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优化了S1核酸酶突变检测体系,结果表明(1)扩增法比杂交法产生异源双链DNA更节省时间;(2)PCR产物可以直接作为突变检测的底物;(3)适当降低反应温度、适当增加反应缓冲液中的NaCl浓度、适当缩短反应时间可提高突变检测效果。 相似文献
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Summary Aseptically cultured shoots of Chinese gooseberry exhibited growth disorder and morphological aberrances, and some died after being exposed to sufficient gamma-ray irradiation. The death rate was dose dependant and the LD50 was 80–90 Gy and 50–60 Gy respectively for cv. Hayward and clone 4. All petiole explants irradiated with gamma-ray could form calli as the control, but the rate of differentiation of adventitious shoots of the petiole explants decreased and was dependant on dose. Sensitivity of the shoot or petiole explants to gamma-ray irradiation varied with species. Gamma-ray irradiation did not deter either the 2-node segments from producing axillary shoots M1, M2, and M3 or the advantitious shoots originating in the petiole explants and the M3 shoots from forming advantitious roots. Therefore, using aseptically cultured axillary or adventitious buds for mutation breeding of Chinese gooseberry is feasible. A bacterium surviving in the explants lessened the efficiency of these two in vitro techniques in mutation breeding of Chinese gooseberry.Abbreviations IAA
3-indole acetic acid
- IBA
-indole butyric acid
- MS
Murashige & Skoog (1962) 相似文献
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狗牙根辐射诱变后代外部性状变异分析 总被引:12,自引:1,他引:12
对1997-2002年经过辐射诱变得到的狗牙根诱变后代的6个坪用性状指标的统计分析结果表明,1)不同的辐射诱变剂量处理的后代间坪用性状有显著差异,尤其在节间直径和密度上,6 800 rads处理的后代节间都比9 000 rads处理的后代粗,而密度是前者低于后者,2种剂量处理都可以显著降低草层高度;2)不同辐射诱变代数的后代间存在着差异,原始种源进行多代的辐射诱变,可以使得叶片显著变窄,草层高度显著降低,节间变细变长; 3)同一材料在相同辐射剂量诱变处理后匍匐茎不同节之间也存在显著的差异. 相似文献
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用0.05%、0.10%、0.15%3种不同浓度秋水仙素和24、48、72 h 3个不同处理时间,对日本矮紫薇种子发芽性状和幼苗生长性状进行研究.结果表明:0.05%、0.10%、0.15%浓度秋水仙素处理日本紫薇种子24、48、72 h,随着秋水仙素浓度的增加和处理时间的延长其种子发芽率、发芽势、出苗率均有降低的现象,0.15%浓度秋水仙素溶液处理的日本矮紫薇种子72 h时出苗率低于10%.随着秋水仙素处理浓度和时间的延长,幼苗生长表现出矮化、根部变短、叶片变厚的趋势.该研究获得了3株矮化的变异植株及2株株型变异的幼苗,对日本矮紫薇突变体产生、筛选及良种选育提供了基础. 相似文献
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Yayoi OTSUKA-YAMASAKI Osamu INANAMI Haruka SHINO Reeko SATO Masahiro YAMASAKI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(2):315
Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family. 相似文献
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XU Wan-yun WANG Hui-min WANG Wen-lun HU Meng-wei YAN Guo LI Jian-hua GAO Jian-feng 《中国畜牧兽医》2016,43(3):568-576
The specific primers were designed according to Ovis aries DRA gene sequence deposited in GenBank and the multiple cloning site of the plasmid pYD1,which was a vector used for protein surface display on Saccharomyces cerevisiae.The gene encoding DRA was amplified by PCR using the genomic RNA of Ovis aries.The 762 bp fragment was cloned and released in GenBank and registration number was KR422362.The PCR product was inserted into the yeast surface display plasmid vector pYD1 by double enzyme digestion.It was indicated that DRA gene was successfully integrated into the genome.Dot mutation was made at both ends of exon 2 in DRA gene for making restriction enzyme cutting site and design the exon 2 specific primers according to mutated Ovis aries DRA gene sequence.Sequenced exon 2 amplification products based on DNA pooling of sheep large sample template was analyzed the polymorphic loci.The polymorphic exon 2 246 bp fragment was obtained by double enzyme digestion and connected to surface display restructuring mutation carriers pYD1-DRA by the same double enzyme digestion,and then we successfully constructed yeast surface display libraries.We transformed it into Saccharomyces cerevisiae EBY100 cell.Yeast monoclone was identified by PCR amplification and sequencing,and we confirmed that DRA gene had been integrated into Saccharomyces cerevisiae genome.After galactose induced,it was detected that DRA gene library had been successfully demonstrated on the yeast cell surface under the fluorescence microscope by immunofluorescence method. 相似文献