首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   25376篇
  免费   1253篇
  国内免费   2405篇
林业   1564篇
农学   2159篇
基础科学   676篇
  3553篇
综合类   9969篇
农作物   1456篇
水产渔业   1058篇
畜牧兽医   5383篇
园艺   2041篇
植物保护   1175篇
  2024年   111篇
  2023年   400篇
  2022年   758篇
  2021年   992篇
  2020年   913篇
  2019年   1128篇
  2018年   708篇
  2017年   1099篇
  2016年   1395篇
  2015年   1253篇
  2014年   1372篇
  2013年   1614篇
  2012年   1971篇
  2011年   1980篇
  2010年   1651篇
  2009年   1670篇
  2008年   1457篇
  2007年   1664篇
  2006年   1321篇
  2005年   955篇
  2004年   794篇
  2003年   611篇
  2002年   489篇
  2001年   404篇
  2000年   368篇
  1999年   340篇
  1998年   204篇
  1997年   193篇
  1996年   186篇
  1995年   186篇
  1994年   124篇
  1993年   130篇
  1992年   106篇
  1991年   95篇
  1990年   84篇
  1989年   66篇
  1988年   60篇
  1987年   45篇
  1986年   36篇
  1985年   25篇
  1984年   14篇
  1983年   8篇
  1982年   5篇
  1981年   6篇
  1980年   7篇
  1978年   5篇
  1977年   4篇
  1962年   6篇
  1956年   12篇
  1955年   3篇
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
1.
A 4‐week feeding trial was conducted using five groups of hybrid grouper (61.15 ± 0.15 g) to explore the potential effects of three methionine (Met) sources. Five isonitrogenous and isolipidic diets were formulated that included a fishmeal (FM) diet; nonsupplemented (NS) diet; or NS diets with the addition of the L‐methionine (L‐Met), DL‐methionine (DL‐Met), or coated Met (Co‐Met) to obtain the same Met level as the FM diet. Fish were randomly distributed into sea cages (30 fish per cage). Weight gain and specific growth rate in the DL‐Met group and FM group were significantly higher than those in all other groups (p < .05). In the proximal and distal intestines, of the 20 gene and time combinations (10 per gene), there were 14 combinations (70%) in which there were no significant differences in gene expression levels between the FM and DL‐Met groups (p > .05). The main reason for the same growth effect between the DL‐Met and FM groups may be attributed to the synchronized absorption at most time points after feeding, which had similar expression patterns of B0AT1 and ASCT2 of the proximal and distal intestines between two groups.  相似文献   
2.
3.
吕珽  陈虹吟  汤承  岳华 《畜牧兽医学报》2021,52(8):2361-2368
旨在调查川西北牦牛哺乳动物正呼肠孤病毒(MRV)的感染情况并分离病毒。采用RT-PCR方法,对采自川西北15个牧场的72份牦牛腹泻粪便样本和其中5个牧场的15份腹泻牦牛血清样本进行MRV检测,阳性样本进一步用分型PCR确定其血清型。结果显示,粪便样本中MRV检出率为20.83%(15/72),血清2型的比例为60%(9/15);血清样本中MRV检出率为40%(6/15),血清2型的比例为83.33%(5/6);未检测到其他血清型。成功地从腹泻粪便中分离到1株MRV血清2型毒株(TCID50为4×10-8.56·mL-1),并获得长度为23 587 bp的分离株全基因组,该分离株与中国猪源毒株的遗传关系最近;与GenBank中所有的MRV S1基因相比,该分离株有4个独特的氨基酸突变。本研究从牦牛中检测到MRV,并分离到1株牛源MRV血清2型毒株,为进一步研究MRV血清2型生物学特性奠定了基础。  相似文献   
4.
Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.  相似文献   
5.
【目的】干旱是严重影响玉米生长发育进程的一个重要因素。挖掘玉米抗旱相关基因,通过转基因功能验证和转录组分析,解析关键基因在响应干旱胁迫过程中的分子调控机制,为抗旱分子育种和遗传改良提供理论依据。【方法】以玉米自交系B104(WT)为背景材料,利用农杆菌介导方法构建过表达ZmIBH1-1转基因株系(ZmIBH1-1-OE);通过对转基因植株进行草铵膦抗性筛选、标记基因和目的基因PCR检测,以及运用实时荧光定量PCR检测目的基因的表达情况,鉴定阳性植株和株系;以WT和ZmIBH1-1-OE转基因株系为材料,通过干旱处理(20% PEG6000),进行表型鉴定和耐旱生理生化指标测定,验证ZmIBH1-1的抗旱功能;通过对干旱胁迫下玉米4叶期转录组的比较分析,鉴定出差异表达的基因(differentially expressed genes,DEGs);结合DAP-seq(DNA affinity purification sequencing)分析,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的下游靶基因,利用基因组可视化软件IGV(integrative genomics viewer)分析ZmIBH1-1蛋白结合候选靶基因的位置,然后通过Dual-Luciferase试验验证ZmIBH1-1蛋白与靶基因的调控关系。【结果】通过玉米遗传转化获得12个转化事件;T3代中,能同时检测到标记基因Bar和目的基因ZmIBH1-1的植株有458个,实时荧光定量PCR检测结果表明,ZmIBH1-1-OE中ZmIBH1-1的表达量显著高于WT,株系3和株系8表达量最高,将其自交获得T4代转基因株系用于后续试验。在干旱胁迫条件下,ZmIBH1-1-OE株系存活率、叶片相对含水量、叶绿素含量、可溶性蛋白含量及其生理生化指标(超氧化物歧化酶、过氧化物酶、过氧化氢酶活性)均显著高于WT,说明玉米中过量表达ZmIBH1-1赋予玉米更高的耐旱性。转录组分析结果表明,WT与ZmIBH1-1-OE株系在干旱胁迫下有1 214个差异表达基因;Gene Ontology(GO)功能富集分析结果表明,差异表达基因主要涉及生物过程、细胞组分和分子功能,如在生物过程中主要涉及到光合作用、应激响应、脱水响应等;KEGG富集分析表明,差异表达基因主要参与植物激素信号传导、新陈代谢等过程。结合转录组显著差异表达基因和DAP-Seq分析所得到ZmIBH1-1蛋白的靶基因,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的11个候选靶基因,包括2个钙信号相关基因、3个半胱氨酸代谢相关基因、1个bHLH转录因子、1个应激响应蛋白、1个谷胱甘肽转移酶、1个氧化还原过程蛋白和2个乙烯响应因子;基因组可视化结果显示ZmIBH1-1蛋白可以结合靶基因启动子区;随后通过Dual-Luciferase试验进一步表明,ZmIBH1-1蛋白可以直接作用于11个候选靶基因,其中,ZmIBH1-1蛋白可以促进ZmCa-MZmSYCOZmbHLH54ZmGlu-r1ZmCLPB3ZmP450-99A2的表达,抑制ZmAGD12ZmCYSZmCYSBZmERF-107ZmEIN3的表达。此外,在干旱胁迫下NAC、WRKY、MYB等转录因子在ZmIBH1-1-OE和WT株系中也存在差异表达。【结论】ZmIBH1-1的过表达可以增强玉米苗期的耐旱性;ZmIBH1-1蛋白通过直接调控乙烯信号通路中的ZmERF-107ZmEIN3的表达提高玉米的耐旱性;ZmIBH1-1蛋白通过直接调控钙信号相关基因ZmCa-MZmAGD12增强玉米的耐旱性;ZmIBH1-1蛋白可能通过间接调控NAC、WRKY、MYB等转录因子响应干旱胁迫。  相似文献   
6.
Street trees provide shade and increase human thermal comfort during hot summer. In this study, we investigated the spatial distribution of shade provision of street trees in Boston, Massachusetts. The sky view factor (SVF), which influences the solar radiation to the ground and affects human thermal comfort, was used to indicate the contribution of street trees on shade provision. Google Street View (GSV) panoramas were used to calculate the photographic method based SVF (SVFP), with the consideration of all kinds of obstructions within street canyons. A building height model was used to calculate the simulation based SVF (SVFs), with consideration of obstruction of building blocks only. Considering the fact that street trees and building blocks are the two major obstructions of radiation within street canyons, therefore, the difference between the two SVF estimation results can be considered as the shade provision of street trees. The results show that street trees help to decrease the SVF by 24.61% in Boston, Massachusetts. The shading level varies spatially in the study area. Generally, the southwestern area has much higher shading level than the north and the east. We further explored the shading variation among different socioeconomic groups in the study area. Result shows that Hispanics tend to live in neighborhoods with lower shading level. This study can help to provide a reference for future urban greening projects for global climate change adaption.  相似文献   
7.
8.
通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。  相似文献   
9.
We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.  相似文献   
10.
CAO Rui-ping  WANG Jiao  WANG Ce 《园艺学报》2018,34(6):1061-1066
AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP+)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP+-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP+ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP+ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP+-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.  相似文献   
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号