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1.
向大豆导入几丁质酶基因的初步研究 总被引:32,自引:8,他引:24
以根癌农杆菌的Ti质粒为介导,将抗真菌病的几丁质酶基因,导入默经江省栽培大豆一东农37号,吉林28号等14个品种。从子叶节和下胚轴诱导出丛生芽,并再生植株,经PCR和DNA分子杂交检测,经PCR和DNA分子杂交检测的113株大豆幼苗中,有7株呈阳性反应,证明几丁质酶基因已导入并整合到大豆的基因组中。 相似文献
2.
Nobuhito Mitani Shozo Kobayashi Yoko Nishizawa Takeshi Kuniga Ryoji Matsumoto 《Scientia Horticulturae》2006
RCC2, cDNA clone encoding rice class-I chitinase, was introduced into trifoliate orange (Poncirus trifoliata Raf.). Chitinase activity of the transformed plants was higher than that of the non-transformed plants. The introduced gene was not detected in the scions grafted onto the transformed plants. The effect of the introduction of foreign gene into rootstocks on the scions was discussed. 相似文献
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4.
A.T. Rodríguez R.M. Cárdenas M.G. Velázquez 《Pesticide biochemistry and physiology》2007,89(3):206-215
Seeds of rice (Oryza sativa L.) were treated with chitosan and hydrolyzed chitosan at 100, 500 and 1000 mg L−1. After 18 days of germination, spore suspension of Pyricularia grisea was applied. The enzyme activity of phenylalanine ammonia-lyase, β-1-3-glucanase, chitinase and chitosanase in leaves of rice seedlings was evaluated after 24, 72, 120 and 168 h of inoculation. Blast affected area (%) was evaluated 7 and 14 days after spraying spore suspension. Chitosan performance to elicit defense response induction was associated with the concentration and type of chitosan. The activity of most of the enzymes tested was induced in leaves of treated seeds with chitosan and hydrolyzed chitosan at 1000 and 500 mg L−1, respectively. The highest enzyme activities were observed with hydrolyzed chitosan after 72 h however, compared to chitosan, the activity was not maintained during the entire post-inoculation period. The highest control (0 = no lesions) of P. grisea in rice seedlings was observed at 1000 mg L−1 in both chitosan and hydrolyzed chitosan treated leaves. Symptoms of infection by P. grisea were evident after 14 days evaluation date, but according to the standard scale proposed by the International Rice Research Institute, these symptoms fell into the resistance category of blast diseases. 相似文献
5.
以短短芽胞杆菌FJAT-0809-GLX基因组DNA为模板,通过PCR扩增得到几丁质酶基因chiD序列,再将chiD序列连接到pMD18-T克隆载体上,形成重组载体pMD18-T-chiD,转化大肠杆菌DH5ɑ并测序,经过核苷酸和氨基酸序列分析获得几丁质酶基因chiD的序列片段为1 524 bp,编码507个氨基酸,理论蛋白质分子量约54.55 ku,其等电点约为5.77,表明该几丁质酶为酸性几丁质酶。将重组质粒pMD18-T-chiD双酶切后与表达载体pET-28a连接,构建重组表达载体pET28a-chiD,并转入大肠杆菌BL21中进行IPTG诱导表达,结果表明:重组蛋白质的分子量约55 ku,与预测的蛋白分子量结果一致。为了提高重组蛋白的表达量,对培养时间、IPTG浓度和温度3个参数进行优化,并将表达产物进行SDS-PAGE分析,最后得出重组载体pET28a-chiD的最佳诱导表达参数分别为8 h、0.5 mmol/L和28℃。这为短短芽胞杆菌来源的几丁质酶的进一步研究奠定了良好基础。 相似文献
6.
The species of an ectomycorrhizal (ECM) community were investigated in a temperate oak forest by morphotyping and ITS rDNA sequencing. Thirty-six ECM morphotypes were found at the site. The niche effect (as organic soil, mineral soil or dead woody debris artificially introduced in the soil) on the ECM community structure and on the potential catabolic activities of the most abundant morphotypes was studied. The morphotypes in each niche were subjected to enzymatic tests developed for hydrolytic and oxidative enzymes involved in the decomposition of organic compounds. The ECM community structure varied widely depending on the soil horizon or habitat patch. The species richness was higher in the A1 horizon than in the other niches. Different ECM species had different activity patterns for the eight enzymatic tests while co-occurring in the same niche. Catabolic activities also changed within species between niches. Dead woody debris were extensively colonized by two saprotrophic fungi (Megacollybia platyphylla and Armillaria sp.) and, in this particular niche, ECM morphotypes predominantly belonged to the genera Lactarius and Tomentella. These morphotypes showed high chitinase activities. This study suggested also that some ECM fungi could obtain nutrients via the chitin degradation of dead or live saprobes. 相似文献
7.
为了解盾壳霉寄生核盘菌的生化机理,采用了紫外分光光度法测定了盾壳霉寄生菌核过程中,葡聚糖酶和几丁质酶的变化,并用电导率仪测定了这个过程中电导率的变化。结果表明:从被盾壳霉寄生的活菌核检测到(处理)葡聚糖酶和几丁质酶活性明显高于活菌核和被盾壳霉寄生的死菌核,其中处理的葡聚糖酶活性从15 d以后就明显高于菌核自身也高于盾壳霉本身的酶活性,处理的几丁质酶活性从10 d起就维持在较高水平,处理的电导率从10 d以后明显高于活菌核。在这个过程中这两种酶活性提高,电导率增大,膜透性增强。 相似文献
8.
几丁质酶广泛存在于植物、动物及微生物细胞和组织中,参与多种生理过程。研究发现许多动物、植物、微生物都可以产生几丁质酶。笔者主要对微生物几丁质酶的特性、功能及几丁质酶在植物真茵病害防治中的应用方面进行了综合论述,并对几丁质酶在植物病害生物防治和抗病基因工程中的应用前景进行了展望。 相似文献
9.
抗逆生防木霉筛选及其相关因子诱导 总被引:1,自引:0,他引:1
对供试各木霉菌株经303、5、404、55、0℃保温24 h后,于28℃培养6~7 d,进行耐高温菌株筛选。结果表明,耐高温生防木霉菌株T-30经热激后可产生抗逆蛋白,且表达量较大。对高温抗逆蛋白进行DEAE-32及Sephadex G-100柱层析得到单一蛋白洗脱峰。SDS-PAGE电泳发现该抗逆蛋白含有2个亚基,分子量分别为30和42 kD。酶学检测发现,该高温抗逆蛋白有几丁质酶活力,其最适pH值为5.0,最适温度为50℃,Km值为0.91 mg/ml,对Na+有较强的耐受性。 相似文献
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