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Single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD) requires high efficiency and accuracy. Allele dropout (ADO), the random amplification failure of one of the two parental alleles, remains the most significant problem in PCR-based PGD testing since it can result in serious misdiagnosis for compound heterozygous or autosomal dominant conditions. A number of different strategies (including the use of lysis buffers to break down the cell and make the DNA accessible) have been employed to combat ADO with varying degrees of success, yet there is still no consensus among PGD centres over which lysis buffer should be used (ESHRE PGD Consortium, 1999 ). To address this issue, PCR amplification of three genes (CFTR, LAMA3 and PKP1) at different chromosomal loci was investigated. Single lymphocytes from individuals heterozygous for mutations within each of the three genes were collected and lysed in either alkaline lysis buffer (ALB) or proteinase K/SDS lysis buffer (PK). PCR amplification efficiencies were comparable between alkaline lysis and proteinase K lysis for PCR products spanning each of the three mutated loci (ΔF508 in CFTR 90% vs 88%; R650X in LAMA3 82% vs 78%; and Y71X in PKP1 91% vs 87%). While there was no appreciable difference between ADO rates between the two lysis buffers for the LAMA3 PCR product (25% vs 26%), there were significant differences in ADO rates between ALB and PK for the CFTR PCR product (0% vs 23%) and the PKP1 PCR product (8% vs 56%). Based on these results, we are currently using ALB in preference to PK/SDS buffer for the lysis of cells in clinical PGD. Copyright © 2001 John Wiley & Sons, Ltd.  相似文献   
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Developing a relationship between pest abundance and damage to crops is essential for the calculation of economic injury levels (EIL) leading to informed management decisions. The crop modelling framework, APSIM, was used to simulate the impact of mouse damage on yield loss on wheat where a long-term dataset on the density of mice was available (1983–2003). The model was calibrated using results from field trials where wheat plants were hand clipped to imitate mouse damage. The grazing effect of mice was estimated using the population density, daily intake per mouse and the proportion of wheat grain and plant tissue in the diet to determine yield loss. The mean yield loss caused by mice was 12.4% (±5.4S.E.; range −0.5 to 96%). There were 7/21 years when yield loss was >5%. A damage/abundance relationship was constructed and a sigmoidal curve explained 97% of variation when accounting for different trajectories of mouse densities from sowing to harvest. The majority of damage occurred around emergence of the crop when mouse densities were >100 mice ha−1. This is the first time that field data on mouse density and a crop simulation model have been combined to estimate yield loss. The model examines the efficacy of baiting and how to estimate EILs. Because the broadscale application of zinc phosphide is cheap and effective, the EIL is very low (<1% yield loss). The APSIM model is highly flexible and could be used for other vertebrate pests in a range of crops or pastures to develop density/damage relationships and to assist with management.  相似文献   
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Chinese softshell turtles (Pelodiscus spp.) are widely distributed, ranging from the Amur and Ussuri Rivers in the Russian Far East through the Korean Peninsula, Japan, and eastern, central, and southern China to southern Vietnam. In East and Southeast Asia, Chinese softshell turtles are traditionally exploited for food and have been farm-bred in China since the Spring and Autumn Period, more than 2400 years ago. Currently, the annual production of Pelodiscus amounts to 340,000 t in China alone. Using mitochondrial DNA (2428 bp) and five nuclear loci (3704 bp), we examined broad sampling of wild and farm-bred Pelodiscus to infer genetic and taxonomic differentiation. We discovered four previously unknown mitochondrial lineages, all from China. One lineage from Jiangxi is deeply divergent and sister to the mitochondrial lineage of Pelodiscus axenaria. The nuclear loci supported species status for P. axenaria and the new lineage from Jiangxi. Pelodiscus maackii and P. parviformis, both harboring distinct mitochondrial lineages, were not differentiated from P. sinensis in the studied nuclear markers. The same is true for two new mitochondrial lineages from Zhejiang, China, represented by only one individual each, and another new lineage from Anhui, Guangdong, Jiangxi and Zhejiang, China. However, Vietnamese turtles yielding a mitochondrial lineage clustering within P. sinensis were distinct in nuclear markers, suggesting that these populations could represent another unknown species with introgressed mitochondria. Its species status is also supported by the syntopic occurrence with P. sinensis in northern Vietnam and by morphology. In addition, we confirmed sympatry of P. axenaria and P. parviformis in Guangxi, China, and found evidence for sympatry of P. sinensis and the new putative species from Jiangxi, China. We also discovered evidence for hybridization in turtle farms and for the occurrence of alien lineages in the wild (Zhejiang, China), highlighting the risk of genetic pollution of native stock. In the face of the large-scale breeding of Pelodiscus, we claim that the long-term survival of distinct genetic lineages and species can only be assured when an upscale market segment for pure-bred softshell turtles is established, making the breeding of pure lineages lucrative for turtle farms. Our findings underline that the diversity of Pelodiscus is currently underestimated and threatened by anthropogenic admixture. We recommend mass screening of genetic and morphological variation of Chinese softshell turtles as a first step to understand and preserve their diversity.  相似文献   
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Meeting the growing demand for food in the future will require adaptation of water and land management to future conditions. We studied the extent of different adaptation options to future global change in the Mediterranean region, under scenarios of water use and availability. We focused on the most significant adaptation options for semiarid regions: implementing irrigation, changes to cropland intensity, and diversification of cropland activities. We used Conversion of Land Use on Mondial Scale (CLUMondo), a global land system model, to simulate future change to land use and land cover, and land management. To take into account future global change, we followed global outlooks for future population and climate change, and crop and livestock demand. The results indicate that the level of irrigation efficiency improvement is an important determinant of potential changes in the intensity of rain-fed land systems. No or low irrigation efficiency improvements lead to a reduction in irrigated areas, accompanied with intensification and expansion of rain-fed cropping systems. When reducing water withdrawal, total crop production in intensive rain-fed systems would need to increase significantly: by 130% without improving the irrigation efficiency in irrigated systems and by 53% under conditions of the highest possible efficiency improvement. In all scenarios, traditional Mediterranean multifunctional land systems continue to play a significant role in food production, especially in hosting livestock. Our results indicate that significant improvements to irrigation efficiency with simultaneous increase in cropland productivity are needed to satisfy future demands for food in the region. The approach can be transferred to other similar regions with strong resource limitations in terms of land and water.  相似文献   
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