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1.
DNA polymorphism patterns linked to the A-globin gene were analyzed in healthy Japanese using four different restriction endonucleases. The chromosomes with the A-globin gene were mapped through an evaluation of the presence of seven different restriction sites (HincII 5 to ; HindIII in G and A; HincII in, and 3 to, 1; AvaII in ; Bam-HI 3 to ). Among 36 chromosomes analyzed, 20 chromosomes had a haplotype of [+–––––+]. Among 55 individuals examined, 7 possessed a homozygous haplotye of [+–––––+]. All Japanese with the AT-globin gene had a subhaplotype of [–++–+] 5 to the -globin gene. Their major haplotypes were [–++–+–+] and [–++–++–]. It was expected that the presence of the AT-globin gene in Japanese may be deduced from subhaplotypes 5 to the -globin gene.  相似文献   

2.
In order to maintain axenic seedstock cultures axenically of thecommercially important red seaweed, Porphyra yezoensis, aprocedure was developed for axenic isolation and culture of conchocelis andmonospores. For axenic isolation of the conchocelis, contaminated microalgaewere most effectively removed by filtering contaminated samples through a100-m mesh after sonication. Removal of bacteria and otheralgaewas accomplished using a mixture of 5 agents (0.02% chitosan, 100 gml–1 GeO2, 10 gml–1 ampicillin, 40 gml–1 kanamycin and 200 gml–1 streptomycin). Axenic single colonies wereisolatedfrom a semi-solid medium prepared from 1% transfer gel. After collectingmonospores from the 40–50% density layer on a percoll-gradient, removalofbacteria and fungi from the monospores was accomplished using a mixture of 5antibiotics (3.5 g ml–1 nystatin, 2 mgml–1 ampicillin, 400 gml–1 kanamycin, 50 gml–1 neomycin and 800 gml–1 streptomycin). Axenic single juvenile blades wereisolated from a semi-solid medium prepared from 0.5% transfer gel.  相似文献   

3.
The present study demonstrates a procedure for the rapid development of a high number of somatic embryos from embryogenic suspension culture. This method might be efficient for mass propagation of Phnix dactylifera L. Embryogenic callus placed in liquid medium with 10–5M ABA yielded an average 72 embryos per 100ml of culture medium within 2months, while those placed on solid medium yielded an average of 33, 20 and 16 embryos per 100ml of culture medium respectively for 10–7, 10–6 and 10–5 M ABA after 4months. The combination of 2,4-DIchlorophenoxyacetic acid (2,4-D) (4.5×10–7M), glutamine (6.7×10–4M), and ABA (10–5M) (L8 liquid medium) showed a beneficial effect on somatic embryos production compared to 2,4-D and glutamine alone, while this combination significantly (p<0.05) increased the accumulation of storage proteins (144 and 138mgg–1 DW respectively for Jihel and Bousthami noir cultivars) in somatic embryos. The somatic embryos which underwent maturation on medium containing only 4.5×10–7M 2,4-D and 10–5M ABA (L6 liquid medium) accumulated more sugars (292 and 265mgg–1 DW respectively for Jihel and Bousthami noir) than those matured on any other liquid medium. Histological studies revealed that somatic embryos (developed in L6 and L8 liquid media) accumulated less reserve compounds (proteins and sugars) than zygotic embryos. The addition of activated charcoal (0.25 and 0.5gl–1) and phytagel® (2.5gl–1) to the germination medium may be useful for enhancing the germination of Phnix dactyliferasomatic embryos.  相似文献   

4.
Three-year-old spruce (Picea abies) saplings were planted and cultivated for 2 years in pots with 3 1 substrate, consisting of a homogenized mixture of sand, peat and forest soil with a high organic content (volume ratio 11.52). This substrate was amended with 10–180 mol Cd [kg soil dry weight (DW)]–1, 50–7500 mol Zn (kg soil DW)–1 (determined with 1 M ammonium acetate extracts) or combinations of both elements. Annual xylem growth rings in stems of plants treated with 50 mol Cd (kg soil DW)–1 or 7500 mol Zn (kg soil DW)–1 were significantly narrower than in control plants. Growth reductions were more pronounced in the second year of the experiment. The contents of Cd and Zn in stem wood and needles were positively correlated with the substrate concentrations. The Mg contents of the spruce needles were inversely correlated with soil concentrations of Cd and Zn. Root development was impeded at moderate concentrations of Cd (50 mol kg–1) or Zn (1000 mol kg–1) in the substrate. The adverse effects of potentially toxic trace elements, like Cd or Zn, on xylem growth of spruce plants are discussed with regard to possible growth reductions in forest trees under field conditions.  相似文献   

5.
The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10–3g l–1h–1) could not adequately account for the rate of sterol uptake (72×10–3g l–1h–1) and the specific growth rate (56×10–3 h–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm–1), the sterol-grown cells had strong surface activity (40mNm–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.  相似文献   

6.
Adventitious shoot regeneration was observed using leaf-petiole explants from shoot-proliferating cultures of Comet red raspberry (Rubus idaeus L.). A maximum regeneration rate of 70% (3.7 shoots/explant) was obtained using 4.5–9.1 M (1–2 mg l–1) N-phenyl-N-1,2,3-thiadiazol-5-ylurea (thidiazuron or TDZ) with 2.5–4.9 M (0.5–1 mg l–1) 1H-indole-3-butanoic acid (IBA) or 2.3 M (0.5 mg l–1) TDZ with 4.9 M (1 mg l–1) IBA in modified Murashige-Skoog medium. TDZ was more effective than N-(phenylmethyl)-1H-purin-6-amine (BA) at promoting regeneration in combinations tested with IBA (maximum 50% regeneration rate; 1.8 shoots/explant). Variation in the agar concentration or incubation temperature, orientation or scoring of the leaf-petiole explants and use of separate leaf or petiole explants had no effect on shoot regeneration. Incubation in the dark for 1, 2 or 3 weeks prior to growth in the light did not influence the percent regeneration rate but depressed the number of adventitious shoots. Explant source, from micropropagated shoots or greenhouse-grown plants, had an effect on shoot regeneration that was genotype dependent. Only 8 of 22 (36%) raspberry cultivars were capable of regeneration from leaf explants derived from greenhouse-grown plants.  相似文献   

7.
Tannase (tannin acyl hydrolase E.C. 3.1.1.20) has been isolated from Aspergillus niger van Tieghem and purified 29-fold. The enzyme had a temperature optimum of 60°C, pH optimum of 6.0 with a second peak at pH 4.5, Km of 0.20mM and Vmax of 5.0mol min–1 mg–1protein.  相似文献   

8.
Wen  Zhao  Shuang-Lin  Dong 《Hydrobiologia》2003,492(1-3):181-190
Primary productivity, biomass and chlorophyll-a of size fractionated phytoplankton (<0.22 m, <3 m, <8 m, <10 m, <40 m, <64 m, <112 m and <200 m) were estimated in 6 ponds and 5 experimental enclosures. The results showed that the planktonic algae less than 10 m are important in the biomass and production of phytoplankton in saline–alkaline ponds. The production of size fractionated phytoplankton corresponding to <112 m, <10 m and <3 m in saline–alkaline ponds were 10.5 ± 6.6 , 8.6 ± 5.4 and 0.33 ± 0.1 mgC l–1 d–1, respectively. Mean community respiration rate was 1.80 ± 0.73, 1.69 ± 0.90 and 1.38 ± 1.12 mgC l–1 d–1, respectively. The average production of phytoplankton corresponding to micro- (10–112 m), nano- (3–10 m) and pico- (<3 m) were 1.61, 8.30 and 0.33 mgC l–1 d–1, respectively. The ratio of those to the total phytoplankton production was 15%, 79% and 3%, respectively. The mean respiration rate of the different size groups was 0.11, 0.31 and 1.38 mgC l–1 d–1; the ratio of those to total respiration of phytoplankton was 6%, 17% and 77%, respectively. The production of size-fractionated phytoplankton corresponding to <200 m, <10 m and <3 m in enclosures was 2.19 ± 1.63, 2.08 ± 1.75 and 0.22 ± 0.08 mgC l–1 d-1, respectively. Mean community respiration rates were 1.25 ± 1.55, 1.17 ± 1.42 and 0.47 ± 0.32 mgC l–1 d–1, respectively. The average production of phytoplankton corresponding to micro- (10–200 m), nano- (3–10 m) and pico- (<3 m) plankton was 0.11, 1.86 and 0.22 mgC l–1 d–1, respectively. The ratio of those to the total production of phytoplankton was 5%, 85% and 10%, respectively. The mean respiration rate of different size groups were 0.08, 0.72 and 0.46 mgC l–1 d–1, the ratio of those to total respiration of phytoplankton was 6%, 57% and 37%, respectively. The concentrations of chlorophyll-a of the phytoplankton in the corresponding size of micro- (10–112 m), nano- (3–10 m) and pico- (<3 m) plankton in the experimental ponds were 19.3, 98.2 and 11. 9 g l–1, respectively. The ratio of those to the total chlorophyll-a was 15%, 76% and 9%, respectively. The concentrations of chlorophyll-a of phytoplankton micro- (10–200 m), nano- (3–10 m) and pico- (<3 m) plankton in enclosures were 1.7, 34.3 and 3.0 g l–1, respectively. The ratio of those to the total chlorophyll-a was 4%, 88% and 8%, respectively.  相似文献   

9.
Synopsis Arsenic persists in Chautauqua Lake, New York waters 13 years after cessation of herbicide (sodium arsenite) application and continues to cycle within the lake. Arsenic concentrations in lake water ranged from 22.4–114.81 g l–1, = 49.0 ag l–1. Well water samples generally contained less than 10 g l–1 arsenic. Arsenic concentrations in lake water exceeded U.S. Public Health Service recommended maximum concentrations (10 g l–1) and many samples exceeded the maximum permissible limit (50 g l–1). Fish accumulated arsenic from water but did not magnify it. Fish to water arsenic ratios ranged from 0.4–41.6. Black crappie (Pomoxis nigromaculatus) contained the highest arsenic concentrations (0.14–2.04 g g–1 ), X = 0.7 g g–1) while perch (Perca flavescens), muskellunge (Esox masquinongy) and largemouth bass (Micropterus salmoides) contained the lowest concentrations (0.02–0.13 g g–1). Arsenic concentrations in fish do not appear to pose a health hazard for human consumers.  相似文献   

10.
A DNA sequence (8–19T) of 2.3 kilobase pairs (kb) of Drosophila melanogaster was localized by in situ hybridization to the extreme ends of polytene chromosomes and to the chromocenter. The relative abundance of this sequence at the ends of polytene chromosomes X2L2R3L3R is 13.41.902.7. This differential distribution is probably due to different copy numbers at the individual telomeric regions. Restriction enzyme analysis of genomic DNA shows that 8–19T sequences are interspersed with other sequences. The clone 8–19T, which contains most of this interspersed repetitive sequence, is itself not internally repetitive but has a complex sequence composition. Some of these sequences are transcribed into poly(A)+RNA. We suggest that the ends of Drosophila chromosomes are of a complex arrangement with some sequences common to all ends.  相似文献   

11.
Protistan community grazing rates upon both bacterioplankton and autotrophic picoplankton were estimated using fluorescently-labeled prey and by measurement of extracellular hydrolysis of 4-methylumbelliferyl (MUF) -N-acetylglucosaminide in a eutrophic reservoir and an oligo-mesotrophic lake during phytoplankton blooms. In addition, enzyme methods were optimized in bacterivorous flagellate cultures by two enzyme assays, based on fluorometric detection of protistan digestive activity, which were compared and calibrated independently against flagellate bacterivory. Enzymatic hydrolyses of MUF -N,N,N-triacetylchitotriose and MUF -N-acetylglucosaminide were measured in cell-free (sonicated) and whole-cell (unsonicated) samples. The hydrolysis of both substrates, using the whole-cell enzyme assay at in situ pH, was correlated significantly with total grazing rate of Bodo saltans. Thus the whole-cell enzyme assay with MUF -N-acetylglucosaminide was used for freshwater samples. High-affinity (K m < 1 mol 1–1) and low-affinity (K m > 100 mol 1–1) enzymes were distinguished kinetically in most samples from both systems studied. Activities (V max ) of the high-affinity enzyme varied from 0.24 to 1.43 nmol 1–1 h–1. Protistan community grazing on bacterioplankton was in the range of 0.15–1.36 g C 1–1 h–1. both for lake and reservoir, the differences being observed in grazing on picocyanobacteria (lake, 0.03-0.22 g C 1–1 h–1. reservoir, 0.35–1.56 g C 1–1) h–1. The enzyme activities were correlated significantly with the protistan grazing both on bacterioplakton (r s = 0.62, P < 0.001) and total procaryotic picoplankton (the sum of organic carbon grazed from bacteria and picocyanobacteria, r s = 0.73, P < 0.001) in the eutrophic reservoir. Weaker relationships (r s = 0.42) with a lower slope were found for the oligo-mesotrophic lake. Ingestion rate studies are time-consuming and the digestive enzyme assay with MUF -N-acetylglucosaminide presents a rapid alternative for estimating total protistan prokaryotic picoplanktivory in freshwaters.  相似文献   

12.
Mercury budget of an upland-peatland watershed   总被引:6,自引:2,他引:6  
Inputs, outputs, and pool sizes oftotal mercury (Hg) were measured in a forested 10 hawatershed consisting of a 7 ha hardwood-dominatedupland surrounding a 3 ha conifer-dominatedpeatland. Hydrologic inputs via throughfall andstemflow, 13±0.4 g m–2 yr–1over the entire watershed, were about doubleprecipitation inputs in the open and weresignificantly higher in the peatland than in theupland (19.6 vs. 9.8 g m–2 yr–1). Inputs of Hg via litterfall were 12.3±0.7g m–2 yr–1, not different in thepeatland and upland (11.7 vs. 12.5 g m–2yr–1). Hydrologic outputs via streamflow were2.8±0.3 g m–2 yr–1 and thecontribution from the peatland was higher despiteits smaller area. The sum of Hg inputs were lessthan that in the overstory trees, 33±3 gm–2 above-ground, and much less than eitherthat in the upland soil, 5250±520 gm–2, or in the peat, 3900±100 gm–2 in the upper 50 cm. The annual flux of Hgmeasured in streamflow and the calculated annualaccumulation in the peatland are consistent withvalues reported by others. A sink for Hg of about20 g m–2 yr–1 apparently exists inthe upland, and could be due to either or bothstorage in the soil or volatilization.  相似文献   

13.
Development of an L6 myoblast in vitro model of moniliformin toxicosis   总被引:1,自引:0,他引:1  
L6 myoblasts were used as an in vitro model to investigate the role of moniliformin and its interaction with monensin in turkey knockdown syndrome and sudden death syndromes in poultry. Cell viability and microscopic and ultrastructural alterations noted in L6 myoblasts cultured in the presence of moniliformin (0.0–0.3 g/l) were compared to those observed in parallel cultures also containing one of the following compounds: selenium (0–0.004 ng/l), thiamine (0–0.3 g/l), or pyruvate (0–0.46 g/l). Marked dilation of the RER, membranous whorls, glycogen deposition, membrane-bound cytoplasmic inclusions and necrosis were observed in myoblasts exposed to 0.03/2-0.30 g moniliformin/l medium. Supplementation of medium with thiamine and pyruvate, or selenium, provided significant protection to cells exposed to 0.0–0.3 g/l or 0.0–0.15 g moniliformin/l, respectively. Dose-dependent differences in protein and ATP production were not detected. Myoblasts grown in medium containing 0–0.15 g moniliformin/l and 7.5–50.0 M A23187, beauvericin or monensin had degrees of cytotoxicity similar to parallel cultures receiving only an ionophore. L6 myoblasts were a useful model of moniliformin toxicosis. The findings of this study suggest cytotoxicity due to moniliformin in L6 myoblasts may be due in part to oxidative damage and altered pyruvate metabolism, and that moniliformin does not predispose myoblasts to ionophore toxicosis. This study supports the results of in vivo investigations in poultry that moniliformin and monensin do not act synergistically to induce knockdown or monensin toxicosis.  相似文献   

14.
Summary Respiration of an undescribed species of soil nematode of the genus Chiloplacus from the Canadian High Arctic was measured at 2°, 5°, 10°, 15°, 20° and 25°C. The corresponding metabolic rates were 0.2697×10-3 l, 0.3406×10-3 l, 0.8408×10-3 l, 0.8539×10-3 l, 1.8420×10-3 l and 2.9360×10-3 l O2 ind-1 h-1, respectively, for a nematode of 1.0 g dry weight. The relationship between respiration and dry weight for Chiloplacus sp. at 10°C is described by the function log R=-3.0693+0.8844 log W. Q10 values for the 2°–5°, 5°–10°, 10°–15°, 15°–20° and 20°–25°C temperature intervals were 2.18, 6.09, 1.03, 4.65 and 2.54, respectively. Chiloplacus sp. showed raised metabolic rates at low tempetatures compared with species from warmer environments. Metabolic rates of representative samples of the soil, nematode fauna (dominated by individuals of the genus Plectus) from the same location were 0.1593×10-3 l, 0.3603×10-3 l and 0.5332×10-3 l O2 ind-1 h-1 at 5°, 10° and 15°C for an average nematode of 0.4297 g dry weight.  相似文献   

15.
This study addresses the temporal distribution of forms of phosphorus in the soil of a temporarily flooded riparian forest of the valley of the river Garonne (Southwest of France). A sequential extraction for forms of phosphorus of increasing chemical stability was used. During the study period (13 months), the forest was flooded a few days during March and May. In winter, resin-Pi concentration was high (26 g g–1) in comparison to spring values (<9 g g–1). NaHCO3-Po, NaHCO3-Pi or NaOH-Pi concentrations increased during winter (up to 74, 124 and 78 g g–1 respectively) and decreased significantly during spring (32, 44 and 32 g g–1 respectively). This pattern was attributed to simultaneous mineralization and plant uptake during the growing season and to the flood events (erosional processes and P-release). During summer and fall, resin-Pi concentration increased significantly (up to 26 g g–1 in October). NaHCO3-Po concentrations remained low during spring and summer (<33 g g–1), and increased significantly in fall (>45 g g–1 NaHCO3-Pi or NaOH-Pi increased in late spring or summer (90 g g–1 and 68 g g–1 respectively). Increasing concentrations of the labile forms during late spring or summer were ascribed to the warm temperature and soil dryness that limited plant growth. HCl-Pi increased regularly after the floods (174 g g–1 before the flood events to 254 g g–1 after the floods). Residual P presented a similar pattern i.e. 214 g g–1 and 279 g g–1 respectively before and after the flood events. This pattern was attributed to a progressive incorporation of flood deposits to the soil.  相似文献   

16.
Summary Measurements were made of the difference in the electrochemical potential of protons ( ) across the membrane of vesicles reconstituted from the ATPase complex (TF 0 ·F 1) purified from a thermophilic bacterium and P-lipids. Two fluorescent dyes, anilinonaphthalene sulfonate (ANS) and 9-aminoacridine (9AA) were used as probes for measuring the membrane potential () and pH difference across the membrane ( pH), respectively.In the presence of Tris buffer the maximal and no pH were produced, while in the presence of the permeant anion NO 3 the maximal pH and a low were produced by the addition of ATP. When the ATP concentration was 0.24mm, the was 140–150 mV (positive inside) in Tris buffer, and the pH was 2.9–3.5 units (acidic inside) in the presence of NO 3 . Addition of a saturating amount of ATP produced somewhat larger and pH values, and the attained was about 310 mV.By trapping pH indicators in the vesicles during their reconstitution it was found that the pH inside the vesicles was pH 4–5 during ATP hydrolysis.The effects of energy transfer inhibitors, uncouplers, ionophores, and permeant anions on these vesicles were studied.  相似文献   

17.
Natural abundance of 15N in tropical plants with emphasis on tree legumes   总被引:6,自引:0,他引:6  
Natural abundance of 15N ( 15N) of leaves harvested from tropical plants in Brazil and Thailand was analyzed. The 15N values of non-N2-fixing trees in Brazil were +4.5±1.9, which is lower than those of soil nitrogen (+8.0±2.2). In contrast, mimosa and kudzu had very low 15N values (–1.4+0.5). The 15N values of Panicum maximum and leguminous trees, except Leucaena leucocephala, were similar to those of non-N2-fixing trees, suggesting that the contribution of fixed N in these plants is negligible. The 15N values of non-N2-fixing trees in Thailand were +4.9±2.0. Leucaena leucocephala, Sesbania grandiflora, Casuarina spp. and Cycas spp. had low 15N values, close to the value of atmospheric N2 (0), pointing to a major contribution of N2 fixation in these plants. Cassia spp. and Tamarindus indica had high 15N values, which confirms that these species are non-nodulating legumes. The 15N values of Acacia spp. and Gliricidia sepium and other potentially nodulating tree legumes were, on average, slightly lower than those of non-N2-fixing trees, indicating a small contribution of N2 fixation in these legumes.  相似文献   

18.
Based on field measurements in two agriculturalecosystems, soil respiration and long-term response ofsoil organic carbon content (SOC) was modelled. Themodel predicts the influence of temperature increaseas well as the effects of land-use over a period ofthirty years in a northern German glacial morainelandscape. One of the fields carried a maizemonoculture treated with cattle slurry in addition tomineral fertilizer (maize monoculture), the otherwas managed by crop rotation and recieved organicmanure (crop rotation). The soils of both fieldswere classified as cambic Arenosols. The soilrespiration was measured in the fields by means of theopen dynamic inverted-box method and an infrared gasanalyser. The mean annual soil respiration rates were 268 (maizemonoculture) and 287 mg CO2 m-2 h-1(crop rotation). Factors controlling soil respirationwere soil temperature, soil moisture, root respirationand carbon input into the soil. Q10-valuesof soil respiration were generally higher in winterthan in summer. This trend is interpreted as anadaptive response of the soil microbial communities.In the model a novel mathematical approach withvariable Q10-values as a result oftemperature and moisture adjustment is proposed. Withthe calibrated model soil respiration and SOC werecalculated for both fields and simulations over aperiod of thirty years were established. Simulationswere based on (1) local climatic data, 1961 until1990, and (2) a regional climate scenario for northernGermany with an average temperature increase of 2.1 K.Over the thirty years period with present climateconditions, the SOC pool under crop rotation wasnearly stable due to the higher carbon inputs, whereasabout 16 t C ha-1 were lost under maizemonoculture. Under global warming the mean annualsoil respiration for both fields increased and SOCdecreased by ca. 10 t C ha-1 under croprotation and by more than 20 t C ha-1 undermaize monoculture. It was shown that overestimationof carbon losses in long-term prognoses can be avoidedby including a Q10-adjustment in soilrespiration models.  相似文献   

19.
Four methoxypolyethylene glycols (MPEG, molecular masses 350, 750, 2000 and 5000 Da), each activated by nitrophenyl chloroformate, were used to modify trypsin. Compared with the native trypsin, the MPEG-modified trypsin was more stable against temperature between 30°C and 70°C, longer chain of MPEG moiety corresponding to higher thermal stability. The T for the native and the modified trypsin (0.4 mg ml–1) was increased from 47°C to 66°C. The stabilization effect caused by MPEG modification was the result of decreasing in both the autolysis rate and the thermal denaturation rate. The thermodynamic analysis of the thermal denaturation process showed that the activation free energy (G*) of the native and the modified trypsin at 60°C was increased from 102.9 to 109.3 kJ mol–1; the activation enthalpy (H*) was increased from 57.4 to 86.9 kJ mol–1; the activation entropy (S*) was increased from –136 to –67 J molK–1. A possible explanation for the decreased thermal denaturation rate caused by MPEG modification was also discussed.  相似文献   

20.
Callus was initiated from immature leaf and stem segments of rose (Rosa hybrida cv. Landora) and subcultured every four weeks on a basal medium of half-strength Murashige & Skoog (1962) salts plus 30 g l-1 sucrose (1/2 MS) and supplemented with 2.2 M BA, 5.4 M NAA and 2.2–9.0 M 2,4-D. Embryogenic callus and subsequently somatic embryos were obtained from 8-week-old callus culture on 1/2 MS+2.2 M BA+0.05 M NAA+0.3 M GA3+200–800 mg l-1 L-proline. Long-term cultures were established and maintained for up to 16 months by repeated subculture of embryogenic callus on L-proline deficient medium. About 12% of cotyledonary stage embryos taken from cultures cold-stored at 8±1°C for 4 days germinated on 1/2 MS+2.2 M BA+0.3 M GA3+24.7 M adenine sulphate.Abbreviations BA benzyladenine - NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - GA3 gibberellic acid  相似文献   

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