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1.
Early morphogenetic events and repetitive embryogenesis from callus culture of betel nut palm (Areca catechu L.) were studied using scanning electron microscopy. On Murashige and Skoog (MS) medium supplemented with 2 mg dm−3 dicamba, callus culture has capacity to form plantlets via somatic embryogenesis and to form secondary embryos for about 4 years. However, various abnormal embryos without differentiation
of the leaf sheath and shoot apical meristem were observed, which showed bell-shaped and then cup-shaped or mushroom-shaped
structures. These abnormal embryos contained distinctive structures, including a disk-shape interior region, surfaces with
grooves and a stalk-like posterior region. During subculture, these abnormal embryos enlarged, became deformed and gradually
lose their shape and then converted into nodular, compact embryogenic callus. It was also found that secondary embryos originated
from interior surfaces or posterior regions of abnormal embryos, and gave rise to the next cycle of normal and abnormal embryos. 相似文献
2.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
3.
Chin-Wen Ho Wei-Ting Jian Hui-Chun Lai 《In vitro cellular & developmental biology. Plant》2006,42(3):240-246
Summary
In vitro seedlings of Lilium × formolongi Hort. evs. Norikula, RaiZen No. 1, RaiZen No. 3, RaiZen Early, and Bailansa were used to induce callus by variously modified
Murashige and Skoog (MS) media, using protocols for flask culture and bioreactor culture. Green embryogenic callus proliferated
from roots near the base of bulblets of five varieties on media containing 0.53–5.3 μM α-naphthaleneacetic acid (NAA), and 28 cell lines were obtained by subcultures on the same medium. For flask culture, the
fresh weight (FW) of embryogenic cell clumps doubled every 4 wk on MS basal salts supplemented with 0.53°M NAA and 30 g l−1 sucrose. The maximum frequency of somatic embryos that developed into plantlets was 76.67±17% when plated onto solid MS basal
medium without plant growth regulators (PGRs). Among the treatments using four types of bioreactors, the best cell growth
and regeneration rate (74±0.14%) of somatic embryos was in a modified 2–1 bioreactor. Cells incubated in the other three bioreactors
furned brown and died. Histological study revealed that regeneration was by somatic embryogenesis. The regenerants showed
normal growth and flowering after 8–9 mo, in the field. A cell line of cv. Norikula has been subcultured in MS basal salts
containing 0.53 μM NAA every 2 mo. for 6 yr. The cell aggregates became more synchronous and many typical embryogenic cells with dense cytoplasm
were observed under a light microscope. The long-term embryogenic cells plated on MS basal medium still gave rise to numerous
somatic embryos and converted into plantlets. 相似文献
4.
Plant regeneration through somatic embryogenesis of Areca catechu L. was established using leaf, root and stem segments as explants. Embryogenic callus was induced and maintained on medium
supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or 3,6-dichloro-2-methoxybenzoic acid (dicamba) at concentrations
2, 4, 6 and 8 mg dm−3 in darkness. Somatic embryos were found on primary callus in the presence of 2 and 4 mg dm−3 dicamba and during subculture on 2 – 8 mg dm−3 2,4-D or 2 – 4 mg dm−3 dicamba-containing media. Plantlet conversion from embryos was successfully achieved on growth regulator-free medium. The
plants grew well when transplanted to containers in shaded greenhouse. 相似文献
5.
Pradeep C. Deo Robert M. Harding Mary Taylor Anand P. Tyagi Douglas K. Becker 《Plant Cell, Tissue and Organ Culture》2009,99(1):61-71
Callus was initiated in three different “esculenta” taro cultivars by culturing corm slices in the dark on half-strength MS
medium supplemented with 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) for 20 days followed by subculture of all corm slices
to half-strength MS medium containing 1.0 mg/l thidiazuron (TDZ). Depending on the cultivar, 20–30% of corm slices produced
compact, yellow, nodular callus on media containing TDZ. Histological studies revealed the presence of typical embryogenic
cells which were small, isodiametric with dense cytoplasms. Somatic embryos formed when callus was transferred to hormone-free
medium and ~72% of the embryos germinated into plantlets on this medium. Simultaneous formation of roots and shoots during
germination, and the presence of shoot and root poles revealed by histology, confirmed that these structures were true somatic
embryos. Plants derived from somatic embryos appeared phenotypically normal following 2 months growth in a glasshouse. This
method is a significant advance on those previously reported for the esculenta cultivars of taro due to its efficiency and reproducibility. 相似文献
6.
Summary Efficient and highly reproducible induction of somatic embryogenesis was obtained in four out of seven selected clones of
neem, Azadirachta indica A. Juss. This was achieved either directly from root and nodal explants or indirectly from callus cultures initiated from
leaf explants excised from 1-yr-old axenic plants. Direct induction of somatic embryogenesis was achieved both from nodal
and root segments within 8 wk of culture on MS1 medium without growth regulators. However, the addition of 2.3–4.5 μM thidiazuron and 0.5 μM 2,4-dichlorophenoxyacetic acid into the medium were necessary to induce somatic embryogenesis via callus phase from leaf
explants. Repetitive embryogenesis was observed within 3–4 wk following transfer of somatic embryos to a plant growth regulator-free
medium. When somatic embryos of nodal and root segments were left on the induction medium without subculturing, approximately
15% of the somatic embryos developed into whole plantlets after passing through a series of developmental stages. Plantlets
thus produced were hardy, lush green, and acclimatized casily under greenhouse conditions. However, somatic embryos derived
from leaf explants showed low conversion rates (<5%). HPLC analysis revealed no detectable levels of azadirachtin in somatic
embryos. 相似文献
7.
Summary Indirect somatic embryogenesis, encapsulation, and plant regeneration was achieved with the rare rhoeophytic woody medicinal
plant Rotula aquatica Lour. (Boraginaceae). Friable callus developed from leaf and internode explants on Murashige and Skoog (MS) medium with 0.45 μM 2,4-dichlorophenoxyacetic, acid (2,4-D) was most effective for the induction of somatic embryos. Subculture of the callus
onto half-strength MS medium with the same concentration of 2,4-D resulted in highly embryogenic callus. Suspension culture
was superior to solid medium culture for somatic embryogenesis. Embryogenic callus.during subsequent transfer to suspension
cultures of half-strength MS medium having 0.23 μM 2,4-D induced the highest number of somatic embryos (a mean of 25.6 embryos per 100 mg callus) and the embryos were grown
up to the torpedo stage. Transfer of embryos to half-strength MS basal solid medium allowed development, of 50% of the embryos
to the cotyledonary stage. Of the cotyledonary embryos, 90% underwent conversion to plantlets on the same medium. Encapsulated
cotyledonary embryos exhibited 100% conversion to plantlets. Ninety-five percent of the plantlets established in field conditions
survived, and were morphologically identical to the mother plant. 相似文献
8.
Segments taken from flower-stalk internodes of Oncidium Sweet Sugar formed somatic embryos and shoot buds directly from wound surfaces or via nodular masses proliferation within
1.5 months, when cultured on a Gelrite-gelled 1/2-MS basal medium supplemented with thidiazuron (0.1–3 mg l−1) in darkness. In light, when subcultured, these nodular masses proliferated into green compact callus, and produced somatic
embryos, shoot buds and/or yellowish abnormal structures spontaneously. Supplementing 0.1–1 mg l−1 NAA enhanced embryo formation, but retarded proliferation of shoot buds and yellowish abnormal structures. Somatic embryos
that directly formed from wound surfaces of flower stalk explants usually developed into abnormal structures, but the callus-derived
embryos could germinate into PLBs and eventually developed to normal plantlets on a hormone-free basal medium for 3–4 weeks.
Both the embryo-and shoot bud-derived regenerants developed into healthly plantlets when potted in sphagnum moss and acclimatized
in the greenhouse.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
9.
Summary An in vitro culture procedure was established for repetitive embryogenesis and plant regeneration from seed-derived protocorms of Phalaenopsis amabilis var. formosa Shimadzu (Orchidaceae). Seed-derived protocorms were cultured on modified half-strength Murashige and Skoog (1962) basal medium (1/2MS) devoid
of plant growth regulators. After 45 d, 28.1% of protocorms formed embryos from their posterior regions. 1-Phenyl-3-(1,2,3-thiadiazol-5-yl)-urea
(TDZ; 0.45, 4.54, and 13.62 μM) promoted direct embryo formation. The best response was at 13.62 μM TDZ, and 100% of the protocorms formed a mean number of 13.5 embryos after 45d of culture. By contrast, naphthaleneacetic
acid (NAA) at 0.54 and 5.37 μM inhibited direct embryo formation. On basal medium devoid of plant growth regulators, 18.8% of primary proliferating embryos
could form more embryos. TDZ (0.45, 4.54, and 13.62 μM) also promoted this process. Proliferating embryos/protocorms were transferred to basal medium devoid of plant growth regulators
for plantlet formation. Plantlets were successfully obtained from the embryos after 4–6 wk. Following subculture every 6 wk
for three passages, the plantlets were transferred to sphagnum moss in a container for acclimatization in the greenhouse.
The survival rate was 100%. 相似文献
10.
Summary A protocol was developed for high frequency somatic embryogenesis and plant regeneration from cotyledon and hypocotyl explants
of Eruca sativa. Explants grown on Murashige and Skoog (MS) medium supplemented with 4.52 μM 2,4-D formed embryogenic callus after 4 wk of culture. Secondary somatic embryos were also produced from primary somatic
embryos on MS medium containing 0.56 μM 2,4-D. Somatic embryos developed into mature embryos on MS medium in the presence of 45 gl−1 polyethylene glycol. After desiccation, somatic embryos developed into plantlets by culturing the mature somatic embryos
on 1/2 x MS medium containing 0.24 μM indole-3-butyric acid. 相似文献
11.
D. H. Tejavathi M. D. Rajanna R. Sowmya K. Gayathramma 《In vitro cellular & developmental biology. Plant》2007,43(5):423-428
Somatic embryogenesis from cultures of shoot apices, cotyledon and young leaves of in vitro shoots of Agave vera-cruz Mill. was studied. Embryogenic callus was obtained when explants were cultured on Murashige and Skoog’s (MS) medium (1962)
supplemented with L2 vitamins, 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-d) or 5.37 μM ∝-naphthalene acetic acid (NAA). Somatic embryos differentiated from this embryogenic callus upon subculture
to maturation/conversion medium containing cytokinin either alone or with auxin and l-glutamine. The best combination of growth regulators for development of somatic embryos was found to be 5.37 μM naphthalene
acetic acid plus 0.91 μM zeatin and 40 g/l sucrose. The conversion frequency of somatic embryos to plantlets varied from 46–50%.
Rooted plantlets were transferred directly to pots containing a soil, sand, and manure mixture without any hardening phase
with 96–98% survival of the plantlets. Based on the histological observations, the potential origin of the somatic embryo
is discussed. 相似文献
12.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
13.
Plantlet regeneration via somatic embryogenesis in anther callus of Vitis latifolia L. 总被引:1,自引:0,他引:1
Anthers of Vitis latifolia L. (wild grape) cultured on Nitsch and Nitsch medium supplemented with 20 μM 2,4-D and 9 μM BAP produced callus after 4–6 weeks. Subculture of callus onto Nitsch and Nitsch medium containing 10 μM NAA produced somatic embryos within 6 weeks. On growth regulator-free Nitsch and Nitsch basal medium somatic embryos converted
to plantlets in 6–8 weeks. One gram of callus produced more than 400 somatic embryos with 13.7% being converted to complete
plantlets, which were subsequently established in soil. Regenerated plants were found to have mixoploid populations of cells,
2n = 38 and n = 19.
Received: 23 May 1998 / Revision received: 21 September 1998 / Accepted: 10 October 1998 相似文献
14.
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
15.
Summary Establishment of fast-growing, highly regenerable callus cultures was examined in Muscari armeniacum Leichtl. ex Bak. in order to develop an efficient genetic transformation system. High-frequency callus formation was obtained
from leaf explants of cv. Blue Pearl on media containing 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid
(NAA) or 4-amino-3,5,6-trichloropicolinic acid (picloram, PIC). Fast-growing, yellowish nodular callus lines and white friable
callus lines containing a few somatic embryos were established on initiation medium supplemented with 4.5 μM 2,4-D and with 54 μM NAA, respectively. The yellowish nodular calluses vigorously produced shoot buds after transfer to media containing 0.44–44
μM 6-benzyladenine (BA), whereas the white friable calluses produced numerous somatic embryos upon transfer to plant growth
regulator-free (PGR-F) medium. Histological observation of shoot buds and somatic embryos indicated that the former consisted
of an apparent shoot meristem and several leaf primordia, and the latter had two distinct meristematic regions, corresponding
to shoot and root meristems. Both shoot buds and somatic embryos developed into complete plantlets on PGR-F medium. Regenerated
plants showed no observable morphological alterations. High proliferation and regeneration ability of these calluses, were
maintained for over 2 yr. 相似文献
16.
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
17.
Summary Seedlings from 11 seed sources (lines) of American ginseng from different geographic regions were evaluated on Murashige and
Skoog medium (MS) containing 10 μM α-naphthaleneacetic acid (NAA) and 9 μM 2,4-dichlorophenoxyacetic acid (2,4-D) for callus development and somatic embryo formation. Leaf and stem explants callused
at a frequency of 18.2–100%, while somatic embryos were produced from these calluses at a frequency of 25–87.5% after 5 mo.
Suspension cultures of nine lines were established by transferring embryogenic callus to MS liquid medium with NAA and 2,4-D
at 2.5 and 2.25 μM, respectively, and maintained by subcultures every 8 wk. Globular somatic embryos from these cultures were germinated on
half-strength MS containing 1% activated charcoal, and roots >5 mm in length developed within 3 wk. A 7-d exposure to 3 μM gibberellic acid and 5 μM 6-benzylaminopurine significantly enhanced shoot development and promoted further root development. The chromosome number,
profiles of the common triterpenoid saponins (ginsenosides), and random amplified polymorphic DNA (RAPD) banding patterns
in plantlets derived from suspension culture were compared to those of zygotic seedlings. The chromosome number in root tip
cells and suspension cultured cells was 48. Patterns of the six major ginsenosides, determined by thin-layer chromatography,
in leaves of tissue culture-derived plantlets were identical to those in seedlings. RAPD patterns among plantlets originating
from the same tissue-cultured line were mostly identical; however, altered patterns were observed in some lines that had been
maintained in suspension culture for almost 4 yr. The results from this study indicate that propagation of desired ginseng
genotypes in suspension culture can be achieved, and that biochemical and molecular markers can be used for authentication
of resulting plantlets. 相似文献
18.
S. B. Thencane S. R. Deodhar S. V. Bhosle S. K. Rawal 《In vitro cellular & developmental biology. Plant》2006,42(3):256-261
Summary Immature seeds of Garcinia indica Choiss, were exeised from immature fruits and cultured on Lloyd and McCown (1980), woody plant medium (WPM) with different
combinations of auxins and cytokinins. Somatic embryos were obtained on the media supplemented with 6-benzy laminopurine (BA;
2.2–22.1 μM) alone or in combination with α-naphthalene acetic acid (NAA; 2.6 μM) with 80% frequency within a period of 2–3 wk. Subculture of embryos on medium containing BA (16.0 μM) supplemented with indole-3-acetic acid (IAA: 2.8–5.7 μM) and/or kinetin (4.6 μM) gave rise to clusters of secondary somatic embryos along with maturation of primary embryos. In subsequent subculture on
hormone-free half-strength WPM, the embryo clusters germinated with an increase in the number of secondary somatic embryos.
About 70% of somatic embryos germinated into complete plantlets, which were successfully established under greenhouse conditions. 相似文献
19.
Summary
Kalopanax pictus (Thunb.) Nakai is a tall tree, and its wood has been used in making furniture, while its stem bark is used for medicinal
purposes. Here, we report on the micropropagation of Kalopanax pictus via somatic embryogenesis. Embryogenic callus was induced from immature zygotic embryos. The frequency embryogenic callus
induction is influenced by days of seed harvest. Callus formation was primarily observed along the radicle tips of zygotic
embryos incubated on Murashige and Skoog (MS) medium with 4.4 μM 2,4-dichlorophenoxyacctic acid (2,4-D). Somatic embryogenesis was observed following transfer of embryogenic callus to MS
medium lacking 2,4-D. Somatic embryos at the cotyledonary stage were obtained after 6 wk following culture. Frequency of conversion
of somatic embryos into plantlets was low (35%) on a hormone-free MS basal medium, but it increased to 61% when the medium
was supplemented with 0.05% charcoal. Gibberellic acid (GA3) treatment markedly enhanced the germination frequency of embryos up to 83%. All plantlets obtained showed 98% survival on
moist peat soil (TKS2) artificial soil matrix. About 30 000 Kalopanax pictus plants were propagated via somatic embryogenesis and grown to 3-yr-old plants. These results indicate that production of
woody medicinal Kalopanax pictus plantlets through somatic embryogenesis can be practically applicable for propagation. 相似文献
20.
Summary
In vitro propagation of Quassia amara L. (Simaroubaceae) was attempted using mature and juvenile explants. Attempts to establish in vitro culture using leaf and internode explants from a plant more than 15yr old were unsuccessful due to severe phenolic exudation.
Plant regeneration through direct and indirect somatic embryogenesis was established from cotyledon explants. Murashige and
Skoog (MS) medium with 8.9 μM N6-benzyladenine (BA) and 11.7 μM silver nitrate induced the highest number (mean of 32.4 embryos per cotyledon) of somatic embryos. Direct somatic embryogenesis
as well as callus formation was observed on medium with BA (8.9–13.3 μM). Semi-mature pale green cotyledons were superior for the induction of somatic embryos. Embryos developed from the adaxial
side as well as from the point of excision of the embryonic axis. More embryos were developed on the proximal end compared
to mid and distal regions of the cotyledons. Subculture of callus (developed along with the somatic embryos on medium with
BA alone) onto medium containing 8.9 μM BA and 11.7 μM silver nitrate produced a mean of 17.1 somatic embryos. Primary somatic embryos cultured on MS medium with 8.9 μM BA and 11.7μM silver nitrate produced a mean of 9.4 secondary somatic embryos. Most of the embryos developed up to early cotyledonary stage.
Reduced concentration of BA (2.2 or 4.4 μM) improved maturation and conversion of embryos to plantlets. Ninety percent of the embryos converted to plantlets. The optimized
protocol facilitated recovery of 30 plantlets per cotyledon explant within 80d. Plantlets transferred to small cups were subsequently
transferred to field conditions with a survival rate of 90%. 相似文献