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1.
A protocol has been developed for the induction of somatic embryogenesis from flower explants of chamomile (Chamomilla recutita L.). The effects of several plant growth regulators [α-naphthylacetic acid (NAA), 2,4-dichlorophenoxyacetic acid, 6-benzyladenine (BA) and kinetin (Kin), alone or in combination]
and the flower type (disk or ray flower) were investigated. Both types of flowers responded to the callus and shoot induction
treatments, but formation of globular somatic embryos took place only on disk-flower-derived explants after 2–4 weeks of culture
on a Murashige and Skoog (MS) medium supplemented either with 8.87 μm BA and 1.07 μm NAA or with 26.8 μm NAA and 11.5 μm Kin. However, fully developed, cotyledonary-stage somatic embryos could be induced only on the NAA/Kin medium, 10 weeks after
culture initiation. Germination of the embryos and plant regeneration took place after subculture for 4–5 weeks onto medium
of the same composition. Plantlets regenerated from embryos flowered in vitro on a MS medium supplemented with 8.87 μm BA and 1.07 μm NAA. The significance of the results with respect to chamomile micropropagation and the utilization of wild populations in
breeding programs is discussed.
Received: 6 April 1998 / Revision received: 12 October 1998 / Accepted: 28 October 1998 相似文献
2.
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
3.
Somatic embryogenesis from mature leaves of rose (Rosa sp.) 总被引:9,自引:0,他引:9
Several plant growth regulators (0.3–53.3 μm 6-benzyladenine, 2,4-dichlorophenoxyacetic acid, gibberellic acid, 3-indoleacetic acid, p-chlorophenoxyacetic acid, kinetin and α-naphthylacetic acid), alone or in combination, and culture conditions were tested for their capacity to induce somatic embryogenesis
from mature leaf and stem explants of rose (Rosa sp.) of four commercial rose cultivars (Baccara, Mercedes, Ronto and Soraya). Somatic embryos were only induced from mature
leaf explants derived from Soraya on Murashige and Skoog (MS) medium supplemented with 53.5 μm
p-chlorophenoxyacetic acid and 4.6 μm kinetin, although satisfactory callus induction rates were obtained from all cultivars. After subculturing on the same medium,
embryos at various developmental stages (globular, heart and torpedo shaped) were transferred for maturation onto a MS medium
supplemented with 5.2 μm 6-benzyladenine and 5.7 μm 3-indoleacetic acid. Germination of mature embryos took place after subculturing them onto medium of the same composition.
Plantlets regenerated from embryos and bearing three to four leaves were transferred to a greenhouse.
Received: 4 February 1997 / Revision received: 28 August 1997 / Accepted: 1 October 1997 相似文献
4.
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
5.
Somatic embryogenesis and in vitro rosmarinic acid accumulation in Salvia officinalis and S. fruticosa leaf callus cultures 总被引:1,自引:0,他引:1
The effect of explant age, plant growth regulators and culture conditions on somatic embryogenesis and rosmarinic acid production
from leaf explants of Salvia officinalis and S. fruticosa plants collected in Greece was investigated. Embryogenic callus with numerous spherical somatic embryos could be induced
on explants derived from both species and cultured for 3 weeks on a Murashige and Skoog (MS) medium supplemented with 1.8–18
μm 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (Kin) or 10.5–21 μm 1-naphthalenacetic acid and 6-benzyladenine. Only explants from young plants (with six to eight leaves) responded to the
culture treatments and, in general, low light intensities (50 μmol m–2 s–1) favoured callus formation and induction of somatic embryos. Somatic embryos were further developed on the same medium. Heart-
and torpedo-shaped embryos (1–2 mm long) were subcultured on a growth-regulator-free MS medium for maturation. Maximum rosmarinic
acid accumulation in S. officinalis and S. fruticosa callus cultured on 4.5 μm 2,4-D and 4.5 μm Kin was 25.9 and 29.0 g/l, respectively.
Received: 17 January 1997 / Revision received: 26 May 1997 / Accepted: 30 June 1997 相似文献
6.
Silvia Karina Vila Hebe Yolanda Rey Luis Amado Mroginski 《Journal of Plant Growth Regulation》2007,26(3):268-277
Factors affecting somatic embryogenesis induction and conversion in paradise tree (Melia azedarach) were evaluated. Somatic embryogenesis was influenced by plant growth regulators, explant stage, carbohydrate source and
concentration, gelling agents, light, and induction times. MS medium with 4.54 μM thidiazuron (TDZ) was optimal for the induction
of embryogenic tissue. Zygotic embryos that were 1-1.5 mm long (torpedo and early cotyledonal stage) had a greater embryogenic
response than smaller or larger embryos and better conversion of somatic embryos into plants. In general, embryos that formed
in medium containing 1% or 5% carbohydrate were hyperhydrics or fused, respectively, whereas those that formed in medium with
a carbohydrate concentration of 3% had better morphology. Raffinose at 3% yielded satisfactory somatic embryo induction with
good morphology and the best values of conversion into plants. Induction and conversion of somatic embryos were superior on
medium solidified with agar A-1296. The explants maintained under 160 μmol m−2 s−1 or 1 week in darkness and later 160 μmol m−2 s−1 produced a significantly higher embryogenic index. Only 4 days of treatment on induction medium, with either raffinose or
sucrose at 3% as a carbohydrate source, were required to induce somatic embryogenesis, but longer exposure, until 18 days,
increased the yield and improved the morphology of somatic embryos. 相似文献
7.
Glória Pinto Helena Valentim Armando Costa Sílvia Castro Conceição Santos 《In vitro cellular & developmental biology. Plant》2002,38(6):569-572
Summary Somatic embryos were obtained from a 60-yr-old Quercus suber L. tree. Leaf explants were cultivated on Murashige and Skoog medium with 30 gl−1 sucrose, 3 gl−1 gelrite, pH adjusted to 5.8, and different growth regulator combinations. Callus induction took place at 24±1°C in the dark
during the first 3 wk. After 3 mo, calluses that showed embryogenic structures were transferred to the same medium without
growth regulators. Somatic embryogenesis was only observed in calluses induced on E3 medium (supplemented with 4.5 μM 2,4-dichlorophenoxyacetic acid and 9.0 μM zeatin). On average, 7.5% of the initial explants formed embryogenic calluses in this medium. Somatic embryo proliferation
was high due to secondary embryogenesis. On average, 10% of the somatic embryos germinated and 40% of these germinated embryos
converted into plants. Plants were elongated on the same medium without growth regulators and acclimated to greenhouse conditions. 相似文献
8.
Thirty-two barley cultivars grown in Spain, 18 of the two-row type and 14 of the six-row type, were screened for plant regeneration
from cultured immature embryos. Although there was much variation in regeneration capacity among the cultivars, plants were
obtained from all cultivars except Almunia. No statistical differences were found in the percentage of regeneration between
two- and six-row types. The influence of the auxins 2,4-dichlorophenoxyacetic acid, dicamba, and picloram on the induction
and maintenance of embryogenesis and regeneration capacity after 3–4 months in culture, were evaluated for cultivars Cobra,
Hop and Reinette. Hop had the highest rates of maintenance of embryogenic capacity and plant regeneration. The medium containing
dicamba gave the best embryogenic callus induction, maintenance and regeneration. Five regeneration media, differing in growth
regulators and micronutrient composition, as well as partial desiccation of the calli before regeneration, were tested. The
regeneration medium containing 10 μm copper sulfate gave the best results. Regeneration frequencies after 3–4 months in culture of cultivar Hop were raised from
59.5 to 93.7% in this medium. Silver nitrate and partial desiccation of the calli also enhanced plant regeneration, but the
medium containing 10 μm of silver nitrate reduced root formation.
Received: 30 October 1997 / Revision received: 3 April 1998 / Accepted: 17 April 1998 相似文献
9.
Embryogenic cultures were induced from pinnae removed from young leaf flushes of mature-phase trees of the endangered cycad
species, Ceratozamia euryphyllidia. Induction media consisted of B5 major salts, Murashige and Skoog minor salts and organics, 400 mg l–1 glutamine, 100 mg l–1 asparagine, 100 mg l–1 arginine, 60 g l–1 sucrose, 2 g l–1 gellan gum, 4.65–13.94 μm kinetin and 4.52–9.05 μm 2,4-dichlorophenoxyacetic acid. Cultures were maintained in darkness. Embryogenic cultures were comprised of precotyledonary
somatic embryos that proliferated by somatic polyembryogenesis following subculture onto medium without plant growth regulators.
Somatic embryo development and maturation occurred spontaneously from proliferating cultures on medium without plant growth
regulators. Somatic embryos were monocotyledonous and mature somatic embryos germinated on semisolid medium without growth
regulators. Subsequent development, which included the elongation of the first leaves, occurred only after subculture onto
semisolid medium without plant growth regulators containing 0.5% (wt/vol) activated charcoal and under low light intensity.
The time period from explanting to plant recovery was approximately 3 years.
Received: 25 September 1997 / Revision received: 16 December 1997 / Accepted: 29 December 1997 相似文献
10.
Ezz Al-Dein Al-Ramamneh Sridevy Sriskandarajah Margrethe Serek 《Plant Cell, Tissue and Organ Culture》2006,84(3):333-342
Somatic embryogenesis was induced from phylloclade explants of Schlumbergera truncata cv. Russian Dancer. Callus developed on phylloclade explants and sub-cultured over a period of 16 months on MS medium containing
mainly cytokinins was superior for the induction of somatic embryos compared to callus grown for a shorter time in the establishment
medium. Sub-culture of callus grown in SH-or MS-based liquid media supplemented with 7.0 μM kinetin and transferred onto solid
MS-based medium with either 0.45 μM 2,4-D or without hormones resulted in the differentiation into somatic embryos. SH-based medium proved better than MS-based medium
when used as the first medium for the induction of somatic embryogenesis. However, somatic embryogenesis, contrary to adventitious
shoot formation, was reduced when 2,4-D was included in the MS-based medium used for final transfer compared to the medium without growth regulators, indicating
that a critical hormonal balance was reached. Somatic embryos developed root and shoot poles when grown on G medium. On this
medium approximately 70% germination was recorded in the embryos that were differentiated earlier from the callus that was
grown for a longer time in the establishment medium. This callus was grown on either SH- or MS-based medium supplemented with
7.0 μM kinetin, and then transferred after 30 days (from SH medium) onto MS medium without hormones or after 40 days (from
MS medium) onto MS medium with 0.45 μM 2,4-D. Furthermore, plants from somatic embryos were successfully potted in soil and showed further growth and formation of a second
set of phylloclades (secondary phylloclades). Histological studies showed that somatic embryos had no detectable connection
with the mother explants and that advanced stages of somatic embryos had a contained vascular system. In addition to the normal
dicotyledonous embryos, anomalous embryos with multiple cotyledons and vase-like embryos were observed. Secondary embryos
were also recorded in this study. 相似文献
11.
M. Capuana G. Petrini A. Di Marco R. Giannini 《In vitro cellular & developmental biology. Plant》2007,43(2):101-110
This is the first report on somatic embryogenesis in common ash (Fraxinus excelsior L.). Experiments on somatic embryogenesis induction were carried out on zygotic embryos at different phases of development
and maturation. The embryo axes were isolated and cultured on media containing different plant growth regulators (PGRs). Embryogenic
tissues were obtained from embryos collected at an incomplete maturation phase and cultured on a modified Murashige and Skoog
medium containing 8.8 μM 2,4-dichlorophenoxyacetic acid and 4.4 μM benzyl-adenine (BA). Embryos isolated from seeds at an
advanced stage of maturation showed only organogenetic phenomena. Embryogenic tissues were successfully subcultured and multiplied
on medium containing a reduced concentration of PGRs. After their isolation, somatic embryos were induced to develop and mature
by transfer to PGR-free medium and subsequent culture on medium containing 0.1 μM BA. Somatic embryos developed completely
and also germinated spontaneously. Embryo germination and conversion were significantly improved when subjected to a period
of storage at 4°C and transplant onto woody plant medium. Plantlets were successfully transferred to soil and acclimatized
in a “misted” greenhouse. 相似文献
12.
Summary Three auxin-type herbicides, namely 2.4-dichlorophenoxyacetic acid (2,4-D), (4-chlorophenoxy)acetic acid 2-(dimethylamino)ethyl
ester (centrophenoxine), and quinolinecarboxylic acid (quinclorac) induced direct somatic embryogenesis in seed-derived zygotic
embryo explants of sweet pepper (Capsicum annuum L.) when added to Murashige and Skoog medium with 200 mM sucrose. Optimum concentrations for embryogenesis induction were 0.40–0.45 mM and 1.15–1.30 μM for 2.4-D and centrophenoxine, respectively (in the presence of 5.0 gl−1 activated charcoal), or 40 μM for quinclorac (in medium without activated charcoal). Somatic embryos emerged from the epidermal and subepidermal tissues
and developed on the surface of the explant. Centrophenoxine- or 2.4-D-mediated embryogenesis was accomplished from 95% of
the explants in about 3 wk and, on average, six embryos were formed per explant. Induction efficieney was lower for quinelorac.
Centrophenoxine-mediated embryognesis was possible in 10 pepper cultivars, the extent of the reponse-being genotype-dependent.
embryos detached from the explant and transplanted onto a growth regulator-free medium germinated; however, the recovered
regenerants were without a shoot, and some of them bore a single deformed cotyledon while others had no cotyledons. Regenerants
lacking a shoot were generated irrespective of the auxin type applied and across all responsive genotypes investigated. Absence
of a shoot, resulting from a failure in the establishment of a normal functioning apical shoot meristem, was the principal
developmental disorder that precluded regeneration of normal plants via direct somatic embryogenesis. Since stem cells of
the shoot meristem become established in globular and heart-stage embryos, we deduce that the absence of a shoot in germinating
embryos could orginate from deviant differentiation at these early stages of embryogeny. 相似文献
13.
Somatic embryos were induced on peanut (Arachis hypogaea) leaflets from aseptically germinated embryo axes. Leaflet size influenced percent somatic embryogenesis; 5–8 mm long cut leaflets were superior to 2–3 mm long uncut leaflets. Maximum embryogenesis of 14.6% was obtained after a 15 d incubation on induction medium (modified MS with B5 vitamins, 30 g/l sucrose, 4 g/l Gel-Gro, 40 mg/l 2,4-D +0.2 mg/l kinetin) followed by transfer to a secondary medium with 5 mg/l 2,4-D+0.2 mg/l kinetin. Primary somatic embryos were fused along the axes with no distinct cotyledons, but secondary embryos had single axes with two cotyledons. Other treatments had lower percent embryogenesis, no secondary embryogenesis, and embryos with single axes with two cotyledons. Some somatic embryos converted into normal plants capable of greenhouse survival.Abbreviations MS
Murashige and Skoog (1962) medium
- B5
Gamborg et al. (1968) B5 medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6benzylaminopurine
- NAA
1-naphthaleneacetic acid 相似文献
14.
Attempts were made to determine the influence of sucrose, mannitol, sorbitol and abscisic acid on accumulation of triglycerides
in peanut (Arachis hypogaea L.) somatic embryos. The results revealed that 0.584 m sucrose in the medium produced increased triglycerides in the embryos compared to the control. At 0.730 m sucrose, embryos were necrotic although the triglyceride content was high. Sorbitol at 0.6 m or abscisic acid at 20 μm were effective in increasing triglycerides in the embryos. The increase in triglycerides on a fresh-weight basis observed
with increasing concentration of osmoticium was not apparent when determined in terms of dry weight. However, an increase
in triglyceride as percent fresh weight observed in the presence of 20 μm abscisic acid remained unaltered when determined in terms of percent dry weight. An increase in storage lipid did not improve
conversion of peanut somatic embryos.
Received: 4 April 1997 / Revision received: 15 February 1998 / Accepted: 2 March 1998 相似文献
15.
Regeneration of Acacia mangium through somatic embryogenesis 总被引:2,自引:0,他引:2
Somatic embryogenesis and whole plant regeneration were achieved in callus cultures derived from immature zygotic embryos
of Acacia mangium. Embryogenic callus was induced on MS medium containing combinations of TDZ (1–2 mg/l), IAA (0.25–2 mg/l) and a mixture of
amino acids. Globular embryos developed on embryogenic callus cultured on the induction medium. Nearly 42% of embryogenic
cultures with globular embryos produced torpedo- and cotyledonary-stage embryos by a two-step maturation phase. The first
stage occurred on 1/2-strength MS basal medium containing 30 g/l sucrose and 5 mg/l GA3 followed by the second stage on 1/2-strength MS basal medium containing 50 g/l sucrose. Of the cotyledonary-stage somatic
embryos, 11% germinated into seedlings that could be successfully transferred to pots. Light- and scanning electron microscopy
showed that the somatic embryos originated from single cells of the embryogenic callus. Further, a single cell layer could
be detected beneath the developing somatic embryos that appeared to be a demarcation layer isolating the somatic proembryonic
structure from the rest of the maternal callus. A suspensor-like structure connected the globular embryos to the demarcation
layer. This is the first successful report of plant regeneration through somatic embryogenesis for this economically important
tropical forest species.
Received: 20 January 2000 / Revision received: 28 September 2000 / Accepted: 29 September 相似文献
16.
In vitro plant regeneration in Melia azedarach L. 总被引:1,自引:0,他引:1
Nodal explants of 3- 6-week-old seedlings cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA)
(17.75 μm) produced multiple shoots. Shoots were isolated and induced to root on 1/2-strength MS medium supplemented with indole-3-butyric
acid (4.92 μm). In-vitro-rooted shoots resumed growth after a short period of acclimatization and resulted in plantlets which were successfully
established in soil. In vitro flowering was observed in some of the nodal explants in the above medium, and also in cotyledonary
leaves and internodal explants on MS medium supplemented with a combination of indole-3-acetic acid (IAA) (0.06 μm)+BA (4.44 μm) and IAA (0.06 μm)+kinetin (4.65 μm).
Received: 25 October 1997 / Revision received: 1 May 1998 / Accepted: 15 May 1998 相似文献
17.
Direct somatic embryogenesis and genetic transformation has been achieved in Lathyrus sativus. The genetic transformation was achieved by Agrobacterium tumefaciens and biolistic methods of gene transfer. Somatic embryogenesis was obtained from immature leaflets and nodal segments by using 2,4-0 in the induction medium. Somatic embryos formed germinated on growth regulator-free medium. The transgenic nature of the transformants was confirmed by GUS expression assay. The transgenic shoots/plantlets were obtained only from tissue transformed with biolistic method of gene transfer. 相似文献
18.
Summary
In vitro regeneration of plants via somatic embryogenesis through cell suspension culture was achieved in horsegram. Embryogenic calluses
were induced on leaf segments on solid Murashige and Skoog (MS) medium with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Differentiation of somatic embryos occurred when the embryogenic calluses were transferred
to liquid MS medium containing 2,4-D. Maximum frequency (33.2%) of somatic embryos was observed on MS medium supplemented
with 7.9 μM 2,4-D. Cotyledonary-torpedo-shaped embryos were transferred to liquid MS medium without growth regulators for maturation
and germination. About 5% of the embryos germinated into plants, which grew further on solid MS medium. The plants were hardened
and established in soil. Effects of various auxins, cytokinins, carbohydrates, amino acids, and other additives on induction
and germination of somatic embryos were also studied. A medium supplemented with 7.9 μM 2,4-D, 3.0% sucrose, 40 mg l−1
L-glutamine, and 1.0 μM abscisic acid was effective to achieve a high frequency of somatic embryo induction, maturation, and further development. 相似文献
19.
Plant regeneration via somatic embryogenesis, and transient gene expression in sweet potato protoplasts 总被引:1,自引:0,他引:1
A method for regenerating plants from petiole protoplasts of the in vitro-raised sweet potato cultivar Jewel is described.
Protoplast yields of 3.0–5.0×106 were obtained following 4–6 h digestion of 1- to 2-cm petioles (1 g fresh weight) with 1% Cellulase-R10, 2% Macerozyme-R10,
and 0.3% Pectolyase Y-23 in a washing solution with 9% mannitol. A plating density of 105 protoplasts/ml was optimal for subsequent division. An initial division frequency of 12–15% was obtained in liquid or agarose-solidified
KP8 culture medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) (0.9 μm), and zeatin (2.3 μm). Colonies consisting of 100–200 cells were formed after 4 weeks in the dark at 24±2°C. The frequency of colony formation
was improved by the gradual addition of fresh liquid KP8 medium of lower osmoticum. Protocalli (1–2 mm in diameter) were formed
after an additional 4–6 weeks under continuous illumination and regular dilution with fresh culture medium. Morphogenic callus
formed globular and heart-shaped embryos that developed into cotyledon stage embryos, following transfer of calli onto medium
containing 2,4-D (11.3 μm) and benzylaminopurine (2.2 μm). Subsequently, embryo conversion to plantlets was obtained on basal medium with 2% sucrose and 3.5 μm gibberellic acid. Regenerated plantlets were successfully transplanted in soil. Mature plants appeared phenotypically normal.
The same petiole protoplast populations showed transient expression of the gusA gene introduced using electroporation.
Received: 10 October 1997 / Revision received: 10 February 1998 / Accepted: 2 March 1998 相似文献
20.
Panax japonicus is one of the important medicinal plants. Here, we established the protocol for plant regeneration of P. japonicus via direct somatic embryogenesis. Somatic embryos were directly obtained from the segments of zygotic embryos on MS medium
with 4.4 μM 2,4-D. Thereafter, somatic embryos were produced by repetitive secondary somatic embryogenesis. The secondary
somatic embryo formation was enhanced by plasmolyzing pretreatment (1.0 M mannitol for 10 h). Frequency of secondary somatic
embryo formation from cotyledon segments was lowered by plasmolyzing pretreatment, but the number of somatic embryos per explants
was greatly increased. Plasmolyzing pretreatment resulted in retardation of embryo growth and required subculture to fresh
medium for further growth of embryos into cotyledonary stage. Without plasmolyzing pretreatment, cotyledonary embryos were
obtained after 8 weeks of culture. All the cotyledonary somatic embryos germinated by 5 μM GA3 treatment, but only 15.3% were germinated on hormone-free medium. After 2 months of culture on 1/2 strength WPM medium, plantlets
produced flowers spontaneously. In the anthers of in vitro flowers, microsporogenesis occurred normally with low number of
pollen grains. 相似文献