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21.
A rapidly growing, maintainable, embryogenic suspension culture of Glycine max L. Merrill. has been generated. The culture consisted almost entirely of clumps of proliferating globular embryos with very little nonembryogenic tissues. The number and size of somatic embryo clumps were used to quantify growth of embryogenic tissues under various conditions. Initiation and proliferation of this embryogenic suspension culture were dependent on the inoculum, method of subculture, and composition of the subculture medium. Twenty to 50 mg of highly embryogenic, early-staged soybean tissue were inoculated into 35 ml of liquid culture medium containing 5 mg 1–1 2,4-D and either 15 mM glutamine or preferably 5 mM asparagine. Suspension cultures were subcultured at the same inoculum density every 4 weeks. The embryos matured and germinated following placement on solid media, resulting in consistent plant regeneration. 相似文献
22.
Alteration of the synthesis of lipoxygenase in the early stages of soybean cotyledon culture 总被引:1,自引:0,他引:1
Xuemin Wang Gerhard Bookjans Mitch Altschuler Glenn B. Collins David F. Hildebrand 《Physiologia plantarum》1988,72(1):127-132
A detailed study of lipoxygenase (EC 1.13.11.12) synthesis in cotyledons of soybean [ Glycine max (L.) Merr. cv. Century] cultured in vitro for up to 40 h showed that synthesis of this protein, measured by in vivo [35S]-methionine labelling in connection with immunological methods and cell-free translation of mRNA, underwent a large transient reduction in the first 4 h of culturing and gradually increased in the following 36 h. Northern blot hybridizations with lipoxygenase cDNA clones showed that the decrease in translational activity was the consequence of a considerable reduction in lipoxygenase mRNA in the cotyledons. From these results we conclude that the transient decline in lipoxygenase synthesis in excised soybean cotyledons is regulated at the RNA level. Similarly judged from the analysis of patterns of uni-dimensional gel electrophoresis, the synthesis of a few other polypeptides decreased during the first 4 h of culture as well, while several others increased; in cotyledons cultured for 20 to 40 h the protein-synthesis pattern had returned to that in freshly excised cotyledons. An acclimation period of ca 1 day seems to be needed for isolated soybean cotyledons to stabilize and to resume regular RNA and protein synthesis. 相似文献
23.
Many soybean [Glycine max (L.) Merr.] genotypes that are grown in solution cultures are highly sensitive to the combination of both salinity and inorganic
phosphate (Pi) in the substrate. This effect has been observed on numerous occasions on plants grown in a saline medium that
contained a substantial amount of Ca (i.e., CaCl2/NaCl=0.5 on a molar basis). Because Ca is important in regulating ion transport and membrane permeability, solution culture
experiments were designed to examine the effects of various concentrations of Pi and ratios of CaCl2/NaCl (0 to 0.5 on a molar basis) at a constant osmotic potential (−0.34 MPa) on this adverse interaction. Four soybean cultivars
(‘Lee’, ‘Lee 74’ ‘Clark’ and ‘Clark 63’) were tested.
No adverse salinity x Pi interaction was found on Lee at any ratio and leaf P and Cl were maintained below 300 and 200 mmol
kg−1 dry wt, respectively. Clark, Clark 63 and Lee 74 soybean plants, on the other hand, were severely injured by solution salinity
(−0.34 MPa osmotic potential) when substrate Pi was ≥0.12 mM. Reduced substrate Ca did not intensify the salinity x Pi interaction. On the contrary, the onset of injury was hastened
and more severe with increased CaCl2/NaCl ratios in isotonic solutions. Shoot and root growth rates decreased as injury increased. Leaf P concentrations from
these cultivars grown in saline solutions with 0.12 mM Pi were excessive (>600 mmol kg−1 dry wt) compared with concentrations commonly found in soybean leaf tissue yet they were independent of the severity of injury.
Since leaf Cl increased wiht increased CaCl2/NaCl ratio, we suspect that the severity of foliar injury was related to the combined effects of excessive P and Cl within
the tissue. Lee 74, the only injured cultivar examined that excluded Cl from its leaves, was less sensitive than either Clark
cultivar and its injury was characteristically different. Other ion interactions were reported that may have played a role
in injury susceptibility. 相似文献
24.
Kucey R. M. N. Snitwongse P. Chaiwanakupt P. Wadisirisuk P. Siripaibool C. Arayangkool T. Boonkerd N. Rennie R. J. 《Plant and Soil》1988,108(1):33-41
Controlled environment and field studies were conducted to determine relationships between various measurements of N2 fixation using soybeans and to use these measures to evaluate a number ofBradyrhizobium japonicum strains for effectiveness in N2 fixation in Thai soils.15N dilution measurements of N2 fixation showed levels of fixation ranging from 32 to 161 kg N ha−1 depending on bacterial strain, host cultivar and location. Midseason measures of N2 fixation were correlated with each other, but not related measures taken at maturity. Ranking ofB. japonicum strains based on performance under controlled conditions in N-free media were highly correlated with rankings based on soybean
seed yields and N2 fixation under field conditions. This study showed that inoculation of soybeans with effectiveB. japonicum strains can result in significant increases in yield and uptake of N through fixation. The most effective strains tested
for use in Thai conditions were those isolated from Thai soils; however, effective strains from other locations were also
of benefit. 相似文献
25.
Egg yolk lipoprotein promoted growth of a wide variety of mammalian cell lines, including plasma-cytomas and epithelial cell lines, in serum-free medium. The lipoprotein was active for cell growth when used with insulin, transferrin, ethanolamine and selenite. The most active lipoprotein fraction (YLP-pI7.5) was purified to give a single peak by chromatofocusing and gel filtration, and was homogeneous on a 0.35% agarose gel electrophoretogram. The lipoprotein was characterised as a very low density lipoprotein with a protein content of only 1.3%. This lipoprotein had an optimal concentration of 300 g/ml (4 g protein/ml). It was easily separable from proteinous molecules secreted into the serum-free medium by the cells, since it floated on the surface of the medium after addition of ammonium sulfate, to precipitate protein, and centrifugation. An associated structure of lipid and protein seemed to be still necessary for the lipoprotein to exhibit a growth promoting activity. 相似文献
26.
A serum free medium was developed, that could be used for the large scale propagation of various cell lines in bioreactors. The medium is based on a 1:1 mixture of Iscove's Modified Dulbecco's Medium and Ham's Medium F12, supplemented with transferrin, insulin and a BSA/oleic acid complex. Several myelomas, hybridomas derived from different myelomas and spleen cells, and other lymphoid and non-lymphoid cell lines were cultivated at growth rates comparable to those observed using serum-supplemented media. There was furthermore no reduction in the formation of products such as monoclonal antibodies or recombinant human interleukin-2.Abbreviations Ag8
Mouse myeloma cell line P3-X63-Ag8.653
- BME
Basal Medium Eagle
- BSA
Bovine Serum Albumin
- DMEM
Dulbecco's Modified Eagle's Medium
- EDTA
Ethylenediaminete-traacetic Acid
- e-PC
Phosphatidyl choline from egg yolk
- FCS
Fetal Calf Serum
- FGF
Fibroblast Growth Factor
- GHL
Glycyl-histidyl-lysine
- HDL
High Density Lipoprotein
- HPL
Human Plasma Lipid
- IF
1:1 mixture of IMDM and Ham's F12
- IMDM
Iscove's Modified Dulbecco's medium
- LDL
Low Density Lipoprotein
- NS1
Mouse myeloma cell line NSI-1-Ag4-1
- PBS
Phosphate Buffered Saline
- s-PC
Phosphatidylcholine from soy beans
- s-PE
Phosphatidylethanolamine from soy beans
- s-lecithin
lecithin from soy beans 相似文献
27.
Terry L. Riss Kenneth P. Karey B. Daniel Burleigh David Farker David A. Sirbasku 《In vitro cellular & developmental biology. Plant》1988,24(11):1099-1106
Summary A serum-free clonal density growth assay was developed for the quantification of the biological activity of human recombinant
insulin-like growth factor I (IGF-I). The assay measures IGF-I stimulated growth of Balb/c 3T3 cells cultured over 4 d on
poly-d-lysine-coated plastic surfaces in a serum-free medium formulation composed of a 1∶1 (vol/vol) mixture of Ham's F12 and Dulbecco's
modified Eagle's media, supplemented with 3.0 ng/ml bovine basic fibroblast growth factor (bFGF), 10 μg/ml human transferrin,
100 μg/ml ovalbumin, and 1.0 μM dexamethanose. Low-temperature trypsinization of serum-supplemented stock cultures combined with the use of poly-d-lysine-coated plates made it unnecessary to use serum or fibronectin to promote cell attachment and survival. Serum-free
growth conditions were optimized with respect to the concentrations of the supplements. Addition of IGF-I resulted in 3.5-fold
more cells than control cultures without IGF-I after 4 d. Deletion of bFGF resulted in no IGF-I stimulation of growth. The
concentrations of various preparations of IGF-I required to achieve one-half maximal stimulation of cell number (ED50), ranged between 1.25 and 4.7 ng/ml. In parallel assays, IGF-I was 6.6 times more potent than human recombinant insulin-like
growth factor II and 32 times more potent than insulin. When cells were seeded into medium containing IGF-I, transferrin,
ovalbumin, and dexamethasone but no bFGF, growth was minimal. Dose-response addition of bFGF showed an ED50, of 0.9 ng/ml. The methods reported are useful to monitor the biological potency of recombinant and natural-source growth
factors as well as providing a new means of studying the multiple growth factor requirements of Balb/c 3T3 cells in cultures.
This work was supported by a contract from IMCERA Bioproducts, Inc. 相似文献
28.
Microtiter micromass cultures of limb-bud mesenchymal cells 总被引:4,自引:0,他引:4
Douglas F. Paulsen Michael Solursh 《In vitro cellular & developmental biology. Plant》1988,24(2):138-147
Summary A method is described for growing high-density micromass cultures of chick and mouse limb mesenchyme cells in 96-well microtiter
plates (μTμM cultures). Rapid quantitative estimates of chondrogenic expression were obtained by automated spectrophotometric
analysis of Alcian-blue-stained cartilage matrix extracts performed in the wells in which the cells had been grown. Quantitative
estimates of myogenic expression were obtained similarly using anti-sarcomere myosin monoclonal antibody and modified ELISA
techniques. This μTμM-ELISA method may be adapted for use with other antigens for which specific antibodies are available.
These methods were used to compare cartilage and muscle differentiation in 1 to 4 d μTμM cultures grown in serum-containing
(SCM) and defined (DM) media. The DM contains minimal additives (insulin, hydrocortisone, and in some cases, ascorbate or
transferrin) and supports both chondrogenesis and myogenesis. The colorimetric analyses agree well with the morphologic appraisal
of chondrogenesis and myogenesis. Similar numbers of cartilage nodules formed in all cultures, but in DM the nodules failed
to enlarge; explaining the reduced matrix synthesis in DM as compared with SCM, and suggesting that nodule enlargement is
a discrete, serum-dependent step. Studies of selected additives to DM show that transferrin enhances myogenesis, ascorbic
acid enhances chondrogenesis, and retinoic acid inhibits chondrogenesis. Together, the μTμM system, in situ colorimetric assays
of chondrogenesis and myogenesis, and DM will allow rapid prescreening of teratogens and screening of various bioactive compounds
(e.g., hormones, growth factors, vitamins, adhesion factors) for effects on limb mesenchymal cell differentiation.
This work was supported by grants RR08006-13 (DFP) and HD05505 and HD18577 (MS) from the National Institutes of Health, Bethesda,
MD. MF-20 hybridoma supernatant was obtained from the Developmental Studies Hybridoma Bank, Department of Biology, University
of Iowa, Iowa City, Iowa 52242 (maintained by NIH grant NO1-HD62915). 相似文献
29.
John F. Foley Byron Th. Aftonomos 《In vitro cellular & developmental biology. Plant》1988,24(9):900-904
Summary Colonies of HeLa cells cultured in media supplemented with human or bovine serum or both can be morphologically described
as three types: diffuse, intermediate, and compact, with their modal distribution depending on the serum or sera added to
the growth medium. We have observed that for a particular medium or serum system, the percentage of compact colonies remains
fairly constant under normal culture conditions, 0.2%, whereas the diffuse and intermediate colonies vary over a much wider
range. The presence of certain substances as trypsin, heparin and Darvan in the medium favor the increase of compact colonies
at the expense of other types. Furthermore, we have discovered that colonial morphology is influenced by cocultivation of
the HeLa cells with human fibroblastlike cells, the compact colonies increasing as the density of the fibroblast element introduced
into the mixed cultures is increased. Subsequent investigation revealed that conditioned medium from confluent fibroblast
and HeLa cell cultures contained a factor(s), that significantly increased the percentage of compact colonies. The factor
is nondialyzable, heat-stable and can be neutralized by serum. Recorded in this presentation are preliminary observations
on the kinetics of colony formation and the interaction among the three HeLa cell colony types, the diffuse, the intermediate,
and the compact. The factor's effect on HeLa cell colonial morphology is time dependent and rapidly reversed if the factor(s)
is removed and fresh medium added. 相似文献
30.
杨景峰 《Acta Botanica Sinica》1992,34(5):352-359
以高粱(Sorghum bicolor)和大豆(U.S.Soybean)幼苗为材料研究了仅植物很部受到热刺激时,其未直接受到温度影响的叶组织细胞的反应。当13天龄的高粱幼苗根部经受45℃4小时热处理时,发现其未直接受到热刺激的叶细胞内合成了一些异常的蛋白质,估测的分子量分别为80kD、70kD、33kD和17kD。最明显的两条蛋白质谱带是70kD和17kD。6天龄的大豆幼苗,当其根部经受40℃3小时热处理时,在其叶细胞内也检测到两条较为明显的蛋白质谱带,其分子量分别为60kD和17kD。观测到的这些异常蛋白质命名为‘热应激效应蛋白’,并与热应激蛋白在分子量大小分布上进行了比较。另外,还报道了利用蛋白质合成抑制剂,亚胺环己酮(cyclohexlmide)探讨了热应激蛋白与植物热耐性方面的可能关联。 相似文献