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11.
12.
自CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9)基因
组编辑技术发现以来,迅速在作物中得到广泛应用。但是,CRISPR/Cas9多基因编辑系统在大豆中的研究尚待开
发。本文利用CRISPR/Cas9介导的多基因编辑系统,分别构建了两个载体,一个载体含6个靶点,编辑7个大豆基因
(4个Glycine max ASYMMETRIC LEAVES1(GmAS1)同源基因和3个GmAS2 同源基因),另一个载体含8个靶点,编辑
11个G. max AGAMOUS 家族同源基因(4个GmAG 同源基因,2个G. max SEEDSTICK(GmSTK)同源基因和5个G. max
SHATTERPROOF1(GmSHP1/2)同源基因)。大豆遗传转化后,经表型鉴定和靶点检测发现,CRISPR/Cas9介导的多
基因编辑系统在大豆中成功实现了多基因编辑。当3个GmAS1 同源基因和3个GmAS2 同源基因同时突变时,导致
大豆叶片向远轴面弯曲、皱缩且叶柄变短的表型。当2个GmSHP1 同源基因和2个GmSTK 同源基因同时突变时,导
致豆荚停止发育的不育表型。 相似文献
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Yuhua Fu Pengyu Fan Lu Wang Ziqiang Shu Shilin Zhu Siyuan Feng Xinyun Li Xiaotian Qiu Shuhong Zhao Xiaolei Liu 《Journal of animal science》2021,99(2)
Despite the broad variety of available microRNA (miRNA) research tools and methods, their application to the identification, annotation, and target prediction of miRNAs in nonmodel organisms is still limited. In this study, we collected nearly all public sRNA-seq data to improve the annotation for known miRNAs and identify novel miRNAs that have not been annotated in pigs (Sus scrofa). We newly annotated 210 mature sequences in known miRNAs and found that 43 of the known miRNA precursors were problematic due to redundant/missing annotations or incorrect sequences. We also predicted 811 novel miRNAs with high confidence, which was twice the current number of known miRNAs for pigs in miRBase. In addition, we proposed a correlation-based strategy to predict target genes for miRNAs by using a large amount of sRNA-seq and RNA-seq data. We found that the correlation-based strategy provided additional evidence of expression compared with traditional target prediction methods. The correlation-based strategy also identified the regulatory pairs that were controlled by nonbinding sites with a particular pattern, which provided abundant complementarity for studying the mechanism of miRNAs that regulate gene expression. In summary, our study improved the annotation of known miRNAs, identified a large number of novel miRNAs, and predicted target genes for all pig miRNAs by using massive public data. This large data-based strategy is also applicable for other nonmodel organisms with incomplete annotation information. 相似文献
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[目的]研究虾肽有机肥(SPOF)与磷肥配施的协同增效作用。[方法]采用田间小区试验,设尿素1.4 kg/株(CK)、尿素1.4 kg/株+农家肥50 kg/株+磷酸二氢钾1.54 kg/株(TT1)、尿素1.4 kg/株+SPOF 30 kg/株+磷酸二氢钾1.54 kg/株(TT2)、尿素1.4 kg/株+SPOF 30 kg/株+普通过磷酸钙0.83 kg/株+磷酸二氢钾1.25 kg/株(TT3)。[结果]各处理的产量TT3>TT2>TT1>CK,且TT3达到了显著水平;TT3的品质指标,即可溶性固形物、维生素C、蔗糖、葡萄糖以及硬度均达到了显著水平。[结论]TT3处理虾肽有机肥和磷肥(SSP+KH2PO4)协同将其效力发挥到最大。 相似文献
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旨在维护国家稳定,为预判粮食生产前景、提高粮食生产效率、保障粮食安全提供理论依据。利用湖南省统计数据,运用灰色关联分析法筛选关联性较强的影响因素,并建立GM(1,N)预测模型预测粮食产量。2008—2017年与湖南省粮食产量关联度最大的影响因素是粮食作物播种面积和农业机械总动力;科技因素是影响2008—2017年湖南省粮食产量的主要因素,其次是自然因素,社会因素;2018—2027年湖南省粮食产量有较小波动,且农业机械总动力和财政农业支出影响较大;农业机械总动力在前后十年对粮食产量都有较重要的影响,越来越占据主导地位。粮食产量受国家政策的影响,受农业机械总动力影响最大,维持产量水平需高度重视农业机械化水平,稳步提高粮食作物播种面积。 相似文献
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AIM:To investigate the effect of HMGA2 down-regulation on apoptosis and Notch signaling pathway in renal tubular epithelial cells exposed to high glucose (HG). METHODS:D-glucose at 5, 10, 20 and 30 mmol/L was used to stimulate human renal tubular epithelial HK-2 cells for 2 h, and D-glucose at 30 mmol/L was used to stimulate the HK-2 cells for 10 min, 60 min and 120 min. The protein expression of HMGA2 was determined by Western blot. The HK-2 cells were divided into normal glucose (NG) group, HG group, HG+si-HMGA2 group and HG+NC group, in which siRNA was transfected by LipofectamineTM 2000 for 48 h. Flow cytometry was used to analyze the apoptotic rate, reactive oxygen species (ROS) assay kit was used to detect ROS content, and Western blot was used to detect the protein levels of Notch1, Hes1 and Bcl-2. The HK-2 cells were treated with the Notch signaling pathway inhibitor DAPT, and then the cells were divided into HG group, HG+DAPT group and HG+si-HMGA2+DAPT group. The apoptotic rate was analyzed by flow cytometry. RESULTS:Exposure of the HK-2 cells to D-glucose at different concentrations for different time significantly increased the expression of HMGA2 (P<0.05). Compared with NG group, the protein expression of HMGA2, Notch1 and Hes1 in HG group was increased, the expression of Bcl-2/Bax was decreased, the apoptotic rate was increased, and the content of ROS was increased obviously (P<0.05). Compared with HG group, the protein expression of HMGA2, Notch1 and Hes1 of HG+si-HMGA2 group was decreased, the expression of Bcl-2/Bax was increased, the apoptotic rate was decreased, and the content of ROS was decreased significantly (P<0.05). The apoptotic rate in HG+DAPT group was significantly lower than that in HG group, while the apoptotic rate in HG+si-HMGA2+DAPT group was significantly lower than that in HG+DAPT group (P<0.05). CONCLUSION:Down-regulation of HMGA2 expression inhibits the apoptosis of renal tubular epithelial cells by regulating Notch signaling pathway and decreasing ROS production. 相似文献
20.
AIM:To study the role of ghrelin in cell protection by up-regulating heat shock protein 70 (HSP70) and inhibiting apoptosis induced by oxidative stress through extracellular regulated protein kinases 1/2 (ERK1/2) signaling pathway in the PC12 cells. METHODS:Sodium nitoprusside (SNP) was used to induce oxidative stress injury in the PC12 cells. The cultured PC12 cells were divided into SNP-injured group (incubated with SNP at 0.5 mmol/L for 6, 12, 18 and 24 h), ghrelin pretreatment group (ghrelin at 100 nmol/L was given 30 min before adding SNP); HSP70 inhibitor group (quercetin at 10 μmol/L was added 60 min before ghrelin treatment), ERK inhibitor group (ERK 1/2 inhibitor PD98059 was added 60 min before ghrelin treatment) and control group (added same amount of culture medium only). The apoptotic rate was detected by flow cytometry. The protein expression was determined by Western blot and immunocytochemistry. RESULTS:Compared with control group, the apoptotic rate of PC12 cells in SNP-injured group was significantly increased (P<0.05). Compared with SNP-injured group, ghrelin (100 nmol/L) pretreatment significantly inhibited SNP-induced apoptosis of PC12 cells (P<0.05), and significantly up-regulated the protein expression of HSP70 (P<0.05). Time-effect analysis showed that ghrelin had the most significant effect at 18 h after SNP injury. Quercetin, an inhibitor of HSP 70, significantly reduced the anti-apoptotic effect of ghrelin (P<0.05). Ghrelin pretreatment promoted the phosphorylation of ERK1/2. ERK1/2 inhibitor PD98059 significantly inhibited the effects of ghrelin on up-regulation of HSP70 expression (P<0.05). CONCLUSION:Ghrelin upregulates the expression of HSP70 and inhibits the apoptosis in the PC12 cells induced by oxidative stress by promoting the phosphorylation of ERK1/2. 相似文献