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1.
A procedure for multiple shoot formation from cotyledonary node explants of Eastern redbud (Cercis canadensis L.) cultured on DKW medium containing benzyladenine (BA) and thidiazuron (TDZ) was developed. Explants on medium with TDZ
in combination with BA produced higher numbers of shoots than with either cytokinin alone. The highest number of shoots (7.8
to 9.8 shoots per explant) was obtained when explants from 4 to 10 day-old seedlings were treated with a combination of 10
or 15 μM BA and 0.5 or 1.0 μM TDZ for 20 days before being transferred to the same medium without TDZ. The number of shoots
formed was increased from 5.8 to 7.2 shoots per explant by cutting through the cotyledonary node prior to culture. Histological
studies indicated that the shoots were formed from actively dividing cells located at the axillary bud region. Shoots formed
roots in half strength woody plant medium (WPM) supplemented with 10 to 200 μM indole-3-butyric acid (IBA) cultured for 15
days prior to transfer to greenhouse medium. 相似文献
2.
H. Kodja I. Robene-Soustrade L. Humeau J. Figier J. Govinden-Soulange A. Gurib-Fakim 《Plant Growth Regulation》1998,25(2):75-80
A micropropagation protocol for Psiadia arguta, an endangered endemic plant from Mauritius is described using 15-day old in vitro seedling explants without the radicle. MS basal medium supplemented with TDZ (0.5–1 mg/l) proved to be the most effective medium for the induction of cotyledonary axillary buds as compared to MS medium containing NAA (0.5 mg/l) or both NAA (0.5 mg/l) and TDZ (0.5–1 mg/l). In fact, after transfer to hormone free MS medium, microshoots were obtained only from seedling explants cultured on media containing only TDZ. Regenerated shoots elongated and rooted when cultured on MS8900 containing IBA (0–1 mg/l). Hormone-free MS8900 was the best medium for rooting and development of plantlets for acclimatization. 相似文献
3.
A procedure for multiple shoot formation from somatic embryo explants of Eastern redbud (Cercis canadensis L.) cultured on DKW medium containing benzyladenine (BA) and thidiazuron (TDZ) was developed. TDZ in combination with BA produced more shoots than either treatment alone. The highest number of shoots (3.3 to 3.4 shoots per explant) was obtained from partially desiccated and wounded explants treated with a combination of 5 or 10 M BA and 0.5 or 1.0 M TDZ for 20 days before being transferred to the same medium without TDZ. The number of shoots formed was increased from 1.5 to 3.2 shoots per explant by cutting through the cotyledonary node prior to culture. In addition, the frequency of explants forming shoots was increased by desiccation of somatic embryo explants to approximately 50% moisture and by using somatic embryos with two well formed cotyledons as explants.Abbreviations ABA
abscisic acid
- BA
benzyladenine
- CRD
Completely randomized design
- DKW
Driver and Kuniyuki medium
- LSD
Least significant differences
- TDZ
thidiazuron 相似文献
4.
Stimulation of in vitro shoot proliferation from nodal explants of cassava by thidiazuron, benzyladenine and gibberellic acid 总被引:2,自引:0,他引:2
Basdeo Bhagwat Luiz G. F. Vieiral Larry R. Erickson 《Plant Cell, Tissue and Organ Culture》1996,46(1):1-7
Multiple shoots were produced from nodal explants of cassava (Manihot esculenta Crantz) by a two-step procedure: a 6- to 8-day exposure to 0.11–0.22 µM thidiazuron (TDZ) in liquid Murashige and Skoog (MS) medium followed by culture on agar-solidified MS medium supplemented with 2.2 µM 6-benzyladenine (BA) and 1.6 M gibberellic acid (GA3). TDZ caused the nodal explants to expand and this expansion (growth) continued during culture with BA and GA3. From this expanded explant, clusters of buds and fasciated stems developed continuously and these gave rise to shoots. The shoot proliferation process was open-ended, yielding an average of 31.5 shoots per nodal explant after 10 weeks of culture with genotype CG 1–56. A positive response was also obtained from seven other genotypes evaluated with this protocol.Abbreviations BA
6-benzyladenine
- BM
basal medium
- DPU
1,3-diphenylurea
- GA3
gibberellie acid
- 2iP
isopentenyladenine
- MSM
multiple shoot medium
- NAA
1-naphthaleneacetic acid
- PGR
plant growth regulator
- TDZ
thidiazuron
- Z
zeatin 相似文献
5.
Murashige and Skoog’s (MS) basal medium with benzylaminopurine (BA), kinetin (KN), zeatin (Z), and thidiazuron (TDZ) were
tested for induction of multiple shoots from mature-tree-derived axillary meristems of Pongamia pinnata. Sprouting of buds was 64% on medium devoid of plant growth regulators (PGR). Incorporation of BA, KN, or Z was ineffective
in enhancing sprouting frequency or induction of multiple shoots. Sprouting was completely suppressed in the presence of TDZ.
Caulogenic buds appeared in nodal meristems of these explants after withdrawal of TDZ. The number of shoot buds was more on
explants precultured in higher concentrations. At higher concentrations of this PGR, a swelling developed at the axil. Multiple
shoot primordia appeared and differentiated from this swelling after culturing these explants on MS medium for six passages
of 2 wk each. Shoots were harvested and cultured on 0.45 μM TDZ for further proliferation. Primary explants after harvesting
of shoots were identified as ‘stump’. Reculturing of stumps on 0.45 μM TDZ produced more shoots. This step was followed for
six cycles to obtain additional shoots in each cycle. Shoots maintained on 0.45 μM TDZ elongated and rooted (70%) on growth
regulator-free medium. Rooted shoots (65%) survived transfer to a sand/soil mixture. This report describes the protocol for
micropropagation of P. pinnata using mature-tree-derived nodal meristems. Recycling of mature stock to produce a stream of useable shoots for subculturing
and eventual stabilization is of great value and can possibly be generalized as an isolation protocol especially for woody
species. Repeated proliferation of caulogenic buds from the same origin may also find application in rescue of endangered
germplasm. 相似文献
6.
Nand Narendra Drew Roderick A. Ashmore Sarah 《Plant Cell, Tissue and Organ Culture》2004,77(2):193-201
Nodal explants from in vitro grown seedlings of Davidsonia pruriens and D. jerseyana, established on MS media were treated with various concentrations of three cytokinins. D. pruriens developed optimum shoot growth in terms of shoot height and number of leaves per shoot when 1.0 µM BA was added to basal MS medium while optimum shoot growth for D. jerseyana was obtained when 0.01 µM 2iP was added to the medium. Optimum root initiation and development was obtained when actively growing axillary shoots were cultured on 1/2MS medium plus 32.2 µM IBA for 3–5 days for D. pruriens and 2–3 days for D. jerseyana before transfer to PGR-free medium containing 10 µM riboflavin. Root initiation of more than 80% was achieved with multiple genotypes of D. pruriens and three genotypes of D. jerseyana using juvenile material. The plantlets were transferred to pots and grown in the greenhouse with a success rate of 60% for D. pruriens and 75% for D. jerseyana. Adult D. jerseyana stem explants produced 2–5 shoots per nodal explant upon treatment with 0.1 µM BA. Side shoots from adult D. jerseyana produced similar results for shoot multiplication as for juvenile material. Protocol for multiplication of adult D. pruriens was achieved with much greater difficulty by using material from the green house. Axillary shoots were initiated when 100 µM TDZ was applied to the stem of an adult pot plant and the resultant side shoots were cultured on MS medium containing 1.0 µM BA and 1.0 µM GA3. 相似文献
7.
In vitro micropropagation of holy basil (Ocimum sanctum L.), an Indian medicinal herb, has been accomplished on Murashige and Skoog (MS) medium utilizing young inflorescence explants. MS supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or thidiazuron (TDZ) produced only non-morphogenetic callus. Direct multiple shoots differentiated within 2--3 weeks when explants were cultured on MS containing 6-benzyl aminopurine (BAP). Of the various levels of BAP tested, MS + BAP (1.0 mgl–1) produced the maximum number of shoots. Incorporation of indole-3-acetic acid (IAA) (0.05 mgl–1) along with BAP (1.0 mgl–1) in the culture medium showed a marked increase in the number of shoots. About 92% of the in vitro regenerated shoots rooted on MS hormone-free medium within 2--3 weeks of culture and 85% of the micropropagated plantlets could be successfully established in soil, where they grew normally. 相似文献
8.
Charlene Chang Ben A. Moll Kathleen B. Evenson Mark J. Guiltinan 《Plant Cell, Tissue and Organ Culture》1996,45(1):61-66
In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron 相似文献
9.
Several culture conditions were examined for promoting efficient plant regeneration from explants of Gentiana. Adventitious shoot regeneration from leaf explants of cv. WSP-3 was very superior on MS medium, compared to B5 medium, supplemented with four cytokinins (TDZ, 4PU-30, BA and zeatin). An auxin / cytokinin combination was required for regeneration. TDZ was the most effective cytokinin, while NAA was more effective than IAA or 2,4-D. Optimum conditions for regeneration from explants (leaf, stem and root) of cv. WSP-3, evaluated in terms of regeneration frequency and number of regenerated shoots per explant, were TDZ and NAA in combination, 5–10 mg/l and 0.1 mg/l for leaf and stem explants, and 10 mg/l and 1 mg/l for root explants, respectively. Application of these conditions to eight other commercial cultivars resulted in 30–100% regeneration from leaf explants. The number of chromosomes in each of ten regenerated plants of each cultivar was diploid, 2n=26. Shoots regenerated in vitro were rooted in phytohormone-free medium and transferred to soil.Abbreviations MS medium
Murashige and Skoog's medium (Murashige and Skoog 1962)
- B5 medium
Gamborg B5 medium (Gamborg et al. 1968)
- BA
6-benzylaminopurine
- TDZ
N-phenyl-N'-1,2,3-thiadiazol-5-yl urea
- 4PU-30
N-(2-chloro-4-pyridyl)-N'-phenylurea
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
1-naphthaleneacetic acid 相似文献
10.
Summary A procedure for micropropagation of pitaya using thidiazuron (TDZ) and naphthaleneacetic acid (NAA) is described. Explants
were excised from young joints of mature plants and cultured on Murashige and Skoog medium (MS) containing 0.5 μM NAA and 0.5 μM TDZ. Shoots produced from these first explants were cut up to produce secondary explants, either by decapitation or by longitudinal
division into three parts. The decapitated and longitudinal explants were cultured on MS supplemented with 0.5 μM NAA and either 0.01, 0.09, 0.5, or 0.9 μM TDZ. Decapitated explants produced more shoots at higher frequency that the longitudinal explants. For both types of secondary
explants, most shoots were developed from the distal parts. Shoots produced from secondary explants were rooted in MS and
then transferred to soil where they produced normal plants. 相似文献
11.
S. Zhihui M. Tzitzikas K. Raemakers M. Zhengqiang R. Visser 《In vitro cellular & developmental biology. Plant》2009,45(6):776-782
Pea (Pisum sativum L. cv. Espace) seeds directly cultured on thidiazuron (TDZ)-containing medium formed high numbers of shoots. The number of
shoots per seedling depended on the concentration and duration of the TDZ treatment. The best treatment was 12-wk incubation
on MS medium supplemented with 4 mg/l TDZ followed by 4-wk culture on MS medium supplemented with 0.5 mg/l benzylaminopurine
(BA) and produced more than 400 shoots/seedling. Isolated shoots rooted at a high frequency on MS medium containing 2–3 mg/l
indole-3-butyric acid and 2 mg/l α-naphthalene acetic acid. In addition to the formation of shoots, bud-containing tissues
(BCT) were formed at the cotyledonary nodes, shoot nodes, tendrils, stipules, and internodes. The BCT from the cotyledonary
nodes and the shoot nodes was maintained in its pure state on MS medium supplemented with 4 mg/l TDZ by repeated culture.
Shoot development was accomplished when the BCT were left on MS medium supplemented with 4 mg/l TDZ without subculture prior
to transfer onto MS medium supplemented with 0.5 mg/l BA. 相似文献
12.
Micropropagation of juvenile sycamore maple via adventitious shoot formation by use of thidiazuron 总被引:4,自引:0,他引:4
Eva Wilhelm 《Plant Cell, Tissue and Organ Culture》1999,57(1):57-60
Zygotic embryos of sycamore maple (Acer pseudoplatanus) were dissected into plumule, hypocotyl and radicle sections. The segments
were placed on MS medium containing 1 μM 6-benzyladenine (BA) and/or 0.02 μM to 0.1 μM thidiazuron (TDZ). Hypocotyl and plumule
explants produced callus, adventitious buds and shoots with increasing plant growth regulator concentrations. Hypocotyls produced
more, but smaller shoots compared to plumule segments. Subculturing excised shoots and calluses on Murashige and Skoog (MS)
media with 1 μM BA and/or 0.04 μM TDZ led to continuous production of shoots. The best proliferation capacity occurred with
0.04 μM TDZ and 1.0 μM BA, both shoots and calluses. This combination showed a stimulatory effect also on length of newly
formed shoots. Calluses performed generally better compared to shoot explants independent of growth regulator treatment. Excised
shoots 2 to 3-cm-long were successfully rooted on MS media either with or without growth regulators (123 μM IBA pulse) followed
by transfer to the greenhouse.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Seemab Mukhtar Naseem Ahmad Md Imran Khan Mohammad Anis Ibrahim M. Aref 《Physiology and Molecular Biology of Plants》2012,18(4):381-386
A comparative performance of two explants types (CN and Nodal) for their
efficiency to induce multiple shoot regeneration in Clitoria
ternatea has been carried out. Thidiazuron (TDZ) in different
concentrations (0.05–2.5 μM) was used as a supplement to the Murashige and Skoog’s
(MS) basal media. Explant type apart, two factors viz. concentration and exposure
duration to TDZ played an important role in affecting multiple shoot regeneration.
Cotyledonary node explants produced the best results at 0.1 μM TDZ, while in nodal
explants the highest rate of shoot formation was achieved on MS medium supplemented
with 1.0 μM TDZ. In both the explants, shoot multiplication increased when the
regenerated shoots were subcultured on hormone free MS medium after 4 weeks of
exposure to TDZ. Among the two, cotyledonary node explants produced considerably
higher number of shoots at a comparatively lower concentration of TDZ than nodal
explants. The regenerated shoots rooted best on MS medium containing 1.0 μM
indole-3-butyric acid (IBA) and were successfully established in pots containing
garden soil with 88 % survival rate. All the regenerated plants showed normal
morphology and growth characteristics. 相似文献
14.
Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant 总被引:1,自引:0,他引:1
T. Dennis Thomas 《Acta Physiologiae Plantarum》2007,29(5):455-461
Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented
with various concentrations of BAP (2–8 μM) or TDZ (2–8 μM) alone or in combination with NAA (0.5 and 1.0 μM) produced low
shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15,
25 or 50 μM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After
pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 μM), TDZ (6 μM),
BAP (4 μM) + NAA (1.0 μM), TDZ (6 μM) + NAA (0.5 μM). Control explants were incubated directly on the medium without any pretreatment.
The pretreatment of explants with 15 μM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum
response was observed on MS medium supplemented with 6 μM TDZ. In this medium, 96 % cultures responded with an average number
of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots
per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ
alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present
investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides. 相似文献
15.
Nodal explants, excised from young shoots from mature trees of Syzygium cuminii, were cultured on MS medium supplemented with different concentrations of BA or kinetin. Among these, BA (0.5 or 1.0 mg l–1) induced greening and opening of the incipient shoot buds, which however did not elongate. Elongation of the shoot buds was facilitated on MS medium with 1.0 mg l–1 BA supplemented with casein hydrolysate (1.5 g l–1) or glutamine (200 mg l–1). Nodal explants (microcuttings), taken from shoots developed in vitro, also developed multiple shoots when cultured on MS with1 mg l–1 BA. These explants did not require an additional supply of reduced nitrogen, for further normal development. Shoots developed from explants from mature trees and microcuttings were rooted by sub-culturing them on Knop's medium supplemented with 2% sucrose and 1 mg l–1 IAA. The plants that developed in vitro were planted in soil and were transferred to the field after an acclimatization period of 7–8 months. These plants have been thriving well for more than three years and have no apparent phenotypic aberrations. 相似文献
16.
Summary A procedure for plant regeneration, flower and plant formation from petiolar and inflorescence nodal explants of culantro
is discribed. Leaf petioles were excised from young leaves of non-flowering plants while nodal explants were excised from
the inflorescence. Explants were cultured in Murashige and Skoog (MS) medium alone or supplemented with 0.5μM naphthaleneacetic acid (NAA) and 0.9, 1.8, 4.5, or 9 μM thidiazuron (TDZ). All explants produced multiple shoots. In addition, nodal explants formed flowers. Shoot number, flower
number and shoot length were influenced by TDZ and NAA. Rooted shoots from both types of explants were transferred to soil
where plants were successfully established. 相似文献
17.
Wongwicha W Tanaka H Shoyama Y Tuvshintogtokh I Putalun W 《Zeitschrift für Naturforschung. C, Journal of biosciences》2008,63(5-6):413-417
Licorice plants, Glycyrrhiza glabra, G. uralensis, and G. inflata, were investigated for callus induction using Murashige and Skoog (MS) medium combined with auxins and cytokinins. After 4 weeks of culture, 33-100% of leaf or stem explants formed calli. Maximum of shoot induction from callus cultures was achieved by G. inflata stem explants cultured on MS medium supplemented with 1 mg/l alpha-naphthaleneacetic acid (NAA) and 0.5 mg/l 6-benzyladenine (BA) (67%) which also gave maximum shoot formation per explant (two shoots per explant). These results indicated that all three Glycyrrhiza species regenerated shoots from callus cultures on MS medium combined with NAA and BA or only thidiazuron (TDZ; 0.1 and 0.5 mg/l). Glycyrrhizin contents of G. uralensis calli induced using MS medium in combination with NAA and BA [(27.60 +/- 8.47) microg/g DW] or TDZ alone [(36.52 +/- 2.45) microg/ g DW] were higher than those found in other combinations. 相似文献
18.
Seabuckthorn (Hippophae rhamnoides) is a multipurpose small tree with unique berries of high nutritional and pharmaceutical values. A clonally propagated plant
originating from a 20-year-old tree of H. r. rhamnoides × mongolica hybrid cultivar Julia and seedling offspring of this cultivar were investigated regarding induction of shoot organogenesis
in leaf explants and in roots of intact seedlings, and induction of direct somatic embryogenesis in explants from shoot tissue.
The highest percentage of leaf explants showing shoot organogenesis was achieved (juvenile explants, 65%; adult explants,
75%) when incubated in Murashige and Skoog (MS) medium supplemented with either 4.5 μM of the phenylurea cytokinin thidiazuron
(TDZ) or 2.25 μM TDZ plus 2.2 μM 6-benzyladenine (BA), for juvenile and adult explants, respectively, both supplemented with
0.53 μM α-naphthaleneacetic acid (NAA). Juvenile explants developed on average 18 shoots per explant in the MS medium supplemented
with 4.5 μM TDZ, a four fold increase over those incubated on the medium supplemented with 2.25 μM TDZ and 2.2 μM BA. Adult
leaf explants grown on medium containing 2.25 μM TDZ and 2.2 μM BA medium produced 12 shoots per explant, while those grown
on medium containing 4.5 μM TDZ produced 5 shoots per explant. Shoot organogenesis was observed in roots of intact seedlings
pre-cultured on plain medium lacking nutrients (PM) or woody plant medium (WPM) salts and then grown on WPM salts supplemented
with 4.4 μM BA, 0.29 μM gibberrelic acid (GA3), and 57.0 μM indoleacetic acid (IAA). The number of shoots formed on each seedling
root system was ten fold higher when the pre-culture was in WPM medium indicating a promoting effect of mineral nutrients
in the pre-culture medium. Somatic embryogenesis was induced in both juvenile and adult leaf explants in 65 and 78% of the
explants, respectively, in MS-based medium supplemented with 2.0 μM N-(2-Chloro-4-pyridyl)-N
1-phenylurea (CPPU), 0.53 μM NAA and varying concentrations of BA. There was an interaction effect between MS salt strength
and BA concentration. The most effective medium for inducing somatic embryogenesis in juvenile explants contained half strength
MS salts and 2.2 μM BA and full strength MS salts and 13.2 μM BA for adult explants. 相似文献
19.
Primary bulb explants of Scilla natalensis were cultured in vitro on modified MS medium. Some of these explants initiated shoots, which provided a sterile source of secondary leaf and bulb explants. The secondary explants responded similarly to various combinations of plant growth regulators. Shoots were initiated spontaneously on medium containing no plant growth regulators. The number of shoots initiated was increased by the addition of kinetin or thidiazuron (TDZ) alone, but was reduced by the addition of indole-3-acetic acid (IAA) or naphthalene acetic acid (NAA) alone. Optimal shoot initiation occurred on medium containing 1 to 2 mg l–1 kinetin and 1 to 2 mg l–1 IAA. These shoots were rooted on medium containing 1 mg l–1 IAA. The plantlets were successfully acclimatised in the misthouse/shadehouse. 相似文献
20.
Summary Protocols for both axillary bud proliferation and shoot organogenesis of Euphorbia pulchurrima Winter RoseTM were developed using terminal buds and leaf tissues. Greenhouse-grown terminal buds were placed on Murashige-Skoog (MS) basal
medium supplemented with various concentrations of either benzlyaminopurine (BA) or thidiazuron (TDZ). Explants produced the
greatest number of axillary buds on media containing between 2.2 and 8.8 μM BA. The number of explants that produced axillary buds increased with increasing BA concentration. TDZ at concentrations
between 2.3 and 23.0 μM caused hyperhydricity of shoots and were not effective in promoting shoot proliferation. The most calluses and shoots were
produced from leaf midvein sections from in vitro grown plants placed on the medium containing 8.8–13.3 μM BA and 17.1 μM indole-3-acetic acid (IAA) for 1 mo. before transferring to the medium containing only BA. Adventitious buds were produced
only from red-pigmented callus, and explants that produced callus continued to produce adventitious shoots in the presence
of IAA. Five-mo.-old shoots derived from shoot culture or organogenesis rooted readily in artificial soil with or without
treatment with indolebutyric acid, and were acclimatized in the greenhouse. 相似文献