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51.
旨在获得转淀粉分支酶反义SBEⅠ基因的‘华南木薯8号’转基因植株,为利用转基因技术改良木薯淀粉品质打下基础。在建立了木薯从胚状体子叶到完整植株的再生体系的基础上,用块根特异表达启动子Sporamin驱动的木薯淀粉分支酶SBEⅠ反义基因,通过农杆菌介导法对‘华南木薯8号’进行遗传转化。共接种‘华南木薯8号’子叶517块,获得7株生长良好的转化再生植株,转化再生频率达到1.35%。经PCR检测,其中5株转化再生植株扩增出目的条带,初步证实木薯淀粉分支酶SBEⅠ反义基因已整合进了‘华南木薯8号’基因组中。通过农杆菌介导法可以将淀粉分支酶SBEⅠ反义基因导入到‘华南木薯8号’基因组中,获得了5株转基因植株。 相似文献
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大豆花粉管通道技术转化雪花莲凝集素(GNA)基因 总被引:11,自引:0,他引:11
采用花粉管通道技术,用雪花莲凝集素基因(Galanthus nivalis agglutinin,GNA)转化吉林省主推品种吉林20号、吉林30号、吉林45号品种大豆。通过接蚜鉴定和PCR鉴定,从所获得的种子苗中筛选出转基因植株。对转基因植株的后代进行分子生物学鉴定:(1)PCR分析,转基因植株97TGR1和97TGR2的T2代表现阳性,第5代表现阳性纯合;97TGR1、97TGR2和98FD1~98FD20的T3代Western blotting检测结果证明,GNA基因在蛋白质水平有表达,最高表达量占总可溶性蛋白的0.7%;97TGR1、98TGR2和99JI45 TGR2的Southern blotting检测结果显示,GNA基因已插入大豆基因组;(2)遗传学分析,97TRG1的T2代呈孟德尔3:1分离,97TGR2的T3代出现种皮颜色不规则分离。经过抗蚜性鉴定和连续的筛选,获得抗性纯系;(3)抗蚜性鉴定,转基因株的T1、T2世代转基因植株可抑制蚜虫繁殖量50%~90%;(4)品系鉴定,转基因大豆的抗蚜性达到农学标准抗(R)和高抗(HR)水平;大面积环境释放试验自然感蚜鉴定,转基因系蚜虫发生的高峰比对照延迟,高峰期过后群体蚜量的下降速度也比对照快。本研究认为,大豆花粉管通道技术可以利用于大豆的转基因研究和应用中,GNA基因在改良大豆的抗蚜性上是可取的。 相似文献
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The overall objective of this work was to improve fruit quality, break alternate bearing and reduce hand thinning using fewer chemicals in fruit crops. A device was constructed for mechanical thinning, which consisted of three independent horizontal rotors with ropes and freely adjustable angles on a frame, mounted on a front three point hitch and powered by the tractor hydraulics. This can be adapted to any fruit tree trained as spindle, Solaxe, (tall) vertical axis or fruit wall (le mur fruitier) irrespective of rootstock employed. Rotor speed varied from 300 to 460?rpm at either 5 or 7.5?km/h tractor speed. Eight-year-old or twelve-old apple trees cvs. ‘Gala’ and ‘Golden Delicious’ were mechanically thinned in 2007 between pink bud and full bloom (flower bud stages 6–8 or F1–F2) near Bonn, Germany; non-thinned and hand-thinned apple trees of the same block and variety served as control. Mechanically thinned flowering branches showed a similar amount of ethylene efflux (0.4–0.6?ppm C2H4/branch) as non-thinned flower branches, preventing potentially unexpected subsequent fruit drop, except for those removed by the rotors. The impact of the horizontal rotors on the branches was from the upper side and removed excessive flowers right to the tree trunk viz. the centre of the tree canopy, where fruits of lesser quality are expected leaving 2–3 flowers per cluster. Leaf damage was less than??10%, even at the fast rotor speed of 420?rpm, which was associated with negligible wood injury. Mechanical thinning induced firmer and sweeter fruit, i.e. tastier apples with longer shelf life, relative to control fruit from non-thinned apple trees. The greatest efficacy in terms of final fruit quality in the grading/sorting was achieved by a rotor speed of 360?rpm at a tractor speed of 5?km/h: Fruit mass increased by up to 20?g and the proportion of fruit larger than 70–75?mm by 10–30% compared with the fruit from non-thinned trees. Mechanical thinning with this newly constructed device led to a 10–20% reduction in yield, but increased returns due to better fruit size and colouration in apple with the potential to overcome alternate bearing. 相似文献
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为解决ABS逻辑门限值方法匹配成本高、周期长的问题,将制动器耗散功率原理应用于匹配中.结合逻辑门限值与制动器耗散功率的各自优点,将制动器耗散功率原理能自适应识别各种路面状况的特点应用于匹配中;根据制动器耗散功率极大值对应的滑移率略小于临界滑移率且相差不大的原理,采用制动器耗散功率极大值对应的滑移率代替峰值附着系数对应的滑移率对ABS进行控制;滑移率与制动器耗散功率曲线获得较为简单,因此可以大大提高匹配效率.经过实车匹配验证,本方法提高了匹配效率和控制质量,节约了匹配成本. 相似文献
60.
Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL−1 of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g−1 tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations. 相似文献