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1.
P. I. Hong J. T. Chen W. C. Chang 《Journal of plant biochemistry and biotechnology.》2008,17(2):149-153
The effects of salicylic and acetylsalicylic acid on direct somatic embryogenesis were investigated using leaf explants of two cultivars of Oncidium on 1/2 MS medium with or without thidiazuron. In cv Gower Ramsey, salicylic acid (1, 5, 10, 15, 20, 50 μM) either alone or in combination with thidiazuron (4.54 μM) retarded and delayed embryogenesis. In contrast, in the presence of 4.54 μM of thidiazuron, acetylsalicylic acid at 0.1 μM concentration promoted embryogenesis. In cv Sweet Sugar, all concentrations of salicylic acid with or without thidiazuron proved inhibitory on embryo induction. However, in the presence of 0.45 μM thidiazuron, 0.1 and 1 μM acetylsalicylic acid promoted embryogenesis. In addition, in the presence of 4.54 μM thidiazuron, 0.01, 0.1 and 1 μM acetylsalicylic acid promoted embryogenesis. 相似文献
2.
A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in cotton (Gossypium hirsutum L.) cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming
frequency (82.86 %) was obtained in protoplast cultures from suspension culture cells in KM8P medium with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 μM kinetin (KIN), 1.5 % glucose and 1.5 % maltose. Protocolonies
formed in two months with plating efficiency of 14 %. However, the callus-forming efficiencies from other two explants were
low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7 % of normal plantlets
were obtained on medium contained 3 % maltose or 1 % of each sucrose + maltose + glucose, 2.46 μM indole-3-butyric acid (IBA)
and 0.93 μM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were
transferred to the soil and the highest survival rate (95 %) was observed in transplanting via a new method. 相似文献
3.
Direct somatic embryogenesis from in vitro-cultured leaf segments of multiple disease-resistant pepper, Piper colubrinum Link is reported. Somatic embryos were initiated on Murashige and Skoog's (1962) basal medium containing 2.2 μM benzyladenine+0.46
μM kinetin and multiplied profusely through secondary embryogenesis on the same medium. Some 91% somatic embryos converted
into plantlets on MS medium supplemented with 4.4 μM benzyladenine+0.23 μM kinetin and plantlets developed on half-strength
MS+2.4 μM indole-3-butyric acid. Plantlets were hardened, transferred to soil, and 100% of plants survived. Various developmental
stages of somatic embryogenesis were studied using histological methods.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
Somatic embryos were obtained from immature zygotic embryos of Cedrela fissilis Well. (Meliaceae), after a culture period of 12 months, with regular subcultures every 6–8 weeks. Callus was developed on explants in 2 months
on Murashige and Skoog (MS) medium containing 2,4 dichlorophenoxyacetic acid (2,4-D) or picloram (PIC). When the calli were
transferred to fresh medium, embryogenic tissue appeared on MS + 45 μM 2,4-D, or 22.5 μM 2,4-D + 0.4 μM 6-benzyladenine (BA),
or 20.7 μM PIC after 6 months. Sub-culture of embryogenic tissue in MS medium supplemented with 4.5 μM 2,4-D resulted in the
differentiation into somatic embryos after further 4 months. Repeated secondary somatic embryogenesis was achieved by regular
subculture on this medium. Maturation and conversion of somatic embryos into plantlets was achieved on MS medium without plant
growth regulators and the conversion frequency was approximately 12.5 %. The plantlets were successfully acclimatized in pots
with soil. Histological studies showed that somatic embryos had no detectable connection with the mother explants and that
somatic embryos in advanced stages were bipolar with shoot and root apical meristems, they contained vascular system and showed
typical characteristics of a somatic dicotyledonous embryo. 相似文献
5.
Mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 on MS or N6 nutrient medium supplemented with various concentrations of 2,4-D (4.5 – 22.5 μM) formed embryogenic callus, which differentiated
into somatic embryos within 5 weeks of culture. The somatic embryos after transfer to hormone-free regeneration medium germinated
and formed plantlets. Of the two nutrient formulations, N6 was relatively better than MS for somatic embryogenesis. A culture for 11 d on 100 μM 2,4-D was essential for the establishment
of an embryogenic callus. Shorter duration, 4-d or 7-d culture on 2,4-D medium, supported some proliferation and subsequent
differentiation into shoot-buds or multiple-shoots, in high-frequency cultures. This is first instance in monocots of a controlled
regeneration response; either somatic embryogenesis or shoot formation.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
6.
M. Bobák J. Šamaj A. Pre ová A. Blehová E. Hlinková M. Ove ka A. Hlava ka Z. Kutar ová 《Acta Physiologiae Plantarum》2004,26(3):353-361
We studied indirect somatic embryogenesis in the callus tissue of Drosera spathulata Labill. originated from isolated leaves. Callogenesis was induced on MS medium (Murashige and Skoog 1962), supplemented with
various concentrations of NAA and BA. Somatic embryos regenerated on half-strength MS medium supplemented with 20 μM of NAA
or without growth regulators. The highest efficiency of somatic embryo production was achieved on hormone-free medium. Globular,
heart-, torpedo- and cotyledonary-shaped embryos were observed in embryogenic clusters. Histological and scanning electron
microscopy analysis verifies somatic embryogenesis. Regenerated plants were transferred to soil and were grown to maturity. 相似文献
7.
G. H. Ma C. X. He H. Ren Q. M. Zhang S. J. Li X. H. Zhang B. Eric 《Biologia Plantarum》2010,54(2):361-365
An efficient propagation system via somatic embryogenesis and shoot organogenesis and plant regeneration system for endangered species Primulina tabacum Hance was established. Thidiazuron (TDZ) was the key plant growth regulator for inducing somatic embryogenesis and kinetin
(KIN) and 6-benzylaminopurine (BAP) were the key cytokinins for inducing shoot organogenesis from leaf explants. TDZ combined
with BAP or KIN in the induction Murashige and Skoog medium induced both somatic embryos and adventitious shoots. Leaf explants
with abaxial site in contact with the medium induced less somatic embryos or adventitious shoots compared to inversely placed
leaf explants and the optimum pH was 6.5–7.0. Secondary somatic embryos or adventitious shoot could be induced from primary
somatic embryos using TDZ and BAP. Shoots developed adventitious roots on rooting medium containing 0.5 μM indole-3-butyric
acid and 0.2 % activated carbon. Over 90 % of plantlets survived following acclimatization and transfer to potting mixture
(sand:Vermiculite:limestone; 1:2:1). 相似文献
8.
Margaret J. Hutchinson T. Senaratna J. M. Tsujita Praveen K. Saxena 《Plant Cell, Tissue and Organ Culture》1997,47(3):293-297
A simple and efficient procedure was developed for regeneration of a tetraploid cultivar ofAlstroemeria (A. pelegrina x A. psittacina) via somatic embryogenesis in liquid cultures. Embryogenic callus induced from mature zygotic embryos, cultured on MS medium
supplemented with 40 μM NAA and 20 μM kinetin, was used as inoculum for liquid cultures. Pre-culture of the callus on MS medium
supplemented with 80 μM NAA for two days was essential for cell proliferation in the liquid medium. Embryogenic cell aggregates,
obtained by sieving through a 750 μm nylon mesh, continued to proliferate in media containing 10 or 20 μM NAA and 10 or 20
μM kinetin. When transferred to a semi-solid half strength MS medium supplemented with casein hydrolysate, cell aggregates
successfully differentiated into plantlets which later grew to maturity under greenhouse conditions. 相似文献
9.
Akella Mahalakshmi Bhumica Singla Jitendra P. Khurana Paramjit Khurana 《Plant Cell, Tissue and Organ Culture》2007,88(2):167-174
Wheat leaf bases cultured for 1 day on 2,4-d (10 μM) display the induction of somatic embryogenesis. The induction of somatic embryogenesis by 2,4-d appears to be calcium-mediated as treatment of leaf bases with the calcium chelator, EGTA, prior to 2,4-d treatment, inhibited the induction of somatic embryogenesis. This sensitivity of auxin to reduced calcium levels can be reversed
by calcium ions alone and not any other divalent cation like magnesium or zinc. Additionally, the expression of the three
calcium-regulated genes, Triticum aestivum calmodulin binding protein kinase, calcium-dependent protein kinase, and putative calcium binding protein was analyzed in
wheat leaf bases which suggest a specific role for Ca2+ in somatic embryogenesis. Application of the calcium ionophore, A23187, either alone or along with 2,4-d, induced somatic embryogenesis. This specificity for calcium was verified both by treatment with the calcium antagonist TMB8,
and the elimination of calcium from the medium, resulting in reduction of somatic embryogenesis by 80%. Treatment with calcium
channel blockers like verapamil and nifedipine, calcium antagonist, lanthanum, and calmodulin inhibitors chlorpromazine and
fluphenazine, prior to the 2,4-d treatment, inhibited induction of somatic embryogenesis. The present study thus provides evidence for the involvement of
calcium–calmodulin in the stimulus–response coupling of auxin-induced somatic embryogenesis in wheat leaf base system. 相似文献
10.
Repetitive Somatic Embryogenesis of Ocotea catharinensis Mez. (Lauraceae): Effect of Somatic Embryo Developmental Stage and Dehydration 总被引:3,自引:1,他引:2
Claudete Santa Catarina Alessandra dos Santos Olmedo Geraldine de Andrade Meyer Jonice Macedo Wagner de Amorim Ana Maria Viana 《Plant Cell, Tissue and Organ Culture》2004,78(1):55-62
Repetitive embryogenesis of Ocotea catharinensis from globular/early cotyledonary somatic embryos was successfully supported by WPM supplemented with 22.7 g l−1 sorbitol, 20 g l−1 sucrose, 400 mg l−1 glutamine and 2 g l−1 Phytagel. The best medium to induce repetitive embryogenesis in cotyledonary somatic embryos was half strength WPM supplemented
with 20 g l−1 sucrose, 400 mg l−1 glutamine, 1.5 g l−1 activated charcoal and 2 g l−1 Phytagel. The mature somatic embryos gradually air dehydrated showed repetitive embryogenesis after subculture on half strength
B5 medium supplemented with 20 g l− sucrose, 20 g l−1 Phytagel, 1.5 g l−1 activated charcoal, 115.6 μM gibberellic acid and 214.8 μM naphthaleneacetic acid. The early cotyledonary, cotyledonary and
mature somatic embryos tolerated respectively 95, 86 and 54% fresh weight losses without losing their repetitive embryogenesis
potential. Cotyledonary and mature somatic embryos gradually air dehydrated in sealed Petri dishes showed 40–41% repetitive
embryogenesis respectively after 20 days and 12 weeks desiccation storage. Repetitive embryogenesis in cotyledonary somatic
embryos was significantly stimulated by chemical dehydration with 0.5 M sorbitol and 56% repetitive embryogenesis was achieved
even after exposure to 2 M sorbitol for 24 h. The cotyledonary somatic embryos when alginate-encapsulated showed 47% repetitive
embryogenesis even after chemical dehydration in 1.5 M sorbitol for 4 days followed by 1 h air dehydration, but failed to
survive to the same dehydration conditions without encapsulation. The optimized repetitive embryogenesis and desiccation protocols
offer the possibility to use in vitro techniques for continuous reliable somatic embryo production and short term germplasm storage. 相似文献
11.
Xingyu Yang Jinfeng Lü Jaime A. Teixeira da Silva Guohua Ma 《Plant Cell, Tissue and Organ Culture》2012,109(2):213-221
Primulina tabacum is a rare and endangered species that is endemic to China. Establishing an efficient regeneration system is necessary for
its conservation and reintroduction. In this study, when leaf explants collected from plants grown in four ecotypes in China
are incubated on Murashige and Skoog (MS) medium containing 5.0 μM thidiazuron (TDZ) for 30 days, then transferred to medium
containing 5.0 μM 6-benzyladenine (BA), adventitious shoots are then observed. Conversely, when leaf explants are incubated
on medium containing 5.0 μM BA for 30 days, then transferred to medium containing 5.0 μM TDZ, somatic embryogenesis is induced.
This indicates that somatic embryogenesis and shoot organogenesis could be switched simply by changing the order of two cytokinins
supplemented in the culture medium. Histological investigation has revealed that embryogenic cells are induced within 30 days
following incubation of explants in medium containing TDZ. Only if embryogenic cells were induced, TDZ could enhance somatic
embryogenesis and BA could stimulate shoot organogenesis. When comparing explants from different ecotypes, leaf explants
from Zixiadong in Hunan Province could induce low numbers (1–2) of either somatic embryos or adventitious shoots on medium
containing either 5.0 μM TDZ or 5.0 μM BA, respectively. Whereas, leaf explants from plants collected from the other three
ecological habitats could induce 50–70 somatic embryos/adventitious shoots per explant. Moreover, somatic embryos could induce
secondary somatic embryogenesis and adventitious shoots on different media. All regenerated shoots developed adventitious
roots when these are transferred to rooting medium, and over 95% of plantlets have survived following acclimatization and
transfer to a potting mixture (1:1, sand:vermiculite). 相似文献
12.
Benjamin Rodríguez-Garay Antonia Gutiérrez-Mora Beatríz Acosta-Duefias 《Plant Cell, Tissue and Organ Culture》1996,46(1):85-87
Plant regeneration through direct somatic embryogenesis of leaf blade explants from in vitro propagated plants of Agave victoria-reginae Moore, is described. Somatic embryogenesis was evident in a 6-week period on agarsolidified MS medium supplemented with L2 vitamins and 2,4-dichlorophenoxyacetic acid (1,4 µM), and germination of somatic embryos was achieved after 8 weeks on half-strength MS medium and 4 weeks on half-strength SH medium, both lacking growth regulators. Hyperhydricity of somatic embryos and plantlets was reduced by the use of vented culture vessel lids during the last 4 weeks on SH medium. Shoot proliferation was obtained, and hyperhydricity was eliminated on a modified MS medium (with NH4NO3 reduced to 5 mN) supplemented with kinetin (4.6 µM) and 1-naphthaleneacetic acid (1.6 µM) and the use of vented culture vessel lids.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- MS
Murashige and Skoog
- LOG-1
MS modified medium by Castro-Concha et al. (1990)
- L2
Phillips and Collins (1979) vitamins
- SH
Schenk and Hildebrandt 相似文献
13.
K. P. Martin A. Shahanaz Beegum C.-L. Zhang A. Slater P. V. Madhusoodanan 《Biologia Plantarum》2007,51(4):769-772
In vitro propagation of an anticancerous drug synthesizing plant, Ophiorrhiza prostrata D. Don, was established through indirect somatic embryogenesis. Friable embryogenic calluses were initiated from O. prostrata leaf and internode explants on Murashige and Skoog (MS) media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) either
alone or in combination with N6-benzyladenine (BA) or kinetin (KIN). Somatic embryos were developed after subculture of the friable calluses onto half strength
MS media containing 0.45 or 2.26 μM 2,4-D alone or in combination with BA or KIN. Medium supplemented with 2.26 μM 2,4-D and
2.22 μM BA was optimal, supporting the production of a mean of 5.8 globular embryos. Subculture of globular embryo-bearing
calluses on half strength MS medium without growth regulators produced the highest embryo frequency, and the majority of them
developing to early torpedo stage. Somatic embryos underwent maturation and converted to plantlets at high frequency (90 %)
on half strength MS medium supplemented with 0.44 μM BA. Somatic embryo-derived plantlets with well-developed roots were established
in field conditions with a 90 % survival rate. 相似文献
14.
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
15.
Indirect somatic embryogenesis was induced on leaf explants of greenhouse-grown Helianthus maximiliani plants. Leaves of the regenerated plants were used as starting explants for the induction of direct somatic embryogenesis.
Another cycle of somatic embryogenesis was induced on the leaves of regenerated plants. In both cases, leaf explants were
cultured on media containing different auxin/cytokinin ratios. The auxin/cytokinin ratio had an influence on the intensity
of embryo formation, germination and the capability to regenerate plants. Somatic embryogenesis was generally more intensive
on the medium with lower concentrations of 6-benzylamino-purine. Further, the percentage of regenerated plants was higher
when embryos were induced on high-cytokinin, low-auxin medium. Secondary somatic embryogenesis was induced on embryos by culture
in liquid hormone-free medium. Similar to direct embryogenesis the efficiency of secondary embryogenesis depended on the medium
used for the induction of the primary embryos. In contrast to the mostly low frequencies of conversion of secondary embryos
into plants that has been observed in other species, the percentage of regenerated plants from secondary embryos of H. maximiliani was quite high, although slightly lower than that obtained in primary embryos.
Received: 28 March 2000 / Revision received: 1 September 2000 / Accepted: 2 October 2000 相似文献
16.
Plant regeneration via somatic embryogenesis in cotton 总被引:6,自引:0,他引:6
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and stem explants
of abnormal seedling as an explant have been developed in Gossypium hirsutum L. Embryogenic callus and somatic embryos have been obtained directly from the explants of cotton abnormal seedlings. Plant
growth regulators influenced the induction of cotton somatic embryogenesis. The optimal medium for direct somatic embryogenesis
was modified MS medium supplemented with 0.1 mg l-1 ZT and 2 g l-1 activated carbon. On this medium, an average of 28.0 and 28.1 matured somatic embryos formed from per leaf and stem explants
respectively. The highest frequency of somatic embryogenesis was 100%. The somatic embryos were converted into normal plantlets
when cultured on modified MS medium supplemented with 0.1 mg l-1 ZT. Upon transfer to soil, plants grew well and appeared normal. Plants could be regenerated within 60–80 days. The system
of cotton somatic embryogenesis and plant regeneration described here will facilitate the application of plant tissue culture
and genetic engineering on cotton genetic improvement.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
17.
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
18.
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
19.
This is the first report where shoot regeneration in strawberry cultivar Chandler has been achieved simultaneously through
both somatic embryogenesis and shoot bud formation. Direct somatic embryogenesis was observed in leaf discs which were cultured
on medium containing MS salts + B5 vitamins + 2% glucose + 18.16 μM thidiazuron (TDZ) and given both chilling and dark treatment for 2 wk at 4 ± 2°C followed
by incubation at 25 ± 2°C under 16-h photoperiod for third wk. After 3 wk, these explants were then subcultured on medium
containing MS salts + B5 vitamins + 2% glucose and incubated under 16-h photoperiod at 25 ± 2°C for further growth and development. Direct regeneration
via de novo shoot bud formation was observed in leaf disks which were given dark treatment and were cultured on medium containing MS
salts + B5 vitamins + 2% glucose supplemented with 9.08 μM TDZ. There was a synergistic effect of photoperiod, dark, and chilling treatments
on somatic embryogenesis, whereas chilling treatment had an inhibitory effect on shoot organogenesis. 相似文献
20.
Brassinolide (BR), which is the most biologically active brassinosteroid, was used to examine the potential effect of hormone
on cotton somatic embryogenesis. Ten-day-old cotton (Gossypium hirsutum L., cv. Cooker) seedlings were used for explant source and hypocotyls were removed and cultured on MS basal medium with B5 vitamins supplemented with 1 mg/L 6-benzylaminopurine + 0.5 mg/L kinetin for callus induction. After one month proliferating
calli pieces were collected and cultured on MS basal medium containing various concentrations of BR (0.1, 0.5, 1.0 μM) with
their controls. BR treatments were negatively effective on the fresh weight of calli when compared to control. Differential
somatic embryogenesis maturation rates due to BR treatment were observed. Somatic embryogenesis was stimulated especially
for transition to cotyledonary phase at 0.5 mg/L BR. Histological preparations from embryogenic calli and somatic embryos
at different stages of development revealed the spontaneous polyploidisation during early somatic embryogenesis on BR-treated
calli. Present results suggest that BR negatively effected calli growth, however, had a stimulating role in maturation of
somatic embryos. 相似文献