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农杆菌介导的蝴蝶兰基因转化系统的建立 总被引:9,自引:1,他引:9
针刺后的蝴蝶兰‘White Hikaru’的类原球茎(PLB),与含绿色荧光蛋白基因( )和潮霉素磷酸转移酶基因(^p£)的pCAMBIA 1300一SmGFP的根瘤农杆菌LBA4404 共培养,培育出了转基因的蝴蝶兰。经绿色荧光蛋白检测和Southern印迹,证实了再生植株中含cop基因和hpt基因。 相似文献
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缺铁胁迫下4种野生梨砧木介质pH值及根系Fe~(3+)还原酶活性的变化 总被引:5,自引:0,他引:5
以川梨、砂梨、豆梨和杜梨为材料,研究了在缺铁胁迫下培养介质的pH值和根系Fe3+还原酶活性的变化。结果表明,缺铁胁迫使4种砧木的介质pH值于处理的第2天就迅速降低,并且一直低于低铁和正常供铁处理;而低铁处理下,介质pH值在最初几天内反而高于正常处理,至4~5d之后才逐渐低于正常供铁处理;同时,NH4+-N对降低介质pH值和缓解缺铁症状有利。4种砧木的根系Fe3+还原酶活性都随供铁浓度的增加而不同程度的增大:无铁处理下,砂梨根系Fe3+的还原能力最小、杜梨最大;而在低铁和正常供铁条件下,砂梨和川梨的还原能力最强,杜梨次之,豆梨最小。还原酶大小与症状表现之间具有一致性。 相似文献
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JIANG Xun ZENG Yao-ying HE Xian-hui XU Li-hui DI Jing-fang FENG Zheng ZHAO Jing-xian WANG Qing WANG Tong SHI Jian-bo 《园艺学报》2004,20(6):924-928
AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37% at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage. 相似文献
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AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis. 相似文献
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Yohei Kato 《Pesticide biochemistry and physiology》2004,79(2):64-73
A hydrophilic form of acetylcholinesterase (AChE) was purified from N-methyl carbamate susceptible (SA) and highly N-methyl carbamate-resistant (N3D) strains of the green rice leafhopper (GRLH), Nephotettix cincticeps Uhler. Both of purified AChE from SA and N3D strains displayed the highest activities toward acetylthiocholine (ATCh) at pH 8.5. In the SA strain, the optimum concentrations for ATCh, propionylthiocholine (PTCh), and butyrylthiocholine (BTCh) were about 1 × 10−3, 2.5 × 10−3, and 1 × 10−3 M, respectively. However, in the N3D strain, substrate inhibition was not identified for ATCh, PTCh, and BTCh to 1 × 10−2 M. The Km value in the SA strain was 51.1, 39.1, and 41.6 μM and that in the N3D strain was 91.8, 88.1, and 85.2 μM for ATCh, PTCh, and BTCh, respectively. The Km value in the N3D strain indicated about 1.80-, 2.25-, and 2.05-fold lower affinity than that of the SA strain for ATCh, PTCh, and BTCh, respectively. The Vmax value in the SA strain was 70.2, 30.5, and 4.6 U/mg protein and that in the N3D strain was 123.0, 27.0, and 14.5 U/mg protein for ATCh, PTCh, and BTCh, respectively. The Vmax value in the N3D strain was 1.75- and 3.15-fold higher for ATCh and BTCh than that in the N3D strain. However, it was 1.13-fold lower for PTCh. The increased activity of AChE in the N3D strain is due to the qualitatively modified enzyme with a higher catalytic efficiency. The bimolecular rate constant (ki) for propoxur was 27.1 × 104 and 0.51 × 104 M−1 min−1 in the SA and N3D strain and that for monocrotophos was 0.031 × 104 and 2.0 × 104 M−1 min−1 in the SA and N3D strain. AChE from the N3D strain was 53-fold less sensitive than SA strain to inhibition by propoxur. In contrast, AChE from the N3D strain was 65-fold more sensitive to inhibition by monocrotophos than AChE from the SA strain. This indicated negatively correlated cross-insensitivity of AChE to propoxur and monocrotophos. 相似文献
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不同温度下圆叶决明降解过程红壤可溶性氮及氮水解酶动态变化研究 总被引:1,自引:0,他引:1
为了探明圆叶决明(Chamaecrista rotundifolia)降解过程果园红壤供氮水平的变化规律,本研究采用模拟培养试验,研究15℃和25℃培养果园红壤硝态氮、铵态氮、可溶性总氮、可溶性有机氮含量的变化及脲酶、蛋白酶、天冬酰胺酶活性的变化。结果表明,圆叶决明降解过程果园红壤4种可溶性氮含量均显著提高,且在培养140 d达到最大值,25℃培养的效果更佳。其中,可溶性总氮、可溶性有机氮和硝态氮含量随时间的动态变化可用三次曲线方程来拟合。圆叶决明降解过程还能显著提高脲酶、蛋白酶和天冬酰胺酶的活性,其中蛋白酶活性和脲酶活性的变化可用指数方程和三次曲线方程来拟合。本研究认为圆叶决明降解过程可提高果园红壤可溶性氮含量,因此可通过翻压圆叶决明提高土壤的供氮水平。 相似文献