全文获取类型
收费全文 | 490篇 |
免费 | 44篇 |
国内免费 | 264篇 |
出版年
2024年 | 1篇 |
2023年 | 17篇 |
2022年 | 23篇 |
2021年 | 18篇 |
2020年 | 21篇 |
2019年 | 31篇 |
2018年 | 22篇 |
2017年 | 18篇 |
2016年 | 18篇 |
2015年 | 26篇 |
2014年 | 42篇 |
2013年 | 36篇 |
2012年 | 54篇 |
2011年 | 51篇 |
2010年 | 39篇 |
2009年 | 40篇 |
2008年 | 47篇 |
2007年 | 42篇 |
2006年 | 45篇 |
2005年 | 21篇 |
2004年 | 23篇 |
2003年 | 24篇 |
2002年 | 27篇 |
2001年 | 30篇 |
2000年 | 18篇 |
1999年 | 16篇 |
1998年 | 8篇 |
1997年 | 2篇 |
1996年 | 3篇 |
1995年 | 3篇 |
1994年 | 1篇 |
1993年 | 8篇 |
1992年 | 1篇 |
1991年 | 5篇 |
1988年 | 5篇 |
1987年 | 1篇 |
1986年 | 1篇 |
1985年 | 2篇 |
1981年 | 1篇 |
1980年 | 1篇 |
1975年 | 1篇 |
1959年 | 1篇 |
1956年 | 1篇 |
1954年 | 1篇 |
1950年 | 1篇 |
1925年 | 1篇 |
排序方式: 共有798条查询结果,搜索用时 281 毫秒
61.
Phosphoinositide-specific phospholipase C (PI-PLC) is an important enzyme, which is a key player involved in eukaryotic signal
transduction pathways. In plants, it plays a key role in growth and development as well as environmental stress. However,
little is known about its roles in signal transduction during sexual reproduction process. In this study, we cloned and characterized
a gene of full-length PI-PLC from ovules of Torenia fournieri, designated as TfPLC1. It was 2,171 bp in length, including an open reading frame encoding a polypeptide of 583 amino acids with molecular mass
of 66.02 kDa. The amino acid sequence deduced from the cDNA sequence shows 40–76% similarity to other plant PI-PLCs and contains
the characteristic X, Y and C2 domains. Northern blot analysis demonstrated it was predominantly expressed in ovules and flowers.
Furthermore, TfPLC1 promoter::GUS transgenic analysis indicated it specifically expressed in ovule, stigma and mature pollen grain. Immunohistochemical
staining showed that, in mature stigma, TfPLC1 protein was principally localized in the cells of stigmatic receptive surface.
Together, our data suggest that TfPLC1 may play an important role in plant sexual reproduction.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
62.
农杆菌介导的水稻双载体共转化法中部分影响因素的研究 总被引:6,自引:1,他引:5
以T-DNA区分别只含有潮霉素选择标记基因(HPT)和GUS报告基因的双元载体pCAMBIA1300和pCAMBIA0301用于农杆菌介导的水稻共转化试验。根据经农杆菌浸染并共培养3d后水稻愈伤组织中的GUS瞬间表达情况及其稳定共转化率,测定了不同农杆菌菌株搭配及其不同浓度配比对共转化效率的影响。结果表明,在两种农杆菌菌液浓度比为1:1的情况下,农杆菌EHA105/pCAMBIA1300与EHA105/pCAMBIA0301组合共转化水稻的效率要高于其他菌株的组合;在以农杆菌EHA105/pCAMBIA1300与EHA105/pCAMBIA0301进行共转化时,两种菌液浓度比为1:2时共转化效率最高。 相似文献
63.
以Fluo-3AM为Ca~(2 )荧光探针,结合激光共聚焦扫描显微技术,观察到在处理后数十秒内,气孔关闭之前,茉莉酸(JA)可引起[Ca~(2 )]cyt的迅速上升;叶照和JA的前体物亚麻酸(LA)几乎不能引起[Ca~(2 )]cyt的明显变化;钙的螯合剂EGTA预处理可完全阻断JA诱导气孔关闭的效应,并且JA不再引起保卫细胞[Ca~(2 )]cyt增加;质膜Ca~(2 )通道的抑制剂硝苯吡啶(nifedipine,NIF)可减弱JA诱导气孔关闭的效应,也使JA诱导保卫细胞[Ca~(2 )]cyt增加的幅度有所下降;胞内Ca~(2 )释放的抑制剂钌红不能明显改变JA诱导气孔关闭的趋势,但使JA引起的保卫细胞[Ca~(2 )]cyt增加有所降低。实验结果表明:Ca~(2 )参与JA诱导气孔关闭的信号转导;推测JA引起的[Ca~(2 )]cyt升高可能主要来源于胞外,但不能完全排除胞内Ca~(2 )的释放。 相似文献
64.
从300条随机引物中筛选出能稳定扩增的26个引物.对黄瓜育成品种“春玉”等21个实验材料进行扩增,在扩增出的173条谱带中,多态性带有80条,比例为46.24%。“春玉”在用引物E13扩增时有特异缺失条带,大小为400bp,可作为特征性指纹图谱,为其产权保护提供分子依据。利用各材料的DNA指纹可将不同参试材料鉴别出来。同时利用MEGA软件进行UPGMA聚类分析,将参试材料在相似系数0.706处分为4个组群。 相似文献
65.
66.
检测伪狂犬病的PCR方法的建立及其在临床诊断中的应用 总被引:19,自引:0,他引:19
根据文献,通过计算机分析设计并合成了1对用于扩增伪狂犬病病毒(pseudorabies virus,PRV)gB基因281bp片段的引物,上游引物(P1)位于gB基因的1827~1851位,下游引物(P2)位于gB基因的2083~2107位.以PRV闽A株细胞培养毒为模板,筛选最佳反应条件和试剂工作浓度,建立了检测PRV的PCR方法.应用该方法对保存的16株伪狂犬病强弱毒株的细胞培养液进行基因扩增,均获得了281bp的特异性目的DNA片段.可是,对正常细胞与其它6种引起猪病毒性疫病相关病毒进行检测,结果均为阴性,没有出现交叉反应.对扩增产物测序,结果序列与文献报道一致,证明PCR扩增产物和方法的特异性.对PRV闽A株细胞毒提取物DNA进行检测,其最低检出量为15.8pg.用病毒分离、双抗体夹心ELISA和PCR等3种方法检测1994~2000年期间送检的临床样品和保存的PRV毒种,对所获得的结果进行χ2分析,证明PCR检出率明显高于前2种方法.对1999~2001年期间广东、福建、海南等省的76个大中型猪场送检的348份病料进行检测,检出阳性病料68份,病料阳性率为19.54%;检出阳性猪场27个,猪场阳性率为35.53%.对27个阳性猪场分析发现,种猪场阳性率为7.41%(2/27),商品猪场阳性率为92.59%(25/27).PRV在自然发病猪体内分布较广,脑、肾、肺、脾、肝、淋巴结均有PRV的存在,PRV检出率最高的组织为脑,其检出率为5/5,依次为肾12/15、肺9/16、脾10/20、肝7/18、淋巴结4/11等.实验结果表明,所建立的PCR技术可用于伪狂犬病的快速诊断和流行病学调查. 相似文献
67.
68.
Abstract Experiments were conducted in laboratory to study the impacts of rice genotypes and nitrogen levels on the preference of the egg predator, Cyrtorhinus lividipennis , for eggs of brown planthopper(BPH), Nilaparvata lugens (Stal), and their relation to the rice volatiles. In two-choice tests, the female predators showed different preference for BPH eggs on different rice genotypes, but not for BPH eggs on rice plants treated with different levels of nitrogen. The olfactory response test revealed that more predator oriented to the odour from healthy rice plants compared with the plain air (control); the predator preferred BPH nymph-damaged plants to healthy plants, and BPH female adult-damaged plants to BPH nymph-damaged plants. The comparative studies between rice genotypes and nitrogen levels indicated that there were obvious differences in attractiveness to the predator between rice genotypes, while there was no significant difference between rice plants treated with different levels of nitrogen. The results showed that rice volatiles played an important role in mediating the foraging behaviour of C. lividipennis . Implications for augmenting the effectiveness of natural enemies by adjusting rice attributes and cultural practices are discussed. 相似文献
69.
YUNSHENG LOU KAZUYUKI INUBUSHI TAKAYUKI MIZUNO TOSHIHIRO HASEGAWA YANHUNG LIN HIDEMITSU SAKAI WEIGUO CHENG KAZUHIKO KOBAYASHI 《Global Change Biology》2008,14(11):2678-2687
A pot experiment was conducted to investigate CH4 emissions from a sandy paddy soil as influenced by rice cultivars and atmospheric CO2 elevation. The experiment with two CO2 levels, 370 μL L−1 (ambient) and 570 μL L−1 (elevated), was performed in a climatron, located at the National Institute for Agro‐Environmental Sciences, Tsukuba, Japan. Four rice cultivars were tested in this experiment, including IR65598, IR72, Dular and Koshihikari. Tiller number, root length and grain yield were clearly larger under elevated CO2 than under ambient CO2. IR72 and Dular showed significantly higher tiller number, root length and grain yield than Koshihikari and IR65598. Average daily CH4 fluxes under elevated CO2 were significantly larger by 10.9–23.8% than those under ambient CO2, and varied with the cultivars in the sequence Dular ≧ IR72>IR65598 ≧ Koshihikari. Dissolved organic C (DOC) content in the soil was obviously higher under elevated CO2 than under ambient CO2 and differed among the cultivars, in the sequence IR72>Dular>Koshihikari>IR65598. The differences in average daily CH4 fluxes between CO2 levels and among the cultivars were related to different root exudation as DOC content, root length and tiller number. This study indicated that Koshihikari should be a potential cultivar for mitigating CH4 emission and simultaneously keeping stable grain yield, because this cultivar emitted lowest CH4 emission and produced medium grain yield. 相似文献
70.
Hong Zhao Lichen Xu Huihui Dong Jianhua Hu Hainv Gao Meifang Yang Xuan Zhang Xiaoming Chen Jun Fan Weihang Ma 《PloS one》2015,10(8)
Vibrio vulnificus is a common gram-negative bacterium, which might cause morbidity and mortality in patients following consumption of seafood or exposure to seawater in Southeast China. We retrospectively analyzed clinical data of patients with laboratory confirmed V. vulnificus infection. Twenty one patients were divided into a survival group and a non-surviving (or death) group according to their clinical outcome. Clinical data and measurements were statistically analyzed. Four patients (19.05%) died and five patients gave positive cultures from bile fluid, and 16 other patients gave positive culture from blood or blisters. Ten patients (47.62%) had an underlying liver disease and marine-related events were found in sixteen patients (76.2%). Patients with heavy drinking habits might be at increased mortality (p = 0.028). Clinical manifestations of cellulitis (47.6%), septic shock (42.9%) and multiple organ failure (28.6%) were statistically significant when comparing survivors and non-survivors (p = 0.035, p = 0.021 and p = 0.003, respectively). The laboratory results, including hemoglobin < 9.0 g/L (p = 0.012), platelets < 2.0×109 /L, prothrombin time activity (PTA) <20%, decreased serum creatinine and increased urea nitrogen were statistically significant (p = 0.012, p = 0.003, p = 0.028 and p = 0.028, respectively). Patients may be at a higher risk of mortality under situations where they have a history of habitual heavy alcoholic drink consumption (p = 0.028, OR = 22.5, 95%CI 1.5–335.3), accompanied with cellulitis, shock, multiple organ failure, and laboratory examinations that are complicated by decreased platelets, hemoglobin and significantly prolonged prothrombin time (PT). 相似文献