supplied with 17.8 μM BA, 2.7 μM NAA
and 250 mg l−1 cefotaxime. The explants are then transferred to a fresh auxin-free medium and given light. Histological studies revealed
that all the regenerated shoots originated from callus. Regenerated shoots were multiplied, rooted and successfully established
in soil.
Received: 2 April 1999 / Revision received: 10 November 1999 / Accepted: 15 November 1999 相似文献
7.
A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and
the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected
with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic
plants of all three cultivars used were obtained with frequencies of 1.6–2.7% when the explants were regenerated in shoot
regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that
transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion.
Received: 15 December 1998 / Revision received: 2 July 1999 · Accepted: 8 July 1999 相似文献
8.
Megaspore germination and sporophyte formation of Marsilea vestitaH. and G. was studied under different light, pH, temperature,and crowding conditions during a 6 d experimental period. Maximumgermination and sporophyte development occurs under relativelylow light intensity. Darkness and high light intensity inhibitsporophyte development. Selected wavelengths of light (red,far-red, green, blue) and darkness reduce both megaspore germinationand sporophyte development as compared to white light. Megasporesand sporophytes show maximum development at 25 ?C in light,whereas their development is reduced at all temperatures indarkness. The optimum pH range for megaspore germination ispH 78 and that for sporophyte development is pH 7. Sporocarpcontents alter the pH of unbuffered acidic and basic media towarda more optimal growth condition (pH 68) for megasporegermination. Megaspore germination and sporophyte developmentvary inversely with conditions of crowding. Root and leaf growthon developed sporophytes is very similar in most treatments. 相似文献
9.
Plant regeneration in Arachis pintoi was obtained via two developmental pathways: organogenesis and somatic embryogenesis. Organogenic callus cultures were initiated
from pieces of leaf on MS medium supplemented with NAA or 2,4-D in combination with BA, KIN or 2iP. The most suitable combination
for plant regeneration through organogenesis was an initial medium composed of 10 mg/l NAA+1 mg/l BA followed by transfer
of the callus to a shoot induction medium (MS+1 mg/l BA). Rooting of regenerated shoots was readily achieved by culture on
MS+0.01 mg/l NAA. Embryogenic callus cultures were initiated from pieces of leaf on MS medium supplemented with PICL in combination
with KIN, ZEA, BA or 2iP, and the most suitable combinations were 20 mg/l PICL+1 mg/l BA or 2iP. When pieces of embryogenic
callus were subcultured on MS+1 mg/l BA, somatic embryos were differentiated and developed further into well-developed plants
in MS+1 g/l AC followed by MS medium devoid of plant growth regulators.
Received: 29 April 1999 / Revision received: 24 November 1999 / Accepted: 18 December 1999 相似文献
10.
Hongyan Dai Zhihong Zhang Xiuwu Guo 《In vitro cellular & developmental biology. Plant》2007,43(1):2-8
Hawthorn (Crataegus spp.) is an important plant with a long history as an ornamental and a source of medicine. A protocol is outlined for adventitious
bud regeneration from leaf and cotyledon explants of Chinese hawthorn (C. pinnatifida Bge. var. major N.E.Br.). Adventitious buds were induced on both the leaves of sprouting winter buds and the leaves of in vitro plants, but the percentage of bud regeneration from leaves of in vitro plants was very low—less than 6%. On N6 medium supplemented with 31.08 μM BA and 9.67 μM NAA, the percentages of bud regeneration
from leaves of sprouting winter buds of cultivars “Liaohong” and “Qiujinxing” were 31.4% and 17.6%, respectively. The regeneration
abilities of three kinds of cotyledon explants, immature cotyledon, mature cotyledon, and cotyledon leaf, were compared. The
percentage of bud regeneration from cotyledon leaves was higher. On MS media supplemented with 4.44 μM BA and 4.54–9.08 μM
TDZ, the percentages of bud regeneration from cotyledon leaves of cultivars “Qiujinxing” and “Xiajinxing” were 27.7 ± 7.8%
and 20.1 ± 4.7%, respectively, and the numbers of buds per explant were 5.9 ± 1.6 and 3.2 ± 0.7, respectively. On B5 medium
supplemented with 2.22 μM BA, 2.32 μM Kn, and 0.57 μM IAA, adventitious buds grew quickly and 80–100% of buds developed into
shoots. The shoots rooted successfully with the two-step rooting method. Ninety days after transplantation, more than 80%
plants were survived. This system of adventitious bud regeneration from leaf and cotyledon explants could be useful for the
genetic transformation and polyploidization of Chinese hawthorn. 相似文献
11.
The regeneration of meristematic tissues from sporophytes of Laminaria digitata was studied by protoplast and tissue culture. Sequential treatment of explants in sterile seawater with 1% Betadine for 5 min,
1% commercial bleach for 1–2 min and 2% antibiotic treatment supplemented with 1 μM GeO2 overnight enabled viable explants as high as 55%. Different morphogenetic responses were observed from tissue culture on
media supplemented with plant growth regulators alone or in combination, mainly filamentous calluses up to 50% according to
the media. Dark green compact calluses were observed on two combinations: 4 μM Pi + 2 μM N-(2-chloro-4-pyridyl)-N’-phenylurea
(CPPU) and 0.04 μM Pi + 0.44 μM 6-benzylaminopurine. Thalloid-like structures comparable to adventitious buds were regenerated
on medium supplemented with 4 μM Pi + 0.45 μM zeatin but at low frequency suggesting a strong genotypic effect. Friable calluses
were developed from protoplasts in enriched medium with polyamines and containing 0.40 μM CPPU + 0.45 μM 2,4-dichlorophenoxyacetic
acid. In order to produce protoplasts, a one-step enzymatic protocol was developed and yields reached 22 × 106 protoplasts per gram of fresh weight. 相似文献
12.
V. Menéndez Y. Abul B. Bohanec F. Lafont H. Fernández 《Acta Physiologiae Plantarum》2011,33(6):2493-2500
The fern Asplenium nidus L. is in great demand as an ornamental plant. The aim of this work was to investigate the influence of phytohormones in promoting
a gametophytic and sporophytic growth in homogenized sporophytes tissue. Exogenous application of 0.5 and 5 μM N
6-benzyladenine, 0.05 and 0.5 μM indole-3-acetic acid (IAA), and 0.3 and 3 μM gibberellic acid (GA3) favoured sporophyte regeneration, whereas gametophyte regeneration took place when plant material was cultured in a hormone-free
liquid MS medium. The endogenous contents of the auxin IAA, the cytokinins trans-zeatin, trans-zeatin riboside, dihydrozeatin, dihydrozeatin riboside, isopentenyladenine and isopentenyladenosine, and the gibberellins
GA1, GA3, GA4, GA7, GA9 and GA20 in growing gametophytes and sporophytes were evaluated. Similar levels of the auxin and cytokinins and qualitative differences
in the gibberellins were found between both generations. 相似文献
13.
Activated charcoal affects morphogenesis and enhances sporophyte regeneration during leaf cell suspension culture of Platycerium bifurcatum 总被引:1,自引:0,他引:1
W.-L. Teng 《Plant cell reports》1997,17(2):77-83
In a leaf cell suspension culture of Platycerium bifurcatum, the incorporation of activated charcoal (AC) greatly increased the number of regenerated sporophytes even in growth regulator-free
medium. The degree of improvement was dependent on cell aggregate sizes and medium composition. The maximal increase was observed
in medium with 5.37 μM NAA and 4.44 μM BA, from 9 to 1520 sporophytes. The qualitative improvement by AC included: (1) regeneration of single sporophytes, which
was relatively less frequent in non-AC media, (2) prevention of the formation of gametophyte clusters prior to sporophyte
regeneration from 30- to 60-μm cells cultured in MS basal medium, (3) prevention of the formation of bud clusters, sporophytes
with multiple bud primordia, `nodule'-like bud clusters in growth regulator containing media, and (4) prevention of the occurrence
of hyperhydricity of regenerated sporophytes.
Received: 9 December 1996 / Revision received: 18 April 1997 / Accepted: 5 July 1997 相似文献
14.
Dr. Smitha Hegde Vipin Kumar Menon Rudolf Noronha L. D’Souza 《In vitro cellular & developmental biology. Plant》2006,42(6):508-513
Summary Callus induction and regeneration studies were carried out on a medicinal fern, Drynaria quercifolia native to Asian countries. It is a seasonal fern that regenerates only during the monsoons. Callus was induced on Knop’s
(1865) medium supplemented with 20 gl−1 sucrose, 8gl−1 agar, and either 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 4-amino-3,5,6-trichloropicolinic
acid (picloram), or indole-3-butyric acid at different concentrations. Morphogenetic callus obtained on 5 mgl−1 2,4,5-T was subcultured onto solid and liquid media (shaken flask and discontinuously stirred bioreactor cultures) for callus
proliferation and regeneration studies. A significant amount of sporophyte regeneration was observed on solid medium containing
10 mgl−1 6-(δ, δ-dimethylallylamino) purine (2iP). Sporophyte regeneration from callus followed an atypical pattern of development.
Leafy structures of single-cell thickness with a microrhizome were formed as sporophyte initials. Prolonged cultures of these
structures resulted in the formation of juvenile sporophytes in vitro. The use of liquid media resulted in increased biomass in culture. The present study is the first report of a successful
system for callus production and regeneration of sporophytes from leafy structures in ferns. The method can be successfully
applied for generation of biomass of D. quercifolia, throughout the year. 相似文献
15.
E. Schiantarelli A. De la Peña M. Candela 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2001,102(2-3):335-341
The Landsberg erecta× Columbia recombinant inbred lines (RILs) of Arabidopsis have been used in order to identify and localize chromosome regions involved in the genetic control of the in vitro regeneration
ability. Callus morphology (CM) and shoot regeneration (SR) traits have been considered for both leaf and root explants. The
MAPMAKER analysis of leaf culture data has revealed at least one chromosome region involved with CM and several with SR, the
29–30 region of chromosome 1 being common for the two traits. Root explants did not segregate for CM but several QTLs have
been detected for SR. The chromosome regions involved with leaf culture regeneration seem to be different from those of root
cultures, although the regeneration of abnormal shoots in leaf explants share two chromosome regions with the regeneration
of normal shoots in root cultures.
Received: 19 April 2000 / Accepted: 12 May 2000 相似文献
16.
The report describes a system for somatic embryogenesis and direct plant regeneration from the embryos of Manihot glaziovii. Somatic embryos were obtained by culturing young leaf lobes (3–6 mm long) adjacent to the apex in Murashige and Skoog medium
containing 18 μm 2,4-dichlorophenoxy acetic acid for 20 days and then transferring them to a maturation medium with 0.5 μm 6-benzylaminopurine. Secondary embryogenesis was induced from cotyledonary segments of somatic embryos by using the same
protocol as that for primary embryogenesis. For regeneration, somatic embryos were cultured in medium supplemented with 10−4 m kinetin and 53.4% of them developed into plantlets. Linamarin and linamarase were not detected in calli or in somatic embryos.
Linamarin content was found to be highest in leaves of regenerated plantlets, followed by stem and root tissues. Levels of
linamarase activity were almost the same in leaves and stem tissues and very low in roots.
Received: 19 April 1999 / Revision received: 11 August 1999 / Accepted: 17 August 1999 相似文献
17.
A protocol for the micropropagation of mature Eucalyptus tereticornis Smith has been developed using regenerated shoots from axillary bud explants. The trees were selected on the basis of their
better growth rate, physical and phenotypic characteristics and freedom from disease. Regeneration was obtained in modified
Murashige and Skoog (1962) medium. Evaluation of explant regeneration throughout the year indicated that the incidence of
browning of explants was maximum during the month of February, while dominance of the microbes in endogenously infected explants
peaked in August–September. Regeneration from primary explants was maximum during the months of March–April. Subcultures were
carried out every 4 weeks. Effects of hormones and media composition on regeneration and growth were studied. Phytagel induced
vitrification, while calcium chloride dihydrate reduced vitrification and induced the elongation of shoots. Best rooting was
obtained with half-strength, modified MS medium supplemented with 1.0 mg/l indolebutyric acid. Plantlets were hardened in
a nonsterile potting mix at high humidity and gradually exposed to the ambient environment over a period of 6 weeks, and upon
transfer to field conditions the survival rate varied from 84% to 100%.
Received: 15 October 1998 / Revision received: 18 June 1999 · Accepted: 12 July 1999 相似文献
18.
Mitsuhiro Takayama 《Ichthyological Research》2002,49(2):165-170
The spawning period of the Far Eastern brook lamprey, Lethenteron reissneri, in the headstream of the Himekawa River is estimated to be between mid-March and late May with the peak of spawning activity
between early April and early May. The sex ratio (female:male) in 1999 ranged from 1 : 2.5 to 1 : 3.0 (mean 1 : 2.8) and in
2000 from 1 : 0.8 to 1 : 4.0 (mean 1 : 2.4). In >90% of the observations of spawning nests, males outnumbered females. The
construction area of spawning nests tended to shift upstream during the spawning period. The nests were constructed at water
depths between 5 and 70 cm, water velocity between 10 and 30 cm/s, and on substrate with pebbles of 5–20 mm in diameter. Lethenteron reissneri constructed nests on substrate similar with Petromyzon marinus, but at shallower points and in areas with a slower water velocity.
Received: April 2, 2001 / Revised: December 12, 2001 / Accepted: December 27, 2001 相似文献
19.
This is the first report where shoot regeneration in strawberry cultivar Chandler has been achieved simultaneously through
both somatic embryogenesis and shoot bud formation. Direct somatic embryogenesis was observed in leaf discs which were cultured
on medium containing MS salts + B5 vitamins + 2% glucose + 18.16 μM thidiazuron (TDZ) and given both chilling and dark treatment for 2 wk at 4 ± 2°C followed
by incubation at 25 ± 2°C under 16-h photoperiod for third wk. After 3 wk, these explants were then subcultured on medium
containing MS salts + B5 vitamins + 2% glucose and incubated under 16-h photoperiod at 25 ± 2°C for further growth and development. Direct regeneration
via de novo shoot bud formation was observed in leaf disks which were given dark treatment and were cultured on medium containing MS
salts + B5 vitamins + 2% glucose supplemented with 9.08 μM TDZ. There was a synergistic effect of photoperiod, dark, and chilling treatments
on somatic embryogenesis, whereas chilling treatment had an inhibitory effect on shoot organogenesis. 相似文献
20.
The cost of sexual reproduction is incurred when the current reproductive episode contributes to a a decline in future plant performance. To test the hypotheses that a trade-off exists between current sexual reproduction and subsequent clonal regeneration and that resources limit reproduction and regeneration, plants of the widespread moss Pterygoneurum ovatum were subjected to induced sporophytic abortion, upper leaf removal, and nutrient amendment treatments. Sexually reproducing plants were slower or less likely to produce regenerative structures (protonemata or shoots) and produced fewer regenerative tissue areas or structures. The ability and the timeline to reproduce sexually and regenerate clonally were unaffected by an inorganic nutrient amendment. However, when leaves subtending the sporophyte were removed, the sporophytes were less likely to mature, tended to take a longer time to mature, and were smaller compared to sporophytes from shoots with a full complement of upper leaves. Our findings indicate that plants investing in sexual reproduction suffer a cost of decreased clonal regeneration and indicate that sporophyte maturation is resource-limited, with upper leaves contributing to the nutrition of the sporophyte. This study represents only the second explicit experimental demonstration of a trade-off between sexual and asexual reproduction in bryophytes. 相似文献