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1.
Murugesan Dhandapani Doo Hwan Kim Seung-Beom Hong 《In vitro cellular & developmental biology. Plant》2008,44(1):18-25
High-frequency plant regeneration of C. roseus cv. ‘little bright eye’ via somatic embryogenesis and organogenesis from five out of six explants was standardized. Two factors
were found to be important for regeneration: (1) the type of explants, and (2) the combination and concentrations of plant
growth regulators. The highest regeneration percentage through somatic embryogenesis was obtained from mature zygotic embryo
in MS medium supplemented with 7.5 μM of thidiazuron (TDZ). The mature embryo also regenerated efficiently via organogenesis
in MS medium supplemented with either 2.5 μM TDZ or 5.3 μM α-naphthalene acetic acid (NAA) and 2.2 μM 6-benzylaminopurine
(BA). Hypocotyl and cotyledon did not induce somatic embryogenesis and organogenesis in TDZ-containing medium but gave a maximum
percentage of shoots in MS medium supplemented with 5.3 μM NAA and 2.2 μM BA. Stem nodes and meristem tips showed better regeneration
via organogenesis in the medium supplemented with NAA and BA and in lower concentrations of TDZ. 相似文献
2.
Ashok Kumar Sahrawat Suresh Chand 《In vitro cellular & developmental biology. Plant》2001,37(1):55-61
Summary An efficient method was established for high-frequency embryogenic callus induction and plant regeneration from 3-,4-, 5-
and 7-d-old coleoptile segments of Indica rice (Oryza sativa L. cv. Kasturi), Compact and friable callus developed from the cut ends and also on the entire length of the coleoptile segments
cultured on Murashige and Skoog (MS) basal medium (1962) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, 4.50–18.0
μM), kinetin (2.32 μM) and sucrose (3%, w/v). High frequency embryogenic callus induction and somatic embryo development was achieved when embryogenic
calluses were transferred to MS medium supplemented with 2.25 μM 2,4-D, 2.32 μM kinetin, 490 μM
L-tryptophan and 3% (w/v) sucrose. Plant regeneration was achieved by transferring clumps of embryogenic callus onto MS medium
containing 2.85 μM indole-3-acetic acid (IAA), 17.77 μM 6-benzylaminopurine (BA) and 3% (w/v) sucrose. Histological observations of embryogenic calluses revealed the presence of
somatic embryos and also plant regeneration via multiple shoot bud formation. Three, 4- and 5-d-old coleoptile segments showed
a significantly (P<0.05) higher frequency of plant regeneration and mean number of plantlets per explant in comparison to 7-d-old coleoptile
segments. The highest frequency (73.5%) of plant regeneration and mean number of plantlets (11.9±1.0) was obtained from 4-d-old
coleoptile segments. Regenerated shoots were rooted on MS basal medium containing 4.92 μM indole-3-butyric acid (IBA) and plants were successfully transferred to soil and grown to maturity. 相似文献
3.
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
4.
Mohammed A. M. Aly Essam A. Amer Wasef A. Al-Zayadneh Alaa E. Negm Eldin 《Plant Cell, Tissue and Organ Culture》2008,95(1):107-114
Inducing somatic embryogensis from jojoba [Simmondsia chinensis (Link) Schneider] explants to produce artificial seeds in the laboratory (in vitro) may prove highly profitable, as the seeds
contain a characteristic liquid wax of economic importance in industry, nutrition and medicine. Thus, there is a need to examine
the effect of the factors involved in the in vitro process on the quality and quantity of the synthesized fatty acids in comparison
with those naturally produced in vivo. Immature zygotic embryos and mature leaf explants were cultured on Murashige and Skoog
basal medium (MS) supplemented with various levels of 2,4-D, BA and sucrose. Embryogenic calluses developed from the zygotic
embryos and leaf explants over a period of 2–4 weeks with the highest response at 0.4 μM 2,4-D, 2.2/4.4 μM BA and 117 mM sucrose
(4%). Following induction, the zygotic embryo derived somatic embryos developed to the globular, heart, torpedo, and cotyledon
stages. Direct somatic embryogenesis was observed with some of the zygotic embryo explants. Leaf-derived embryogenic calluses
did not mature on any of the maturation/germination media examined up to 4 weeks of culture. Analysis of fatty acids indicated
that the mature seeds are characterized with long chain saturated fatty acids C22:0 behenic Acid. The zygotic embryo-derived
somatic embryos (SE-Z) and leaf-derived somatic embryos (SE-L) are characterized with the induction of the essential polyunsaturated
fatty acid C18:2 (omega-6) linoleic acid, (omega-3) alpha-linolenic acid (ALA), with higher values of long chain saturated
fatty acids C16:0 palmitic acid and monounsaturated fatty acid C18:1 oleic acid. These results indicate that manipulating
the growth regulators in the induction media influenced the fatty acids synthesis and hence the fatty acids profile in jojoba
somatic embryos. 相似文献
5.
T. H. Lan P. I. Hong C. C. Huang W. C. Chang C. S. Lin 《In vitro cellular & developmental biology. Plant》2009,45(1):44-47
Whole plants were regenerated from excised leaves of Drimiopsis kirkii Baker (Lily of the Valley) through direct somatic embryogenesis. An initial exposure to a low level of 2,4-dichlorophenoxyacetic
acid (2,4-D, 0.45 μM) in the medium was essential in inducing the direct formation of somatic embryos. A high concentration
of 2,4-D (4.52 μM) in the proliferation medium reduced embryogenesis and enhanced callus formation. The presence of kinetin
in the medium enhanced the somatic-embryogenesis-inducing effect of 2,4-D (0.45 μM). The maximum embryogenesis rate (4,026
somatic embryos per gram of leaf) was obtained in explants cultured for 30 d in medium supplemented with 2.33 μM kinetin and
0.45 μM 2,4-D (embryo induction medium). Kinetin (4.65 μM) also enhanced embryo germination (97.6%), but the presence of α-naphthalene
acetic acid in the medium drastically reduced embryo germination. Following conversion, the regenerated plantlets were transferred
to soil and showed normal morphological characteristics. 相似文献
6.
A protocol was developed for the induction, maturation and germination of somatic embryos from leaf tissue of jojoba [Simmondsia chinensis (Link) Schneider]. Explants were placed on their adaxial sides in Petri dishes and maintained in darkness on half-strength
Murashige and Skoog basal medium (MS/2). Combinations of 2,4-dichlorophenoxyacetic acid (1.35–4.52 μM) with 6-benzylaminopurine
(1.33–4.43μM) and 2 synthetic cytokinins, N-(2-chloro-4pyridyl)-N′-phenylurea (1.21–4.03μM) or (E)-6-[3-(trifluoromethyl)-but-2-enylamino]
purine (1.11–3.71μM) resulted in formation of embryogenic cultures and somatic embryos. After two 30-day subcultures, embryogenic
cultures were transferred onto MS/2 medium supplemented with different auxins and cytokinins. Somatic embryo maturation, germination
and plantlet formation were achieved using 1-naphthaleneacetic acid (3.75μM) or indole-3-butyric acid (3.44μM) in combination
with BA (0.44 or 1.33μM) or F3iP (0.37 or 1.11μM). Histology confirmed each stage of development.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
Summary Regeneration of plants via somatic embryogenesis was achieved from zygotic embryo explants isolated from mature seeds of Schisandra chinensis. Merkle and Sommer's medium, fortified with 2,4-dichlorophenoxyacetic acid (2,4-D; 9.04 μM) and zeatin (0.09 μM), was effective for induction of embryogenic callus. The development of a proembryogenic mass and somatic embryos occurred
on Murashige and Skoog medium (MS) free of plant growth regulators. The embryogenic callus induced on Merkle and Sommer's
medium supplemented with 2,4-D (9.04 μM) and zeatin (0.09 μM) showed development of the maximum number of somatic embryos when transferred to MS medium free of plant growth regulators.
The maximum maturation and germination of cotyledonary somatic embryos (46.3%) occurred on MS medium supplemented with 2,4-D
(0.45 μM) and N6-benzyladenine (1.11 μM). The somatic embryo-derived plants were successfully hardned, with a survival rate of approximately 67%, and established
in the field. 相似文献
8.
Influence of boron on somatic embryogenesis in papaya (Carica papaya L.) cv. Honey Dew was investigated. Immature zygotic embryos were grown in the induction medium containing Murashige and
Skoog basal salts, with B5 vitamins, picloram (1 mg dm−3) or 2,4-dichlorophenoxy acetic acid (2 mg dm−3) and different concentrations of boric acid (30 to 500 mg dm−3). Maximum somatic embryo initiation was observed at 62 mg dm−3 boric acid irrespective of the growth regulator used. The cotyledonary stage somatic embryos were germinated on MS basal
medium devoid of growth regulators. The regenerated plantlets were hardened under greenhouse conditions and transferred to
field.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
9.
Wang Xiao Xue-Lin Huang Xia Huang Ya-Ping Chen Xue-Mei Dai Jie-Tang Zhao 《Plant Cell, Tissue and Organ Culture》2007,90(2):191-200
A protocol for plant regeneration from protoplasts of Musa acuminata cv. Mas (AA) via somatic embryogenesis was developed. Viable protoplasts were isolated from embryogenic cell suspensions
at a yield of 1.2 × 107 protoplasts/ml packed cell volume (PCV). Liquid and feeder layer culture systems with medium-A and medium-B were used for
protoplast culture. In liquid culture system, medium-B was more efficient for inducing cell division (17.5% at 14 days) and
colony formation (6.7% at 28 days) than medium-A. However, all protoplast-derived cell colonies (PDCC) obtained from liquid
culture system could not develop further. In feeder layer culture system, there was no significant difference between medium-A
and medium-B on cell division and colony formation of the cultured protoplasts, and the cell division frequency at 14 days
and colony formation frequency at 28 days were 24.5% and 11.2%, respectively, in medium-B. Comparative study on the effects
of BAP (2.2 μM, 4.4 μM, 8.8 μM), zeatin (0.4 μM, 0.8 μM, 1.2 μM) and TDZ (0.2 μM, 0.4 μM, 0.6 μM) on embryo formation of PDCC
from feeder-layer culture indicated that TDZ was best. TDZ at 0.4 μM induced 7906 mature embryos per ml PCV PDCC, which was
4-fold the frequency as with BAP at 4.4 μM, 7.5-fold as with zeatin at 0.8 μM and 150-fold as control medium (no mentioned
cytokinins) after 45 days on M3 medium. About 44% of the mature embryos were converted into plantlets with poor root system
after subculture on M4 medium. Root further development of regenerated plantlets was promoted by addition of activated charcoal
(AC) to MS basal medium. 相似文献
10.
Marie-Anne Lelu-Walter Michèle Bernier-Cardou Krystyna Klimaszewska 《Plant Cell, Tissue and Organ Culture》2008,92(1):31-45
Several factors affecting somatic embryogenesis (SE) in Pinus sylvestris from self- and cross-pollinated seed families were studied with the aim of producing large quantities of clonal plants. Somatic
embryogenesis initiation from zygotic embryos was improved on a medium with lower than standard concentrations of 2,4-dichlorophenoxyacetic
acid (2.2 vs. 9.5 μM) and 6-benzyladenine (2.2 vs. 4.5 μM). On this medium, initiation rates of four controlled crosses, including
one self-cross, varied from 3% to 25%. Among the maturation factors tested, the concentration of abscisic acid (ABA 80, 120 μM)
had no significant effect on the production of mature somatic embryos when the medium contained 0.1 M sucrose. When sucrose
concentration was 0.2 M, however, 1.4 times more mature somatic embryos were produced on medium with 80 μM compared with 120 μM
ABA. Under our best maturation conditions, mature somatic embryos accumulated amounts of storage proteins that were similar
to the amounts in mature zygotic embryos. Activated charcoal exerted a beneficial effect on mature somatic embryo production
of 24-week-old cultures; there was no evidence of such an effect in 8-week-old cultures. Thirty-seven embryogenic lines from
a self-cross and an out-cross were chosen for clonal plant production. Highly embryogenic lines produced mature somatic embryos
that were more likely to convert to plants than those from less embryogenic lines. After 4 months of growth in a shade house,
plantlet survival rates exceeded 70% for 31 lines out of 35. This report describes an improved method for accelerated production
of large quantities of Scots pine for clonal tests. 相似文献
11.
Suresh Chand Ashok Kumar Sahrawat 《In vitro cellular & developmental biology. Plant》2002,38(1):33-38
Summary An efficient plant regeneration protocol has been developed from root explants of Psoralea corylifolia L., an endangered medicinally
important herbaceous plant species belonging to the family Fabaceae. Nodular embryogenic callus was initiated from young root
segments cultured on Murashige and Skoog (MS) medium (1962) supplemented with α-naphthaleneacetic acid (NAA; 2.68–13.42 μM)
or 2,4-dichlorophenoxyacetic acid (2.4-D; 2.25–11.25 μM) in combination with 6-benzylaminopurine (BA: 2.2. μM). thiamine HCl
(2.9 μM), L-glutamine (342.23 μM) and sucrose (3.0% w/v). The highest frequency (95.2%) of embryogenic calluses was obtained on MS medium
supplemented with the growth regulators NAA (10.74 μM) and BA (2.2 μM). Development and maturation of somatic embryos was
achieved after transfer of embryogenic calluses to MS medium supplemented with 1.34 μM NAA or 1.12 μM 2,4-D and 4.4–13.2 μM
BA. The maximum number (13.8±1.34) of cotyledonary stage somatic embryos was obtained on MS medium containing 1.34 μM NAA
and 13.2 μM BA. Germination of somatic embryos occurred on MS medium without any growth regulators and also on MS medium enriched
with BA (1.1–8.8 μM), although the maximum germination frequency (76.1%) was obtained on 4.4 μM BA plus 1.45 μM gibberellic
acid (GA3). Plant regeneration without complete somatic embryo maturation was also achieved by transferring clumps of nodular embryogenic
calluses onto MSO medium or MS medium supplemented with NAA (1.34 μM) and BA (2.2–8.8 μM). The highest frequency of plant
regeneration (93.3%) and mean number of plantlets (15.4±0.88) were obtained on MS medium containing 1.34 μM NAA and 4.4 μM
BA. Regenerated plants with well-developed root systems were transferred to pots where they grew vigorously, attained maturity
and produced fertile seeds. 相似文献
12.
Somatic embryos of myrtle (Myrtus communis L.) were induced from mature zygotic embryos cultured in MS medium supplemented
with several concentrations of 2,4-D (2.26 μM – 18.98 μM) or Picloram (2.07 μM – 16.5 μM) combined with 0.087 M or 0.23 M
sucrose. For all the concentrations of 2,4-D or Picloram tested, 0.087 M sucrose proved to be more effective than 0.23 M.
The best frequencies of induction were obtained in a medium containing 2.26 μM 2,4-D in which 97.3% of the explants produced
somatic embryos. Although most embryos were produced from the adaxial side of the cotyledons, some of them differentiated
from the hypocotyl. Secondary somatic embryos were often seen arising from the periphery of the former somatic embryos. Somatic
embryo development was not synchronous but practically all the embryos germinated well after being transferred to media containing
GA3 (0.29, 0.58 and 1.44 μM) alone. When benzyladenine was combined with gibberellic acid, germinating somatic embryos produced
adventitious shoot buds which contributed to an increase in plantlet regeneration. Histological observations suggested that
somatic embryos arise from the upper surface of the cotyledons probably from peripheral cells. Polyphenol-rich cells were
usually seen in association with meristematic-like cells from which somatic embryos originate or with earlier steps of somatic
embryo differentiation. Regenerated plants were phenotypically normal, showing a diploid (2n = 22) set of chromosomes.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Laura Y. Solís-Ramos Sara Nahuath-Dzib Antonio Andrade-Torres Felipe Barredo-Pool Tomas González-Estrada Enrique Castaño de la Serna 《Biologia》2010,65(3):504-511
Capsicum chinense is recalcitrant in in vitro morphogenesis. No efficient, reproducible somatic embryogenesis regeneration system exists for this species, impeding regeneration
from transformed cells. An indirect somatic embryogenesis protocol is developed using mature C. chinense zygotic embryo segments (ZES). The ZES cultured in semi-solid Murashige-Skoog (MS) medium supplemented with 8.9 μM naphthaleneacetic
acid, 11.4 μM indoleacetic acid and 8.9 μM 6-benzylaminopurine, developed an embryogenic callus and 8% of the calli developed
somatic embryos. Torpedo-stage somatic embryos were detached from the callus and subcultured in semi-solid MS medium without
growth regulators, producing a 75% conversion rate to plantlets with well-formed root tissue. Histological analysis showed
the developed structures to have no vascular connection with the callus and to be bipolar, confirming that this protocol induced
formation of viable somatic embryos from mature C. chinense ZES. All acclimated plantlets survived under greenhouse conditions. This protocol will facilitate regeneration of genetically
transformed plants using either biolistics or Agrobacterium tumefaciens approach. 相似文献
14.
In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production
of β-carboline alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium
supplemented with 5.0 μM 6 benzyl adenine (BA) and 2.5 μM α-naphthaleneacetic acid (NAA). The embryogenic callus was maintained
on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was
at a maximum (18.1 ± 0.9 per g of callus) on MS medium containing 5.0 μM BA and 2.5 μM NAA together with 75 mg l−1 casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation
of β-carboline alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus
culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus
accumulated equivalent amounts of harmaline (66.4 ± 0.5 μg/g dry weight), harmine (82.7 ± 0.6 μg/g dry weight), and diosgenin
(170.7 ± 1.0 μg/g dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals. 相似文献
15.
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
16.
A simple and efficient system was developed for rapid somatic embryogenesis from leaf explants of Merwilla plumbea, a traditional but threatened medicinal plant in South Africa. Friable embryogenic callus (FEC) was obtained from leaf explants
on embryogenic callus induction medium containing agar-solidified Murashige and Skoog (MS) salts and vitamins, 8.3 μM picloram,
2.3 μM thidiazuron (TDZ) and 20 μM glutamine. FEC was subsequently incubated in embryogenic callus proliferation medium containing
4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.1 μM picloram for 7 days before it was transferred to liquid somatic embryo
medium (SEML) containing MS medium supplemented with 0.4 μM picloram and 0.9 μM TDZ. In SEML supplemented with 150 mg L−1 haemoglobin, 5.4–35.6 somatic embryos per settled cell volume of 500 mg FEC were obtained. These embryos were at globular
to cotyledonary developmental stages. Embryo maturation, germination and plant formation rate was 94.4% following transfer
of SEs to half-strength MS medium supplemented with 1.4 μM gibberellic acid. Plantlets transferred into soil acclimatized
in the misthouse and established successfully in the greenhouse (100%). This is the first report on induction of Merwilla plumbea somatic embryogenesis. The protocol developed offers controlled vegetative propagation by alleviating extinction threats,
ensures germplasm conservation and provides a system for physiological, biochemical, molecular and cellular studies of embryo
development. 相似文献
17.
Somatic embryos were induced on roots excised from in vitro plants of Prunus avium× pseudocerasus `Colt'. On medium containing 6-benzylamino purine (BAP, 1.5 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D, 15 μM), a mean
of 25 (s.e. ± 2.0) somatic embryos were produced on intact root systems and 15 (s.e. ± 1.7) on roots systems cut into 10 mm
pieces. Most somatic embryos were formed directly on intact roots and indirectly (from callus) on sectioned roots. A mean
of 2.5 (s.e. ± 0.25) secondary embryos per primary embryo were formed directly on primary embryos after they were transferred
to medium containing BAP (1.5 μM), indole-3-butyric acid (10 μM) and 2,4-D (5 μM). After transfer to a medium containing BAP
(2 μM) and gibberellic acid (GA3, 3 μM), shoots developed in 75% (s.e. ± 7.3) of the embryos. Somatic embryos were not induced on explants of in vitro roots or shoots of P. avium, and were induced infrequently on zygotic embryos, although a wide range of media were tested. Possible reasons for the contrasting
embryogenic ability of `Colt' and P. avium are discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
P. I. P. Perera D. M. D. Yakandawala V. Hocher J-L. Verdeil L. K. Weerakoon 《Plant Cell, Tissue and Organ Culture》2009,96(2):171-180
The effect of growth regulators on induction of androgenesis in coconut was investigated using seven different growth regulators
at various concentrations and combinations. Three auxins (1-naphthalene acetic acid—NAA, indoleacetic acid—IAA, picloram)
and three cytokinins (2-isopentyl adenine-2-iP, kinetin, zeatin) were tested either alone or in combination with 2,4-dichlorophenoxyacetic
acid (2,4-D), using modified Eeuwens Y3 liquid medium as the basal medium. Among the tested auxins, 100 μM NAA in combination
with 100 μM 2,4-D enhanced the production of calli/embryos (123) whereas IAA and picloram showed negative and detrimental
effects, respectively, for androgenesis induction over 100 μM 2,4-D alone. Kinetin and 2-iP enhanced the production of calli/embryos
when 100 μM 2,4-D was present in the culture medium. Both cytokinins at 10 μM yielded the highest frequencies of embryos (113
and 93, respectively) whereas zeatin (1 or 2.5 μM) had no impact on microspore embryogenesis. When calli/embryos (produced
from different treatments in different experiments) were sub-cultured in somatic embryo induction medium (Y3 medium containing 66 μM 2,4-D), followed by maturation medium (Y3 medium without growth regulators) and germination medium (Y3 medium containing 5 μM-6-benzyladenine—BA and 0.35 μM gibberellic acid—GA3), plantlets were regenerated at low frequencies (in most treatments ranging from 0% to 7%). 相似文献
19.
Jin Cui Jianjun Chen Richard J. Henny 《In vitro cellular & developmental biology. Plant》2009,45(1):34-43
Plant regeneration through direct somatic embryogenesis in Aeschynanthus radicans ‘Mona Lisa’ was achieved in this study. Globular somatic embryos were formed directly from cut edges of leaf explants and
cut ends or on the surface of stem explants 4 wk after culture on Murashige and Skoog (MS) medium supplemented with N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA), TDZ with 2,4-dichlorophenoxyacetic acid (2,4-D),
or 6-benzylaminopurine (BA) or kintin (KN) with 2,4-D. MS medium containing 9.08 μM TDZ and 2.68 μM 2,4-D resulted in 71%
of stem explants producing somatic embryos. In contrast, 40% of leaf explants produced somatic embryos when induced in medium
containing 6.81 μM TDZ and 2.68 μM 2,4-D. Somatic embryos matured, and some germinated into small plants on the initial induction
medium. Up to 64% of stem explants cultured on medium supplemented with 9.08 μM TDZ + 2.68 μM 2,4-D, 36% of leaf explants
cultured on medium containing 6.81 μM TDZ and 2.68 μM 2,4-D had somatic embryo germination before or after transferring onto
MS medium containing 8.88 μM BA and 1.07 μM NAA. Shoots elongated better and roots developed well on MS medium without growth
regulators. Approximately 30–50 plantlets were regenerated from each stem or leaf explant. The regenerated plants grew vigorously
after transplanting to a soil-less substrate in a shaded greenhouse with more than a 98% survival rate. Three months after
their establishment in the shaded greenhouse, 500 plants regenerated from stem explants were morphologically evaluated, from
which five types of variants that had large, orbicular, elliptic, small, and lanceolate leaves were identified. Flow cytometry
analysis of the variants along with the parent showed that they all had one identical peak, indicating that the variant lines,
like the parent, were diploid. The mean nuclear DNA contents of the variant lines and their parent ranged from 4.90 to 4.99 pg
2C−1, which were not significantly different statistically. The results suggest that the regenerated plants have a stable ploidy
level, and the regeneration method established in this study can be used for rapid propagation of ploidy-stable Aeschynanthus radicans. 相似文献
20.
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