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51.
    
Genome editing, which is an unprecedented technological breakthrough, has provided a valuable means of creating targeted mutations in plant genomes. In this study, we developed a genomic web tool to identify all gRNA target sequences in the coffee genome, along with potential off-targets. In all, 8,145,748 CRISPR guides were identified in the draft genome of Coffea canephora corresponding to 5,338,568 different sequences and, of these, 4,655,458 were single, and 514,591 were covering exons. The proof of concept was established by targeting the phytoene desaturase gene (CcPDS) using the Agrobacterium tumefaciens transformation technique and somatic embryogenesis as the plant regeneration method. An analysis of the RNA-guided genome-editing events showed that 22.8% of the regenerated plants were heterozygous mutants and 7.6% were homozygous mutants. Mutation efficiency at the target site was estimated to be 30.4%. We demonstrated that genome editing by the CRISPR/Cas9 method is an efficient and reliable way of knocking out genes of agronomic interest in the coffee tree, opening up the way for coffee molecular breeding. Our results also showed that the use of somatic embryogenesis, as the method for regenerating genome-edited plants, could restrict the choice of targeted genes to those that are not essential to the embryo development and germination steps.  相似文献   
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Interactions within and between species sharing the same resources are characterised by competition or facilitation, and can be influenced by factors such as larval numbers and phenotypic plasticity of the interactions. The effect of larval density on the survival and relative growth rate of the stemborers Busseola fusca (Fuller) and Sesamia calamistis Hampson (both Lepidoptera: Noctuidae), and Chilo partellus (Swinhoe) (Lepidoptera: Crambidae) were studied, as well as the temporal plasticity of their competitive interactions. These stemborers attack maize crops (Zea mays L.) (Poaceae) in sub‐Saharan Africa. Experiments were conducted in the laboratory under controlled conditions at the optimum development temperature (25 °C) for the three species. Surrogate stems filled with artificial diet were intra‐ and interspecifically infested with larvae of each species. The effect of larval density on competition was studied at low (six larvae) and high (12 larvae) levels of infestation, whereas the temporal plasticity of competition was evaluated at 7, 14, 21, or 28 days after infestation. The two experiments involved single‐ and multi‐species infestation treatments. Larval numbers and wet mass in each artificial stem were recorded in each experiment. Survival and relative growth rate of the three species were significantly higher at low‐infestation levels when facing either intra‐ or interspecific competition. The intensity of competition was also temporally plastic among the species and increased as the duration of competition increased. These results are discussed in terms of general infestations of cereal crops by borers.  相似文献   
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According to mitochondrial data, the common mole, Talpa europaea, is paraphyletic. This could be explained by either an ancient introgression of mtDNA from the Iberian blind mole T. occidentalis to T. europaea, or the existence of a differentiated taxonomic entity in northern Spain that needs to be described. In this study, we combined mitochondrial (Cytb) and nuclear (HDAC2) data to investigate these two alternative hypotheses. Based on both mitochondrial and nuclear data and an extensive geographical sampling (399 sequenced individuals), we show that the populations of T. europaea from Spain and south‐western France (south of the Loire River) are phylogenetically closer to T. occidentalis than to T. europaea. The Spanish–French lineage has some morphological characters resembling more to T. occidentalis (e.g. eyes) and others resembling more to T. europaea (external measurements, mesostyle of the first upper molar). It also seems to have several distinctive dental characters, suggesting that it should be recognized as a new species. Within the three lineages, we found a marked phylogeographical pattern, with several allopatric or parapatric lineages, dating from the Pleistocene. Our genetic data combined with species distribution models support the presence of several putative glacial refugia during glacial maxima for each species.  相似文献   
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How the variation in phenotypic traits like cell size and motility impacts predator-induced cellular aggregation is not known. Furthermore the genetic composition of cell groups in mixed populations of Chlamydomonas has not been investigated. An examination of these two questions will not only enhance our understanding of Chlamydomonas ecology, but also shed light on the primordial steps before integrated multicellular groups were established. Group living comes with viability and reproductive costs and it is not known how these are shared if groups are genetically heterogeneous. We observed that the natural predator Peranema trichophorum (Euglenoidea) induced clumping in Chlamydomonas. When co-cultured with P. trichophorum cells protected themselves by forming facultative groups (reverting back to a unicellular lifestyle once predators were removed). The dynamics of group formation in different Chlamydomonas species and strains correlated with cell size and swimming speed. Small or less motile strains aggregated more readily than large, fast-swimming ones. Interestingly, Chlamydomonas groups were both intra-species and inter-species chimaeric. This suggests that the predator-induced group formation in Chlamydomonas involved cells coming together rather than staying together and during aggregation cells showed little or no discrimination between self and non-self. These data demonstrate that the dynamics of cell aggregation, in unicellular volvocines at least, depends on phenotypic traits like cell size and motility and high genetic relatedness is not mandatory at this initial stage. These findings further our understanding of aggregation in mixed Chlamydomonas populations and have implications for understanding the very first steps on the road to simple multicellularity.  相似文献   
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Tropospheric ozone (O3) produces harmful effects to forests and crops, leading to a reduction of land carbon assimilation that, consequently, influences the land sink and the crop yield production. To assess the potential negative O3 impacts to vegetation, the European Union uses the Accumulated Ozone over Threshold of 40 ppb (AOT40). This index has been chosen for its simplicity and flexibility in handling different ecosystems as well as for its linear relationships with yield or biomass loss. However, AOT40 does not give any information on the physiological O3 uptake into the leaves since it does not include any environmental constraints to O3 uptake through stomata. Therefore, an index based on stomatal O3 uptake (i.e. PODY), which describes the amount of O3 entering into the leaves, would be more appropriate. Specifically, the PODY metric considers the effects of multiple climatic factors, vegetation characteristics and local and phenological inputs rather than the only atmospheric O3 concentration. For this reason, the use of PODY in the O3 risk assessment for vegetation is becoming recommended. We compare different potential O3 risk assessments based on two methodologies (i.e. AOT40 and stomatal O3 uptake) using a framework of mesoscale models that produces hourly meteorological and O3 data at high spatial resolution (12 km) over Europe for the time period 2000–2005. Results indicate a remarkable spatial and temporal inconsistency between the two indices, suggesting that a new definition of European legislative standard is needed in the near future. Besides, our risk assessment based on AOT40 shows a good consistency compared to both in‐situ data and other model‐based datasets. Conversely, risk assessment based on stomatal O3 uptake shows different spatial patterns compared to other model‐based datasets. This strong inconsistency can be likely related to a different vegetation cover and its associated parameterizations.  相似文献   
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A total of 1347 weaned lambs from eight genotypes were tested over five consecutive years: Romanov (ROM) and Lacaune (LAC) pure breeds, the two F1 crossbreeds (RL and LR) and the offspring of ewes from these four genotypes sired with Berrichon-du-Cher rams (BCF). The lambs were individually exposed to three challenging tests involving novelty, human contact and social isolation. Ten synthetic variables were used to express social reactivity (i.e., active vs. passive strategy), exploratory activity and reactivity to humans. BCF crossbreds were more active (i.e., high bleats, locomotion and attempts to escape) than purebreds and F1. In contrast, ROM expressed more passive responses (i.e., low bleats and vigilance postures) than LAC and BCF crossbreds. In addition, ROM approached a motionless human less and had longer flight distances to an approaching human than did LAC and BCF crossbreds. When restrained, ROM, and to a lesser extent B×ROM and B×LR, avoided human contact more than did LAC, RL and B×LAC. Most of these differences were explained by direct additive genetic effects while maternal influences or heterosis effects were rarely significant. The highest heritability was for high bleats (h2 = 0.48). Females were more active and avoided human contact more than did males.  相似文献   
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