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感染高粱花叶病毒甘蔗叶片cDNA文库构建及评价 总被引:1,自引:0,他引:1
为了研究甘蔗花叶病与甘蔗寄主致病的互作分子机制,利用SMART技术成功构建了感染高梁花叶病毒甘蔗叶片的cDNA文库,用于后续酵母双杂交互作蛋白筛选试验.采用Omega公司Plant RNA Kit提取感染高粱花叶病毒甘蔗叶片总RNA,经过Oligotex纯化获得mRNA,将其反转录成cDNA第一链,再在DNA聚合酶作用下,通过长距离PCR扩增双链cDNA.经SfiI酶切并去除短片段后,连接到pGADT7-SfiI载体上,成功获得初级cDNA文库,最后以初级文库100万克隆为基数扩增,得到扩增文库并提取质粒.经检测构建的文库容量为1.6×106 cfu,文库滴度2.2× 106 cfu/mL,文库cDNA插入片段长度主要分布在700~2 000 bp,文库重组率约为96%.结果表明,该文库质量较好,为筛选分离抗病功能基因及开展寄主与病毒互作的研究奠定了基础. 相似文献
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以甘蔗(Saccharum officinarum Linn)为材料,利用SMART技术构建甘蔗叶片全长cDNA文库,从甘蔗叶片中提取总RNA并分离mRNA,用分离到的微量mRNA合成cDNA第1链,通过长距离PCR扩增得到足量的双链cDNA,同时比较了不同双链cDNA与载体的连接效率和转化效率。结果表明,所构建的甘蔗叶片cDNA文库库容和重组百分比分别是1.8×106个克隆和98.6%,插入片段长度主要集中在0.6-2.5kb,平均插入片段长度约为1.2kb。本研究为国内首次报道甘蔗全长cDNA文库,所构建的文库拥有较好的质量,为项目组进一步开展甘蔗功能基因组学研究提供了平台。 相似文献
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全长cDNA文库构建是改良胡椒性状的分子机理研究基础,有助于重要相关基因的克隆、功能分析、调控及其利用.以性状优良并具代表性的海南蒟(Piper hainanense)为试材,采用优化的In-Fusion SMARTerTM Directional cDNA Library Construction Kit技术,构建叶片材料的全长cDNA文库.结果表明:文库的滴度为2.0×106 cfu/mL,文库容量为1.3×106 cfu.从文库中随机挑取20个克隆进行质粒DNA提取、PCR及酶切鉴定,结果显示:插入片段大小为1.0~2.0 kb,重组率为100%,表明该文库质量较高,可满足后续研究需要. 相似文献
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花生蛴螬中肠组织cDNA文库的构建 总被引:1,自引:0,他引:1
利用Qiagen试剂盒提取花生害虫华北大黑鳃金龟幼虫蛴螬中肠总RNA,并纯化mRNA,然后按照Stratagene试剂盒合成双链cDNA,经过凝胶柱层析,收集符合要求的cDNA片段,加工后与Uni-ZAP XR Vector连接,经Gigapack Ⅲ Gold体外包装,即获得蛴螬的cDNA文库,未扩增文库滴度1.95×106pfu/mL,重组率为99.7%,扩增后文库滴度达1.34×109pfu/mL,插入cDNA片段的长度平均为1.34kb,表明成功构建了高质量的蛴螬中肠cDNA文库。 相似文献
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Lakshman Lal 《Potato Research》1987,30(2):329-334
Summary The effects of the leaves of five plant species, one from each of the generaAmbrosia, Anemone, Eupatorium, Eucalyptus andLantana, on potato tuber moth were investigated under indigenous storage conditions at the Central Potato Research Station, Shillong
(1800 m above sea level). Their action was compared with that of a biological insecticide (spores ofBacillus thuringiensis), a chemical insecticide (carbaryl), and an untreated control. The data collected after six months storage on tuber damage,
sprout damage and the rotting indicated that the leaves ofLantana aculeata provided most protection to the tubers, reducing damage from over 70% in the check to below 5%, and sprout damage from over
45% to below 3%. Next best wasEucalyptus globulus followed byB. thuringiensis. They may be used on tubers stored for table use or for seed as they had no adverse effect on germination or on the yield
of a subsequent crop. 相似文献
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《Journal of Crop Improvement》2013,27(2):81-114
Summary The efficiency of N fertilizers is usually poor; often less than 50% of the applied N is taken up by the crop. This review focuses on various N fertilizers with respect to the significance of different N loss pathways, namely (i) ammonia volatilization, (ii) dinitrogen and nitrogen oxide emissions, and (iii) nitrate leaching. Further, the significance of biological N immobilization, ammonium fixation and, finally, the impact of nitrate vs. ammonium uptake on crop yield are also discussed. The reviewed literature shows that N fertilizers may differ markedly in their susceptibility to losses. There is, however, considerable scope to improve N efficiency of each N source by proper N management practices. 相似文献
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M. I. Siri P. Villanueva M. J. Pianzzola L. Franco Fraguas G. Galván M. Acosta F. Ferreira 《Potato Research》2004,47(3-4):127-138
Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results
for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin
presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear
relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins
and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds. 相似文献
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Alexander N. Shikov Elena V. Flisyuk Ekaterina D. Obluchinskaya Olga N. Pozharitskaya 《Marine drugs》2020,18(11)
Marine organisms represent an excellent source of innovative compounds that have the potential for the development of new drugs. The pharmacokinetics of marine drugs has attracted increasing interest in recent decades due to its effective and potential contribution to the selection of rational dosage recommendations and the optimal use of the therapeutic arsenal. In general, pharmacokinetics studies how drugs change after administration via the processes of absorption, distribution, metabolism, and excretion (ADME). This review provides a summary of the pharmacokinetics studies of marine-derived active compounds, with a particular focus on their ADME. The pharmacokinetics of compounds derived from algae, crustaceans, sea cucumber, fungus, sea urchins, sponges, mollusks, tunicate, and bryozoan is discussed, and the pharmacokinetics data in human experiments are analyzed. In-depth characterization using pharmacokinetics is useful for obtaining information for understanding the molecular basis of pharmacological activity, for correct doses and treatment schemes selection, and for more effective drug application. Thus, an increase in pharmacokinetic research on marine-derived compounds is expected in the near future. 相似文献
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Novel food and non-food uses for sorghum and millets 总被引:4,自引:3,他引:4
Sorghum and millets have considerable potential in foods and beverages. As they are gluten-free they are suitable for coeliacs. Sorghum is also a potentially important source of nutraceuticals such antioxidant phenolics and cholesterol-lowering waxes. Cakes, cookies, pasta, a parboiled rice-like product and snack foods have been successfully produced from sorghum and, in some cases, millets. Wheat-free sorghum or millet bread remains the main challenge. Additives such as native and pre-gelatinised starches, hydrocolloids, fat, egg and rye pentosans improve bread quality. However, specific volumes are lower than those for wheat bread or gluten-free breads based on pure starches, and in many cases, breads tend to stale faster. Lager and stout beers with sorghum are brewed commercially. Sorghum's high-starch gelatinisation temperature and low beta-amylase activity remain problems with regard to complete substitution of barley malt with sorghum malt . The role of the sorghum endosperm matrix protein and cell wall components in limiting extract is a research focus. Brewing with millets is still at an experimental stage. Sorghum could be important for bioethanol and other bio-industrial products. Bioethanol research has focused on improving the economics of the process through cultivar selection, method development for low-quality grain and pre-processing to recover valuable by-products. Potential by-products such as the kafirin prolamin proteins and the pericarp wax have potential as bioplastic films and coatings for foods, primarily due to their hydrophobicity. 相似文献
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Résumé
Fusarium culmorum et, dans une moindre mesure,F. avenaceum, F. sambucinum, Colletotrichum coccodes etCylindrocarpon sp. ont été isolés de tubercules entreposés en atmosphère contr?lée (AC) et atteints de pourriture grave. Des expériences
d'inoculation de tubercules avec 10 espèces ou variétés fongiques et conservation en AC à 8 et 10°C ont montré que: 1) les
champignons non pathogènes en atmosphère normale (ex.:Cylindrocarpon sp.) le devenaient en AC, 2) ceux qui étaient faiblement pathogènes en atmosphère normale étaient nettement plus virulents
en AC (ex.:F. culmorum), 3) les espèces ou variétés virulentes en conditions d'air normales (Fusarium sulphureum etPhoma exigua var.foveata) ne provoquaient pas de pourriture plus grave en AC et leur activité pouvait même être légérement freinée.
Summary The extensive rotting of tubers which occurred during storage trials in controlled atmospheres has led to a mycological and pathological study of the rots. The most frequently isolated species wasFusarium culmorum althoughF. avenaceum, F. Sambucinum, Coletotrichum coccodes and aCylindrocarpon sp. were also present. Tubers of cvs Bintje and Ulla were inoculated with 19 isolates of 10 fungal species or varieties associated with rotting of stored tubers (Table 1). The tubers were incubated at 8 or 10°C in normal or controlled (2% O2:1% CO2, 2% O2:6% CO2, 2% O2:12% CO2, 6% O2:6% CO2) atmospheres and the extent of rotting was determined a month later (Tables 2 and 3 and Fig. 2). Species which were weakly virulent in a normal atmosphere (variousFusarium spp. andPhoma exigua var.exigua) were clearly more aggressive in the controlled atmosphere and this was especially evident in the case ofF. culmorum. Species ot pathogenic in a normal atmosphere under experimental conditions (Colletotrichum coccodes, Cylindrocarpon sp.,Phoma eupyrena) caused rotting in the controlled atmosphere. The two species which were virulent in a normal atmosphere (F. sulphureum andP. exigua var.foveata) were also virulent in the controlled atmosphere and in some cases the extent of rotting was even less. As a general rule, rotting caused by weakly virulent fungi was greater in both cultivars in the controlled than in a normal atmosphere.
Zusammenfassung Eine starke F?ulnis von in Kontrotrollierter Atmosph?re gelagerten Kartoffelknollen veranlasste eine mykologische und pathologische Untersuchung. Der am h?ufigsten isolierte Pilz warFusarium culmorum. F. avenaceum, F. sambucinum, Colletotrichum coccodes undCylindrocarpon sp. wurden ebenfalls bestimmt. Neunzehn St?mme von 10 Pilzsorten oder-Arten welche bei Lagerf?ulnis vorkommen (Tab. 1) wurden auf Knollen der Sorten Bintje und Ulla inokuliert. Die Lagerung wurde bei 8 oder 10°C under Normalbedingungen oder kontrollierter Atmosph?re durchgeführt (2% O2:1% CO2, 2% O2:6% CO2, 2% O2:12% CO2, 6% O2:6% CO2). Der F?ulnisgrad (Abb. 1) wurde nach einem Monat bonitiert. Die Ergebnisse sind in den Tabellen 2 und 3 dargestellt und in der Abb. 2 illustriert. Unter in Normalbedingungen nur schwach virulente Arten (verschiedeneFusarien undPhoma exigua var. exigua) waren deutlich aktiver in kontrollierter Atmosph?re. Dies was besonders der Fall fürF. culmorum. Pilze, die im Versuch unter Normalbedingungen nicht pathogen waren (Colletotrichum coccodes, Cylindrocarpon sp., Phoma eupyrena), haben unter kontrollierter Atmosph?re Sch?den verursacht. Hingegen werden die beiden unter Normalbedingungen sehr pathogenen Arten (F. sulphureum undP., exigua var.foveata), durch kontrollierte Atmosph?re wenig beeinflusst. Es wurde sogar in einigen F?llen eine gewisse Verringerung des F?ulnisbefalls beobachtet. Ganz allgemein haben die getesteten kontrollierten Bedingungen die Entwicklung der Pilzf?ulnis an beiden Kartoffelsorten gef?rdert, speziell durch Pilzarten, welche unter Normal-bedingungen wenig Sch?den verursachen.相似文献
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Alejandro Carlos Tozzini María Fernanda Ceriani María Verónica Saladrigas H. Esteban Hopp 《Potato Research》1991,34(3):317-324
Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets
that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and
slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging
toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta.
Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X1 (also called RXact). 相似文献
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Alejandro M. S. Mayer Abimael D. Rodríguez Orazio Taglialatela-Scafati Nobuhiro Fusetani 《Marine drugs》2013,11(7):2510-2573
The peer-reviewed marine pharmacology literature from 2009 to 2011 is presented in this review, following the format used in the 1998–2008 reviews of this series. The pharmacology of structurally-characterized compounds isolated from marine animals, algae, fungi and bacteria is discussed in a comprehensive manner. Antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral pharmacological activities were reported for 102 marine natural products. Additionally, 60 marine compounds were observed to affect the immune and nervous system as well as possess antidiabetic and anti-inflammatory effects. Finally, 68 marine metabolites were shown to interact with a variety of receptors and molecular targets, and thus will probably contribute to multiple pharmacological classes upon further mechanism of action studies. Marine pharmacology during 2009–2011 remained a global enterprise, with researchers from 35 countries, and the United States, contributing to the preclinical pharmacology of 262 marine compounds which are part of the preclinical pharmaceutical pipeline. Continued pharmacological research with marine natural products will contribute to enhance the marine pharmaceutical clinical pipeline, which in 2013 consisted of 17 marine natural products, analogs or derivatives targeting a limited number of disease categories. 相似文献