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1.
When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus.
A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM
NAA, and 6 μM giberrellic acid (GA3). However, adding 24 μM silver nitrate (AgNO3) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length
of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm)
were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal.
Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l−1 sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with
globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants
to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred
to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when
these were transferred to MS medium was supplemented with 60 g l−1 sucrose. 相似文献
2.
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
3.
G. Vengadesan N. Selvaraj R. Prem Anand V. Gaba A. Ganapathi 《In vitro cellular & developmental biology. Plant》2005,41(6):789-793
Summary Suspension culture of cucumber (Cucumis sativus L.) has been an inefficient method for production of somatic embryos owing to problems with embryo maturation and conversion.
Embryogenic callus of cv. Green Long was induced on semisolid Murashige and Skoog (MS) medium containing 6.8 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.2 μM 6-benzylaminopurine (BA). A large number of globular somatic embryos were obtained on transfer of the callus to MS liquid
medium supplemented with 87.6 mM sucrose, 1.1 μM 2,4-D, and improved by the addition of 342.4 μM
l-glutamine. MS medium supplemented with 87.6 mM sucrose was more effective in somatic embryo production than other sugars. Subsequent development led to the formation of
heart-and torpedo-shaped embryos. Maturation of somatic embryos occurred on plant growth regulator-free MS semi-solid medium
containing 175.2 mM sucrose and 0.5 gl−1 activated charcoal. Conversion of embryos into plants was achieved on half-strength MS semi-solid medium containing 87.6
mM sucrose and 1.4 μM gibberellic acid (GA3) in a 16h photoperiod. Twenty-seven percent of embryos were converted into normal plants. 相似文献
4.
P. Jha C. B. Yadav V. Anjaiah V. Bhat 《In vitro cellular & developmental biology. Plant》2009,45(2):145-154
An efficient in vitro plant regeneration protocol through somatic embryogenesis and direct shoot organogenesis has been developed for pearl millet
(Pennisetum glaucum). Efficient plant regeneration is a prerequisite for a complete genetic transformation protocol. Shoot tips, immature inflorescences,
and seeds of two genotypes (843B and 7042-DMR) of pearl millet formed callus when cultured on Murashige and Skoog (MS) medium
supplemented with varying levels of 2,4-dichlorophenoxyacetic acid (2,4-D; 4.5, 9, 13.5, and 18 μM). The level of 2,4-D, the
type of explant, and the genotype significantly effected callus induction. Calli from each of the three explant types developed
somatic embryos on MS medium containing 2.22 μM 6-benzyladenine (BA) and either 1.13, 2.25, or 4.5 μM of 2,4-D. Somatic embryos
developed from all three explants and generated shoots on MS medium containing high levels of BA (4.4, 8.8, or 13.2 μM) combined
with 0.56 μM 2,4-D. The calli from the immature inflorescences exhibited the highest percentage of somatic embryogenesis and
shoot regeneration. Moreover, these calli yielded the maximum number of differentiated shoots per callus. An efficient and
direct shoot organogenesis protocol, without a visible, intervening callus stage, was successfully developed from shoot tip
explants of both genotypes of pearl millet. Multiple shoots were induced on MS medium containing either BA or kinetin (4.4,
8.8, 17.6, or 26.4 μM). The number of shoots formed per shoot tip was significantly influenced by the level of cytokinin (BA/kinetin)
and genotype. Maximum rooting was induced in 1/2 strength MS with 0.8% activated charcoal. The regenerated plants were transferred
to soil in pots, where they exhibited normal growth. 相似文献
5.
Summary An in vitro culture procedure was established for somatic embryogenesis and plant regeneration from callus cultures of the important
palm ‘betel nut’ (Areca catechu L.). Segments of zygotic embryos of Areca catechu L. were cultured on Murashige and Skoog basal medium supplemented with dicamba (9.05, 18.1, 27.15, and 36.2 μM). After 7–8 wk in darkness, wounded regions of explants formed callus with yellow, soft, glutinous structures. Proliferation
and maintenance of callus was on the same dicamba-containing medium. With regular subculture every 8 wk, the callus showed
pale yellow, compact and nodular structures. During subculture, somatic embryos were formed spontaneously from nodular callus
tissues within 2–4 mo. The embryos developed into plantlets after 10 wk of culture on basal medium free of plant growth regulators.
After subculturing every month for 3 mo., the plantlets were transferred to containers for acclimatization in the greenhouse.
The survival rate was 24%. 相似文献
6.
Jing Li Yang Bo Zhao Eun Soo Seong Myong Jo Kim Won Hee Kang Na Young Kim Chang Yeon Yu Cheng Hao Li 《Plant biotechnology reports》2010,4(4):261-267
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver
nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when
petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously
from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing
1.0 mg l−1 BA and 1.0 mg l−1 NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg l−1 GA3 had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully
acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the
primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction
of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained
by 1% sucrose. Most secondary embryos (87.2–94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary
secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose. 相似文献
7.
Pradeep C. Deo Mary Taylor Robert M. Harding Anand P. Tyagi Douglas K. Becker 《Plant Cell, Tissue and Organ Culture》2010,100(3):283-291
Embryogenic callus was initiated by culturing in vitro taro corm slices on agar-solidified half-strength MS medium containing
2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for 20 days followed by transfer to 1.0 mg/L thidiazuron (TDZ). Callus was
subsequently proliferated on solid medium containing 1.0 mg/L TDZ, 0.5 mg/L 2,4-D and 800 mg/L glutamine before transfer to
liquid medium containing the same components but with reduced glutamine (100 mg/L). After 3 months in liquid culture on an
orbital shaker, cytoplasmically dense cell aggregates began to form. Somatic embryogenesis was induced by plating suspension
cells onto solid media containing reduced levels of hormones (0.1 mg/L TDZ, 0.05 mg/L 2,4-D), high concentrations of sucrose
(40–50 g/L) and biotin (1.0 mg/L). Embryo maturation and germination was then induced on media containing 0.05 mg/L benzyladenine
(BA) and 0.1 mg/L indole-3-acetic acid (IAA). Histological studies of the developing embryos revealed the presence of typical
shoot and root poles suggesting that these structures were true somatic embryos. The rate of somatic embryos formation was
500–3,000 per mL settled cell volume while approximately 60% of the embryos regenerated into plants. 相似文献
8.
High-frequency plant regeneration via somatic embryogenesis and organogenesis and in vitro flowering of regenerated plantlets in Panax ginseng 总被引:4,自引:0,他引:4
W. Tang 《Plant cell reports》2000,19(7):727-732
The morphogenesis ability of light yellowish globular callus derived from cotyledons of mature zygotic embryos of Panax ginseng was investigated. The optimal media for somatic embryogenesis and shoot organogenesis were MS medium containing 0.5 mg l–1 2,4-dichlorophenoxyacetic acid, 0.1 mg l–1 6-benzyladenine (BA), and 500 mg l–1 lactoalbumin hydrolysate, and SH medium supplemented with 0.5 mg l–1 α-naphthaleneacetic acid, 0.1 mg l–1 BA, and 500 mg l–1casein hydrolysate. The influences of glucose, mannose, fructose, and sorbose in the media on somatic embryogenesis and shoot
organogenesis were revealed as differences in the numbers of somatic embryos and adventitious shoots per gram of morphogenic
callus. The best regeneration of somatic embryos was obtained on medium containing glucose, with a mean of 8.7 somatic embryos
per gram of callus. The best regeneration of shoots was observed on medium containing fructose, with an average of 12.2 adventitious
shoots per gram of callus. Of the somatic embryos 95% were converted into regenerated plantlets, and 100% of adventitious
shoots rooted to form regenerated plantlets. Regenerated plants were successfully established in soil. Flowering was observed
in 5.7% of the regenerated plants derived from shoot organogenesis and in 1.4% of the regenerated plants derived from somatic
embryogenesis.
Received: 1 December 1998 / Revision received: 13 September 1999 / Accepted: 20 September 1999 相似文献
9.
A. Othmani C. Bayoudh N. Drira M. Marrakchi M. Trifi 《Plant Cell, Tissue and Organ Culture》2009,97(1):71-79
Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year
old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth
regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months
on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping
and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with
1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus
was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and
306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments
was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots.
The growth of regenerated somatic plants was also monitored in the field. 相似文献
10.
Mya Thuzar Apichart Vanavichit Somvong Tragoonrung Chatchawan Jantasuriyarat 《Acta Physiologiae Plantarum》2011,33(1):123-128
An efficient and rapid plant regeneration system through somatic embryogenesis was developed using 13-week-old zygotic embryos
of oil palm (Elaeis guineensis Jacq.) cv. ‘Tenera’. Zygotic embryos were cultured on MS and N6 media supplemented with 2.0 mg L−1 picloram, 2,4-D and dicamba. The highest embryogenic callus formation (32%) was observed on N6 medium with 2,4-D after 3 month
culture on callus induction medium. Somatic embryos were continuously formed from nodular calli on embryo maturation medium
[N6 + 0.1 mg L−1 2,4-D, 0.16 g L−1 putrescine, 0.5 g L−1 casein amino acids and 2.0 g L−1 activated charcoal(AC)] for 3–5 months. Histological analysis confirmed that embryo development occurred via somatic embryogenesis.
For plant regeneration, modified N6 medium (MN6) with AC (0.5 g L−1) without growth regulators, induced both shoot and root formation simultaneously with the highest regeneration rate of 56%.
This combined shoot and root induction protocol shortened the culture time to 9–12 months. Furthermore, after acclimatization,
more than 85% of transferred plants from our protocol developed successfully in the soil. 相似文献
11.
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
12.
Perera PI Hocher V Verdeil JL Doulbeau S Yakandawala DM Weerakoon LK 《Plant cell reports》2007,26(1):21-28
Unfertilized ovaries isolated from immature female flowers of coconut (Cocos nucifera L.) were tested as a source of explants for callogenesis and somatic embryogenesis. The correct developmental stage of ovary explants and suitable in vitro culture conditions for consistent callus production were identified. The concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) and activated charcoal was found to be critical for callogenesis. When cultured in a medium containing 100 μM 2,4-D and 0.1% activated charcoal, ovary explants gave rise to 41% callusing. Embryogenic calli were sub-cultured into somatic embryogenesis induction medium containing 5 μM abscisic acid, followed by plant regeneration medium (with 5 μM 6-benzylaminopurine). Many of the somatic embryos formed were complete with shoot and root poles and upon germination they gave rise to normal shoots. However, some abnormal developments were also observed. Flow cytometric analysis revealed that all the calli tested were diploid. Through histological studies, it was possible to study the sequence of the events that take place during somatic embryogenesis including orientation, polarization and elongation of the embryos. 相似文献
13.
M. Muruganantham S. Amutha A. Ganapathi 《In vitro cellular & developmental biology. Plant》2010,46(1):34-40
The regeneration of plants via somatic embryogenesis liquid shake culture of embryogenic calluses was achieved in Vigna mungo (L.) Hepper (blackgram). The production of embryogenic callus was induced by seeding primary leaf explants of V. mungo onto Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) medium supplemented (optimally) with 1.5 mg/l 2,4-dichloro-phenoxyacetic acid. The embryogenic callus was then transferred
to liquid MS medium supplemented (optimally) with 0.25 mg/l 2,4-dichloro-phenoxyacetic acid. Globular, heart-shaped, and torpedo-shaped
embryos developed in liquid culture. The optimal carbohydrate source for production of somatic embryos was 3% sucrose (compared
to glucose, fructose, and maltose). l-Glutamine (20 mg/l) stimulated the production of all somatic embryo stages significantly. Torpedo-shaped embryos were transferred
to MS (Physiol Plant 15:473–497, 1962) liquid medium containing 0.5 mg/l abscisic acid to induce the maturation of cotyledonary-stage embryos. Cotyledonary-stage
embryos were transferred to 1/2-MS semi-solid basal medium for embryo conversion. Approximately 1–1.5% of the embryos developed
into plants. 相似文献
14.
Maurecilne Lemes da Silva Daniela Lopes Paim Pinto Miguel Pedro Guerra Eny Iochevet Segal Floh Cláudio Horst Bruckner Wagner Campos Otoni 《Plant Cell, Tissue and Organ Culture》2009,99(1):47-54
The objective of the present work was to induce somatic embryogenesis from zygotic embryos of Passiflora cincinnata Masters. Zygotic embryos formed calli on media with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and
4.5 μM benzyladenine (BA) after 30 days of in vitro culture. A concentration of 18.1 μM 2,4-D resulted in the largest number
of somatic embryos. Embryogenic calli were yellowish and friable, forming whitish proembryogenic masses. Morphologically,
embryogenic cells were small and had large nuclei and dense cytoplasm, whereas non-embryogenic cells were elongated, with
small nuclei and less dense cytoplasm. Calli cultured under white light on basal Murashige and Skoog’s medium with activated
charcoal produced embryos in all developmental stages. There were differences among the treatments, with some leading to the
production of calli with embryos and some only to callus formation. Some abnormalities were associated with somatic embryos,
including fused axes, fused cotyledons and polycotyledonary embryos. Production of secondary somatic embryos occurred in the
first cycle of primary embryo development. Secondary embryos differentiated from the surface of the protodermal layer of primary
embryos with intense cell proliferation, successive mitotic divisions in the initial phase of embryoid development, and a
vascular system formed with no connection to the parental tissue. This secondary embryogenic system of P. cincinnata is characterized by intense proliferation and maintenance of embryogenic competence after successive subcultures. This reproducible
protocol opens new prospects for massive propagation and is an alternative to the current organogenesis-based transformation
protocol. 相似文献
15.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
16.
Plant regeneration through somatic embryogenesis was achieved using immature zygotic embryos (ZE) of Sorbus pohuashanensis as explants. Over 50% of immature ZEs from immature seed collected at 30 days after pollination produced direct somatic embryos
(SEs) on Murashige and Skoog (MS) medium supplemented with 0–0.44 μM 6-benzyladenine (BA) in combination with 5.73 μM naphthaleneacetic
acid (NAA) or with 0.91–2.26 μM 2,4-dichlorophenoxyacetic acid (2,4-D) alone. Fourteen to 23 SEs per explant were regenerated
on MS medium supplemented with BA 0.44 μM in combination with NAA 5.73 μM. SE formation decreased when sucrose concentrations
were higher than 40 g L−1. Repetitive embryogenesis occurred following culture on solid MS medium containing 12 μM abscisic acid, 75 g L−1 polyethylene glycol, and 20 g L−1 sucrose at 25 ± 1°C under a 16-h photoperiod with a light intensity of 40 μmol m−2 s−1. Over 40% of the mature SEs germinated on solid MS medium under light condition described previously. Up to 40% of the regenerated
plantlets were successfully acclimatized under greenhouse conditions. Plantlets derived from SEs grew vigorously with similar
morphology as those germinated from ZEs. Histological studies of explants at various developmental stages of somatic embryogenesis
revealed that SEs passed through globular, heart, torpedo, and mature stages. Similar to ZE suspensors, similar structures
of SE degenerated in later stages of embryo development. ZE and SE are a effective means of regenerating tissue culture plantlets
for S. pohuashanesis. 相似文献
17.
P. I. P. Perera V. R. M. Vidhanaarachchi T. R. Gunathilake D. M. D. Yakandawala V. Hocher J. L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,99(1):73-81
Coconut is a cross pollinating palm, propagated only by seeds. Tissue culture is the only vegetative propagation method available
for coconut. Consistent callogenesis was obtained by culturing unfertilised ovaries at -4 stage in CRI 72 medium containing
100 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1% activated charcoal. Callusing was improved by application of 9 μM thidiazuron
(TDZ). Embryogenic calli were subcultured onto somatic embryogenesis induction medium containing 66 μM 2,4-D. Stunted growth
was observed in the somatic embryos after subculture onto CRI 72 medium containing abscisic acid (ABA). Maturation of somatic
embryos could be achieved in Y3 medium without growth regulators. Conversion of somatic embryos was induced by adding gibberellic acid (GA3) to conversion medium containing 5 μM 6-benzyladenine (BA) while 2-isopentyl adenine (2iP) increased the frequency of plant
regeneration. A total of 83 plantlets was produced from 32 cultured ovaries. 相似文献
18.
Mehdi Bakhshaie Mesbah Babalar Masoud Mirmasoumi Ahmad Khalighi 《Plant Cell, Tissue and Organ Culture》2010,101(2):229-235
A somatic embryogenesis (SE) protocol was established for the regeneration of Lilium ledebourii (Baker) Boiss. whole plants using new vegetative bulblet microscales and transverse thin cell layers (tTCLs) of young bulblet
roots as the explant sources. Bulblets were induced from bulb scale explants cultured for at least 3 months in the dark on
Murashige and Skoog (MS) medium containing 3% sucrose, 0.8% agar, and different concentrations of α-naphthaleneacetic acid
(NAA), 6-benzyladenine (BA), and thidiazuron. Embryo-like structures were obtained from tTCL explants of 3-month-old bulblets
(excised from bulb scale explants) following culture on solid MS medium containing 3% sucrose and various concentrations of
NAA and BA for 3 months in the dark. Both the explant source and the type of plant growth regulators affected the differentiation
of somatic embryos. The highest percentage (65.55%) of embryogenesis was obtained from bulblet microscale tTCLs cultured on
solid MS medium containing 0.54 μM NAA and 0.44 μM BA. Plants with normal shoots and roots were obtained following a 3-month
culture of embryos on growth regulator-free MS medium at 25 ± 1°C under a 16/8-h light/dark photoperiod (light intensity 40 μmol m−2 s−1, cool-white fluorescent light). The plants were successfully acclimatized in the growth chamber. 相似文献
19.
In the present study, the procedures for induction of somatic embryogenesis (SE) in an in vitro culture of the tulip have been developed. SE was initiated on flower stem explants isolated from “Apeldoorn” bulbs during
their low-temperature treatment. Bulbs had not been chilled or had been chilled for 12 or 24 weeks at 5°C. The explants were
cultured with exogenous auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (Picloram), α-naphthaleneacetic
acid (NAA) at 1–100 μM and cytokinins: benzyladenine (BA) and zeatin (ZEA) at 0.5–50 μM. Increase in auxin concentrations
caused an intensive enlargement of the explant parenchyma, which changed into homogenous colorless callus. On the same media,
vein bundles developed into yellowish, nodular callus. Picloram was more efficient in inducing the formation of embryogenic
nodular callus than 2,4-D, whereas the latter stimulated formation of colorless callus. The base of the lower part of the
flower stem isolated from bulbs chilled for 12 weeks proved to be the best explant for callus formation. The highest number
of somatic embryos was produced on medium with 25 μM Picloram and 0.5 μM BA. Development of adventitious roots was noticed
in the presence of 2,4-D. Globular embryos developed into torpedo stage embryos under the influence of BA (5 μM) and NAA (0.5 μM).
Morphological and anatomical data describing development of callus and somatic embryos are presented. 相似文献
20.
S. Paul A. Dam A. Bhattacharyya T. K. Bandyopadhyay 《Plant Cell, Tissue and Organ Culture》2011,105(2):271-283
A reproducible protocol for direct and indirect somatic embryogenesis was established in a small aromatic tree, Murraya koenigii. Embryogenic callus was obtained from 90% zygotic embryonic axis (ZE) and 70% cotyledon (COT) explants in Murashige and Skoog
(MS) basal medium supplemented with 8.88 μM 6-benzyladenine (BA) and 2.675 μM α-naphthaleneacetic acid (NAA). Globular somatic
embryos were induced and further matured from such embryogenic callus by subsequent culture on the same basal media containing
thidiazuron (TDZ) (2.27–9.08 μM). The highest frequency of somatic embryos (14.58 ± 0.42) was recovered from ZE-derived callus
after 6 weeks. The age and type of explant and concentration of TDZ played an important role in the development of somatic
embryos. Explants excised from 60-day-old seed differentiated from 96.67% of ZE explants and 86.67% from COT explants when
cultured on MS basal medium supplemented with 4.54 and 9.08 μM TDZ, respectively, after 4 weeks. The best result obtained
for the average frequency of somatic embryos (11.28 ± 0.32) was from ZE explants, which was significantly higher than COT
explants (7.34 ± 0.97). Most of the somatic embryos (above 95%), irrespective of their origin, germinated after 4 weeks in
1/2 MS basal media containing 2.32 μM kinetin (KN) and 1.07 μM NAA. Well-rooted plantlets were successfully acclimatized.
Histological analysis and scanning electron micrographs confirmed the initiation, development, and germination of somatic
embryos from both explants. 相似文献