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   5528篇
  免费   296篇
  国内免费   457篇
林业   224篇
农学   693篇
基础科学   15篇
  351篇
综合类   2323篇
农作物   391篇
水产渔业   546篇
畜牧兽医   1124篇
园艺   304篇
植物保护   310篇
  2024年   13篇
  2023年   46篇
  2022年   95篇
  2021年   129篇
  2020年   118篇
  2019年   135篇
  2018年   105篇
  2017年   161篇
  2016年   219篇
  2015年   170篇
  2014年   244篇
  2013年   246篇
  2012年   342篇
  2011年   423篇
  2010年   377篇
  2009年   399篇
  2008年   402篇
  2007年   418篇
  2006年   368篇
  2005年   291篇
  2004年   259篇
  2003年   225篇
  2002年   175篇
  2001年   160篇
  2000年   154篇
  1999年   114篇
  1998年   83篇
  1997年   78篇
  1996年   58篇
  1995年   71篇
  1994年   55篇
  1993年   26篇
  1992年   41篇
  1991年   33篇
  1990年   18篇
  1989年   13篇
  1988年   4篇
  1987年   5篇
  1985年   2篇
  1983年   1篇
  1980年   1篇
  1962年   1篇
  1955年   3篇
排序方式: 共有6281条查询结果,搜索用时 62 毫秒
31.
为了研究SCoT分子标记技术对甜菜种质资源鉴定的可行性。利用80条SCoT引物对48份甜菜种质资源进行鉴别,同时对种质资源进行了遗传多样性分析和亲缘关系的鉴定。结果表明,80条SCoT引物中有6条能够扩增出清晰、且多态性高的条带,分别为SCoT1、SCoT12、SCoT13、SCoT14、SCoT17和SCoT23,其中引物SCoT1、SCoT12、SCoT14和SCoT23单独使用均可鉴别全部的48份种质资源,引物SCoT13和SCoT17共同使用可以鉴别48个种质资源;聚类分析结果表明,在遗传距离0.15处,95.8%的种质资源均聚为一类,从分子角度上表明甜菜种质资源遗传距离较小。本研究为利用SCoT分子标记技术鉴别甜菜种质资源、对种质资源进行亲缘关系鉴定、杂交组合亲本选配以及分子标记辅助育种等提供相关科学依据。  相似文献   
32.
Quality control and screening of active substances in traditional Chinese medicines have been performed using fingerprint analysis. The spectrum-effect relationship between chromatography fingerprints and efficacy of herbal drugs is considered as a potentially useful method for determining active ingredients in complex mixtures. The study was designed to develop a method for determining the bioactive components of a compound Chinese medicine called Tiefeng based on spectrum-effect relationships between high-performance liquid chromatography (HPLC) fingerprints and anticoccidial activities. Four peaks of the established HPLC fingerprint indicate the main bioactive components of this medicine. In addition, pharrnacodynamic atlas was defined and used to assess the anticoccidial activity of Tiefeng from different sources for the first time. We found that the level of anticoccidial activity of Tiefeng was consistent with the degree of similarity between the pharmacodynamic atlas and chromatogram of any sample. Furthermore, effect of this medicine was related with the main active constituents, along with the origin and the harvesting time.  相似文献   
33.
In the Gilgel Gibe catchment in Ethiopia, local farmers intensify land use on Planosols by adjusting a traditional soil burning practice known as guie. The burning practice used to be applied in a cycle of shifting cultivation. However, more recently, farmers burn small plots to make fertile seedbeds for Eucalyptus seedlings in the first year before these trees are transplanted to larger plots. The purpose of this research was to assess the physico‐chemical properties of Planosols that have been subjected to burning over the last 10 yrs and evaluate the contribution of guie to land‐use intensification of these soils. Transect studies and interviews of local farmers, followed by chemical, physical and micromorphological analyses of samples from selected plots were used to compare the soil properties of recently (0–2 yrs) and formerly (3–10 yrs) burnt Planosols with those of unburnt Planosols. The analytical results show that the burning practice improved nutrient availability in the first 2 yrs after guie. Increased amounts of exchangeable aluminium (Al) were reported in the long term. Charge fingerprints illustrate that the nutrient‐buffering capacity of the soil was high shortly after the practice but subsequently decreased with time. Given the population pressure on the formerly extensively used Planosols, it is argued that the current application of guie on small, localized plots for raising Eucalyptus seedlings is well adapted to the local socio‐economic context and promotes land‐use intensification on the Planosols. The increased exchangeable Al content of former Eucalyptus seedbeds merits further in‐depth research into the biophysical sustainability of the burning practice.  相似文献   
34.
In order to develop a promising vaccine candidate utilizing a combined approach to induce both antibody production and T-cell activity, the DNA fragment containing MA of HCV with five conserved epitopes was synthesized. Two types of HCV vaccine candidates (the DNA type and DNA/polymers) were constructed using MA. PLA-PEG-PLA and PLGA-PEG-PLGA were synthesized and used as micelles with encapsulated plasmid pcDNA3.1(+)-MA. The preparation of copolymers, the cloning and analysis of recombinant plasmid DNA, in vitro expression, and immunogenicity in transgenic mice were evaluated in detail. The results indicated that even single immunization and oral immunization with DNA/polymers achieved satisfying immune responses in vivo tests. As biodegradable and nontoxic triblock copolymers, the novel copolymers demonstrated a great advantage, as they made long-term and single-immunizing vaccines possible; in addition, the copolymers showed a better adjuvant effect and scarcely any side effects.  相似文献   
35.
建立菠萝 DNA 甲基化水平的 HPLC 测定方法,分析菠萝愈伤组织 DNA 甲基化水平变化,为进一步研究菠萝 体细胞无性系变异机理奠定基础。通过对流动相和水解温度等条件的优化,建立菠萝 DNA 甲基化水平的检测方法。结 果表明,分离 C 和 5m-C 的最佳流动相为甲醇∶磷酸二氢钾∶三乙胺为 10∶90∶0.2(V/V),pH 3.0,DNA 的最佳水解 温度为 90 ℃。利用此体系分析菠萝愈伤组织和胚性愈伤组织的 DNA 甲基化变化,结果表明,菠萝愈伤组织在分化过 程中 DNA 总甲基化水平呈动态变化,变化范围为 5.14%~96.86%。此外,胚性愈伤组织甲基化水平低于非胚性愈伤组 织。推测 DNA 甲基化影响菠萝愈伤组织的分化及胚性愈伤组织的形成。  相似文献   
36.
Soils encompass a huge diversity of organisms which mostly remains to be characterized due to a number of methodological and logistical issues. Nonetheless, remarkable progress has been made in recent years toward developing strategies to characterize and describe soil biodiversity, especially thanks to the development of molecular approaches relying on direct DNA extraction from the soil matrix.Metabarcoding can be applied to DNA from any environment or organism, and is gaining increasing prominence in biodiversity studies. This approach is already commonly used to characterize soil microbial communities and its application is now being extended to other soil organisms, i.e. meso- and macro-fauna.These developments offer unprecedented scientific and operational opportunities in order to better understand soil biodiversity distribution and dynamics, and to propose tools and strategies for biodiversity diagnosis. However, these opportunities also come with challenges that the scientific community must face. Such challenges are related to i) clarification of terminology, (ii) standardisation of methods and further methodological development for additional taxonomic groups, (iii) development of a common database, and (iv) ways to avoid waste of information and data derived from metabarcoding. In order to facilitate common application of metabarcoding in soil biodiversity assessment, we discuss these opportunities and challenges and propose solutions towards a more homogeneous framework.  相似文献   
37.
38.
To characterize the DNA rearrangement of both the T-DNA region and the genomic insertion site during T-DNA insertion, the Genomewalker strategy was used to isolate the junctions between the inserted DNA and the plant genomic DNA in six rapeseed events as well as the genomic DNA at the sites before integration. During transformation in each of the six events, portions of both the right border(RB) and left border(LB) regions of the T-DNA were deleted, ranging from a 7 nucleotide deletion of the LB repeats in event RF1 to a 207 bp deletion of the LB region in event RF2. For the six events, T-DNA integration resulted in a deletion at the target site spanning less than 100 bp. Sequence analysis indicated that the T-DNA was integrated into the coding region of various native rapeseed genes in events RF1 and RF2. Duplications of the genomic DNA target site were observed in events RF2, RF3 and Topas 19/2. And multimerization of transgenes was found in event Topas 19/2, in which, the T-DNA was integrated as a head-to-head(RB-to-RB) concatemer into the recipient genome. In event MS1, chromosomal translocation or a large target-site deletion may have occurred during T-DNA integration, which was identified due to a failure to amplify the presumptive insertion site based on the flanking rapeseed DNA sequences. Our results provide comprehensive data concerning transgene organization and the genomic context of the T-DNA in six rapeseed events, which can aid in the developing of insert fingerprinting and the monitoring of long-term genetic stability and potential unintended effects of transgenic events.  相似文献   
39.
Legislation limiting the use of chlorpropham (CIPC), the major potato sprout suppressant, has led to a need for new technologies to extend storage life of tubers. Ultra violet C (UV-C) has been used postharvest to reduce disease incidence on many crops, yet its use and efficacy as a sprout suppressant has not been investigated. The aim of this project was to identify the optimum dose and treatment timing of UV-C treatment on potato tubers as an alternative method of sprout suppression to reduce the dependence on chemical sprout suppressants. Up to six potato cultivars over two seasons were treated with varying doses of UV-C ranging from 0 to 30 kJ m−2 either at harvest or at first indication of dormancy break. The tubers were stored at 9 °C and sprout growth and incidence assessed. Treatment with moderate UV-C doses (5–20 kJ m−2) suppressed sprout length and sprout incidence in a range of cultivars. Periderm DNA damage and programmed cell death were not detected in response to any of the UV-C doses. The inactive ABA metabolite, ABA-GE, increased in response to 10 or 20 kJ m−2 within 72 h of treatment. Multivariate analysis showed a negative relationship between ABA metabolites and sprout growth/incidence during storage. This study found that UV-C reduced sprout growth in potato with no deleterious effects on tuber quality. This suggests potential for further development as an alternative or supplement to conventional sprout suppressant technologies.  相似文献   
40.
不同的样本特性和提取方法对获得微生物总DNA的质量有重要影响。文章基于高含固率木质纤维素厌氧发酵物腐殖酸、酚类物质含量高、质地均一性差、微生物浓度低的特点,研究了4种方法提取不同高含固率粪秸厌氧发酵物中微生物总DNA的效果。结果表明,常规的十二烷基磺酸钠法(sodium dodecyl sulfate,SDS)、十二烷基磺酸钠和溴化十六烷基三甲铵结合法(sodium dodecyl sulfate and cetyltrimethyl ammonium bromide,SDS-CTAB)和商业的粪便试剂盒法提取的DNA质量均较差,SDS法和试剂盒法未能获得聚合酶链式反应(polymerase chain reaction,PCR)扩增目的条带,SDS-CTAB法得到的条带较模糊;改进SDS-CTAB法获得的DNA杂质少、纯度高,具有较好的稳定性,A260/A280和A260/A230值分别为1.74~1.86和1.65~1.86,每克样品的DNA浓度在50 ng·μL^-1以上,电泳条带单一齐整、清晰明亮,PCR扩增的目的条带清晰度高,适宜后续分子生物学技术的分析。林格氏液洗脱、聚乙烯吡咯烷酮-40(Polyvinyl Pyrrolidone-40,PVP-40)洗涤液除杂以及裂解液和多种酶联合破壁是改进SDS-CTAB法获得该类专一性样本高质量微生物总DNA的关键步骤。  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