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1.
Mature zygotic embryos of balloon flower (Platycodon grandiflorum) formed embryogenic calluses at a frequency of 43% when cultured on Murashige and Skoog medium supplemented with 4.52 μM
2,4-dichlorophenoxyacetic acid (2,4-D). Cell suspension cultures were established from embryogenic calluses using MS liquid
medium with 4.52 μM 2,4-D. Following transfer to solid MS basal medium, cell suspension cultures gave rise to somatic embryos,
which then developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
2.
Simões-Gurgel Claudia Cordeiro Lívia da Silva de Castro Tatiana Carvalho Callado Cátia Henriques Albarello Norma Mansur Elisabeth 《Plant Cell, Tissue and Organ Culture》2011,106(3):537-545
The effects of different levels of Murashige and Skoog (MS) basal medium, 2,4-dichlorophenoxyacetic acid (2,4-D), and sucrose
on anthocyanin production and biomass accumulation of cell suspension cultures of Cleome rosea were investigated. Cultures were established in liquid MS medium containing 30 g l−1 sucrose and supplemented with 0.90 μM 2,4-D. Proliferating cell suspension cultures achieved the highest growth capacity,
a fourfold increase in biomass accumulation, following subculture at the exponential growth phase, 14–18 days of culture.
Moreover, the presence of 2,4-D was essential for anthocyanin production and biomass accumulation. On the other hand, increasing
levels of sucrose above 30 g l−1 resulted in a drastic reduction in biomass accumulation. Anthocyanin production was highest in cell suspension cultures grown
on half-strength MS medium (1/2 MS), 30 g l−1 sucrose, and 0.45 μM 2,4-D. These cell suspension cultures were mainly composed of small aggregates of spherical cells with
similar morphology observed in anthocyanin-producing and non-producing cultures. Moreover, microscopic analysis of anthocyanin-producing
cultures showed the presence of mixtures of non-pigmented, low-pigmented, and high-pigmented cells. 相似文献
3.
Summary Efficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth
regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and
Skoog (MS) medium supplemented with 4.52μM2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior
and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52μM 2,4-D to suspension cultures of half- or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 μM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous
with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength
MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets
acclimatized under field conditions with 90% survival. 相似文献
4.
Summary
Cupressus macrocarpa and C. arizonica were examined for callus and cell culture production in vitro. Both species produced callus on agar-solidified MSCY medium supplemented with vitamins, antioxidants, 0.14 μM kinetin (KIN), and 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension cultures of both species were established in liquid MSCY medium. Seiridin
(SE) and iso-seiridin (ISE), two phytotoxic butenolides produced by Seiridium cardinale, S. cupressi, and S. unicorne, the causal agents of many canker diseases of cypress, were tested on callus or cell suspension cultures. In the medium without
other plant growth regulators (PGR), SE promoted cell proliferation of cypress better than ISE, for callus initiation, callus
maintenance, and cell suspension cultures. The growth rates of cypress callus tissues and suspension cultures of both cypress
species on media containing 50–150 μM SE or ISE were measured. At concentrations of 50 μM and higher, growth rates increased exponentially with the SE concentration. A comparison with KIN and 2,4-D indicated that
50 μM SE promoted growth of callus tissues and cell suspension cultures more than 100 μM ISE. SE can also interact with, or counteract, KIN and 2,4-D. It was demonstrated that SE could replace KIN in the medium
for C. arizonica. SE could be involved in cell enlargement and proliferation processes. The less susceptible cypress species (C. arizonica) had a high content of terpenoids than that of the more susceptible species (C. macrocarpa). SE could be a useful tool as a phytohormonal-like regulator to manipulate physiological changes at the cellular level and
as an elicitor of sensitivity or tolerance of cypress germplasm to the phytotoxin. 相似文献
5.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
6.
By means of co-culture in growth regulator-free medium we analysed whether factors secreted into the medium of Daucus carota (carrot) somatic embryo cultures would be able to overcome the developmental arrest of globular Arabidopsis thaliana somatic embryos. Instead of Arabidopsis embryogenesis being promoted the development of carrot somatic embryos was inhibited at the globular stage in the presence
of Arabidopsis suspension culture aggregates with attached globular embryos. Several experiments showed that this was due to the release
of previously accumulated 2,4-D by the Arabidopsis cultures. (1) In addition to arresting carrot embryogenesis, co-culture with Arabidopsis cell suspensions also induced callus formation on Arabidopsis root segments. (2) Both effects only occurred with Arabidopsis suspensions grown in the presence of 2,4-D and not with those grown in the presence of NAA, demonstrating that Arabidopsis is not segregating a “general” inhibiting factor. (3) Both effects could be prevented by either binding 2,4-D to active charcoal
or by washing it away by changing the medium daily. (4) Uptake of 2,4-D into Arabidopsis cells during culture in 2,4-D containing medium and subsequent release of 2,4-D after transfer to growth regulator-free medium
was measured. (5) These low levels of released 2,4-D (0.2– 0.5 μm) could mimic the observed effects. Taken together these data suggest that the high intracellular 2,4-D content of Arabidopsis cultures may interfere with Arabidopsis somatic embryo development beyond the globular stage.
Received: 13 November 1997 / Revision received: 2 February 1998 / Accepted: 16 November 1998 相似文献
7.
Culture conditions for high frequency plant regeneration via somatic embryogenesis in cell suspension cultures of Chelidonium
majus var. asiaticum are described. Immature ovules formed embryogenic calluses at a frequency of 40% when cultured on Murashige
and Skoog (MS) medium supplemented with 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The optimum ovule size for embryogenic
callus formation ranged from 1 to 1.5 mm in length. Cell suspension cultures were established from embryogenic calluses using
MS liquid medium containing 4.52 μM 2,4-D. Upon plating onto MS basal medium, cell aggregates from cell suspension cultures
produced somatic embryos which then developed into plantlets. Regenerated plantlets were transplanted to potting soil and
grown to maturity in a growth chamber.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
8.
Summary Seedlings from 11 seed sources (lines) of American ginseng from different geographic regions were evaluated on Murashige and
Skoog medium (MS) containing 10 μM α-naphthaleneacetic acid (NAA) and 9 μM 2,4-dichlorophenoxyacetic acid (2,4-D) for callus development and somatic embryo formation. Leaf and stem explants callused
at a frequency of 18.2–100%, while somatic embryos were produced from these calluses at a frequency of 25–87.5% after 5 mo.
Suspension cultures of nine lines were established by transferring embryogenic callus to MS liquid medium with NAA and 2,4-D
at 2.5 and 2.25 μM, respectively, and maintained by subcultures every 8 wk. Globular somatic embryos from these cultures were germinated on
half-strength MS containing 1% activated charcoal, and roots >5 mm in length developed within 3 wk. A 7-d exposure to 3 μM gibberellic acid and 5 μM 6-benzylaminopurine significantly enhanced shoot development and promoted further root development. The chromosome number,
profiles of the common triterpenoid saponins (ginsenosides), and random amplified polymorphic DNA (RAPD) banding patterns
in plantlets derived from suspension culture were compared to those of zygotic seedlings. The chromosome number in root tip
cells and suspension cultured cells was 48. Patterns of the six major ginsenosides, determined by thin-layer chromatography,
in leaves of tissue culture-derived plantlets were identical to those in seedlings. RAPD patterns among plantlets originating
from the same tissue-cultured line were mostly identical; however, altered patterns were observed in some lines that had been
maintained in suspension culture for almost 4 yr. The results from this study indicate that propagation of desired ginseng
genotypes in suspension culture can be achieved, and that biochemical and molecular markers can be used for authentication
of resulting plantlets. 相似文献
9.
Natasha Maurmann Carina M. B. De Carvalho Andréia Loviane Silva Arthur G. Fett-Neto Gilsane L. von Poser Sandra B. Rech 《In vitro cellular & developmental biology. Plant》2006,42(1):50-53
Summary
Valeriana glechomifolia is an endemic species of southern Brazil, capable of accumulating, in all of its organs, the terpene derivatives known as
valepotriates, the presumed sedative components of the roots of pharmaceutically used species of Valeriana. In vitro cultures of the plant were established and the accumulation of acevaltrate, didrovaltrate, and valtrate in callus, cell suspension,
and untransformed root cultures was studied. Leaves of in natura plants and roots of micropropagated plantlets were used as the explants for callus induction and root culture establishment,
respectively, on Gamborg B5 basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or with kinetin (KIN).
Culture growth and secondary metabolite yields were enhanced with 2,4-D (4.52μM) and KIN (0.93μM). Maximum valepotriate contents, quantified by HPLC, of acevaltrate (ACE) 2.6mg g−1 DW, valtrate (VAL) 10.2mgg−1 DW, and didrovaltrate (DID) 2.9mg g−1 DW were observed in root cultures after 7–8wk of culture. 相似文献
10.
Culture conditions for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Ranunculus kazusensis are described. Zygotic embryos formed white nodular structures and pale-yellow calluses at a frequency of 84.9% when cultured
on half-strength Schenk and Hildebrandt (SH) medium supplemented with 0.1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). However, the frequency of white nodular structure and off-white callus formation
decreased with an increasing concentration of 2,4-D up to 10 mg l−1, when the frequency reached 25%. Cell suspension cultures were established from zygotic embryo-derived pale-yellow calluses
using half-strength SH medium supplemented with 0.1 mg l−1 of 2,4-D. Upon plating onto half-strength SH basal medium, over 90% of cell aggregates gave rise to numerous somatic embryos
and developed into plantlets. Regenerated plantlets were successfully transplanted to potting soil and grown to maturity at
a survival rate of over 90% in a growth chamber. The plant regeneration system established in this study can be applied to
mass propagation and conservation of this species. 相似文献
11.
R. Prem Anand A. Ganapathi V. Ramesh Anbazhagan G. Vengadesan N. Selvaraj 《In vitro cellular & developmental biology. Plant》2000,36(6):475-480
Summary Embryogenic callus was induced from primary leaves of Vigna unguiculata (L.) Walp. in MS medium (Murashige and Skoog, 1962) containing 2,4-dichlorophenoxyacetic acid (2,4-D). Greenish-white, friable
embryogenic calluses were used to establish suspension cultures. A shaking speed of 90 rpm and 0.4 ml packed cell volume per
25 ml medium were found to be optimal for maintaining suspension cultures. Globular, heart-shaped and torpedo-shaped embryos
were developed in suspension culture containing 4.52 μM 2,4-D. Maturation of cotyledonary-stage somatic embryos was achieved on 0.05 μM 2,4-D, 5 μM abscisic acid and 3% mannitol. Twenty-two percent of the embryos were converted into plants and survived; survival in the
field was 8–10%. 相似文献
12.
Summary Indirect somatic embryogenesis, encapsulation, and plant regeneration was achieved with the rare rhoeophytic woody medicinal
plant Rotula aquatica Lour. (Boraginaceae). Friable callus developed from leaf and internode explants on Murashige and Skoog (MS) medium with 0.45 μM 2,4-dichlorophenoxyacetic, acid (2,4-D) was most effective for the induction of somatic embryos. Subculture of the callus
onto half-strength MS medium with the same concentration of 2,4-D resulted in highly embryogenic callus. Suspension culture
was superior to solid medium culture for somatic embryogenesis. Embryogenic callus.during subsequent transfer to suspension
cultures of half-strength MS medium having 0.23 μM 2,4-D induced the highest number of somatic embryos (a mean of 25.6 embryos per 100 mg callus) and the embryos were grown
up to the torpedo stage. Transfer of embryos to half-strength MS basal solid medium allowed development, of 50% of the embryos
to the cotyledonary stage. Of the cotyledonary embryos, 90% underwent conversion to plantlets on the same medium. Encapsulated
cotyledonary embryos exhibited 100% conversion to plantlets. Ninety-five percent of the plantlets established in field conditions
survived, and were morphologically identical to the mother plant. 相似文献
13.
Soluble polyvinylpyrrolidine and bovine serum albumin adsorb polyphenols from soybean suspension cultures
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Soybean (Glycine max [L.] Merrill, cv. Dare) suspension cultures grown in Gamborg B5 medium became discolored and the cells began aggregating after 1 week in culture, especially in the absence of 2,4-dichlorophenoxyacetic acid (2,4-D). The addition of either soluble polyvinylpyrrolidine (PVP) or bovine serum albumin (BSA) to cultures grown in Gamborg B5 medium with 2,4-D prevents discoloration and cell aggregation by adsorbing excess polyphenols from the cells. Transfer of the PVP-treated cultures to fresh medium without 2,4-D stimulated the recurrence of excess polyphenols. Cultures pretreated with BSA did not develop excess polyphenols when transferred to fresh 2,4-D-free medium. Addition of either PVP or BSA to cultures grown in the absence of 2,4-D was found to inhibit growth. 相似文献
14.
Summary Callus cultures were established from pith tissue of Coryphantha elephantidens (Lem.) Lem. on Murashige and Skoog (MS) basal medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin. Highest shoot regeneration frequency was observed on a medium containing 6.9 μM kinetin and 2.3 μM 2,4-D under 30 μE m−2 s−1 light intensity with a 16-h photoperiod. Calluses retained organogenic potential throughout several passages of subculture
(18 mo.). Shoots were rooted on MS medium without plant growth regulators. All (100%) plantlets transplanted to soil survived
acclimatization. Regenerated plants showed good overall growth and were morphologically similar to the mother plants. 相似文献
15.
Santos-Gomes PC Seabra RM Andrade PB Fernandes-Ferreira M 《Journal of plant physiology》2003,160(9):1025-1032
Sage (Salvia officinalis L.) calli were established by culturing internodal segments, excised from aseptic seedlings, on MS basal medium gellied with agar and supplemented with 0.05 mg/L dichlorophenoxyacetic acid (2,4-D) in presence of benzyladenine (BA) or zeatin (ZEA) or kinetin (KIN), at 1.5 mg/L. Suspended cells were established by transferring one callus to 50 mL of liquid MS basal medium devoid of agar and containing the same type of hormonal supplementation used in respective calli growth. The highest growth of calli and suspensions occurred with 1.5 mg/L ZEA. However, with this cytokinin supplementation, as well as with 1.5 mg/L KIN, both in presence of 0.05 mg/L 2,4-D, suspensions differentiated small root shaped structures. Well shaped, majority single cell suspensions were formed under the effect of 0.05 mg/L 2,4-D and 0.5 mg/L KIN. Calli grown with 0.05 mg/L 2,4-D and 1.5 mg/L BA and suspended cells grown with 0.05 mg/L 2,4-D and ZEA or KIN at 1.5 mg/L, or KIN at 0.5 mg/L, were searched for phenolics production. Twelve phenolic compounds were identified in calli: gallic acid, 3-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, caffeic acid, rosmarinic acid, hesperetin, epirosmanol, hispidulin, genkwanin, carnosol, carnosic acid, and methyl carnosate. With the exception for genkwanin and epirosmanol all of these phenolic compounds were also produced by the sage suspension cultures grown in the presence of 1.5 or 0.5 mg/L KIN. Genkwanin was the only phenolic absent in the suspensions grown with 1.5 ZEA. Suspended cells, grown with 0.5 mg/L KIN, and calli cultures showed the highest specific accumulation of the total phenolics, with rosmarinic acid representing 94-97 percnt;. 相似文献
16.
Myung Jin Oh Hye Ryun Na Hong-Keun Choi Jang Ryol Liu Suk Weon Kim 《Plant biotechnology reports》2008,2(1):87-92
An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension
cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when
cultured on half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly
with increasing concentrations of 2,4-D up to 3 mg l−1, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryo-derived white friable callus
were established using half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos
and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by
up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting
soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high
frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield,
which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could
be useful for the mass propagation and transformation of selected elite lines. 相似文献
17.
Bang Jae W. In Dong S. Chung Sung H. Liu Jang R. 《Plant Cell, Tissue and Organ Culture》1998,55(2):151-154
Hypocotyl segments of Bupleurum falcatum L. formed embryogenic calluses when cultured on Murashige and Skoog's (MS) medium
supplemented with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension cultures were initiated by placing calluses into
medium with 0.45 μM 2,4-D. Protoplasts were enzymatically isolated from suspension cultures. They were plated at a density
of 5 × 104 protoplasts per ml on MS medium supplemented with 9% mannitol, 9.0 μM 2,4-D, 4.4 μM BA, 4.6 μM kinetin, and 0.6%
Seaplaque agarose. After four weeks of culture, microcalluses were formed and subsequently transferred to MS solid medium
with 18.1 μM 2,4-D. Upon transfer to MS basal medium, microcalluses gave rise to somatic embryos at a frequency of approximately
10%. They subsequently developed into plantlets. The regenerants were successfully transplanted to potting soil and grown
to maturity in a greenhouse. The regenerants had the normal chromosome number of 2n=2x=20 and did not show morphological aberrancy.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
19.
A procedure is outlined for the establishment of a proliferating cell suspension culture and subsequent plant regeneration
of the latex-producing plant,Calotropis gigantea (Linn.) R. Br. Friable calluses were obtained by culturing hypocotyl explants on modified Murashige and Skoog medium with
2.69 μM α-naphthaleneacetic acid and 4.44 μM 6-benzyl-aminopurine. Friable calluses were transferred to modified Murashige
and Skoog liquid medium containing 500 mg l−1 casein hydrolysate, 5% (v/v) coconut water and 5% (w/v) sucrose to initiate suspension cultures. Suspensions were subcultured
every 10–12 days and supplemented with 13.56 μM 2,4-dichlorophenoxyacetic acid (2,4-D). After 3–4 subcultures, suspensions
consisted of small, highly cytoplasmic cell clumps and large vacuolate cells. Plating the suspension clumps on medium containing
4.52 μM 2,4-dichlorophenoxyacetic acid and culturing in darkness produced macroscopic calluses, which subsequently produced
a high number of shoots when placed in light and supplemented with 2.22 μM 6-benzyl-aminopurine and 0.45 μM 2,4-dichlorophenoxyacetic
acid. Shoots were rooted using Bonner's solution containing 0.49 μM indole-3-butyric acid, and the plants successfully transferred
to soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
20.
Glycine max (L.) Merr. (soybean) andGlycine soja Sieb. and Zucc. cell suspension cultures were grown and used as inoculum sources for growing callus on agar-solidified nutrient
media. Concentrations and chemical forms of the growth regulators in liquid and solidified media were altered in an attempt
to achieve in vitro plant regeneration. Numerous embryoids, particularly ofG. soja, were produced on basal nutrient media supplemented with 100 ppm casein hydrolysate, 0.1 μM abscisic acid, 2.25 μM 2,4-dichlorophenoxyacetic acid, and 15 μM adenine or 0.46 μM kinetin. Often the roots of the embryoids elongated. This was enhanced in the presence of an inhibitor of gibberellin synthesis
(1 to 20 μM Amo 1618). Callus recovered from aG. soja suspension culture produced one shoot structure when grown on a solid medium containing 0.2 μM Amo 1618 and 80 μM glutathione. The shoot structure consisted of two distinct buds, one producing two leaves. The shoot did not develop into
a plant. Although regeneration of soybean plants was not achieved, these observations suggest that it may be achievable.
The investigations reported in this paper (no. 81-3-100) were performed in connection with a project of the Kentucky Agriculture
Experimental Station and the paper is published with the approval of the Director. 相似文献