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91.
    
To establish a suitable and effective protocol of protein extraction for two-dimensional gel electrophoresis (2-DE) analysis in kenaf leaf tissues, three extraction methods (trichloroacetic acid/acetone, urea/thiourea, and phenol extraction methods) were applied to the extraction of kenaf leaf protein. The results were compared in regard to protein extraction efficiency, sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and 2-DE gels. Furthermore, the 2-DE system was optimized for four aspects: the pH range of IPG (immobilized pH gradient) stripes, sampling methods, sample volumes, and concentration of polyacrylamide gels. The data presented showed that the phenol extraction method is the best method to perform 2-DE analysis of kenaf leaf protein. The protein extracted from phenol extraction method reached the purity of (26.40±0.859)%, showed (25.67±1.53) protein bands in one dimension SDS-PAGE gels, and (1 374±54.44) protein spots on 2-DE gels. The research also indicates that kenaf leaf protein spots were distributed mainly within the pH range of 4-8. More clear background with a better distribution effect and many protein spots could be obtained on 2-DE gels under the conditions of active rehydration loading, 24 cm IPG strips (linear pH gradient of 4-7), 1.4 mg samples, and 12% SDS-PAGE gels. Abstract To establish a suitable and effective protocol of protein extraction for two-dimensional gel electrophoresis (2-DE) analysis in kenaf leaf tissues, three extraction methods (trichloroacetic acid/acetone, urea/thiourea, and phenol extraction methods) were applied to the extraction of kenaf leaf protein. The results were compared in regard to protein extraction efficiency, sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and 2-DE gels. Furthermore, the 2-DE system was optimized for four aspects: the pH range of IPG (immobilized pH gradient) stripes, sampling methods, sample volumes, and concentration of polyacrylamide gels. The data presented showed that the phenol extraction method is the best method to perform 2-DE analysis of kenaf leaf protein. The protein extracted from phenol extraction method reached the purity of (26.40±0.859)%, showed (25.67±1.53) protein bands in one dimension SDS-PAGE gels, and (1 374±54.44) protein spots on 2-DE gels. The research also indicates that kenaf leaf protein spots were distributed mainly within the pH range of 4-8. More clear background with a better distribution effect and many protein spots could be obtained on 2-DE gels under the conditions of active rehydration loading, 24 cm IPG strips (linear pH gradient of 4-7), 1.4 mg samples, and 12% SDS-PAGE gels.
相似文献   
92.
    
Thirty-two Wistar rats were divided randomly into four groups of eight with six females and two males in each group. The rats were exposed to high fluoride drinking water (45 mg F- L-1 from 100 mg NaF L-1), low dietary iodine (0.0855 mg kg-1), or both together in order to assess the effects of these three regimens on the thyroid function of the offspring rats. After the animal model was established, the offspring rats were bred and 10-, 20-, 30-, 60-, and 90-d-old rats were used for the experiment. The treatments for the offspring rats were the same as those of their parents. In comparison with control rats, the relative thyroid glands were changed by three regimens, but the mean values of thyroid weight in the experimental groups saw no marked difference. Serum TT3 levels were increased in all stages in the low iodine (LI) group. In the high fluoride (HiF) group, increase in TT3 levels was observed except in 20-d-old rats. Decrease in TT3 at 20- and 90-d and increase in TT3 at 30- and 60-d were found in HiF+LI group. Serum TT4 levels first saw an increase, and then dropped in the LI and HiF+LI group. However, an increase in TT4 was found in the HiF group. The levels of TSH in serum rocketed at d 20, and then dropped in the next stages in experimental groups. The results suggested that thyroid disorder could be induced by high flroride in drinking water, low iodine diet, or both of them. Exposure time to fluoride or low iodine diet was one of the important factors that fluoride can induce the development of thyroid dyfunction. Abstract Thirty-two Wistar rats were divided randomly into four groups of eight with six females and two males in each group. The rats were exposed to high fluoride drinking water (45 mg F- L-1 from 100 mg NaF L-1), low dietary iodine (0.0855 mg kg-1), or both together in order to assess the effects of these three regimens on the thyroid function of the offspring rats. After the animal model was established, the offspring rats were bred and 10-, 20-, 30-, 60-, and 90-d-old rats were used for the experiment. The treatments for the offspring rats were the same as those of their parents. In comparison with control rats, the relative thyroid glands were changed by three regimens, but the mean values of thyroid weight in the experimental groups saw no marked difference. Serum TT3 levels were increased in all stages in the low iodine (LI) group. In the high fluoride (HiF) group, increase in TT3 levels was observed except in 20-d-old rats. Decrease in TT3 at 20- and 90-d and increase in TT3 at 30- and 60-d were found in HiF+LI group. Serum TT4 levels first saw an increase, and then dropped in the LI and HiF+LI group. However, an increase in TT4 was found in the HiF group. The levels of TSH in serum rocketed at d 20, and then dropped in the next stages in experimental groups. The results suggested that thyroid disorder could be induced by high flroride in drinking water, low iodine diet, or both of them. Exposure time to fluoride or low iodine diet was one of the important factors that fluoride can induce the development of thyroid dyfunction. 相似文献   
93.
    
γ-Gliadins are an important component of wheat seed storage proteins. Four novel γ-gliadin genes (Gli-ng1 to Gli-ng4) were cloned from wheat (Triticum aestivum) and Aegilops species. The novel γ-gliadins were much smaller in molecular size when compared to the typical γ-gliadins, which was caused by deletion of the non-repetitive domain, glutamine-rich region, 3´ part of the repetitive domain, and 5´ part of the C-terminal, possibly due to illegitimate recombination between the repetitive domain and the C-terminal. As a result, Gli-ng1 and Gli-ng4 only contained two and three cysteine residues, respectively. Gli-ng1, as the representative of novel γ-gliadin genes, has been sub-cloned into an Escherichia coli expression system. SDS- PAGE indicated that the both cysteine residues of Gli-ng1 could participate in the formation of intermolecular disulphide bonds in vitro. Successful cloning of Gli-ng1 from seed cDNA of T. aestivum cv. Chinese Spring suggested that these novel γ-gliadin genes were normally transcribed during the development of seeds. Phylogenic analysis indicated that the four novel γ-gliadin genes had a closer relationship with those from the B (S) genome of wheat. Abstract γ-Gliadins are an important component of wheat seed storage proteins. Four novel γ-gliadin genes (Gli-ng1 to Gli-ng4) were cloned from wheat (Triticum aestivum) and Aegilops species. The novel γ-gliadins were much smaller in molecular size when compared to the typical γ-gliadins, which was caused by deletion of the non-repetitive domain, glutamine-rich region, 3´ part of the repetitive domain, and 5´ part of the C-terminal, possibly due to illegitimate recombination between the repetitive domain and the C-terminal. As a result, Gli-ng1 and Gli-ng4 only contained two and three cysteine residues, respectively. Gli-ng1, as the representative of novel γ-gliadin genes, has been sub-cloned into an Escherichia coli expression system. SDS- PAGE indicated that the both cysteine residues of Gli-ng1 could participate in the formation of intermolecular disulphide bonds in vitro. Successful cloning of Gli-ng1 from seed cDNA of T. aestivum cv. Chinese Spring suggested that these novel γ-gliadin genes were normally transcribed during the development of seeds. Phylogenic analysis indicated that the four novel γ-gliadin genes had a closer relationship with those from the B (S) genome of wheat. 相似文献   
94.
    
Here, we investigated the effect of dietary arginine (Arg) supplementation on innate immunity and the antioxidant ability of broiler chickens. The experiment was designed as a single-factorial arrangement (n=8 cages/treatment, six birds/cage), and we used four dietary Arg concentrations (10.0, 15.0, 20.0 or 25.0 g kg–1). On day 21, the birds were killed to obtain spleen, cecal tonsil and liver samples to determine the gene expression and antioxidant characteristics. Increasing the Arg concentration linearly decreased (P<0.05) the mRNA expression of splenic interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α). Dietary Arg supplementation quadratically decreased (P<0.05) the expression of interleukin-1b (IL-1b) and interferon-γ (IFN-γ) mRNA in the spleen. Increasing Arg concentrations linearly and quadratically reduced the expression of IL-18 mRNA in the spleen. Meanwhile, increasing dietary Arg supplementation linearly and quadratically increased the lymphotactin mRNA (P<0.05) expression, and linearly increased the macrophage inflammatory protein-1β (MIP-1β) and toll-like receptor 15 (TLR15) mRNA expression in the cecal tonsils. Dietary Arg supplementation linearly (P<0.05) increased the glutathione peroxidase (GSH-Px), catalase (CAT), and lysozyme (LZM) activities in the liver. However, the malondialdehyde (MDA) activity in the liver was not influenced by the dietary Arg concentration (P>0.05). No significant (P>0.05) effect was found on the activity of superoxide dismutase (SOD) in the liver. Thus high levels of Arg supplementation (>20.0 g kg–1) may potentially suppress the innate immunity of broiler chickens, and dietary Arg supplementation enhances the antioxidant activity in broiler chickens. Keywords: arginine innate immunity antioxidant ability broiler > Received: 09 November 2015 >>Accepted: > Fund: This study was supported by the National Key Technology R&D Program of China (2012BAD39B01) and the Agricultural Science and Technology Innovation Program of CAAS (ASTIP-IAS07) in China.  相似文献   
95.
    
In this study,four genotypes(Acva-1,Acva-2,Acva-3 and ZM-2) of Actinidia germplasm resources were grown in different NaCl concentrations(0,0.4,0.8 and 1.2 g L–1).The growth,physiological and biochemical indicators were measured,and a graded scale was developed as the salt damage index(SDI) according to different damage symptoms in leaves.The results showed SDI increased gradually,and average number and length of new shoot decreased significantly.Three antioxidant enzymes(superoxide dismutase,peroxidase and catalase) and two osmotic adjustment substances(soluble sugar and proline) showed different changes in old and new leaves of four genotypes.SPAD values exhibited a decreased trend in the whole except in the new leaves of Acva-2.Malonaldehyde contents increased and root activity decreased with the increasing salt concentrations.Principal component analysis was used to assess the salt tolerance,and the results showed Acva-3,from Actinidia valvata Dunn.,had the strongest tolerance to salt,and could be a potential resistant resource to the salt-tolerance dedicated rootstock breeding of kiwifruit.  相似文献   
96.
    
Abstract Conventional soil maps contain valuable knowledge on soil–environment relationships. Such knowledge can be extracted for use when updating conventional soil maps with improved environmental data. Existing methods take all polygons of the same map unit on a map as a whole to extract the soil–environment relationship. Such approach ignores the difference in the environmental conditions represented by individual soil polygons of the same map unit. This paper proposes a method of mining soil–environment relationships from individual soil polygons to update conventional soil maps. The proposed method consists of three major steps. Firstly, the soil–environment relationships represented by each individual polygon on a conventional soil map are extracted in the form of frequency distribution curves for the involved environmental covariates. Secondly, for each environmental covariate, these frequency distribution curves from individual polygons of the same soil map unit are synthesized to form the overall soil–environment relationship for that soil map unit across the mapped area. And lastly, the extracted soil–environment relationships are applied to updating the conventional soil map with new, improved environmental data by adopting a soil land inference model (SoLIM) framework. This study applied the proposed method to updating a conventional soil map of the Raffelson watershed in La Crosse County, Wisconsin, United States. The result from the proposed method was compared with that from the previous method of taking all polygons within the same soil map unit on a map as a whole. Evaluation results with independent soil samples showed that the proposed method exhibited better performance and produced higher accuracy.
相似文献   
97.
    
Received 3 March, 2018 Accepted 2 April, 2018 Abstract The cereal cyst nematodes (Heterodera avenae, Heterodera filipjevi, Heterodera latipons) are considered to be one of the most important plant parasitic nematodes attacking most cereals and can cause significant crop losses (Sikora 1988). In China, H. filipjevi (Madzhidov 1981) Stelter, 1984, was first reported from Henan province (Peng et al. 2010) and a few years later in Anhui province and Xinjiang Uygur Autonomous Region (Peng et al. 2016, 2018) . In December 2017, a survey for cereal cyst nematodes on winter wheat was conducted in Shandong Province, China. A total of 79 samples that including roots and rhizosphere soil were collected. Cysts and second-stage juveniles (J2s) were isolated from each soil sample using the sieving-decanting method. Wheat roots were stained with acid fusion to observe the development of cereal cyst nematodes. One sample collected from Yangzhuan Village in Huanggang Town, Shan County of Heze City (GPS 34°38´23.10´´N, 116°05´42.95´´E), Shandong Province, was found that the wheat roots were heavily parasitized by cyst nematodes, and most of the nematodes in roots had developed to fourth-stage (J4) in mid-December of 2017. The morphological and molecular studies of cyst and J2s were carried out to confirm the identification of H. filipjevi in one winter wheat field soil and root sample from Shan County. The cysts were lemon shaped with prominent vulval cone, brown to black in colour. Cuticle with irregular zig-zag pattern. Neck prominent, vulval cone bifenestrate with horseshoe-shaped fenestra, bullae and underbridge strongly developed. The main morphometrics of cysts (n=8) were length (including neck) (688 to 948 μm, mean=794 μm, standard deviation=87 μm), width (465 to 620 μm, mean=529 μm, standard deviation=63 μm), neck length (71.5 to 126.3 μm, mean=86.5 μm, standard deviation=9.2 μm), fenestra length (43.8 to 71.3 μm, mean=58.0 μm, standard deviation=15.1 μm), fenestra width (19.8 to 32.0 μm, mean=25.0 μm, standard deviation=3.9 μm), length of vulval slit (8.1 to 9.7 μm, mean=9.1 μm, standard deviation=0.5 μm) and length of underbridge (64.5 to 101.3 μm, mean=82.6 μm, standard deviation=12.8 μm). Measurements of J2s (n=10); body length (556.7 to 617.0 μm, mean=584.3 μm, standard deviation=23.2 μm); stylet (22.8 to 24.1 μm, mean=23.3 μm, standard deviation=0.4 μm), tail (59.6 to 68.6 μm, mean=65.8 μm, standard deviation=3.5 μm) and hyaline tail terminus (35.9 to 41.1 μm, mean=38.6 μm, standard deviation=2.1 μm). Genomic DNA was isolated from single cysts (n=6), and the internal transcribed spacer regions were amplified with primers TW81 (5´-GTTTCCGTAGGTGAACCTGC-3´) and AB28 (5´-ATATGCTTAAGTTCAGCGGGT-3´) (Joyce et al. 1994) and 28S rDNA-D2/D3 regions were amplified with primers D2A (5´-ACAAGTACCGTGAGGGAAAGTTG-3´) and D3B (5´-TCGGAAGGAACCAGCTACTA-3´) (Subbotin et al. 2006). The obtained internal transcribed spacer regions (ITSs) sequences (GenBank accession MG859977) is 99% identical to those of H. filipjevi from Turkey (KR704292.1 and KR704304.1), the United States (KP878490.1 and GU079654.1) and China (KY448473.1 and KY448473.1). The obtained 28S rDNA-D2/D3 sequences (GenBank accession MG859980) also to be 99 to 100% identical to those of H. filipjevi from China (GU083597.1, KT314235.1, GU083592.1). The species-specific primers of H. filipjevi (HfF1, 5´-CAGGACGAAACTCATTCAACCAA-3´; HfR1, 5´-AGGGCGAACAGGAGAAGATTAGA-3´) were also used to identify this population (Peng et al. 2013), the specific band was obtained species-specific primers of H. filipjevi. Based on the morphological and molecular data, the species of the cyst-forming nematode was identified as H. filipjevi. As far as we know, this is the first report of H. filipjevi in Shandong Province, China. The population density of H. filipjevi were found much higher than those of other CCN, it can serious infect winter wheat at seedling stage which often cause economically damaging to wheat, so the spread of H. filipjevi would be a risk for the cereal production of Shandong province. 相似文献   
98.
99.
设计了一条绿色高效地提取纯化大豆异黄酮,并联产制备大豆分离蛋白和低聚糖的工艺.首先通过单因素实验和响应面(RSM)优化,确定了超声辅助提取大豆异黄酮的工艺条件:超声功率250 W,70%乙醇,提取温度70℃,提取时间120 min,大豆异黄酮提取量为2104.25 μg·g-1,提取率85.34%;并通过三步简单的分离纯化从豆粉中得到1 838.00,μg·g-1、纯度为62.09%的异黄酮,最终收率74.54%;最后以提取异黄酮后的剩余豆渣为原料,联产制备了大豆分离蛋白(216.25 mg·g-1)和低聚糖(73.75 mg·g-1),全过程无废液废渣排出.  相似文献   
100.
本试验旨在通过4个剂量-反应试验,研究30~38周龄产蛋鸡对赖氨酸(Lys)、蛋氨酸(Met)、色氨酸(Trp)和苏氨酸(Thr)4种必需氨基酸(AA)的需要量及其比例.每个试验均采用360只海兰灰产蛋鸡,设5个处理,分别采食5个AA梯度水平的试验饲粮,每个处理6个重复,每个重复12只鸡.采用蛋公鸡回肠食糜法测定饲料原料的标准回肠可消化(SID)值.试验预试期14 d,正试期56 d.结果表明:海兰灰产蛋鸡的产蛋率、蛋重、日产蛋量和料蛋比与SIDLys、SID Met、SID Trp和SID Thr的摄入水平均有较好的折线回归关系;以日产蛋量为自变量(x),AA摄入水平为依变量(Y)分别建立的回归方程为:YSID Lys=51.60-0.041 6(682.5-x),R2=0.96;YsID Met=51.43-0.085(314.1-x),R2 =0.95;YSID Trp =51.17-0.155 4(163.0-x),R2=0.97;YsID Thr=52.85-0.057 6(499.5-x),R2=0.93.在饲粮粗蛋白质12.8%的条件下,以日产蛋量为参照指标确定的30 ~ 38周龄产蛋鸡的SID Lys、SID Met、SID Trp和SID Thr需要量分别为684、313、171和506 mg/(只·d).以日产蛋量为指标建立的30~38周龄产蛋鸡理想SID AA模式为:Lys:Met:Trp:Thr=100∶46∶24∶73.  相似文献   
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