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1.
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
2.
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
3.
Xueping Shi Xigang Dai Guofeng Liu Junwei Zhang Guogui Ning Manzhu Bao 《In vitro cellular & developmental biology. Plant》2010,46(2):117-125
An efficient protocol for secondary somatic embryogenesis in camphor tree is reported. Secondary somatic embryos (SSEs), initially
obtained from the primary embryos of a nascent embryogenic culture in 2002, were proliferated and maintained for more than
4 yr via cyclic secondary somatic embryogenesis. Throughout this period, the embryo populations retained a high level of competence
for plant regeneration. SSEs were produced on the surfaces of the cotyledons and radicular ends of maternal somatic embryos
(MSEs). Histological observations of the various stages of secondary embryo development revealed four typical stages, namely,
globular, heart-shaped, torpedo, and cotyledonary. The process of secondary embryogenesis continued in a cyclic way, with
each newly formed embryo producing a subsequent generation of secondary embryos. In order to progress developmentally beyond
proliferation cycles, cotyledonary embryos from one of embryogenic lines (L14) were cultured on Murashige and Skoog (MS) medium
with 0.1–3.0 mg l−1 abscisic acid (ABA) or 0.05–1.0 mg l−1 thidiazuron (TDZ) in darkness for 2 mo to achieve maturation. Matured embryos were then transferred to MS-based germination
medium containing either 0.1 mg l−1 TDZ, 0.2 mg l−1 indole-3-butyric acid (IBA), and 0.5 mg l−1 6-benzylaminopurine (BA) or 0.1 mg l−1 TDZ and 0.2 mg l−1 IBA and were cultured in light for germination. Over 50% of embryos matured in the presence of 0.5 mg l−1 ABA were able to germinate with shoots and poor root system. Frequencies of embryos germinating normal shoots among different
genotypes did not change significantly. A total of 93% of the shoots from the germinated embryos converted to plantlets on
half strength MS medium with 0.5 mg l−1 IBA by 3 wk. Plantlets acclimatized successfully to ex vitro conditions and developed as field-grown plants with normal appearance. 相似文献
4.
Kourosh Vahdati Shima Bayat Hassan Ebrahimzadeh Maryam Jariteh Masoud Mirmasoumi 《Plant Cell, Tissue and Organ Culture》2008,93(2):163-171
Low efficiency of embryo maturation, germination and conversion to plantlets is a major problem in many species including
Persian walnut. We studied the effects of abscisic acid (ABA) and sucrose, on the maturation and germination of Persian walnut
(Juglans regia) somatic embryos. Individual globular somatic embryos were grown on a maturation medium supplemented with different combinations
of ABA and sucrose for ca. 1 month, until shoot meristems and radicles had developed. White and opaque embryos in late cotyledonary
stage were subjected to desiccation after the culture period on maturation media. The number of germinated somatic embryos
was influenced by the concentrations of ABA in the maturation medium. The best treatment for germination, in which both shoot
and root were developed contained 2 mg l−1 ABA and resulted in 41% conversion of embryos into plantlets. Regeneration was reduced at higher levels of ABA. While ABA
always reduced the rate of secondary embryogenesis, treatments containing 4.0% sucrose significantly increased the number
of secondary embryos. On the other hand, sucrose had little influence on maturation. Normal and abnormal embryos were verified
anatomically. 相似文献
5.
Summary The effects of abscisic acid (ABA) (0, 0.09 μM, 0.19 μM, 0.28 μM, and 0.38 μM) or ancymidol (0, 0.98 μM, 1.95 μM, 2.93 μM, 3.90 μM) in embryo germination medium on the conversion of primary embryos to plantlets and secondary embryogenesis were evaluated
for asparagus. ABA and ancymidol each significantly enhanced both responses. ABA was more effective than ancymidol in promoting
the conversion of primary embryos to plantlets, while the converse was true for the production of secondary embryos. The most
effective treatments for embryo conversion were 0.19 and 0.28 μM ABA; 75–77% bipolar and 55–57% globular embryos converted to plantlets. For secondary embryogenesis, the most effective treatments
were 1.95 and 2.93 μM ancymidol; 99–101 and 84–86 somatic embryos were produced from 10 globular and 10 bipolar embryos, respectively. Bipolar
embryos generally converted to plantlets better than globular embryos, but more secondary embryos were produced from globular
embryos than from bipolar embryos in all treatments. ABA and ancymidol also affected the morphology of the plantlets produced.
The plantlets from the embryos incubated on the medium with ancymidol had strong and thick shoots and roots, while those on
the medium with ABA had long, thin shoots and short thin roots. 相似文献
6.
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and petiole expiants has been developed in sweetpotato [Ipomoea batatas L. (Lam.)]. The optimal somatic embryogenic response was obtained in the genotype PI 318846-3 with a two-step protocol: (1) stage I-incubation of expiants in the dark for 2 weeks on Murashige Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l) and 6-benzylaminopurine (0.25 mg/l) and, (2) stage II-culture in the light on MS medium with abscisic acid (ABA) (2.5 mg/l). The addition of ABA was critical for enhanced production of somatic embryos. Secondary somatic embryos were produced from the primary embryos cultured on MS medium with 2,4-D at 0.2 mg/l. The somatic embryos were converted into normal plantlets when cultured on basal MS medium. Upon transfer to soil, plants grew well and appeared normal with no mortality. The system of somatic embryogenesis described here will facilitate tissue culture, germplasm conservation and gene transfer research of sweetpotato due to its rapidity (6 to 10 weeks), prolific plant production by direct embryogenesis, ease of secondary somatic embryo production and reproducibility.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine, 2,4-D-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- KIN
kinetin
- MS
medium of Murashige and Skoog (1962)
- NAA
1-naph-thaleneacetic acid
- PIC
picolinic acid
- TDZ
thidiazuron 相似文献
7.
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
8.
A protocol has been developed for achieving somatic embryogenesis and plant regeneration from petiole-derived callus of Heracleum candicans Wall. Callus was initiated on MS medium supplemented with 0.5 mg l–1 2,4-D and 0.5 mg l–1 BAP and subcultured on a medium containing double strength MS macrosalts, 1 mg l–12,4-D and 0.25 mg l–1 Kn. Numerous globular embryos were formed on the surface of the callus upon transfer to auxin-rich MS medium that lacked
cytokinins. The globular embryos differentiated into mature embryos only when 2,4-D was removed from the medium. Mature embryo
formation was significantly influenced by the pH of the medium and the addition of AgNO3 and ABA. Eighty-five percent of the somatic embryos were converted into plantlets when transferred to a medium supplemented
with 0.01 mg l–1 BAP and 0.01 mg l–1 IBA. The regenerated plants have been established in soil and appear to be identical to the parent plants in morphology and
chromosome number.
Received: 5 November 1997 / Revision received: 9 February 1998 / Accepted: 19 February 1998 相似文献
9.
Efficient somatic embryogenesis (SE) and in vitro flowering and fruiting were achieved in Saposhnikovia divaricata (Turcz.) Schischk. Friable embryogenic callus developed from the root, internode, and leaf explants on Murashige and Skoog
medium (MS) with 2.26 μM 2,4-dichlorophenoxyacetic acid (2,4-D), and subsequently developed into somatic embryos on MS medium
containing 4–5% sucrose, 1.74 μM naphthaleneacetic acid (NAA), 4.44 μM 6-benzylaminopurine (BA), and 1.90 μM abscisic acid
(ABA). Then the mature embryos were separated and transferred onto MS with 3% sucrose and 0.6% agar for further development
and conversion to plantlets. In vitro flowering and fruiting were obtained when the subcultures were carried out for over
15 months. Paclobutrazol (PP333) or ethephon (ETH) at low levels promoted flowering significantly. Also, abnormal rootless
somatic embryos of S. divaricata could form flowers and fruits in vitro. 相似文献
10.
Diego Muñoz-Concha Sean Mayes Gracia Ribas Michael R. Davey 《Plant Cell, Tissue and Organ Culture》2012,109(1):123-130
The endangered Chilean tree Gomortega keule (Mol.) Baillon produces edible fruit, making it a potential crop. However, its cultivation from seed or cuttings is extremely
difficult. This paper reports the induction of somatic embryogenesis and the initiation of liquid cultures in this species.
Callus was induced from zygotic embryos and field-collected shoots. Somatic embryogenesis on zygotic embryos occurred at a
low frequency. Induction of somatic embryogenesis was accomplished on micropropagated shoots after 6.5 months on semi-solid
Murashige and Skoog (MS) medium with 30 g/l sucrose, 1.0 mg/l 2,4-dichlorophenoxyacetic acid and 1.0 mg/l 6-(γ,γ-dimethylallylamino)
purine (2iP). Liquid cultures of compact callus and small aggregates were obtained and showed optimum proliferation in MS
medium with 20 g/l sucrose, 0.01 mg/l α-naphthaleneacetic acid and 0.1 mg/l 2iP. The proliferation of friable embryogenic
callus was observed in liquid medium and will allow the propagation of selected genotypes of this tree on a large scale. Genetic
variation in two embryogenic genotypes cultured in vitro was not detected in an assessment using microsatellites; this approach
is suitable for tracing genotypes. 相似文献
11.
Trabelsi El Bahri Bouzid Sadok Bouzid Monji Elloumi Nedra Belfeleh Zina Benabdallah Abdallah Ghezel Rachida 《Journal of Plant Biology》2003,46(3):173-180
We induced somatic embryogenesis from the cotyledon segments ofOlea europaea (L) cvs. ‘Chetoui’, ‘Chemleli’, and ‘Arbequina’. Calli were established from all three cultvars on OMc media supplemented
with IBA and 2i-R The greatest success was obtained with media that contained zero or low concentrations of growth regulators.
High levels of hormones (i.e.,>0.5 mgL-1 IBA and 2i-P) inhibited embryogenesis. Embryos at different maturation stages were observed with continuously proliferating
secondary embryogenesis. Abnormally shaped embryos and teratoma were also noted. Four weeks was the optimal incubation period
for inducing embryogenesis on the auxin-containing medium. In addition, 30 to 40 gL-1 sucrose was more effective than glucose in stimulating the growth and maturation of somatic embryos. Embryogeic efficiency
was also higher when multivariate combinations of nitrogen sources (inorganic and organic nitrogen forms) were used. The plantlets
that were derived from our germinating somatic embryos were similar to those obtained from axillary buds. 相似文献
12.
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
13.
Yong Wook Kim So Young Park In Sun Park Heung Kyu Moon 《Plant biotechnology reports》2007,1(4):237-242
We have tested plantlet formation by somatic embryogenesis using immature seeds of Magnolia obovata. Seed collection date appeared to be critical for embryogenic cell induction. The optimal collection date was 3–4 weeks postanthesis.
The embryogenic cells proliferated, formed somatic embryos, and were subsequently converted into normal plantlets under optimized
culture conditions. Somatic embryo formation from the embryogenic calli was better on sucrose medium than on glucose medium.
The optimum level of sucrose appeared to be 3% with an average of 28 somatic embryos per plate. About 25% of somatic embryos
were converted into normal plantlets in 1/2 MS medium containing gibberellic acid (GA3). During somatic embryo germination, secondary embryogenesis was frequently observed in the lower part of the hypocotyl or
radicle ends of germinating somatic embryos. Finally, about 85% of converted plantlets survived in an artificial soil mixture,
were transferred to a nursery, and have grown normally. 相似文献
14.
High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.) 总被引:5,自引:0,他引:5
A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to “torpedo” were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species. 相似文献
15.
An efficient protocol was established for regeneration of Desmodium motorium via somatic embryogenesis. Embryogenic calli were induced from cotyledon segments (6 mm, 16 days old) lacking embryo axis,
excised from seedlings grown in vitro on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA) (2.9 μM)
in combination with 6-benzyladenine (BA) (4.44 and 8.88 μM). Differentiation of embryogenic calli into globular and heart-shaped
somatic embryos was achieved on transfer to hormone-free MS medium. When incubated for 4 days on MS medium supplemented with
BA (8.88 μM), 95% of the globular and heart-shaped somatic embryos matured into torpedo and cotyledonary stages with minimum
(10%) abnormalities. Modified MS basal medium without hormones and containing half-strength macronutrients and 0.88 M sucrose
was suitable for germination of mature somatic embryos. Regenerated plantlets were successfully transferred to earthen pots
with survival rate of 50%. Secondary embryogenesis was observed when pre-existing somatic embryos at globular and heart-shaped
stages were cultured on MS medium supplemented with various concentrations of BA, adenine sulphate (AdS) and abscisic acid
(ABA) individually. 相似文献
16.
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
17.
Ying Bao Guofeng Liu Xueping Shi Wen Xing Guogui Ning Juan Liu Manzhu Bao 《Plant Cell, Tissue and Organ Culture》2012,109(3):411-418
This study describes a plant regeneration system for Rosa hybrida ‘Samantha’, a mainstream commercial cultivar, via direct somatic embryogenesis. Somatic embryogenesis was induced from in
vitro-derived leaf explants, achieving a frequency of 7.5% following 8 weeks of culture on a Murashige and Skoog (MS) medium
supplemented with 3.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 30 g/L glucose. We evaluated the effects of various
types and concentrations of carbohydrates and plant growth regulators (PGRs) on the proliferation and germination of secondary
somatic embryos. MS medium containing 1.0 mg/L 2,4-D, 0.05 mg/L 6-benzyladenine (BA) and 60 g/L glucose was found to be the
most effective in promoting the proliferation of somatic embryos. The highest germination rate (56.2%) of somatic embryos
was observed on MS medium containing 1.0 mg/L Thidiazuron (TDZ) and 0.5 mg/L BA. Whole plantlets were obtained by culturing
germinated shoots on rooting medium comprising 1/2-strength MS salts supplemented with 0.05 mg/L α-Naphthalene acetic acid
(NAA). Plantlets grew vigorously with normal vegetative and flowering characteristics. 相似文献
18.
Summary Sodium chloride-tolerant plantlets of Dendrocalamus strictus were regenerated successfully from NaCl-tolerant embryogenic callus via somatic embryogenesis. The selection of embryogenic
callus tolerant to 100 mM NaCl was made by exposing the callus to increasing (0–200 mM) concentrations of NaCl in Murashige and Skoog medium having 3% (w/v) sucrose, 0.8% (w/v) agar, 3.0 mg l−1 (13.6 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5mg l−1 (2.3μM) kinetin (callus initiation medium). The tolerance of the selected embryogenic callus to 100 mM NaCl was stable through three successive transfers on NaCl-free callus initiation medium. The tolerant embryogenic callus
had high levels of Na+, sugar, free amino acids, and proline but a slight decline was recorded in K+ level. The stable 100 mM NaCl-tolerant embryogenic callus differentiated somatic embryos on maintenance medium [MS medium +3% sucrose +0.8% agar +2.0
mg l−1 (9.0 μM) 2,4-D+0.5 mg l−1 (2.3 μM) kinetin] supplemented with different (0–200 mM) concentrations of NaCl. About 39% of mature somatic embryos tolerant to 100 mM NaCl germinated and converted into plantlets in germination medium [half-strength MS+2% sucrose+0.02 mg l−1 (0.1 μM) α-naphthaleneacetic acid +0.1 mg l−1 (0.49 μM) indole-3-butyric acid] containing 100 mM NaCl. Of these plantlets about 31% established well on transplantation into a garden soil and sand (1:1) mixture containing
0.2% (w/w) NaCl. 相似文献
19.
Yongxue Yang Guofeng Liu Manzhu Bao 《In vitro cellular & developmental biology. Plant》2006,42(6):520-524
Summary A protocol of somatic embryogenesis and plant regeneration from petiole segments of Parthenocissus tricuspidata Planch. has been developed. Embryogenic tissue was induced on B5 (Gamborg) basal medium supplemented with 2.25–9.0 μM 2,4-dichlorophenoxyacetic acid, 500 mg l−1 casein hydrolysate (CH), and 0.1 gl−1 activated charcoal. Somatic embryos were induced on B5 medium containing various concentrations of benzyladenine (BA) (4.44,
6.66, and 8.88 μM) and α-naphthaleneacetic acid (NAA) (0, 0.54, and 1.61 μM) plus 500 mg l−1 CH. Ninety percent of normal somatic embryos were converted into plantlets directly on Murashige and Skoog (MS) medium free
of plant growth regulators. Shoots could be induced from abnormal somatic embryos on MS medium containing 4.44 μM BA, 0.05 μM NAA, and 500 mg l−1 CH. Genotypic differences were found in the process of somatic embryogenesis and plant regeneration. Histological analysis
confirmed the process of somatic embryogenesis. Regenerated plantlets with well-developed roots were successfully acclimatized
in greenhouse and all plants showed normal morphological characteristics. 相似文献
20.
Ana Margarida Capelo Sónia Silva Gina Brito Conceição Santos 《Plant Cell, Tissue and Organ Culture》2010,103(2):237-242
We describe a protocol for somatic embryogenesis (SE) induction from an adult wild olive tree (Olea
europaea ssp. europaea var. sylvestris. The protocol used confirms for the first time that there is no need to use juvenile or rejuvenated material for SE induction.
For SE induction, petiole and leaf (proximal, intermediary and distal zones) explants were grown on Murashige and Skoog (MS)
or Olive Medium (OM) media with different combinations of plant growth regulators (PGR): α- naphthaleneacetic acid (NAA),
Zeatin (Zea), indole-3-butyric acid (IBA), 2-isopentyl adenine (2iP), thidiazuron (TDZ) and 6-benzylaminopurine (BAP). All
media had 30 g/l sucrose and 7 g/l agar, and the pH was adjusted to 5.8. Cultures were incubated in the dark and, after 3 months,
they were transferred to MS medium without PGR for expression. Petiole explants gave the highest callus production, while
for SE induction and expression distal blade leaf and petiole explants gave the highest rates. The best medium for SE induction
was MS with 12.25 μM IBA plus 4.56 μM Zea. Histological analyses confirmed the individuality of globular somatic embryos.
This is the first report of SE expression in explants without rejuvenation in Olea genus, and opens perspectives for using this strategy in SE protocols both for this wild genotype and for commercial genotypes. 相似文献