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1.
Tissue culture and plant regeneration of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. Ex Steud
Gerardo Armando Aguado-Santacruz José Luis Cabrera-Ponce Víctor Olalde-Portugal M. A. Rosario Sánchez-González Judith Márouez-Guzmán Luis Herrera-Estrella 《In vitro cellular & developmental biology. Plant》2001,37(2):182-189
Summary As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H. B. K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated
from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid
(2,4-D), Picloram (4-amino-3, 5,6-trichloropicolinic acid), N6-benzyladenine (BA) or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations
containing 1 mg l−1 (4.52 μM) 2,4-D plus 0.5 mg l−1 (2.22 μM) BA. and 2 mg l−1 (8.88 μM) BA plus 1 mg l−1 (4.14 μM) Picloram with or without 40 mg l−1 (296.08 μM) adenine. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient
treatments for induction of embryogenic callus contained 2 mg l−1 (9.05 μM) 2,4-D combined with 0.25 (1.11 μM) or 0.50 mg l−1 (2.22 μM) BA, or 1 mg l−1 (4.52 μM) 2,4-D with 0.50 mg l−1 (2.22 μM) BA. Regeneration was achieved in hormonefree Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus
1 mg l−1 (1.44 μM) gibberellic acid. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from 9 for 2 mg
l−1 (9.05 μM) 2,4-D to 62.2 for induction medium containing 2 mg l−1 (8,28 μM) Picloram, 1 mg l−1 (4.44 μM) BA and 40 mg l−1 (296.08 μM) adenine. Regnerated plants grown in soil under greenhouse conditions reached maturity and produced seeds. 相似文献
2.
A. Śliwińska O. Olszowska M. Furmanowa A. Nosov 《In vitro cellular & developmental biology. Plant》2008,44(2):69-77
Efficient plant regeneration through somatic embryogenesis was achieved in Polyscias filicifolia. Embryogenic calluses were induced on Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l−1 benzylaminopurine (BAP; type I callus) and on MS medium with 2.0 mg l−1 2,4-D and 0.01 mg l−1 kinetin (type II callus) from leaf explants of a 2-yr-old plant. Primary somatic embryos (PSEs) developed after four passages
of suspension culture established from embryogenic callus when cultured in liquid half-strength MS medium (1/2 MS) without
growth regulators. PSEs in the cotyledonary stage were multiplied by adventitious embryogenesis. Single secondary somatic
embryos (SSEs) or their clusters developed at the base of PSE hypocotyls and regenerated into plantlets in a one-step process
on plant growth regulator-free 1/2 MS medium. Low sucrose concentration of 15 g l−1 promoted development of normal SSEs. All SSEs regenerated into single, well-rooted plantlets on a Nitsch and Nitsch medium
supplemented with 0.5 mg l−1 kinetin, 0.1 mg l−1 indole-3-butyric acid, and 10 mg l−1 adenine sulfate. Subsequent two subculture cycles on the same medium were necessary to obtain plantlets sufficiency developed
to allow successful transfer to the soil. Rooted plantlets were established in a peat mixture with 90% survival, with the
plants showing normal morphological characteristics. 相似文献
3.
Wei Tang Zhongchen Guo Fan Ouyang 《In vitro cellular & developmental biology. Plant》2001,37(5):558-563
Summary Mature zygotic embryos of eight (open-pollinated) families of loblolly pine (Pinus taeda L.) were cultured on eight different basal salt formulations, each supplemented with 36.2 μM 2,4-dichlorophenoxyacetic acid, 17.8 μM 6-benzyladenine, 18.6 μM kinetin, 500 mg l−1 casein hydrolysate, and 500 mg l−1
l-glutamine for 9 wk; embryogenic tissue was formed on cotyledons, hypocotyls, and radieles of mature zygotic embryos. Callus
was subcultured on the callus proliferation medium, the same as the induction medium but with one-fifth concentration of auxin
and cytokinin for 9 wk. On this medium a white to translucent, glossy, mucilaginous embryogenic callus containing embryogenic
suspensor masses (ESMs) was obtained. The highest frequency of explants forming embryogenic tissue, 17%, occurred on a modified
Murashige and Skoog salts basal medium containing the concentration of KNO3, Ca(NO3)2·4H2O, NH4NO3, KCl, ZnSO4·7H2O, and MnSO4·H2O, 720, 1900, 400, 250, 25.8, and 25.35 mg l−1, respectively. Embryogenic suspension cultures were established by culturing embryogenic callus in liquid callus proliferation
medium. Liquid cultures containing ESMs were transferred to medium containing abscisic acid, polyethylene glycols, and activated
charcoal for stimulating the production of cotyledonary somatic embryos. Mature somatic embryos germinated for 4–12 wk on
medium containing indole-butyric acid, gibberellic acid, 6-benzyladenine, activated charcoal, and reduced sucrose concentration
(15 g l−1). Two hundred and ninety-one regenerated plantlets were transferred to a perlite:peatmoss:vermiculite (1∶1∶1) mixture, then
the plants were transplanted to soil in the earth, and 73 plantlets survived in the field. 相似文献
4.
The effects of growth regulators, cold-pretreatment of flower buds, ovule (embryo sac) developmental stage and genotype on
induction of gynogenesis in unpollinated ovule cultures were assessed in niger (Guizotia abyssinica (L. f.) Cass.). Indirect callus-mediated gynogenesis occurred in cvs JNC-6 and Ootacamund when the ovules were cultured on
MS medium supplemented with 30 g l−1 sucrose and 2,4-D either alone (0.5–2.0 μM) or in combination (2.0 μM) with different cytokinins, such as adenine, BA, 2iP
and kinetin (0.5–2.0 μM). An optimum induction of gynogenesis was fostered on medium supplemented with 2.0 μM 2,4-D and 1.0 μM
kinetin. Cold-pretreatment of flower buds had no stimulatory effect, but ovules collected one day before anthesis were most
responsive to gynogenesis. The results showed significant variations in genotypic competence for gynogenesis with cv. Ootacamund
being the most responsive (12.5%) and cv. IGP-76 the least (2.5%). Gynogenic embryos differentiated and matured on media (30 g l−1 sucrose) supplemented with 0.5 μM NAA plus 1.0 μM kinetin, and 0.5 μM ABA, respectively. The haploidy (2n = 1x = 15) of gynogenic plants was confirmed by cytological analysis. 相似文献
5.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
6.
B. L. Lad S. Jayasankar F. Pliego-Alfaro P. A. Moon R. E. Litz 《In vitro cellular & developmental biology. Plant》1997,33(4):253-257
Summary Embryogenic nucellar cultures were established on B5 major salts, MS minor salts and organics, 400 mg/l−1 glutamine, 60 g/l−1 sucrose, 2 g/l−1 gellan gum, and 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D). There was no clear relationship between developmental age of the nucellar explants
and induction of embryogenic cultures. The temporal requirements for culture initiation and for induction of embryogenic competence
from nucellar explants were determined by pulsing the cultures for 0, 7, 14, 21, 28, 35, 42, 49, 56, and 63 d. Culture initiation
required a minimum 7–14 d pulse with 2,4-D, and was maximum after a 56-d pulse; however, embryogenic competence was optimum
after a minimum of 28 d exposure to 2,4-D. Somatic embryogenesis occurred directly from the nucellar explants at low frequencies.
Somatic embryo maturation only occurred following plating of suspensions onto semisolid medium, and was stimulated by 2.4–4.8
μM kinetin and 4.4 μM 6-benzyladenine. 相似文献
7.
Jameel M. Al-Khayri Christine E. Shamblin Edwin J. Anderson 《In vitro cellular & developmental biology. Plant》1996,32(4):227-232
Summary This study was conducted to establish and optimize a regeneration system for adapted U.S. rice genotypes including three commercial
rice cultivars (LaGrue, Katy, and Alan) and two Arkansas breeding lines. Factors evaluated in the study were genotype, sugar
type, and phytohormone concentration. The system consisted of two phases, callus induction and plant regeneration. In the
callus induction phase, mature caryopses were cultured on MS medium containing either 1% sucrose combined with 3% sorbitol
or 4% sucrose alone, and 0.5 to 4 mg·L−1 (2.26 to 18.10 μM) 2,4-D with or without 0.5mg·L−1) (2.32 μM) kinetin. In the plant regeneration phase, callus was transferred to 2,4-D-free MS medium containing 0 or 2 mg·L−1 (9.29 μM) kinetin combined with 0 or 0.1 mg·L−1 (0.54 μM) NAA. Callus induction commenced within a week, independent of the treatments. Callus growth and plant regeneration, however,
were significantly influenced by interactions among experimental factors. Generally, the greatest callus growth and plant
regeneration were obtained with 0.5 mg·L−1 (2.26 μM) 2,4-D and decreased with increasing 2,4-D concentrations. Kinetin enhanced callus growth only when combined with 0.5 mg·L−1 (2.26 μM) 2,4-D, and 4% sucrose. Inducing callus on kinetin-containing medium generally enhanced regeneration capacity in the presence
of sucrose but not with a sucrose/sorbitol combination. Media containing sucrose alone generally supported more callus proliferation,
but the sucrose/sorbitol combination improved regeneration of some cultivars. NAA and kinetin had little effect on regeneration. 相似文献
8.
A three-stage procedure for embryogenesis in Trachyspermum ammi was developed from cotyledon and cotyledonary node explants cultured in Murashige and Skoog (MS) liquid medium supplemented
with 0.2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Globular somatic embryos without intervening callus phase developed in 4 wk. The
development of embryos to heart and torpedo stages required second-stage subculture of the explants (along with developing
embryos) in liquid medium with lower concentrations of 2,4-D. Further development of embryos required a third-stage subculture
in hormone-free liquid medium supplemented with 100 mg l−1 casein hydrolysate. Regeneration of complete plantlets occurred after the fully developed somatic embryos were transferred
to solidified half-strength MS medium supplemented with 1 mg l−1 gibberellic acid. 相似文献
9.
A. V. Loskutov G.-Q. Song K. C. Sink 《In vitro cellular & developmental biology. Plant》2008,44(4):239-245
Callus selection (CS) and the flamingo-bill explant (FB) methods were evaluated for efficacy in transformation for celery.
Agrobacterium tumefaciens strains EHA105 and GV3101, each with the bar gene under the promoters NOS (pGPTV-BAR) or 35S (pDHB321.1), were used. Leaf explants were inoculated and co-cultivated for
2 d in the dark. Calluses emerged on the explants on callus medium (C), Murashige and Skoog (MS) medium + 2,4-Dichlorophenoxyacetic
acid (2,4-D) (2.3 μM) + kinetin (2.8 μM) + timentin (300 mg·l−1). Calluses 4- to 6-wk-old were selected for glufosinate (GS) resistance by a two step method. First, calluses were transferred
to C medium + GS 0.35, 0.5, 1, 2, 5, or 10 mg·l−1; calluses formed only with 0, 0.35 and 0.5 mg·l−1 GS. All growing calluses from 0 and 0.35 mg·l−1 and a few from 0.5 mg·l−1, were divided and placed back on C + GS 0.35–0.5 mg·l−1 for another 5–6 wk. Second, tolerant clones were again divided and placed on C + GS 1–50 mg·l−1. When cultivar XP85 was inoculated with both strains, using pGPTVBAR, 19 glufosinate resistant (GR) callus clones were selected,
but shoots regenerated only for strain EHA105 inoculations. When both of the strains (each with pDHB321.1) were inoculated
on cv. XP166, 3 and 12 GR calluses occurred for EHA105 and GV3101, respectively. Using CS, a total of 34 GR callus clones
were selected, and shoots were regenerated from over 50% of them on Gamborg B5 medium + 6-(γ, γ-dimethylallylamino) purine
2ip (4.9 μM) + naphthaleneacetic acid (NAA; 1.6 μM) and rooted on MS in 5–6 mo total time. Conversely, using FB with inoculation
by GV3101/pDHB321.1 on cv. XP166 yielded putative transgenic celery plants confirmed by polymerase chain reaction (PCR) in
just 6 wk. Transformation of the bar gene into celery was confirmed by PCR for 5 and 6 CS and FB lines, respectively. Southern blot analyses indicated 1–2 copies
in CS lines and 1 copy in FB lines. Herbicide assays on whole plants with 100 and 300 mg·l−1 glufosinate indicated a range of low to high tolerance for lines derived by both methods. The bar gene was found to be Mendelian inherited in one self-fertile CS derived line. 相似文献
10.
Qin-Mei Wang Feng-Zhan Gao Xiang Gao Fan-Yu Zou Xin Sui Meng Wang Yue-Jun Hui Li Wang 《Plant Cell, Tissue and Organ Culture》2012,109(2):191-200
An efficient in vitro micropropagation system for Clivia miniata Regel was developed using basal tissues of young petals and young ovaries as explants. For callus induction, explants were
incubated on Murashige and Skoog (MS) medium containing either 2.22 μM 6-benzyladenine (BA) and 4.52 μM 2,4-dichlorophenoxyacetic
acid (2,4-D) or 4.44 μM BA, 5.37 μM α-naphthaleneacetic acid (NAA), and 9.05 μM 2,4-D. Moreover, callus was induced from young
ovaries when these were incubated on MS medium containing 8.88 μM BA, 10.74 μM NAA, and 9.05 or 18.10 μM 2,4-D. Subsequently,
callus was transferred to MS medium supplemented with kinetin (KT) and NAA for shoot organogenesis. Frequency of shoot regeneration
from petal-derived callus was highest when callus was transferred to medium containing 2.69 μM NAA with either 9.29 or 13.94 μM
KT. Shoot regeneration frequency from ovary-derived callus was highest when this callus was transferred to medium containing
9.29 μM KT and 10.74 μM NAA. Overall, different explant types exhibited different organogenic capacities wherein, young petals
had higher shoot regeneration frequencies than young ovaries. The highest rooting frequency (98.25 ± 3.04%) was obtained when
shoots were transferred to half-strength MS medium without plant growth regulators. Regenerated plantlets were transplanted
to soil mix and acclimatized, yielding a 96.80% survival frequency. Only 0.6% of regenerated plantlets exhibited morphological
changes. The diploid status (2n = 22) of regenerated plantlets was determined using chromosome counts of root-tips. Moreover, inter-simple sequence repeats
were used to assess the genetic fidelity of regenerated plantlets. Overall, regenerated plants shared 90.5–100.0% genetic
similarities with mother plants and 89.0–100.0% similarities with each other. 相似文献
11.
Natasha Maurmann Carina M. B. De Carvalho Andréia Loviane Silva Arthur G. Fett-Neto Gilsane L. von Poser Sandra B. Rech 《In vitro cellular & developmental biology. Plant》2006,42(1):50-53
Summary
Valeriana glechomifolia is an endemic species of southern Brazil, capable of accumulating, in all of its organs, the terpene derivatives known as
valepotriates, the presumed sedative components of the roots of pharmaceutically used species of Valeriana. In vitro cultures of the plant were established and the accumulation of acevaltrate, didrovaltrate, and valtrate in callus, cell suspension,
and untransformed root cultures was studied. Leaves of in natura plants and roots of micropropagated plantlets were used as the explants for callus induction and root culture establishment,
respectively, on Gamborg B5 basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or with kinetin (KIN).
Culture growth and secondary metabolite yields were enhanced with 2,4-D (4.52μM) and KIN (0.93μM). Maximum valepotriate contents, quantified by HPLC, of acevaltrate (ACE) 2.6mg g−1 DW, valtrate (VAL) 10.2mgg−1 DW, and didrovaltrate (DID) 2.9mg g−1 DW were observed in root cultures after 7–8wk of culture. 相似文献
12.
Embryogenic cultures were induced from immature avocado zygotic embryos representing different botanical races and complex
hybrids. The optimum induction medium consisted of B5 major salts, MS minor salts, 0.4 mg l−1 thiamine HCl, 100 mg l−1 myo-inositol, 30 g l−1 sucrose, 0.41 μM picloram and 8 g l−1 TC agar. Somatic embryogenesis occurred directly from the explants on induction medium, and secondary embryos and proembryonic
masses proliferated in liquid and on semisolid maintenance medium. Embryogenic culture maintainance was optimized in liquid,
filter-sterilized MS medium, supplemented with 30–50 mg l−1 sucrose, 4 mg l−1 thiamine HCl and 0.41 μM picloram. Two types of embryogenic cultures were recognized: –genotypes that proliferated as proembryonic
masses in the presence of auxin (PEM-type) and; –genotypes in which the heart stage and later stages of somatic embryos developed
in the presence of auxin(SE-type). Embryogenic suspension cultures became increasingly disorganized over time, and this was
associated with progressive loss of embryogenic potential.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Callus was initiated from in vitro grown immature leaf and ex vitro grown mature leaf and rhizome explants of Agave sisalana Perr. ex. Engelm, on MS medium containing 2,4-D (9.05 M) and kinetin (4.6 M) or 2,4-D (9.05 M), kinetin (4.6 M) and CH (1000 mg l–1) or mod. MS (NH4NO3, 1500 mg l–1) containing 2,4-D (9.05 M) and kinetin (4.6 M). Light was essential for callus formation which, however, was different in three types of explants on three different media compositions. Increasing NH4
+had a negative impact while addition of CH had a positive impact on callus formation. Shoot regeneration from callus from CH-supplemented medium only was achieved for rhizome and immature leaf tissues. The highest rate of regeneration was obtained with BA (26.6 M) as the sole hormone. Shoot buds g–1 callus varied according to BA concentrations. Shoot proliferation rate increased on half-strength MS medium containing BA (8.9 M). Microshoots developed on MS medium containing BA (2.22 M) and GA3 (1.44 M) and finally rooted on MS medium containing IAA (11.42 M). Acclimatized rooted plantlets are growing satisfactorily in ex vitro. This is the first report on plant regeneration via organogenesis of A. sisalana. 相似文献
14.
Myung Jin Oh Hye Ryun Na Hong-Keun Choi Jang Ryol Liu Suk Weon Kim 《Plant biotechnology reports》2008,2(1):87-92
An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension
cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when
cultured on half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly
with increasing concentrations of 2,4-D up to 3 mg l−1, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryo-derived white friable callus
were established using half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos
and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by
up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting
soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high
frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield,
which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could
be useful for the mass propagation and transformation of selected elite lines. 相似文献
15.
Summary Callus cultures were established from pith tissue of Coryphantha elephantidens (Lem.) Lem. on Murashige and Skoog (MS) basal medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin. Highest shoot regeneration frequency was observed on a medium containing 6.9 μM kinetin and 2.3 μM 2,4-D under 30 μE m−2 s−1 light intensity with a 16-h photoperiod. Calluses retained organogenic potential throughout several passages of subculture
(18 mo.). Shoots were rooted on MS medium without plant growth regulators. All (100%) plantlets transplanted to soil survived
acclimatization. Regenerated plants showed good overall growth and were morphologically similar to the mother plants. 相似文献
16.
Mei-Chun Lu 《Plant Cell, Tissue and Organ Culture》2004,78(1):93-96
High frequency plant regeneration was induced from protocorm-derived callus cultured on half-strength of Murashige—Skoog medium
with 2,4-dichlorophenoxyacetic acid (2,4-D, 0–5 mg l−1) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl, 0–1 mg l−1) urea (TDZ) in the dark. Twelve totipotent callus lines were selected within 76 callus lines regenerated on half-strength
of Murashige—Skoog (MS) medium with 0.5 mg l−1 TDZ. The proliferation rate was 4–5-fold in fresh weight after 30 days of culture on half-strength MS medium containing 5
mg l−1 2,4-D and 0.5 mg l−1 TDZ in the dark. The maximum number of shoot buds generated by 0.01 g callus explant was 134 after 4 months of culture. These
calli were regenerated to plantlets via protocorm-like bodies (PLBs) after 75–150 days of culture. The shoots, with two true
leaves, were transferred to hormone-free medium, rooting and eventually formed plantlets. Totipotent callus lines of Pleione formosana Hayata have been successfully established in this study. 相似文献
17.
An in vitro protocol for efficient plant regeneration has been developed from mature embryo explants of highland barley (Hordeum vulgare L. var. nudum Hk. f.) under endosperm-supported culture. Embryos with (endosperm-supported culture, ES) or without endosperm
(non-endosperm-supported culture, NES) were excised from mature seeds and cultured on MS medium supplemented with various
concentrations of 2,4-D (1–5 mg l−1) for callus induction. The percentage of callus induction from ES explants was significantly (P < 0.05) lower than that from NES. The highest frequency (97.6%) of callus induction was obtained from NES explants on MS
medium containing 3 mg l−1 2,4-D. When the primary calli were maintained at a reduced concentration of 2,4-D (0.5 mg l−1) for 3 weeks, embryogenic calli were formed. The embryogenic calli were then transferred to MS medium supplemented with different
concentrations of BA (1–5 mg l−1) and 500 mg l−1 casein hydrolysate (CH) for shoot regeneration. However, the capacity of plant regeneration from ES explant-derived calli
was significantly (P < 0.05) higher than that from NES. The best response (81.3%) was observed from ES explant-derived calli on MS medium containing
2 mg l−1 BA. Regenerated plantlets with well-developed root systems were transferred to pots where they grew well, attained maturity
and produced fertile seeds. This method could be employed for genetic manipulation studies. 相似文献
18.
Embryogenic cultures were induced from leaflets from new vegetative flushes of mature ‘Brewster’ litchi trees on B5 medium
containing 400 mg l−1 glutamine, 200 mg l−1 casein hydrolysate, 30 g l−1 sucrose, 4.52 μM 2,4-D, 9.30 μM kinetin and 3 g l−1 gellan gum in darkness. Embryogenic cultures consisting of proembryonic cells and masses were maintained either on semi-solid
MS medium supplemented with 4.52 μM 2,4-D and 0.91 μM zeatin or as embryogenic suspension cultures in liquid medium of the
same composition. Maturation of somatic embryos occurred on semi-solid MS medium with 5–20% (v/v) filter-sterilized coconut
water in darkness. Recovery of plants from somatic embryos was improved with 14.4 μM GA3 on half-strength MS medium with 0.2 g l−1 activated charcoal under a 16 h photoperiod provided by cool white fluorescent lights (60–80 μmol s−1 m−2). Plants have been successfully acclimatized in the greenhouse. 相似文献
19.
Summary Callus of Phalaenopsis Nebula was induced from seed-derived protocorms on 1/2 Murashige and Skoog (MS) basal medium plus 0–1.0 mg l−1 (0–4.52 μM) N-phenyl-N′-1,2,3,-thiadiazol-5-yl urea (TDZ) and/or 0–10 mg l−1 (0–45.24 μ M) 2,4-dichlorophenoxyacetic acid (2,4-D). Protocorms 2 mo. old performed better than 1-mo.-old protocorms for callus induction.
More calluses formed on 1/2 MS basal medium supplemented with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. These calluses could be maintained by subculturing every month with basal medium supplemented with 0.5 mg l−1 (2.27 μM) TDZ and 0.5 mg l−1 (2.26 μM) 2,4-D. Protocorm-like bodies were formed, and plants regenerated from these calluses on 1/2 MS basal medium alone or supplemented
with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. Plantlets were then potted on sphagnum moss in the greenhouse and grew well. No chromosomal abnormalities were found
among the root-tip samples of 21 of the regenerated plantlets that were successfully acclimatized. 相似文献
20.
Hippolyte Kodja Isabelle Robene-Soustrade Jacques Figier 《Acta Physiologiae Plantarum》1997,19(3):359-366
Callus cultures of Tabernaemontana persicariaefolia, (Apocynaceae), an endangered species endemic to the Mascarene Islands, were established from leaf explants on MS medium containing either
5 mg·l−1 2,4-D and 0.5 mg·l−1 BA or 5 mg·l−1 2,4-D, 0.5 mg·l−1 BA and 200 mg·l−1 DFMO. Histological studies showed regenerating nodules resembling globular embryos in calli after 4 weeks on the DFMO medium.
Green shoot formation was achieved by sequential subculture of the induced calli on media with gradually decreasing 2,4-D
concentrations (5→1→0 mg·l−1). Regeneration was greatly stimulated in the presence of DFMO. The first emergence of shoots occured 3 weeks earlier than
in untreated callus cultures. 相似文献