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1.
Adventitious shoot buds were induced from leaf and stem explants of Bacopa monnieri on Murashige and Skoog medium supplemented with benzyladenine or kinetin. The source of the explants as well as different
gelling agents in the medium were found to influence shoot induction and eventual shoot growth. The best response was obtained
in leaf explants taken from shoot cultures grown in medium supplemented with 2 μM benzyladenine and gelled with 0.2% gelrite. A transverse section of the leaf explant incubated in this medium showed several
shoot primordia emerging from the leaf surface. This system exhibited a potential for repeated harvesting of the shoots from
the original leaf explant as the latter continued to expand and regenerate new shoots, upon repeated periodical subculturing
onto fresh medium. However, the callusing response of the plant was very low. Qualitative TLC studies of the regenerated shoots
revealed a phytochemical profile similar to that of the field grown-plants.
Received: 20 March 1998 / Revision received: 1 December 1998 / Accepted: 12 December 1998 相似文献
2.
Summary A viable protocol has been developed for direct and indirect shoot regeneration of Vernonia cinerea. To establish a stable and high-frequency plant regeneration system, leaf and stem explants were tested with different combinations
of α-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), and benzylaminopurine (BA). Lateral buds on nodal explants
grew into shoots within 2 wk of culture in Murashige and Skoog (MS) basal medium supplemented with 20.9 μM BA. Excision and culture of nodal segments from in vitro-raised shoots on fresh medium with the same concentration of BA facilitated development of more than 15 shoots per node.
Similarly leaf, nodal, and internodal explants were cultured on MS basal medium supplemented with different concentrations
of BA, NAA, and IAA either alone or in combinations for callus induction and organogenesis. Shoot buds and/or roots were regenerated
on callus. Shoot buds formed multiple shoots within 4 wk after incubation in induction medium. Adventitious buds and shoots
proliferated when callus was cut into pieces and subcultured on MS basal medium containing 20.9 μM BA and 5.3 μM NAA. This combination proved to be the best medium for enhanced adventitious shoot bud multiplication, generating a maximum
of 50 shoots in 4 wk. This medium was also used successfully for shoot proliferation in liquid medium. Root formation was
observed from callus induced in medium containing 8.05–13.4 μM NAA. Regenerated shoots exhibited flowering and root formation in MS basal medium without any growth regulators. Plantlets
established in the field showed 85% survival and exhibited identical morphological characteristics as the donor plant. 相似文献
3.
F. Scaramuzzi G. Apollonio S. D’Emerico 《In vitro cellular & developmental biology. Plant》1999,35(3):217-221
Summary Leaf explants of Sinningia speciosa were cultured in vitro on Murashige and Skoog (MS) basal medium with various growth substances in order to regenerate shoots.
On MS medium supplemented with indoleacetic acid (IAA) and kinetin, 80% of the explants produced green callus and 25 to 30
shoots with roots per explant. On MS supplemented with IAA and N6 benzyladenine (BA), 80% of the explants produced green callus and 40 to 50 shoots per explant but lacked roots. After 3–4
mo., these shoots were removed from the initial explants and transferred separately onto MS supplemented with indolebutyric
acid for their elongation and successive rooting (3 mo.). Histological studies showed that the callus was associated with
mesophyll cell layers, primarily with the spongy parenchyma. The shoots regenerated at the callus surface and were associated
with newly differentiated vascular areas. Recurrent regenerations were obtained from leaf explants or apical meristems excised
from shoots of the previous subcultures. These explants, as compared to initial cultures, had a high frequency of regeneration
and also produced more shoots per explant. Chromosome numbers of root tip cells of the mother plant and of all in vitro-regenerated
plants remained constant: 2n=26. 相似文献
4.
Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration 总被引:7,自引:0,他引:7
The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of citrange (Citrus sinensis L. Osbeck×Poncirus trifoliata L. Raf.) have been investigated. Factors such as cocultivation period, preculture of explants, use of acetosyringone or feeder
plates during cocultivation, cocultivation on a medium rich in auxins, postcultivation in darkness, and different kanamycin
concentrations for selection were assessed. A 3-day cocultivation on a medium rich in auxins improved transformation frequencies,
since it increased the number of dividing cells competent for transformation, at the cut ends of the explants. Exposure of
explants to darkness for 4 weeks on selection medium resulted in further callus development and increased the regeneration
frequency of transgenic shoots. Furthermore, this treatment drastically reduced the number of regenerated escape shoots. A
transformation efficiency of 41.3% was achieved using the optimized transformation procedure.
Received: 4 November 1997 / Revision received: 7 January 1998 / Accepted: 13 February 1998 相似文献
5.
Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants 总被引:11,自引:0,他引:11
R. Ghorbel J. Juárez L. Navarro L. Peña 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(1-2):350-358
The green fluorescent protein (GFP) from Aequorea victoria has been introduced into three different citrus genotypes [Citrus aurantium L., C. aurantifolia (Christm.) Swing. and C. sinensis L. Osbeck×Poncirus trifoliata (L.) Raf.] which are considered recalcitrant to transformation, mainly due to low transformation frequencies and to the regeneration
of escape shoots at high frequencies from the Agrobacterium-inoculated explants. High-level GFP expression was detected in transgenic cells, tissues and plants. Using GFP as a vital
marker has allowed us to localize the sites of transgene expression in specific cells, always occurring in callus tissue formed
from the cambium of the cut ends of explants. Whereas green fluorescent shoots regenerated in all cases from this callus,
most escapes regenerated directly from explants with almost no callus formation. Thus, development of callus from cambium
is a prerequisite for citrus transformation. Furthermore, in vivo monitoring of GFP expression permitted a rapid and easy
discrimination of transgenic and escape shoots. The selection of transgenic shoots could be easily favored by eliminating
the escapes and/or by performing shoot-tip grafting of the transgenic buds soon after their origin. GFP-expressing shoots
have also been observed in citrus explants co-cultivated with Agrobacterium but cultured in a medium without the selective agent kanamycin. This opens the possibility to rescue the transgenic sectors
and to regenerate transgenic plants without using selectable marker genes conferring antibiotic or herbicide resistance, which
is currently a topic of much discussion for the commercialization of transgenic plants.
Received: 28 October 1998 / Accepted: 28 November 1998 相似文献
6.
This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium
supplemented with 0.5–20 μM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing
percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained
on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin
(2–8 μM) alone or in combination with a low level (0.5 μM) of 2,4-D or NAA. The best regeneration of shoots from root explants
was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained
with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were
achieved from cotyledonary explants at 4 μM BA and 0.5 μM NAA. The presence of 0.5 μM or higher levels of 2,4-D in shoot induction
medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully
acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction. 相似文献
7.
A liquid culture protocol was developed to regenerate shoots from cotyledons of germinating seeds of jute (Corchorus capsularis L.). Reproducibility of the protocol was tested in three genotypes of jute. Highest bud differentiation rates into normal
shoots (via shoot-forming explants) were obtained on modified Murashige and Skoog's liquid medium containing 2.7 μM α-naphthaleneacetic acid and 4.4 μM benzylaminopurine. Regenerated shoots were excised, and the best root formation could be induced in medium with 2.5 μM indole-3-butyric acid and 1.5% sucrose. Bud primordia were formed directly on the cut surface of the cotyledons. Scanning
electron micrographs and histological studies confirmed the organogenic nature of the regenerated shoots. The physical condition
of the culture medium and the age of the explants played crucial roles in the induction of shoot development using shoots;
2-day-old explants being optimal. Approximately 70% of the shoots were successfully established in soil after hardening.
Received: 20 October 1997 / Revision received: 4 October 1998 / Accepted: 27 October 1998 相似文献
8.
Nodal explants from selected trees of gum karaya (Sterculia urens Roxb.) in the adult growth phase cultured on Murashige and Skoog (MS) medium supplemented with 6.62 μm N6-benzylaminopurine (BAP) produced an average of six adventitious shoots in 30 days. Shoots were rooted in vitro on 1/4-strength
MS medium containing 9.82 μm indole-3-butyric acid. Nodulated callus was produced from hypocotyl explants cultured on MS medium supplemented with 4.52
μm 2,4-dichlorophenoxyacetic acid and 8.90 μm BAP. Somatic embryos developed when the nodulated callus was transferred to MS medium containing 0.45 μm thidiazuron (TDZ). TDZ treatment for 2 days gave the optimum response. Over 30% of the somatic embryos developed into plantlets
when transferred to 1/4-strength MS basal medium without any growth regulators. Plantlets produced from adventitious shoots
and somatic embryos were acclimatized to ex vitro conditions and established in the field.
Received: 26 November 1997 / Revision received: 14 April 1998 / Accepted: 11 May 1998 相似文献
9.
Jesús Arellano Sara Isabel Fuentes Patricia Castillo-España Georgina Hernández 《Plant Cell, Tissue and Organ Culture》2009,96(1):11-18
A protocol for in vitro regeneration via indirect organogenesis for Phaseolus vulgaris cv. Negro Jamapa was established. The explants used were apical meristems and cotyledonary nodes dissected from the embryonic
axes of germinating seeds. Several auxin/cytokinin combinations were tested for callus induction. The best callus production
was obtained with medium containing 1.5 μM 2,4-dichlorophenoxyacetic acid. After 2 weeks of growth calli were transferred
to shooting medium containing 22.2 μM 6-benzylaminopurine. Shoots regenerated with a frequency of approximately 0.5 shoots
per callus, and upon transfer to rooting medium these shoots produced roots with 100% efficiency. Histological analyses of
the regeneration process confirmed the indirect organogenesis pattern. Greenhouse grown regenerated plants showed normal development
and were fertile. The protocol was reproducible for other nine P. vulgaris cultivars tested, suggesting a genotype independent procedure. 相似文献
10.
Boon Chin Tan Chiew Foan Chin Peter Alderson 《Plant Cell, Tissue and Organ Culture》2011,105(3):457-463
An indirect in vitro plant regeneration protocol for Vanilla planifolia has been established. Juvenile leaf and nodal segments from V. planifolia were used as explants to initiate callus. Nodal explants showed better callus initiation than juvenile leaf explants, with
35.0% of explants forming callus when cultured on Murashige and Skoog (MS) basal medium supplemented with 2.0 mg/l 1-naphthylacetic
acid (NAA) and 1.0 mg/l 6-benzyladenine (BA). Almost 10.0% of juvenile leaf explants were induced to form callus on the MS
basal medium containing 2.0 mg/l NAA and 2.0 mg/l BA, whereas no callus formed in the presence of any concentrations of 2,4-dichlorophenoxyacetic
acid (2,4-D) and BA. After 8 weeks, callus generated was transferred to MS basal medium containing 1.0 mg/l BA and 0.5 mg/l
NAA. A mean number of 4.2 shoots per callus was produced on this medium, with a mean length of 3.8 cm after 8 weeks of culture.
Roots formed on 88.3% of plantlets when they were cultured on MS medium supplemented with 1.0 mg/l NAA, with a mean length
of 4.4 cm after 4 weeks of culture. Of the rooted plantlets, 90.0% survived acclimatisation and were making new growth after
4 weeks. 相似文献
11.
Two plant regeneration methods applicable to Leucaenaleucocephala were developed. In the first method, involvingorganogenesis via callus formation, cotyledon, hypocotyl and root segments wereinitiated on MS medium containing different concentrations ofN6-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), andnaphthaleneacetic acid (NAA). Both compact (type I) and friable (type II) calliwere obtained from the cotyledon and hypocotyl explants treated with differentconcentrations of the growth regulators. Shoots were generated only from thefriable calli formed from the cotyledon explants. The calli formed from thehypocotyl explants did not generate shoots and the root explants died withoutforming callus. Cotyledon explants from 3–4 day old seedlings showedmaximum callus induction compared to those from older seedlings. In a secondmethod involving direct organogenesis, excised cotyledons were cultured on 1/2MS medium containing 10–35 mg l–1N6-benzyladenine (BA) for 7–14 days. Transfer of thecotyledonsto regeneration medium containing low BA resulted in callus formation andsubsequent shoot regeneration from the base of the excised cotyledon explants,with up to 100% frequency. Regenerated shoots rooted best on a basal mediumcontaining no growth regulators. 相似文献
12.
Bimal Kumar Ghimire Chang Yeon Yu Ill-Min Chung 《Plant Cell, Tissue and Organ Culture》2012,108(3):455-464
A simple and efficient procedure was developed for in vitro propagation of Solanum aculeatissimum Jacq. using leaf and petiole explants cultured on Murashige and Skoog (MS) medium supplemented with α-naphthalene acetic
acid (NAA) and 6-benzyladenine (BA). Effects of various plant growth regulators, explant types, carbohydrates, and basal salts
on induction of adventitious shoots were also studied. Leaf explants appeared to have better regeneration capacity than petiole
explants in the tested media. The highest regeneration frequency (79.33 ± 3.60%) and shoot number (11.33 ± 2.21 shoots per
explant) were obtained in leaf explants in MS medium containing 3% sucrose and 0.8% agar, supplemented with 0.1 mg/l NAA and
2.0 mg/l BA, whereas petiole explants were more responsive to 0.1 mg/l NAA and 1.0 mg/l thiadiazuron. Developed shoots rooted
best on MS medium with 1.0 mg/l indole acetic acid (IAA), producing 18.33 ± 2.51 roots per shoot. Histological investigation
showed that the shoot buds originated mainly from epidermal cells of wounded tissues, without callus formation. The regenerated
plantlets were successfully acclimatized in a greenhouse, where over 90% developed into morphologically normal and fertile
plants. Results of flow cytometry analysis on S. aculeatissimum indicated no variation in the ploidy levels of plants regenerated via direct shoot formation and showed almost the same phenotype
as that of mother plants. This adventitious shoot regeneration method may be used for large-scale shoot propagation and genetic
engineering studies of S. aculeatissimum. 相似文献
13.
Rubén Mallón Juan Rodríguez-Oubiña María Luz González 《Plant Cell, Tissue and Organ Culture》2011,106(3):523-530
In vitro culture is currently used to produce plant material for ex situ conservation of endangered species. In this study,
an efficient protocol for shoot regeneration from leaves and roots was developed for Centaurea ultreiae, a critically endangered species. Organogenesis from leaf and root explants was promoted by incubating these explants on
half-strength Murashige and Skoog (MS) medium in the presence of one of four different cytokinins [6-benzyladenine (BA), zeatin,
kinetin or N6-(2-isopentenyl) adenine (2iP)], each provided at five different levels. Shoot organogenesis was induced in both explants.
The best response, 90% of leaf explants producing a mean of 2.48 shoots per explants and 94.3% of root explants producing
a mean of 5.60 viable shoots per explants, was observed when explants were incubated on a medium containing 0.55 μM BA. Histological
studies revealed connectivity between vascular tissues of regenerated shoots and cambial cells of leaf explants. Moreover,
adventitious shoots were derived from pericycle cells of root explants and parenchymatic cells of callus tissues. 相似文献
14.
Manuel L. Robert J. L. Herrera F. Contreras Keith N. Scorer 《Plant Cell, Tissue and Organ Culture》1987,8(1):37-48
In vitro plant regeneration of Agave fourcroydes Lem. (Agavaceae) is described. Results suggest that the NO3
-:NH4
+ balance in the culture medium is a key factor controlling callus growth and organogenesis in rhizome cultures. Stem callus showed limited organogenic capacity, but high cytokinin concentrations induced adventitious shoot formation on stem explants. When these shoots were excised and subcultured, new callus formed at their base from which new shoots arose. The shoots from stem explants and rhizome callus formed extensive root systems in vitro and were transferred to pot culture with a 90% survival rate. 相似文献
15.
Abstract Callus production, shoot formation via organogenesis and rooting of the regenerated shoots are reported in an Egyptian variety of Pisum sativum L. Calli were initiated from hypocotyl, leaf, root and mature embryo explants when cultured on MS medium containing B5 vitamins and supplemented with 2 mg/l 2,4-D+1 mg/l kin. Among the different types of explants, hypocotyl showed best potential for callus proliferation. Hypocotyl, leaf and immature cotyledon explants were used for shoot organogenesis. The best results of shoot formation were achieved when hypocotyl explants were cultured on MS-medium supplemented with 2 mg/l BA+1 mg/l NAA. However, immature cotyledon explants showed the highest frequency of shoot formation with 1 mg/l BA. Data of in vitro rooting showed that maximum root frequency occurred on culture medium containing half strength of MS salts, 40 g/l sucrose and 2 mg/l NAA. 相似文献
16.
The nodal and internodal explants excised from the orthotropic shoots of Sesbania sesban var. bicolor elicited the development of shoots directly from the explants as well as via an intervening callus phase on Nitsch (N) medium. On benzyladenine (BA) supplemented media, the adventitious shoot buds developed
involving a callus phase. An average of 8.9 ± 4.1 shoots developed per nodal explant on N medium containing 0.5 mg dm−3 BA in 95 % cultures, whereas 65 % cultures of internodal explants developed shoots with an average of 5.9 ± 3.6 shoots per
explant on N medium supplemented with 1.0 mg dm−3 BA. On kinetin (Kn) supplemented medium shoots developed directly from the surface of both the explants at all the concentrations
tried. Nodal explants on N medium supplemented with 1.5 mg dm−3 Kn developed an average of 12.5 ± 7.9 shoots per explant in 100 % cultures, while internodal explants induced an average
of 11.6 ± 7.4 shoots per explant in 75 % explants at 0.5 mg dm−3 Kn. The in vitro regenerated shoots developed roots when implanted on N medium supplemented with 2 mg dm−3 indole-3-butyric acid (IBA), after 30 d of inoculation. The in vitro developed plantlets were initially acclimatized under controlled conditions for four months, prior to their transfer to the
field.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
17.
Callus was produced on cotyledon, shoot tip, hypocotyl and root explants of twoCorchorus species on several media. Cytokinin was necessary for callus production on cotyledon explants. BothC.olitorius genotypes produced most callus on media with zeatin and either NAA or IAA, and theC.capsularis genotype produced most callus on media with IAA and either zeatin or BA. High frequencies of regenerated shoots were obtained
from shoot tip explants of both species, from the apical meristem and from callus. Media with 2.0 mg 1−1 BA were superior for both species, and media with zeatin were equally good forC.capsularis only. More regeneration was obtained for all genotypes after subculture of callus on media with 2.0 mg 1−1 zeatin. Cotyledon callus produced less regeneration, also with differences between genotypes; explants of both genotypes
ofC.olitorius produced regeneration on a medium with NAA and zeatin, and theC.capsularis genotype produced regeneration on a medium with IAA and BA. Limited regeneration from root explant callus was obtained forC.capsularis only on medium with BA and IAA. Regeneration was not obtained from hypocotyl callus. Further regeneration of shoots of both
species was obtained from secondary callus after subculture, and from nodal segments of regenerated shoots and of seedling
shoots cultured on basic MS medium without growth hormones. Roots were produced on about 80% of all shoots after transference
to medium with 0.2 mg 1−1 IBA, and rooted plantlets survived and flowered normally after transference to compost. 相似文献
18.
Summary In researching the application of genetic transformation to lily breeding, callus formation from cultured explants and plant
regeneration from induced calluses were examined in 33 Lilium genotypes, 21 species, three Asiatic hybrids, two LA hybrids, two Longiflorum hybrids, three Oriental hybrids, and two Trumpet
hybrids. Seed, bulb scale, leaf, or filament explants were placed on a medium containing 4.1 μM 4-amino-3,5,6-trichloropicolinic acid (picloram; PIC) and cultured in the dark. After 2 mo., callus formation was observed
in 30 genotypes, and a formation frequency of more than 50% was obtained in 24 genotypes. Bulb scale and filament explants
showed great ability to form calluses, whereas seeds had poor ability. Most of the induced calluses were yellow and had a
nodular appearance. When subcultured onto the same fresh medium, twofold or more increases in callus mass were obtained in
1 mo. for 15 genotypes. Callus lines showing sustained growth 1 yr after the initiation of subculture were examined for their
ability to produce shoots on a medium without plant growth regulators (PGRs) and a medium containing 22 μM 6-benzyladenine (BA). Shoot regeneration was observed in all genotypes examined, and a regeneration frequency of over 80%
was obtained in 20 genotypes. Initial explants used for callus induction and callus type (nodular or friable) had no effect
on shoot regeneration. Most of the regenerated shoots developed into complete plantlets following their transfer to a PGR-free
medium. 相似文献
19.
L. V. Hiregoudar H. N. Murthy J. G. Bhat A. Nayeem B. P. Hema E. J. Hahn K. Y. Paek 《Biologia Plantarum》2006,50(2):291-294
This report describes in vitro shoot induction and plant regeneration from mature nodal explants of Vitex trifolia L. on Murashige and Skoog (MS) medium fortified with benzylaminopurine (BAP), kinetin (KN), thidiazuron (TDZ), adenine (ADE),
and 2-isopentenyladenine (2-iP) (0.25 – 10.0 μM). Multiple shoots differentiated directly without callus mediation within
3 weeks when explants were cultured on medium supplemented with cytokinins. The maximum number of shoots (9 shoots per explant)
was developed on a medium supplemented with 5.0 μM BAP. Shoot cultures was established repeatedly subculturing the original
nodal explant on the same medium. Rooting of shoots was achieved on half strength MS medium supplemented with 0.5 μM naphthaleneacetic
acid (NAA). Rooted plantlets transferred to pots containing autoclaved soil and vermiculite mixture (1:1) showed 90 % survival
when transferred to outdoor. 相似文献
20.
A rapid and efficient in vitro plant regeneration method was developed for Matteuccia struthiopteris (L.) Todaro (Ostrich fern). Side shoots, originating in meristems of sectioned rhizomes, were used as explant material. A
very high rate of meristem multiplication was achieved by culturing the explants in half-strength MS liquid medium supplemented
with 2.0 mg/l N-(4-Pyridyl)-N′-phenylurea (4-PU) and 0.5 mg/l thidiazuron (TDZ). Multiplication of the shoot primordia was
faster in suspension culture than on solid medium. Rhizogenesis and growth of regenerants were best achieved on hormone-free
one-quarter-strength MS solid medium amended with 0.4% agar and 1.0% activated charcoal. Regenerated plantlets continued to
grow after transfer to soil in a phytotron.
Received: 19 March 1998 / Revision received: 17 July 1998 / Accepted: 3 August 1998 相似文献