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A polymerase chain reaction (PCR)-based method was developed to detect DNA of Fusarium solani f. sp. glycines , the cause of soybean sudden death syndrome. Two pairs of primers, Fsg1/Fsg2 designed from the mitochondrial small subunit ribosomal RNA gene, and FsgEF1/FsgEF2 designed from the translation elongation factor 1-α gene, produced PCR products of 438 and 237 bp, respectively. Primer specificity was tested with DNA from 82 F. solani f. sp. glycines , 55 F. solani non-SDS isolates, 43 isolates of 17 soybean fungal pathogens and the oomycete Phytophthora sojae , and soybean. The sensitivity of primer Fsg1/Fsg2 was 10 pg while that of FsgEF1/FsgEF2 was 1 ng when using F. solani f. sp. glycines total genomic DNA or down to 103 macroconidia g−1 soil. Nested PCR increased the sensitivity of the PCR assay 1000-fold to 10 fg using primers Fsg1/Fsg2, and 1 pg using primers FsgEF1/FsgEF2. F. solani f. sp. glycines DNA was detected in field-grown soybean roots and soil by PCR using either single pairs of primers or the combination of two pairs of primers. The occurrence of F. solani f. sp. glycines was determined using nested PCR for 47 soil samples collected from soybean fields in 20 counties of Illinois in 1999. F. solani f. sp. glycines was detected in soil samples from all five Illinois Agricultural Statistic Districts including 100, 89, 50, 92 and 50% of the samples from East, Central, North-east and West Districts, respectively.  相似文献   
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The continuous use of plowing for grain production has been the principal cause of soil degradation. This project was formulated on the hypothesis that the intensification of cropping systems by increasing biomass‐C input and its biodiversity under no‐till (NT) drives soil restoration of degraded agro‐ecosystem. The present study conducted at subtropical [Ponta Grossa (PG) site] and tropical regions [Lucas do Rio Verde, MT (LRV) site] in Brazil aimed to (i) assess the impact of the continuous plow‐based conventional tillage (CT) on soil organic carbon (SOC) stock vis‐à‐vis native vegetation (NV) as baseline; (ii) compare SOC balance among CT, NT cropping systems, and NV; and (iii) evaluate the redistribution of SOC stock in soil profile in relation to soil resilience. The continuous CT decreased the SOC stock by 0·58 and 0·67 Mg C ha−1 y−1 in the 0‐ to 20‐cm depth at the PG and LRV sites, respectively, and the rate of SOC sequestration was 0·59 for the PG site and ranged from 0·48 to 1·30 Mg C ha−1 y−1 for the LRV site. The fraction of C input by crop residues converted into SOC stock was ~14·2% at the PG site and ~20·5% at the LRV site. The SOC resilience index ranged from 0·29 to 0·79, and it increased with the increase in the C input among the NT systems and the SOC sequestration rates at the LRV site. These data support the hypothesis that NT cropping systems with high C input have a large potential to reverse the process of soil degradation and SOC decline. Copyright © 2013 John Wiley & Sons, Ltd.  相似文献   
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Landscape Ecology - Anthropogenic land use and cover changes impact biodiversity worldwide. However, ecological groups are differently affected by landscape composition and configuration....  相似文献   
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Several Ponto‐Caspian gobiid species have recently expanded their ranges in Europe and North America. This is the first study to demonstrate passive downstream dispersal (drift) of the round goby (Neogobius melanostomus) and tubenose goby (Proterorhinus semilunaris) in newly colonised areas (River Dyje, Danube basin), a factor that could play an important role in their dispersal. Drift duration (i.e. number of dates on which the species was caught) in round goby was longer than that of both native species and tubenose goby, providing a possible advantage. Size of drifting fish was restricted to a narrow range of 6–8 mm and 5–8 mm for round and tubenose goby, respectively. Drift in both species occurred almost completely during hours of darkness (<1 lux) and fish size did not vary throughout the night. In both species, drift density increased significantly during the first hour after dusk. Round goby density gradually decreased up to dawn, while tubenose goby density varied throughout the night with no clear pattern. Drift of early life stages appears to be an important phenomenon that has not received adequate attention in studies of round and tubenose goby ecology and dispersal.  相似文献   
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In this study the sensitivity of three methods of isolation of udder pathogens from milk samples from subclinical mastitis cases was compared. For analysis 1827 quarter milk samples were selected. Milk was cultured using a standard culture technique (0.01 ml of fresh milk streaked on a sheep blood agar plate and on Edward's medium). In addition, an inoculum of 0.01 ml of the original milk sample was incubated for 24h at 37 degrees C in broth, followed by culture using the standard culture technique. In the third method, the whole milk sample was frozen for 24h, and then incubated for 24h at 37 degrees C, followed by culture using standard culture technique. The isolation percentage of Staphylococcus aureus was 4.7% for standard culture technique, 14.2% for incubation in broth, and 21.5% for the combination of freezing plus incubation. Isolation percentage of Streptococcus dysgalactiae and Streptococcus agalactiae was highest using the standard culture technique, while isolation rate of Streptococcus uberis was not different among the three methods used. With increasing somatic cell count, the likelihood of S. aureus, S. dysgalactiae and S. uberis isolation increased.Based on the relative sensitivity, defined as the isolation rate using a single technique compared to the isolation rate of the three techniques together, a combination of standard culture technique and freezing plus incubation was most attractive for achieving a high isolation rate of S. agalactiae and S. dysgalactiae. Relative sensitivity of S. uberis isolation was highest using the standard culture technique and incubation in broth, while S. aureus was most often isolated using a combination of incubation in broth and freezing plus incubation. A combination of the three methods increased the isolation rate for S. dysgalactiae, S. uberis and S. aureus. The standard culture technique, together with the combination of freezing plus incubation, can be recommended for isolating major udder pathogens. If S. aureus is the pathogen of main interest, using incubation in broth together with the combination of freezing plus incubation performed best.  相似文献   
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ABSTRACT Diaporthe phaseolorum and Phomopsis longicolla isolates from soybean were examined using traditional mycological characteristics and molecular methods. Cultural characteristics including types of fruiting bodies and conidia were assessed for isolates collected from soybean stems and seeds. Cultures were identified as P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, or D. phaseolorum var. sojae. Molecular markers for these groups were developed and analyzed using polymerase chain reaction restriction fragment length polymorphisms (PCR-RFLP) and DNA sequencing in the internal transcribed spacer (ITS) and the 5.8S ribosomal DNA. The ITS(4) and ITS(5) primers amplified PCR products for all isolates studied. Gel electrophoresis of undigested PCR products and DNA sequencing produced various fragment lengths including 604 bp for P. longicolla, 602 and 603 bp for D. phaseolorum var. caulivora, 603 bp for D. phaseolorum var. meridionalis, and from 597 to 609 bp for D. phaseolorum var. sojae. Digestion of these PCR products with enzymes AluI, HhaI, MseI, RsaI, and ScrFI resulted in distinct bands for identification of P. longicolla and the varieties of D. phaseolorum I. All P. longicolla, D. phaseolorum var. caulivora, and D. phaseolorum var. meridionalis isolates were distinguished using AluI and HhaI with RsaI or ScrFI. The banding patterns of D. phaseolorum var. sojae isolates were complex and were separated into 11 subgroups after digestion with AluI, HhaI, MseI, RsaI, and ScrFI. Phylogenetic analysis of 20 isolates of D. phaseolorum and P. longicolla based on the DNA sequence of the ITS region resolved six clades termed A, B, C, D, E, and F. Clade A included all sequenced D. phaseolorum var. caulivora isolates, two from Italy and one from the United States. Isolates in clade B were exclusively associated with D. phaseolorum var. meridionalis. Clades A and B formed a well-supported monophyletic group. Isolates in clades C, D, E, and F were morphologically defined as isolates of P. longicolla, D. phaseolorum var. sojae, and Diaporthe spp. The ITS sequences similarity of seven geographically diverse P. longi-colla isolates illustrated that P. longicolla isolates have a similar genetic background, with some affiliations to some D. phaseolorum var. sojae isolates. Morphological characteristics of the isolates along with the terminal clades of the ITS phylogeny suggest that P. longicolla is an individual species, D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis are varieties of D. phaseolorum, and D. phaseolorum var. sojae is either several varieties of D. phaseolorum or possibly several distinct species.  相似文献   
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