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1.
Summary Success has been achieved in developing a complete protocol for mass propagation of Anogeissus pendula and A. latifolia, two important forest species found in India. Seeds cultured on plant growth regulator-free, semisolid Murashige and Skoog
(MS) medium germinated within 5–6 wk and formed 4–6-cm long shoots. The shoots multiplied on MS+4.4 μM benzyladenine (BA)+5.7 μM indoleacetic acid (IAA) + casein hydrolysate (100 mgl−1) + ascorbic acid (50 mgl−1) + sucrose (3%) + agar (0.8%). A majority of the genotypes rooted with more than 90% efficiency when 5–6 cm individual shoots
were cultured on 1/2MS (only major salts reduced to half strength)+2.3 μM IAA+2.5 μM indolebutyric acid (IBA) + sucrose (3%)+agar (0.8%) for 15 d. Those 10% (approx.) genotypes that did not root well on the
above medium could be rooted with ease by increasing the concentration of IAA in the rooting media from 2.3 to 5.7 μM. The in vitro-raised plants were successfully transferred to the soil with a success rate of over 85%. Using this protocol, over 560 000
tissue-cultured plants of these two species have been produced and dispatched to various state forest departments for field
trials and routine plantations. 相似文献
2.
Summary Sodium chloride-tolerant plantlets of Dendrocalamus strictus were regenerated successfully from NaCl-tolerant embryogenic callus via somatic embryogenesis. The selection of embryogenic
callus tolerant to 100 mM NaCl was made by exposing the callus to increasing (0–200 mM) concentrations of NaCl in Murashige and Skoog medium having 3% (w/v) sucrose, 0.8% (w/v) agar, 3.0 mg l−1 (13.6 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5mg l−1 (2.3μM) kinetin (callus initiation medium). The tolerance of the selected embryogenic callus to 100 mM NaCl was stable through three successive transfers on NaCl-free callus initiation medium. The tolerant embryogenic callus
had high levels of Na+, sugar, free amino acids, and proline but a slight decline was recorded in K+ level. The stable 100 mM NaCl-tolerant embryogenic callus differentiated somatic embryos on maintenance medium [MS medium +3% sucrose +0.8% agar +2.0
mg l−1 (9.0 μM) 2,4-D+0.5 mg l−1 (2.3 μM) kinetin] supplemented with different (0–200 mM) concentrations of NaCl. About 39% of mature somatic embryos tolerant to 100 mM NaCl germinated and converted into plantlets in germination medium [half-strength MS+2% sucrose+0.02 mg l−1 (0.1 μM) α-naphthaleneacetic acid +0.1 mg l−1 (0.49 μM) indole-3-butyric acid] containing 100 mM NaCl. Of these plantlets about 31% established well on transplantation into a garden soil and sand (1:1) mixture containing
0.2% (w/w) NaCl. 相似文献
3.
Summary
In vitro methods were applied to the only remaining plant of the Meelup Mallee (Eucalyptus phylacis), a critically endangered species from the southwest of Western Australia. Shoot explants were initiated into culture using
a 1/2 MS [Murashige and Skoog basal medium (BM) for all experiments] liquid medium supplemented with 1% (w/v) activated charcoal,
which was replenished twice daily, followed by transfer of explants to agar medium supplemented with 0.5 μM zeatin. Explants were cultured under low intensity lighting (PPFD of 5–10 μmol m−2s−1) to minimize blackening of tissues, and some explants were induced to produce nodular green calluses in response to BM supplemented
with 5 μM thidiazuron. Nodular green calluses were induced to form adventitious shoots following transfer to medium supplemented with
0.5 μM zeatin and 1 μM gibberellic acid, A4 isomer (GA4). Development of shoots was completed on 1 μM zeatin + 0.1 μM 6-benzylaminopurine (BA) in vented culture tubes. Regenerated shoots were sequentially cultured on medium containing 0.5
μM zeatin + 0.2 μM indoleacetic acid (IAA) followed by either 0.5 μM zeatin + 1μM GA4 for shoot elongation or 1 μM zeatin + 0.5 μM IAA to optimize shoot growth. Rooted microshoots were produced after 4 weeks on 5 μM indolebutyric acid (IBA) and survived acclimatization and transfer to potting mixture. 相似文献
4.
Summary Short-term culture of rainbow trout (Onchorhynchus mykiss) hepatocytes was used to examine the effect of dexamethasone (DEX) on microsomal CYP 1A1 protein content and 7-ethoxyresorufin-O-deethylase (EROD) activity in vitro. Hepatocytes prepared by controlled collagenase digestion and plated at a density of
0.25 × 106 cells/cm2 in plastic culture dishes precoated with trout skin extract (7.6 μg skin protein/cm2) to facilitate cell attachment were maintained at 16° C. Cells were treated with DEX (10−9 to 10−7
M) or vehicle (dimethyl sulfoxide, DMSO) at 24 h. Microsomal CYP 1A1 protein content and EROD activities were measured at 72
h. Both CYP 1A1 protein as measured by Western blots using CYP 1A1 specific anti-sera and EROD activity were significantly
lower in DEX (10−8 to 10−7
M)-treated hepatocytes compared to untreated (control) or DMSO-treated cells. The effect was dose dependent in that a gradual
decrease of CYP 1A1 protein and EROD activities were seen with increasing doses of DEX (10−8 to 10−7
M). DEX at 10−9
M was ineffective. Concomitant addition of 10−6
M RU486, a type II specific glucocorticoid receptor antagonist, to hepatocytes treated with 10−7
M DEX abolished the DEX effect. RU486 at 10−8
M was ineffective. Spironolactone (10−8 to 10−6
M), a type I specific glucocorticoid receptor antagonist, did not counteract the DEX effect. RU486 or spironolactone (10−6
M) alone had no effect on CYP 1A1 under similar conditions. DEX thus down regulates CYP 1A1 in fish cultured hepatocytes and
this regulation is mediated through the type II glucocorticoid receptor(s). 相似文献
5.
Summary Somatic embryogenesis was induced in callus cultures derived from nucellar tissue of cashewnut (Anacardium occidentale L.). Callus was obtained from nucellar tissue after 3 wk of culture on semisolid Murashige and Skoog (MS) basal medium supplemented
with 2,4-dichlorophenoxyacetic acid (2,4-D, 5 μM)+gibberellic acid (GA3, 15 μM)+N6-benzyladenine (BA, 5 μM). This callus gave rise to an embryogenic mass after 9 wk on maintenance medium containing 2,4-D (10 μM)+GA3 (15 μM)+4% sucrose +0.5% activated charcoal +10% coconut water (CW) +0.05% casein hydrolysate (CH). The embryogenic mass, after
transfer to medium supplemented with 2,4-D (5 μM)+GA3 (30 μM)+4% sucrose +0.5% activated charcoal +10% CW +0.05% CH, gave rise to somatic embryos. The developmental stages of somatic
embryos were observed using light and stereo microscopes. Histological study of somatic embryo development was also carried
out. The present study would be useful for clonal propagation, and variety improvement in cashewnut, which is essential due
to its increasing demand and export potential. 相似文献
6.
Ming-Sound Tsao Judith D. Smith Joe W. Grisham 《In vitro cellular & developmental biology. Plant》1985,21(5):249-253
Summary The ability of a normal rat liver epithelial cell line with phenotypic characteristics of “oval” cells to grow in calcium-poor
medium has been investigated. The growth of these cells could be arrested in medium containing 0.03 mM Ca2+, a concentration below which cell necrosis began to occur 24 h postexposure. With increasing calcium concentration, progressive
cell proliferation was observed. Epithelial growth factor (EGF) (10 ng/ml) increased the survival and proliferation of cells
in calcium-poor medium and the response was inversely correlated with the extracellular calcium concentration. In contrast,
phenobarbital (0.2 to 2 mM), 12-0-tetradecanoylphorbol-13-acetate (0.01 to 1 μg/ml), or retinoic acid (0.001 to 0.1 μg/ml) depressed growth of cells
in calcium-poor medium. The results confirm the ability of EGF to lower the calcium requirement for proliferation of normal
cells, but such an effect does not seem to be a universal property of tumor promoters.
This research was supported by National Institutes of Health Grant CA 29323. 相似文献
7.
ATP-dependent Sr2+ transport was examined in vitro using basolateral membrane (BLM) vesicles isolated from rat renal cortex to clarify the discrimination
mechanisms between strontium (Sr) and calcium (Ca) in renal tubules during reabsorption. ATP-dependent Sr2+ uptake and Ca2+ uptake were observed in renal BLM vesicles and were inhibited by vanadate. Hill plots indicate similar kinetic behavior for
Ca2+ and Sr2+ uptake. The apparentK
m andV
max of ATP-dependent Sr2+ uptake were both higher than those for Ca2+ uptake. ATP-dependent Sr2+ uptake by BLM vesicles diminished in the presence of 0.1 μM Ca2+ and was more markedly inhibited by 1 μM Ca2+. Hill plots of Sr2+ uptake data with and without 0.1 μM Ca2+ showed that the cooperative behavior of Sr2+ uptake was not changed by Ca2+. In the presence of 0.1 μM Ca2+, the affinity of the transport system for Sr2+ and the velocity of Sr2+ uptake in the BLM were both decreased. However, the rate of Ca2+ uptake was not diminished by Sr2+ concentrations of <1.6 μM. These results suggest that Ca2+ is preferentially transported in the renal cortex BLM when Ca2+ and Sr2+ are present at the same time. 相似文献
8.
Summary Micropropagation of Arnica montana L. using Murashige and Skoog (MS) medium supplemented with N6-[2-isopentenyl]adenine (2iP), zeatin and α-naphthaleneacetic acid (NAA) in different concentrations does not ensure the formation
of a high number of regenerated plants; a maximum of 3.2 neoplantlets per explant were obtained. After 4 wk of culture on
medium with zeatin (4.5 μM) and NAA (5.3 μM), plants were 3.06 cm in length. The following step was to improve the clonal propagation of this species. Micropropagation
of Arnica montana L., initiated from nodal segments using semisolid media (4 g l−1 agar), was obtained. Explants were inoculated on MS medium supplemented with NAA (5.3 μM), 2iP (5.0 μM), maize extract (1.0 ml l−1), phloroglucinol (0.6 mM) or adenine sulfate (0.2 mM). Only 3 wk after the inoculation, plant multiplication as well as induction of roots were obtained, the optimal variant
being that containing NAA (5.3 μM), 2iP (5.0 μM) and maize extract (1.0 ml l−1). Six weeks after the inoculation plants were transferred to Perlite, with 80% plant survival being obtained. By isoesterase
pattern we concluded that we have obtained the clonal propagation of Arnica montana, because the pattern of several individuals belonging to different clones was the same. Only one region with esterase activity
that is present in all individuals has been identified. 相似文献
9.
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
10.
J. H. Carver E. P. Salazar M. G. Knize 《In vitro cellular & developmental biology. Plant》1983,19(9):699-706
Summary Reducing serum concentrations in media provides a potential cost advantage. To determine whether such media could be used
for applied mutagenesis assays, we measured cloning efficiency and growth parameters in suspension of Chinese hamster ovary
cells cultured in reduced serum with or without additives (1 μg/ml insulin, 3×10−7
M linoleic acid, 1×10−8
M H2SeO3) or bovine serum albumin (BSA, 1% wt/vol). With the additives and ≤0.5% fetal bovine serum (FBS), Ham’s F12 medium (without
hypoxanthine and thymidine) was more optimal than alpha Eagle’s minimum essential medium (MEM) (without ribosides and deoxyribosides)
for low density cloning and high density suspension growth. Acceptable cloning efficiencies were obtained with 2% FBS plus
BSA without additives in either medium; the addition of BSA resulted in improved colony size and more compact colony morphology.
In alpha MEM, satisfactory growth rates and maximum saturation densities in suspension culture were obtained only with 5%
FBS; in Ham’s F12, 1% FBS+deoxycytidine+BSA yielded satisfactory suspension growth. Spontaneous mutant frequencies were compared
for each medium containing 10% dialyzed FBS (DFBS), 1% FBS plus BSA, or 2% FBS plus BSA. The spontaneous frequency of azaadenine-resistant
phenotypes (mutant at theaprt locus) in 1% FBS plus BSA was significantly lower than the frequency observed in 2% FBS plus BSA or 10% DFBS. Frequencies
of spontaneous mutants resistant to thioguanine (hgprt locus) or fluorodeoxyuridine (tk locus) were similar with 10% DFBS, 1% FBS plus BSA, or 2% FBS plus BSA. Compared to alpha MEM with 10% DFBS, frequencies
of drug-resistant mutants induced by ethyl methanesulfonate or mitomycin C (MMC) were not significantly lower in alpha MEM
with 2% FBS plus BSA; observed mutant frequencies induced by dimethyl-nitrosamine or benzo(a)pyrene seemed to be decreased at lower survival levels.
This work was performed under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory
under Contract W-7405-ENG-48 and supported by the Environmental Protection Agency under Interagency Agreements IAG-D5-E681-AN
and AO. 相似文献
11.
T. Chakbavarty J. G. Norcini J. H. Aldrich R. S. Kalmbacher 《In vitro cellular & developmental biology. Plant》2001,37(5):550-554
Summary Creeping bluestem (Schizachyrium scoparium (Michx.) Nash var. stoloniferum (Nash) J. Wipff) embryogenic callus growing on solid medium was used to establish a cell suspension culture in Murashige
and Skoog (MS) basal medium supplemented with 1.5 mg l−1 (6.8 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), 0.2 mg l−1 (0.88 μM) 6-benzylaminopurine (BA), 0.5 mg l−1 (1.4 μM) zeatin, 0.2 mg l−1 (0.58 μM) gibberellic acid (GA3), and 10% (v/v) of coconut water (CW). Pro-embryos from suspension culture matured on semi-solid MS medium in about 18 wk,
and were then cultured on semi-solid MS medium without growth regulators for 2–3 wk. Shoots were regenerated on MS basal medium
supplemented with 3.0 mg L−1 (13.6 μM) 2,4-D, 1.0 mg l−1 (4.4 μM) BA, 1.0 mg l−1 (2.9 μM) GA3, 0.5 mg l−1 (2.7 μM) 1-naphthaleneacetic acid (NAA), 500 mg l−1 easein hydrolysate, and 10% (v/v) CW. Rooted plantlets were successfully accelimatized to greenhouse and outdoor conditions.
Using this protocol, it would be possible to produce at least 1300 fully acclimatized plantlets annually. 相似文献
12.
David Kirk Susumu Kagawa Gudrun Vener K. Shankar Narayan Y. Ohnuki Lawrence W. Jones 《In vitro cellular & developmental biology. Plant》1985,21(3):165-171
Summary A serum-free medium (HMRI-2) has been developed for the outgrowth and subculture of epithelial cells from normal adult human
ureter and bladder. Medium HMRI-2 consists of Ham’s MCDB 152 with double the amounts of the essential amino acids in Stock
1, low Ca2+ (0.06 mM) and is supplemented with epithelial growth factor, 5 ng/ml; transferrin, 5 μg/ml; insulin, 5 μg/ml; ethanolamine and phosphoethanolamine,
0.1 mM each; hydrocortisone, 2.8×10−6
M; and bovine pituitary extract, 126 μg protein/ml. The cultured cells showed ultrastructural markers of epithelial cells (prekeratin
fibers, tonofilaments, surface microvilli with glycocalyx), exhibited ABO antigens, and had a normal human diploid karyotype.
Primary cultures could be subcultured and also cryopreserved in HMRI-2 in liquid nitrogen. Cells in mass cultures showed a
population doubling time of 40.5±4.5 h and had a maximum in vitro life span of 20 to 25 population doublings. It was observed
that primary outgrowths, secondary cultures, and even cryopreserved cells all retained the capacity to respond to high Ca2+ and serum by differentiation and desquamation. This study has resulted in the availability of easily obtainable serum-free
epithelial cultures from normal adult human ureter and bladder. The useful in vitro life span of these cultures may be important
in future studies of carcinogenesis.
This work was supported by a grant from the National Cancer Institute (R01CA25089), Bethesda, MD. 相似文献
13.
Summary An efficient protocol has been developed for the regeneration of plantlets from leaf explants of witloof chicory (Cichorium intybus L.). Regeneration via callus was obtained on modified Murashige and Skoog semisolid medium (MS) containing 2.0 μM indole-3-acetic acid +5.0 μM 6-furfurylaminopurine (kinetin), and 1000 mgl−1 casein hydrolyzate. At least five or more shoots regenerated from each callus. The shoots were rooted on MS +0.2 μM indole-3-butyric acid. The plantlets thus obtained were successfully established in soil after bardening. Esculin accumulation
was recorded in plant tissues at different stages of differentiation in in vitro cultures and compared with in vivo-grown, plants. The esculin accumulation was higher in in vitro plants. 相似文献
14.
Summary Some native species produce seeds with a low frequency of germination accompanied with a period of dormancy. These features
make it difficult to produce new phenotypes through sexual propagation. Maclura tinctoria has been considered an endangered species due to extensive use of its wood and low frequency of seed germination. The objective
of the present study is to establish an in vitro propagation system for this species. Organogenic friable callus formation from nodal segments has been obtained using woody
plant medium (WPM) supplemented with 10.74 μM 1-naphthaleneacetic acid (NAA)+4.43 μM 6-benzylaminopurine (BA). Results indicate that the highest frequency of shoot formation is observed when WPM supplemented
with 4.03 μM NAA+4.43 BA is used. For root formation, the use of WPM medium (pH adjusted to 7.0) supplemented with 23.62 μM indole-3-butyric acid (IBA) and 4.7gl−1 activated charcoal is recommended. For acelimatization, subjecting rooted plantlets to 70%, 50%, and 30% mesh screen, each
successively for a period of 7 d, has resulted in 97% plantlet survival. 相似文献
15.
Monica Moura 《In vitro cellular & developmental biology. Plant》1998,34(3):244-248
Summary One of the first Azorean endemic vascular taxa chosen for the development ofin vitro multiplication techniques wasHypericum foliosum Aiton, due to its colonizing ability (Sj?gren, 1984), a loss of seed germination capacity after only 1 yr of storage (Maciel,
1994), and the populations' generally low number of individuals. The following culture media were tested usingHypericum foliosum's single node cuttings: Murashige and Skoog (1962), Roest and Bockelmman (1973), Lloyd and McCown (1980), C?rte and Mendon?a
(1985), and Cellárová et al. (1992). Further experiments were performed on CM medium supplemented with four different growth
regulators: α-naphthaleneacetic acid (NAA), N6-benzyladenine (BA), γ, γ-(dimethylallyl) aminopurine (2iP), and kinetin (KIN). The acclimatization stage was carried out
in Jiffy 7? pots and in a 2∶1 or 1∶1 peat/perlite mixture. We found that micropropagation ofHypericum foliosum is possible on CM medium and that the best results with growth regulators were achieved with the following supplements: 0.1
mg/l (0.4 μM) BA and 0.5 mg/l (2.6 μM) NAA+1.0 mg/l (4.4 μM) BA (in the initiation stage), and 0.1 mg/l (0.4 μM) BA (in the elongation stage). As for culture multiplication, 0.1 mg/l (0.4 μM) BA (in the initiation stage) and 0.5 mg/l (2.6 μM) NAA+1.0 mg/l (4.4 μM) BA (both in the initiation and elongation stages), proved to be the most efficient concentrations. The acclimatization stage
was successfully performed in Jiffy 7? pellets. 相似文献
16.
Gerald Audesirk David Shugarts Gina Nelson Julie Przekwas 《In vitro cellular & developmental biology. Plant》1989,25(12):1121-1128
Summary Neurons from brains of chick embryos and pond snails (Lymnaea stagnalis) were cultured for 3 to 4 d in the presence of no toxins, inorganic lead (PbCl2), or organic lead (trielthyl lead chloride). In chick neurons, inorganic lead reduced the percentage of cells that grew neurites
(IC50=270μM total lead, approximately 70 nM free Pb2+) but did not reduce the number of neurites per cell or the mean neurite length. Triethyl lead reduced the percentage of cells
that grew neuites (IC50=0.24 μM) and the mean neurite length (extrapolated IC50=3.6 μM) but did not reduce the number of neurites per cell. InLymnaea neurons, inorganic lead reduced the percentage of cells that grew neurites (IC50=13 μM total lead; approximately 10 nM free Pb2+). Triethyl lead reduced the percentage of cells that grew neurites (IC50=0.4 μM) and exerted significant toxicity at 0.2 μM. The two forms of lead affected neurite growth in qualitatively different ways, which suggests that their mechansms of action
are different.
These experiments were supported by grants from the Environmental Protection Agency, Washington, DC, and the National Institutes
of Environmental Health Science, Research Triangle Park, NC. 相似文献
17.
Vicky P. Kalfakakou Angelos M. Evangelou Jacques Benveniste Bernard Arnoux 《Biological trace element research》1993,38(3):289-299
Isolated guinea pig hearts were perfused, by the Langendorff technique, with 30, 15, 7.5, and 1.5 μM Zn2+ in Chenoweth solution. Contractile force, coronary flow, and heart rate were recorded by means of Narco IV Physiograph. Calcium
inhibitor (Verapamil 1 μM) and anion inhibitor (DIDS: 0.1, 1, and 5 μM) were used subsequently in the perfusing solutions in order to distinguish some of the possible mechanisms that Zn2+ uses to exert its action on cardiac myocytes. Isomolar to zinc concentration of Pb (II) and Co (II) were used to elucidate
whether zinc effects on heart are specific for this metal. All hearts were used to estimate their zinc and calcium content
by means of AAS (Atomic Absorption Spectrometry).
Our findings suggest that the higher the Zn2+ concentration, the more toxic effects on heart are expressed by rapid reversible contractile force reduction and reversible
specific changes of heart rate and flow. Zinc 1.5 μM in the perfusing solution benefits heart performance, but not significantly. Furthermore, the metal exerts specific effects
on guinea pig heart, and it is rather possible that these effects on cardiac myocytes are held through cell membrane receptors. 相似文献
18.
Summary Callus of Phalaenopsis Nebula was induced from seed-derived protocorms on 1/2 Murashige and Skoog (MS) basal medium plus 0–1.0 mg l−1 (0–4.52 μM) N-phenyl-N′-1,2,3,-thiadiazol-5-yl urea (TDZ) and/or 0–10 mg l−1 (0–45.24 μ M) 2,4-dichlorophenoxyacetic acid (2,4-D). Protocorms 2 mo. old performed better than 1-mo.-old protocorms for callus induction.
More calluses formed on 1/2 MS basal medium supplemented with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. These calluses could be maintained by subculturing every month with basal medium supplemented with 0.5 mg l−1 (2.27 μM) TDZ and 0.5 mg l−1 (2.26 μM) 2,4-D. Protocorm-like bodies were formed, and plants regenerated from these calluses on 1/2 MS basal medium alone or supplemented
with 0.1–1.0 mg l−1 (0.45–4.52 μM) TDZ. Plantlets were then potted on sphagnum moss in the greenhouse and grew well. No chromosomal abnormalities were found
among the root-tip samples of 21 of the regenerated plantlets that were successfully acclimatized. 相似文献
19.
Rakhi Chaturvedi S. P. Bhatnagar 《In vitro cellular & developmental biology. Plant》2001,37(2):255-258
Summary A protocol for in vitro shoot regeneration from cotyledon explants of Citrullus lanatus (Thunb.) Matsum. & Nakai cv. Sugar Baby is described. The cotyledons excised from 7-d-old aseptic seedlings showed the highest
percentage of shoots on Murashige and Skoog (MS) + N6-benzyladenine (BA; 3.0 μM) + N6-[2-isopentenyl] adenine (2iP; 3.0 μM) and MS + BA (3.0 μM) + indole-3-acetic acid (IAA; 3.0 μM). Whereas the latter medium induced shoot regeneration after the callusing of the explant, the former stimulated direct shoot
formation. The regenerated shoots were rooted and the resulting plants were established in earthen pots with 55% success. 相似文献
20.
Terramani TT Eton D Bui PA Wang Y Weaver FA Yu H 《In vitro cellular & developmental biology. Animal》2000,36(2):125-132
Summary The purpose of this study is to identify optimal culture conditions to support the proliferation of human macrovascular endothelial
cells. Two cell lines were employed: human saphenous vein endothelial cells (HSVEC) and human umbilical vein endothelial cells
(HUVEC). The influence of basal nutrient media (14 types), fetal bovine serum (FBS), and mitogens (three types) were investigated
in relation to cell proliferation. Additionally, a variety of extracellular matrix (ECM) substrate-coated culture dishes were
also tested. The most effective nutrient medium in augmenting cell proliferation was MCDB 131. Compared to the more commonly
used M199 medium, MCDB 131 resulted in a 2.3-fold increase in cell proliferation. Media containing 20% FBS increased cell
proliferation 7.5-fold compared to serum-free media. Among the mitogens tested, heparin (50 μg/ml) and endothelial cell growth
supplement (ECGS) (50μg/ml) significantly improved cell proliferation. Epithelial growth factor (EGF) provided no improvement
in cell proliferation. There were no statistical differences in cell proliferation or morphology when endothelial cells were
grown on uncoated culture plates compared to plates coated with ECM proteins: fibronectin, laminin, gelatin, or collagen types
I and IV. The culture environment yielding maximal HSVEC and HUVEC proliferation is MCDB 131 nutrient medium supplemented
with 2 mM glutamine, 20% FBS, 50 μg/ml heparin, and 50 μg/ml ECGS. The ECM substrate-coated culture dishes offer no advantage. 相似文献