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1.
Foot-and-mouth disease (FMD) is a highly contagious aphthoviral infection of cloven-hoofed animals, inducing vesiculopustular stomatitis, pododermatitis, and thelitis. Vesicular fluid represents a major pathway of virus excretion, but bovine milk is another important source of virus shedding. We describe here the time course of FMD virus (FMDV) excretion in the milk and characterize associated lesions in the mammary gland. Three dairy cows were infected by nasopharyngeal instillation of FMDV and monitored over 12 d. Autopsy was performed at the end of the study, and specimens were collected for histopathology, IHC, and RT-qPCR. All 3 cows developed fever, drooling, vesiculopustular stomatitis, interdigital dermatitis, and thelitis. FMDV RNA was detectable in whole milk until the end of the trial, but only transiently in saliva, nasal secretions, and blood serum. Although histology confirmed vesiculopustular lesions in the oral and epidermal specimens, the mammary glands did not have unequivocal evidence of FMDV-induced inflammation. FMDV antigen was detectable in skin and oral mucosa, but not in the mammary gland, and FMDV RNA was detectable in 9 of 29 samples of squamous epithelia but only in 1 of 12 samples of mammary gland.  相似文献   
2.
【背景】新烟碱类杀虫剂的作用靶标是昆虫神经系统中的乙酰胆碱受体,由于其良好的内吸性及对人畜低毒性,使其在农业生产上获得了广泛应用,然而这也使得其在植物体内仍然具有较低的残留,而这种亚致死剂量残留仍可对访花昆虫如蜜蜂的行为和神经系统造成不利影响。【目的】明确亚致死剂量新烟碱类杀虫剂吡虫啉对中华蜜蜂(Apis cerana cerana,简称中蜂)神经生理和代谢系统的影响。【方法】首先以两个亚致死浓度梯度剂量5和10 μg·L-1吡虫啉处理工蜂10 d(3个生物学重复),提取总RNA后,以RNA-seq方法对所得文库进行高通量测序,利用生物信息学技术对序列进行从头组装、注释,并对亚致死剂量吡虫啉处理后的差异表达基因进行聚类和富集等分析,最后利用实时荧光定量PCR(RT-qPCR)技术对部分与中蜂神经和代谢系统相关的差异表达基因进行验证。【结果】从两个吡虫啉浓度梯度和对照组数据中共获得9个测序文库,测序有效数据比例超过94.45%,从获得的37 364个unigenes中鉴定出571个差异表达基因。经GO和KEGG富集分析发现这些差异表达基因主要与蛋白质翻译、氧化还原、氧化磷酸化和核糖体等多个通路有关,表明亚致死剂量的吡虫啉对中蜂多个生理过程和代谢通路造成影响。挑选了与昆虫神经信号传递和代谢功能有关的上调或下调差异表达基因,如神经肽F、神经肽SIFamide受体、3-磷酸肌醇依赖性蛋白激酶、激酶(PRKA)锚蛋白1、碳酸酐酶、超氧化物歧化酶、NADH脱氢酶亚基、表皮蛋白和气味结合蛋白17共9个差异表达基因进行了qPCR验证,其表达规律与转录组结果完全一致。【结论】亚致死剂量的吡虫啉能对中蜂神经信号转导、细胞呼吸、免疫反应、内环境稳态的维持和嗅觉感受等多方面造成影响。  相似文献   
3.
Eukaryotic translation elongation factor 1A (eEF1A) in plant is a multifunctional protein. In our previous study, we identified a 651-bp EST sequence of eEF-1α from a suppression subtractive hybridization (SSH) library of Chinese jujube (Ziziphus jujuba Mill.), which was established under jujube witches' broom (JWB) phytoplasma stress. Here we isolated the eEF1A gene (ZjeEF-1α, JF488063) from Chinese jujube by single oligonucleotide nested (SON)-PCR and rapid amplification of cDNA ends (RACE). ZjeEF-1α contains an open reading frame (ORF) of 1344 bp, encoding a polypeptide of 447 amino acids with an estimated molecular mass of 49.29 kDa and a theoretical pI of 9.10. ZjeEF-1α shared over 92% similarity with eEF-1α proteins from other 24 eukaryotes including plants and animals. Search Pfam predicted that ZjeEF-1A contains three typical eEF1A functional domains. Moreover, the corresponding genomic sequence of ZjeEF-1α was also sequenced, in which a 115-bp intervening sequence with splicing site GT-AG was found. Real-time quantitative PCR (RT-qPCR) and western blotting assays revealed that the expression of ZjeEF-1α was enhanced in resistant cultivar ‘Xingguang’ and decreased in sensitive cultivar ‘Zanhuangdazao’ at early stages of phytoplasma infection. It is proved that ZjeEF-1A expression is early response to phytoplasma infection and its possible functions in resistant process to JWB phytoplasma are discussed.  相似文献   
4.
Several treatments were applied in order to delay postharvest degreening in broccoli florets and investigate their effects on genes associated with chlorophyll catabolism. Degradation of chlorophylls is the most evident visual manifestation of broccoli postharvest deterioration, occurring rapidly due to the immature stage in which the material is harvested. Treatments such as storage in modified atmosphere, exposure to hot air, UV-C and white lamps were employed in the current work to induce a delay in degreening and chlorophyll degradation. Expression of genes possibly related to chlorophyll catabolism was analyzed in these samples and discussed. Chlorophyllases, the enzymes traditionally believed to remove the phytol side chain from chlorophyll appear to have a gene expression that was not regulated by postharvest treatments. Pheophytinase, a recently discovered new enzyme in this metabolic pathway, correlated chlorophyll loss accurately in heat, UV-C and white-light treatments, but not in modified atmospheres. Results presented in this work indicate that postharvest treatments that delay chlorophyll degradation have a higher effect on the expression of pheophytinase rather than on chlorophyllase genes.  相似文献   
5.
BACKGROUND: Recent reports indicate increased amounts of mRNA from inflammation-related genes in the prodromal stage of laminitis. HYPOTHESIS: Cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) undergo distinct patterns of expression in equine laminae in the developmental stage (DEV) and acute clinical stage (LAM) of laminitis. ANIMALS: Horses selected from an outbred population were placed into 1 of 4 groups: DEV (n = 5), CON-3h (control group for DEV, n = 5), LAM (n = 5) and CON-10h (control group for LAM, n = 5). METHODS: Laminar and skin samples were obtained from (1) animals either undergoing leukopenia (DEV) or the onset of clinical signs of laminitis (LAM) after black walnut extract (BWE) administration and (2) animals either 3 (CON-3h) or 10 (CON-10h) hours after administration of water. Real-time quantitative polymerase chain reaction (RT-qPCR), immunoblotting, and immunohistochemical analysis were performed for COX-1 and COX-2. RESULTS: Upon immunohistochemical analysis of all 4 groups, COX-2 was expressed by most viable epithelial cells in both laminae and skin. COX-1 exhibited similar epithelial expression to COX-2 in skin epidermis, but was expressed exclusively in the basal layer of laminar epidermis. COX-1 protein was not detectable in dermal vasculature of equine skin or laminae, whereas COX-2 was present in endothelial and vascular smooth muscle cells of dermal vasculature in both skin and laminae in all groups. A marked increase in laminar COX-2 protein concentrations was detected on immunoblotting in the DEV group, although a lesser increase was observed in the LAM group. CONCLUSIONS AND CLINICAL IMPORTANCE: COX-2 protein expression is markedly increased in the resident laminar cell types in the developmental stage of BWE-induced laminitis.  相似文献   
6.
对GenBank已公布的小反刍兽疫病毒N基因序列进行分析,设计引物与探针,建立PPRV通用型与Ⅱ系疫苗毒特异型二重实时荧光RT-PCR方法,同时建立针对PPRVIV系强毒株的特异型荧光PCR方法。特异性试验和灵敏度试验表明:建立的二重荧光RT-PCR方法可特异性检测PPRV病毒,其HEX信号通道(通用型)和TAMRA信号通道(Ⅱ系疫苗毒特异型)的检测灵敏度分别可达10^1 TCID50/mL和10^2 TCID50/mL,完全满足PPRV的检测要求。用二重荧光RT-PCR方法对西藏采集的14份羊血清样品进行检测,并用建立的Ⅳ系强毒株特异型荧光PCR方法对二重荧光RT-PCR的检测效果进行评估,结果显示,该方法可有效甄别PPRV强毒株和疫苗毒株,避免假阳性结果的出现。  相似文献   
7.
【目的】为筛选苹果实时荧光定量PCR实验中最适内参基因,【方法】应用实时荧光定量PCR技术,分析5个传统内参基因18SrRNA、ACTB、GAPDH、UBQ、TUB在苹果不同基因型、不同组织、果实不同发育时期的mRNA表达差异情况。供试的6个不同基因型苹果分别为:新疆野生苹果、八棱海棠、丽江山荆子、‘津轻’、‘国庆’、‘金冠’;6种不同组织为果皮、果肉、叶片、愈伤组织、花瓣、种子;5个果实发育不同时期为花后28、50、74、95、115 d。【结果】经geNorm程序分析发现5种内参基因的表达稳定性各异,UBQ在果实不同基因型和不同发育时期的基因表达分析中最稳定;ACTB和UBQ在6种不同组织中表达均稳定。【结论】UBQ在参试样品中表达均比较稳定,是研究苹果基因表达分析中理想的内参基因。  相似文献   
8.
团花树α扩展蛋白基因的克隆及表达分析   总被引:1,自引:0,他引:1  
采用扩增保守区域、基因组步移及3'RACE技术,在团花树中克隆到15个α-expansin基因的cDNA序列,命名为AcEXPA2-16(GenBank注册号JF922686-JF922700),其相对应的基因组序列GenBank注册号为JF922701-JF922715.包括已知的AcEXPA1,这16个EXPA蛋白的大小及序列高度保守,包括N端的信号肽、3个外显子和2个内含子.通过氨基酸序列比对和系统进化树分析,AcEXPA1-16分为4个亚族,分别为A、B、C和D亚族.定量PCR分析显示AcEXPA1-16基因在同一组织中的表达存在冗余现象,而单个基因在不同的组织中又存在特异表达,尤其AcEXPA8在形成层中的表达水平极高.这表明AcEXPA8在木质部发生的过程中可能起到重要的作用.研究结果为在团花树中研究α-expansin基因与木质部的生长和材质的关系提供信息,并最终为林木分子育种提供潜在的候选基因.  相似文献   
9.
It has been reported that cyanidin 3-O-glucoside (C3G) is an inhibitor of Helicobacter pylori toxin secretion. C3G is classified as an anthocyanin and is a major component of black rice extract (BRE). The present study aimed to identify a new functional food material to prevent H. pylori infection in Mongolian gerbil model. Toxicity in the liver and kidney were not detected after BRE administration (10 or 50 mg/kg). BRE treatment reduced bacterial colonization in animal gastric tissue, as well as infection signs as observed on the analysis of the hematological data. It was also found that the relative mRNA levels of the inflammatory cytokines were reduced in BRE-treated groups. These findings suggest that BRE acts as a potent inhibitor of H. pylori infection and pathogenesis in Mongolian gerbils. We propose that BRE may be used to manage gastroduodenal diseases caused by H. pylori infection.  相似文献   
10.
试验旨在了解在鸡睾丸中高表达的1个长链非编码RNA (long non-coding RNA,lncRNA)及其预测靶基因的时空表达规律,研究二者在鸡弱精子症中的调控作用。根据弱精子症和正常北京油鸡公鸡睾丸转录组测序筛选到的1个高表达的lncRNA (MSTRG.15568.9),采用顺式(cis)作用模式预测其潜在靶基因SPAG4(sperm-associated antigen 4),进一步采用实时荧光定量PCR方法进行表达量分析。分别选择3只0、5、20、30、45、60周龄正常北京油鸡公鸡,检测MSTRG.15568.9与SPAG4基因在不同周龄公鸡睾丸中的表达量差异;选择30周龄3只正常公鸡,采集睾丸、肝脏和脾脏等8个部位组织样品,检测MSTRG.15568.9与SPAG4基因在不同组织间的表达规律;选择45周龄弱精子症公鸡和正常公鸡各3只,对比MSTRG.15568.9与SPAG4基因在睾丸的表达量差异。结果显示,MSTRG.15568.9与SPAG4存在明显的时空表达差异,且二者表达趋势基本一致。在不同周龄的鸡睾丸组织中,MSTRG.15568.9和SPAG4的表达趋势相近,MSTRG.15568.9在20周龄的表达量显著高于0、5、30、45、60周龄(P<0.05),0和5周龄表达量显著低于20、30、45和60周龄(P<0.05);SPAG4在45周龄表达量最高,其次是20周龄(P<0.05)。MSTRG.15568.9和SPAG4在睾丸和肝脏中的表达量均显著高于脾脏、肾脏等组织(P<0.05);在正常睾丸组织中的表达量均显著高于弱精子症睾丸组织(P<0.05)。综上所述,MSTRG.15568.9与SPAG4基因具有较明显的组织表达特异性,且MSTRG.15568.9可能调控SPAG4基因的表达,参与精子发生与精子活力调控;但其具体作用机制需要进一步探索。本研究可为鉴定与鸡弱精子症调节机制相关的功能基因提供参考。  相似文献   
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