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1.
2.
A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm. 相似文献
3.
Pyrophosphate (140 mM, pH 7.1) extracts of two arable soils and one pasture soil were ultrafiltrated separating the extracted material into three fractions: AI with nominal molecular weight (nmw) > 100 kD, AII with nmw between 10 kD and 100 kD and R with nmw < 10 kD. Protease activity was determined in the fractions by using three different substrates: N-benzoyl-l-argininamide (BAA), specific for trypsin; N-benzyloxy-carbonyl-l-phenylalanyl l-leucine (ZPL), specific for carboxypeptidases; and casein, essentially a non-specific substrate. The derivative fractions were also analysed for their amino acid N and humic (HA) and fulvic (FA) acid contents. The organic matter of extracts and derivative fractions obtained from the pasture soil was analysed by isoelectric focusing (IEF) and that of fractions analysed by pyrolysis gas chromatography (Py-GC). Activities of the extract were monitored for their thermal stability and those of the extract and derivative fractions for their optimal pH.Due to the mechanical disintegrating action of sodium pyrophosphate over the humic substances during the fractionation process the amount of total organic C and FA in the fractions was ranked as R > AII > AI. The lowest amino acid N/organic C was found in the R fraction, whereas AII fraction was rich in humic acids, carbohydrates and amino acid N and AI fraction showed the lowest carbohydrate content. At least 70% of the total BAA- and ZPL-hydrolysing activity was associated to particles with nmw higher than 10 kD and at least 30% of these activities were present in particles with nmw higher 100 kD. Casein-hydrolysing activity was quite evenly distributed among the three fractions (AI, AII and R). The extracted protease-organic complexes were resistant to thermal denaturation and some of them showed optimal activity at pH values higher than 10 as a result of the polyanionic characteristics of the humic material surrounding enzyme molecules and of the presence of alkaline protease. Comparison of data obtained in Py-GC analyses and in protease activity suggests that BAA-hydrolysing activity was associated to a highly condensed humic matter and ZPL-hydrolysing activity to less resistant humic substances, while at least some of the extracted casein-hydrolysing activity was present as glyco-proteins not associated to humus. BAA-hydrolysing activity was probably inhibited by fresh organic matter of carbohydrate origin whereas lignin derived organic matter probably inhibited ZPL- and casein-hydrolysing activity. 相似文献
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A review of cyanobacterial odorous and bioactive metabolites: Impacts and management alternatives in aquaculture 总被引:2,自引:0,他引:2
Juliette L. Smith Greg L. Boyer Paul V. Zimba 《Aquaculture (Amsterdam, Netherlands)》2008,280(1-4):5-20
Increased demand has pushed extensive aquaculture towards intensively operated production systems, commonly resulting in eutrophic conditions and cyanobacterial blooms. This review summarizes those cyanobacterial secondary metabolites that can cause undesirable tastes and odors (odorous metabolites) or are biochemically active (bioactive metabolites) in marine and freshwater, extensive and intensive aquaculture systems. For the scope of this paper, biochemically active metabolites include (1) toxins that can cause mortality in aquaculture organisms or have the potential to harm consumers via accumulation in the product (hepatotoxins, cytotoxins, neurotoxins, dermatoxins, and brine shrimp/molluskal toxins), (2) metabolites that may degrade the nutritional status of aquaculture species (inhibitors of proteases and grazer deterrents) or (3) metabolites that have the potential to negatively affect the general health of aquaculture species or aquaculture laborers (dermatoxins, irritant toxins, hepatotoxins, cytotoxins). Suggestions are made as to future management practices in intensive and extensive aquaculture and the potential exposure pathways to aquaculture species and human consumers are identified. 相似文献
6.
Soil enzymes are linked to microbial functions and nutrient cycling in forest ecosystems and are considered sensitive to soil
disturbances. We investigated the effects of severe soil compaction and whole-tree harvesting plus forest floor removal (referred
to as FFR below, compared with stem-only harvesting) on available N, microbial biomass C (MBC), microbial biomass N (MBN),
and microbial biomass P (MBP), and dehydrogenase, protease, and phosphatase activities in the forest floor and 0–10 cm mineral
soil in a boreal aspen (Populus tremuloides Michx.) forest soil near Dawson Creek, British Columbia, Canada. In the forest floor, no soil compaction effects were observed
for any of the soil microbial or enzyme activity parameters measured. In the mineral soil, compaction reduced available N,
MBP, and acid phosphatase by 53, 47, and 48%, respectively, when forest floor was intact, and protease and alkaline phosphatase
activities by 28 and 27%, respectively, regardless of FFR. Forest floor removal reduced available P, MBC, MBN, and protease
and alkaline phosphatase activities by 38, 46, 49, 25, and 45%, respectively, regardless of soil compaction, and available
N, MBP, and acid phosphatase activity by 52, 50, and 39%, respectively, in the noncompacted soil. Neither soil compaction
nor FFR affected dehydrogenase activities. Reductions in microbial biomass and protease and phosphatase activities after compaction
and FFR likely led to the reduced N and P availabilities in the soil. Our results indicate that microbial biomass and enzyme
activities were sensitive to soil compaction and FFR and that such disturbances had negative consequences for forest soil
N and P cycling and fertility. 相似文献
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Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability
to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases
and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an
intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS.
The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system.
The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365. 相似文献
10.
Based on biochemical assays and electrophoretical methods, the inhibitory effects of three plant meals (soybean meal, wheat meal, winged bean meal) on digestive alkaline proteases of discus were investigated. Casein assays revealed that increasing levels of soybean meal caused a linear inhibitory effect on activity of protease. SDS-PAGE images revealed that trypsin and chymotrypsin were the inhibited enzymes. Soybean showed the lowest inhibition rate followed by wheat meal and raw winged bean. There was a quadratic relationship between wheat meal levels and its inhibition of protease acitivity. The highest inhibitory effect was obtained with the winged bean meal with inhibition of caseinolytic activities ranging from 3.6–98.6%. Results from this study showed the potential of both soybean meal and wheat meal as ingredients for practical diet for discus, while demonstrating the need for further improvement in processing method for winged bean meal. 相似文献