共查询到20条相似文献,搜索用时 31 毫秒
1.
Abdullah M.A. Ali A.M. Marziah M. Lajis N.H. Ariff A.B. 《Plant Cell, Tissue and Organ Culture》1998,54(3):173-182
Morinda elliptica (Rubiaceae) cell suspension cultures were established in shake flask system for the production of anthraquinones.
The optimized medium formulation for cell growth and anthraquinone production is proposed. Murashige and Skoog's basal medium
(MS) was found to be the best medium, used in combination with 0.5 mg l-1 NAA and 0.5 mg l-1 kinetin. At the range of sucrose
concentration tested (3–8% w/v), 8% was the best in enhancing both cell growth and anthraquinone production. A strategy to
formulate growth and production medium by manipulating culture age and inoculum age, the type of medium formulation used to
grow inoculum, incubation temperature and light intensity was established. By using 18 month old culture and 7 day old inoculum
at incubation temperature of 27 ± 3 °C, anthraquinone yield of 2.9 g l-1 and 4.5 g l-1, under illumination of 1200 lux and
in the dark was obtained, respectively.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
2.
Archana Kumari Gaurav Sharma Sumangala Bhat Ramesh S. Bhat P. U. Krishnaraj M. S. Kuruvinashetti 《Plant Cell, Tissue and Organ Culture》2011,107(2):215-224
An α-amylase signal peptide from rice was synthesized and fused with endochitinase (ech42) gene cloned from Trichoderma virens. The chimeric gene was designated as PSPα-amyech42, and this was transferred to a plant transformation vector, referred to as pMASGK. Leaf explants of tobacco cv White Burley
were co-cultivated with Agrobacterium tumefaciens strain LBA4404 carrying ech42 with its own signal peptide(ech42SP) and PSPα-amyech42(pMASGK) separately. Putative transformants were selected on Murashige and Skoog (MS) medium, supplemented with 1 mg/l bezyladenine
(BA), 0.5 mg/l naphthalene acetic acid(NAA), and containing 200 mg/l kanamycin and 200 mg/l cefotaxime. Transformation was
further confirmed by PCR with specific primers and Southern blot hybridization. Endochitinase secretion was quantified in
1-week-old cell suspension cultures obtained from 3-week-old callus cultures of transformants carrying PSPα-amyech42, transformants with ech42 and of control (untransformed) plant. Callus cultures of PSPα-amyech42 showed higher endochitinase activity (9–12 times) than those carrying ech42SP (7–8 times) in both medium and cell extracts. Media collected (200 μg of total protein) from PSPα-amyech42 suspension cultures in Potato Dextrose Agar plates showed growth inhibition of 73 and 53% against Sclerotium rolfsii and Rhizoctonia bataticola, respectively, whereas media collected (200 μg of total protein) from ech42SP suspension culture showed inhibition of 14 and 24% against Sclerotium rolfsii and Rhizoctonia bataticola, respectively. 相似文献
3.
Production of solasodine by hairy root,callus, and cell suspension cultures of Solanum aviculare Forst. 总被引:5,自引:0,他引:5
Kittipongpatana Nisit Hock Rick S. Porter John R. 《Plant Cell, Tissue and Organ Culture》1998,52(3):133-143
The production of the steroidal alkaloid solasodine, an alternative to diosgenin as a precursor for the commercial production
of steroid drugs, was studied in hairy root, callus, and cell suspension cultures of Solanum aviculare Forst. through manipulation
of culture medium. The individual and combined effects of medium components on the growth index and the production of solasodine
were analyzed using factorial analysis of variance. Solasodine content was optimized to 6.2 mg g−1in the hairy root, 1.4 mg g−1callus, and 0.7 mg g−1in cell suspension cultures (dry weight). An improved isocratic reversed phase high performance liquid chromatographic method
provided selective determination of the solasodine content of these samples. Analysis of growth and solasodine content of
hairy root cultures and callus cultures demonstrated that the production of solasodine was shown to be growth-dependent in
hairy root cultures but not in callus cultures.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
The effect of osmotic stress on cell growth and phenylethanoid glycosides (PeGs) biosynthesis was investigated in cell suspension
cultures of Cistanche deserticola Y. C. Ma, a desert medicinal plant grown in west region of China. Various initial sucrose concentrations significantly affected
cell growth and PeGs biosynthesis in the suspension cultures, and the highest dry weight and PeGs accumulation reached 15.9 g l−1-DW and 20.7 mg g−1-DW respectively at the initial osmotic stress of 300 mOsm kg−1 where the sucrose concentration was 175.3 mM. Stoichiometric analysis with different combinations of sucrose and non-metabolic
sugar (mannitol) or non-sugar osmotic agents (PEG and NaCl) revealed that osmotic stress itself was an important factor for
enhancing PeGs biosynthesis in cell suspension cultures of C. deserticola. The maximum PeGs contents of 26.9 and 23.8 mg g−1-DW were obtained after 21 days at the combinations of 87.6 mM sucrose with 164.7 mM mannitol (303 mOsm kg−1) or 20 mM PEG respectively, which was higher than that of C. deserticola cell cultures grown under an initial sucrose concentration of 175.3 mM after 30 days. The stimulated PeGs accumulation in
the cell suspension cultures was correlated to the increase of phenylalanine ammonium lyase (PAL) activity induced by osmotic
stress. 相似文献
5.
Mario Arias Zabala Mónica Angarita Juan M. Restrepo Luis A. Caicedo Margarita Perea 《In vitro cellular & developmental biology. Plant》2010,46(3):233-238
Thevetia peruviana is a small tree that produces several compounds with pharmaceutical application, among which peruvoside could be highlighted.
However, these compounds are produced in low concentration in the plant, making it important to develop strategies such as
plant cell culture and elicitation to obtain higher quantities of the desired product. In this work, cell suspension cultures
of T. peruviana were established in four different culture media: Murashige–Skoog (MS), half Murashige–Skoog (half MS), Schenk–Hildebrandt
(SH), and Gamborg (B5) to study their effect on cell growth. Cell growth kinetics were studied in SH medium, and the extracellular
peruvoside production during the culture time was determined. The best culture medium for the establishment of cell suspension
cultures was MS with a growth index of 3.17 ± 0.2 g g−1 inoculum. The cell growth kinetics showed the four characteristic growth phases of a cell culture (lag, exponential, stationary,
and death), and during none of these phases was it possible to observe peruvoside production. The elicitor effect of methyl-jasmonate
(MeJ) was studied in cell suspension cultures established in SH medium. The effect of MeJ concentration and the time in which
it should be applied were determined. The best results were obtained at a concentration of 100 mg l−1 of MeJ applied at the beginning of the culture, which induced a peruvoside production of 8.93 mg l−1 medium. The current results are the first report of an in vitro peruvoside production system. 相似文献
6.
N Kurano C Leist F Messi C Gandor S Kurano A Fiechter 《Journal of biotechnology》1990,16(3-4):245-258
Four sublines of Chinese hamster ovary (CHO) cells were selected or cloned on a 10% fetal calf serum supplemented MEM-alpha medium. Three of them were monolayer cultures and could proliferate by 2000 times a week (mu = 1.1 d 1) in T-flasks. The other subline, S1, could grow in suspension even in static T-flask cultures. The stability in chromosome number of these cell lines was investigated. By evaluating the kinetic growth parameters, i.e. the specific rates of growth, glucose consumption and lactic acid production, and the yields of cells and lactic acid from glucose, the S1 cells were considered to be the most suitable subline for the bioreactor suspension culture. The S1 cells reached the greatest maximum of cell concentration among all cell lines tested because of their efficient glucose utilization. Observed nutrient limitations in the S1 cell culture was overcome by modification of the medium composition, that is addition of 10 mg l-1 hypoxanthine, 1 mg l-1 FeSO4.7H2O, and 0.1 mg l-1 sodium putrescine, elimination of glutamine, supplementation of 6 mM asparagine and double amount of isoleucine, leucine, methionine and vitamins other than ascorbic acid, cyanocobalamin and biotin, increase of NaHCO3 concentration from 26 to 40 mM, and finally decrease of NaCl concentration from 122 to 100 mM. With this modified medium, 7.2 X 10(6) ml-1 of the maximum cell concentration was observed in a glucose fed-batch culture, the cell concentration which was twice as much as in batch cultures with the original medium. 相似文献
7.
Establishment of in vitro plants, cell and tissue cultures from Oldenlandia affinis for the production of cyclic peptides 总被引:1,自引:1,他引:0
In vitro cultured plants from Oldenlandia affinis were established from seeds and grown on a hormone-free medium. In vitro plants produced the cyclic peptide kalata B1 in concentrations of 0.67 mg g−1 dry weight after growth of 30 days. This was approximately 50% of the concentration analysed in green house plants (shoot tips), where different concentrations have been determined in leaves (1.82 mg g−1), shoot tips (1.36 mg g−1), stems (0.36 mg g−1), and in flowers (0.16 mg g−1). Callus and cell suspension cultures could be initiated from aseptic root, stem and leaf explants of O. affinis seedlings and plants. Different light intensities were shown to affect culture growth as well as chlorophyll synthesis. The friable callus was then used for the establishment of a cell suspension culture. Fresh and dry weight measurements showed that growth was optimal on MS medium supplemented with 0.4 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-d). Leaf suspensions cultured on this medium showed a 4-fold increase of biomass by the first week of incubation. No quantifiable amounts of kalata B1 were produced under these conditions. Morphological differentiation seems to be essential for cyclic peptide production. Therefore, several undifferentiated as well as organised cell lines of O. affinis have been developed. These cell lines will constitute a worthwhile starting point for the optimisation of kalata B1 synthesis in liquid media to the objective of producing cyclic peptides under controlled and defined conditions in bioreactors. 相似文献
8.
Jolanta Biesaga-Kocielniak Janusz Kocielniak Maria Filek Anna Janeczko 《Plant Cell, Tissue and Organ Culture》2008,94(2):139-147
The aim of this study was to produce suspension cultures of winter wheat directly from immature embryos bypassing the callus
stage, and to determine their capacity for growth and regeneration in comparison to suspension cultures produced from callus.
The study was carried out using Polish winter wheat varieties: ‘Grana’ and ‘Rosa’. Immature embryos were isolated, homogenized
and transferred directly to liquid medium supplemented with 2,4-D. Actively dividing cell cultures were obtained within 2 months
after the cultures were started. Suspension cultures from callus of immature embryos was also produced. With both cultivars,
faster growth was observed in the suspension cultures produced directly from embryos than in the suspensions produced from
callus. Metabolic activity was higher in the suspension culture produced directly from embryos than in the suspension derived
from callus only in ‘Grana’. The production of 1-amiocyclopropane-1-carboxylic acid (ACC), an ethylene precursor, was lower
in the suspension cultures produced directly from embryos than in the suspensions produced from callus. Morphogenic capacity
was significantly higher in aggregates derived directly from embryos than in aggregates derived from callus. With ‘Rosa’,
about one third of the aggregates derived directly from embryos regenerated shoots. Production of ACC was lower in ‘Rosa’
cell culture that regenerated then in other cell cultures that did not. Photosystem II reactions were more efficient in dark
green aggregates than in light green or pale green aggregates which were unable to regenerate. With the method presented,
wheat cell suspension cultures with a regeneration potential can be produced in 2 or 3 months less time than with traditional
methods. 相似文献
9.
Carimi Francesco Tortorici Maria Concetta De Pasquale Fabio Crescimanno Francesco Giulio 《Plant Cell, Tissue and Organ Culture》1998,54(3):183-189
Somatic embryogenesis was induced and plant regeneration was obtained in 11 different genotypes of sweet orange navel group
[Citrus sinensis (L.) Osb.] from cultures of stigma/style explants and undeveloped ovules. Explants were cultured on 3 different
modifications of Murashige and Skoog medium: 500 mg l-1 malt extract; 500 mg l-1 malt extract and 4.6 μM kinetin; and 500
mg l-1 malt extract and 13.3 μM 6-benzylaminopurine. Sucrose (146 mM) was used as carbon source. Somatic embryogenesis occurred
1–3 months after culture initiation from undeveloped ovule and stigma/style cultures of all the genotypes tested. Somatic
embryos developed into plantlets with a high frequency (74%) after transfer to Murashige and Skoog medium supplemented with
146 mM sucrose and 500 mg l-1 malt extract. Plants were successfully transferred to soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
10.
The isolation of a new type of thrombin inhibitor, called triabin, from the saliva of the hematophagous bug Triatoma pallidipennis,
has recently been described. In the in vitro platelet aggregation inhibition assay triabin has a similar potency as the thrombin
inhibitor hirudin now in phase III clinical trials. However, in another in vitro assay using a low molecular weight substrate
for thrombin, triabin does not inhibit thrombin completely even at 6 fold higher molar doses in comparison with hirudin. This
means that triabin has a novel mode of action towards thrombin making triabin into an interesting candidate as a therapeutic
agent. Recently it has been shown that a recombinant baculovirus can be efficiently used for the triabin production in insect
cells and that the yields in adherent cultures of High Five™ cells (approx. 20 mg l-1) were about 7 fold higher than in adherent
cultures of Sf9 cells (approx. 3 mg l- 1). To optimize the triabin yield from the baculovirus/insect cell expression system,
experiments were performed with suspension adapted cultures of High Five™ cells to investigate the effects of the state of
the host cell, of the multiplicity of infection, of the cell density at the time of infection and of supplementation of the
medium with nutrients and oxygen. Triabin yields of up to 200 mg l-1, as determined by an activity assay, could finally be
obtained here.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
11.
Embryogenic cell suspension cultures were established from calliderived from young leaves of sugarcane (Saccharum officinarumL.) by placing them in liquid medium containing 5 per cent coconutwater (CW), 23 mg 11 2, 4-D and 500 mg 11casein hydrolysate (CH). The cultures were maintained by transferring2.55.0 ml of the suspension to 35 ml of fresh mediumevery 45 days. Organized structures resembling the earlystages of embryogeny were formed when 2, 4-D in the medium waslowered (0.11.0 mg 11) but these did not developbeyond the globular or early scutellar stages. High levels ofsucrose (610 per cent) promoted the formation of proembryoids.Plating of the suspension on MS agar medium supplemented with0.252.0 mg 11 2, 4-D, 5 per cent CW, 500 mg 11CH, with or without activated charcoal, resulted in the formationof embryogenic calli. A large number of embryoids were formedin media containing lower 2, 4-D concentrations. Transfer ofembryoids to half-strength MS medium with 6 per cent sucroseestablished plantlets which were successfully transferred tosoil. Saccharum officinarumL, sugarcane, suspension culture, embryogenesis, regeneration 相似文献
12.
In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of
Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl
and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with 0.5 mg l−1 2,4-D and 1.0 mg l−1 Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences
in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of
cell aggregates was implanted on MS agar medium supplemented with Kin (0.0–2.0 mg l−1), GA3 (0.0–2.0 mg l−1) and AS (80.0 mg l−1). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on GA3-free medium, but the best morphological quality of embryos was observed in the presence of 0.5–1.0 mg l−1 Kin, 0.5 mg l−1 GA3 and 80.0 mg l−1 AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of
aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension
and 100% of uniformity for cotyledon suspension. 相似文献
13.
Cell suspension cultures of Gymnema sylvestre treated with four different elicitors, methyl jasmonate (MJ), yeast extract, chitin and pectin were studied for the production
of gymnemic acid as gymnemagenin equivalent, that was analyzed by high performance liquid chromatography (HPLC). All the four
tested elicitors induced gymnemic acid production in cell suspension cultures. Highest gymnemic acid content was achieved
following treatment with yeast extract (100.47 ± 0.28 mg/l), this was followed by MJ (70.43 ± 0.26 mg/l), pectin (64.19 ± 0.23 mg/l)
and chitin (62.72 ± 0.13 mg/l). The addition of elicitors has shown a significant influence on cell growth that affected cell
growth compared to respective controls. The highest gymnemic acid production was obtained after 20 days of elicitation in
cultures treated with 0.5 g l−l yeast extract, it was 5.25-folds greater than in control. These results suggest that the addition of an elicitor to Gymnema sylvestre cell suspension cultures could stimulate and enhance gymnemic acid production. In our present study we could able to overproduce
gymnemic acid up to 51.97 ± 0.26 mg l−l (dry weight basis) in yeast extract treated cell suspension cultures. 相似文献
14.
Callus and suspension cultures of Theobroma cacao L., initiatedfrom immature cotyledons of beans from pods harvested 120130days after pollination were established. A modified B-5 or MurashigeSkoogagar medium sustained growth of callus without loss of vigourafter each sub-culture. A 15-fold weight increase occurred duringthe 4 week culture periods at 30 ± 1 °C. Coconutwater improved callus growth substantially. The optimum hormonalconcentrations for growth of suspensions were 0.5 mg 11of 2, 4-dichlorophenoxyacetic acid and 0.1 mg I1 of kinetinin a MurashigeSkoog basal medium liquid medium. The optimumtemperature for growth of suspensions was 2530 °C.The cell number and cell mass of suspensions increased 20-foldin 14 days. No organogenesis or embryogenesis was observed. Theobroma cacao L., acao, cell culture, suspension culture, tissue culture. 相似文献
15.
Two lines of the red and pale yellow cell suspension cultures, prepared fromPrunus x yedoensis Matsum. callus induced by Murashige and Skoog's (1962) basal medium supplemented with 2, 4-dichlorophenoxyacetic acid (2,
4-D, 1.0 mg/l), kinetin (0.1 mg/l) and sucrose (30 g/l), were maintained on Schenk and Hildebrandt medium as modified by Mitchell
and Gildow (1975). The red cell suspension culture produced cyanidin 3-monoglucoside, 5, 4′-dihydroxy-7-methoxyisoflavone
4′-glucoside (prunetrin), isoquercitrin, catechin, epicatechin, and procyanidin B-1, B-2, B-3 and B-4, while the pale yellow
cells produced only a small amount of catechin and epicatechin as main flavonoids. These flavonoid compounds found in the
red cell culture were present also in maturePrunus leaves.
Maximum growth and maximum amount of total phenol and proanthocyanidin (procyanidins) were obtained with 0.3 mg/l of both
2,4-D and kinetin. Maximum concentration of anthocyanin was also obtained with 0.3 mg/l 2, 4-D regardless of kinetin concentration.
Accumulation of proanthocyanidin was markedly stimulated by low concentrations of phosphate, which reduced growth by about
half, and also by high concentrations of inorganic nitrogen. Production of both anthocyanin and proanthocyanidin was reduced
by lowered nitrogen levels. Cell growth and production of all phenolics were inhibited when ammonium ion replaced nitrate
in the medium. 相似文献
16.
Callus and cell suspension cultures from the little known Andean crop Mirabilis expansa were developed and maintained on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (1 mg/l) and
kinetin (0.1 mg/l). Callus and cell suspension cultures were screened with antibodies raised against ME1 (27.5 kDa) and ME2
(27 kDa), two ribosome-inactivating proteins (RIPs) previously found in roots of M. expansa. A 29-kDa protein found in callus and cell suspensions reacted strongly with ME1 antibodies. The 29-kDa protein, named MEC,
was purified to homogeneity by ammonium sulfate precipitation and cation exchange perfusion chromatography. Amino acid N-terminal
sequencing revealed close homology between MEC and ME1. The MEC amino acid sequence examined was highly conserved among RIPs
from widely different sources. This new RIP was immunolocalized to the cell walls of callus and cell suspension cultures.
Received: 20 August 1999 / Revision received: 10 December 1999 / Accepted: 21 December 1999 相似文献
17.
De Klerk Geert-Jan Brugge Jolanda Ter Marinova Svetla 《Plant Cell, Tissue and Organ Culture》1997,50(1):39-44
The production of an antifungal spirostanol saponin designated SC-1 has been detected in cell suspension cultures of the Mexican
species Solanum chrysotrichum. Batch cultures of a cell suspension obtained from hypocotyl derived calluses of this species
were grown for 25 days in shake flasks containing Murashige & Skoog (MS) medium. Throughout the growth cycle, fresh and dry
weight, SC-1 yield, and uptake of sucrose, glucose and fructose were determined. The effects of inoculum size and sucrose
concentration on the biomass accumulation and synthesis of the active metabolite, were studied. The maximum SC-1 production,
above 14 mg.g−1 (which was fifty times that of field grown plants), was reached after 20 days using a 2% inoculum and complete MS medium
supplemented with 2 mgl−1 2,4-D, 2 mg l−1kinetin, and sucrose between 30 and 45 gl−1.
.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
Babu KR Swaminathan S Marten S Khanna N Rinas U 《Applied microbiology and biotechnology》2000,53(6):655-660
Escherichia coli TG1 transformed with a temperature-regulated interferon-α expression vector was grown to high cell density in defined medium
containing glucose as the sole carbon and energy source, utilizing a simple fed-batch process. Feeding was carried out to
achieve an exponential increase in biomass at growth rates which minimized acetate production. Thermal induction of such high
cell density cultures resulted in the production of ∼4 g interferon-α/l culture broth. Interferon-α was produced exclusively
in the form of insoluble inclusion bodies and was solubilized under denaturing conditions, refolded in the presence of arginine
and purified to near homogeneity, utilizing single-step ion-exchange chromatography on Q-Sepharose. The yield of purified
interferon-α was ∼300 mg/l with respect to the original high cell density culture broth (overall yield of ∼7.5% active interferon-α).
The purified recombinant interferon-α was found by different criteria to be predominantly monomeric and possessed a specific
bioactivity of ∼2.5 × 108 IU/mg based on viral cytopathic assay.
Received: 8 October 1999 / Received revision: 8 December 1999 / Accepted: 12 December 1999 相似文献
19.
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
20.
María del Pilar Nicasio-Torres Mariana Meckes-Fischer Lucía Aguilar-Santamaría María Luisa Garduño-Ramírez Víctor Manuel Chávez-Ávila Francisco Cruz-Sosa 《Acta Physiologiae Plantarum》2012,34(1):307-316
The antidiabetic properties of Cecropia obtusifolia are attributed to chlorogenic acid (CGA) and isoorientin (ISO) phenolic compounds; both compounds possess hypoglycemic, hypolipidemic,
and antioxidant properties. As a potential strategy for an adequate supply of authentic plant raw material, the aim of this
study was to establish in vitro conditions for the development of cell suspension cultures that produce these bioactive compounds.
Callus cultures of leaf explants from acclimatized tree and in vitro plantlets were set up using different auxin levels; treatments
with 2,4-dichlorophenoxyacetic acid (2,4-D) and α-naphthalene acetic acid (NAA) to 8.92 μM with 6-benzylaminopurine (BAP)
at 2.22 μM stimulate highest callus production. Seedling cotyledon, hypocotyl, leaf, and stem explants developed calli bearing
roots with 2,4-D. With NAA, hypocotyl, cotyledon, and leaf explants developed morphogenic calli; 75% of stem explants formed
calli, and the remaining calli developed shoots. Determined CGA concentrations in calli were similar to those detected in
the leaves of wild trees, and ISO was not produced. Cell suspension cultures were established from leaf explants friable calli
with 8.92 μM 2,4-D in combination with 2.22 μM BAP, employing 4 and 5% inocula in fresh weight; CGA levels were maintained
and ISO was produced only at the end of logarithmic growth. On diminishing nitrate content in Murashige and Skoog (MS) medium
to 8.0 mM, maximum cell biomasses diminished, CGA production is increased and twice with 16.0 and, instead of CGA production
is tripled and quadrupled with 16.0 and 8.0 mM nitrates, respectively, and ISO synthesis was induced earlier and for a longer
time period, increasing its levels at the end of culture. Two compounds with ultraviolet spectra similar to those of caffeic
and ferulic acids were formed. Our results offer a protocol of cell suspension cultures for C. obtusifolia bioactive production and hypoglycemic property conservation. 相似文献