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1.
使用电喷雾质谱检测了糖用甜菜水提取液和乙醇提取液,并且采用串联质谱技术分析了这些提取液中化合物的碎裂规律。实验结果表明水提取液和乙醇提取液都适合在电喷雾正离子模式和负离子模式下检测,说明甜菜中化学成分复杂,种类较多。在串联质谱中,这些成分大多丢失水、二氧化碳、葡萄糖等中性碎片,说明这些化合物含有羟基、羧基、葡萄糖基,这些基团在化合物抗氧化性能中都起到重要作用,这些与文献中的研究结果相同。说明使用电喷雾质谱技术分析甜菜的抗氧化性能十分可靠,甜菜提取液不需经过复杂的前处理就可以检测,操作简单、快速,灵敏度高,可以用于复杂体系抗氧化性的快速筛选。 相似文献
2.
Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues 总被引：2，自引：0，他引：2
Anna Lytovchenko Romina Beleggia Nicolas Schauer Tal Isaacson Jan E Leuendorf Hanjo Hellmann Jocelyn KC Rose Alisdair R Fernie 《Plant methods》2009,5(1):4-11
Background
The concept of metabolite profiling has been around for decades and technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. However, studies are generally confined to polar compounds alone. Here we describe a simple method for lipophilic compounds analysis in various plant tissues. 相似文献3.
Background
Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents. 相似文献4.
Background
Kinome profiling aims at the parallel analysis of kinase activities in a cell. Novel developed arrays containing consensus substrates for kinases are used to assess those kinase activities. The arrays described in this paper were already used to determine kinase activities in mammalian systems, but since substrates from many organisms are present we decided to test these arrays for the determination of kinase activities in the model plant species Arabidopsis thaliana. 相似文献5.
Alfredo J Ibáñez Judith Scharte Philipp Bones Alexander Pirkl Stefan Meldau Ian T Baldwin Franz Hillenkamp Engelbert Weis Klaus Dreisewerd 《Plant methods》2010,6(1):14
Background
Successful defence of tobacco plants against attack from the oomycete Phytophthora nicotianae includes a type of local programmed cell death called the hypersensitive response. Complex and not completely understood signaling processes are required to mediate the development of this defence in the infected tissue. Here, we demonstrate that different families of metabolites can be monitored in small pieces of infected, mechanically-stressed, and healthy tobacco leaves using direct infrared laser desorption ionization orthogonal time-of-flight mass spectrometry. The defence response was monitored for 1 - 9 hours post infection. 相似文献6.
Bennett Young Raymond Wightman Robert Blanvillain Sydney B Purcel Patrick Gallois 《Plant methods》2010,6(1):27
Background
Programmed cell death (PCD) is an essential process for the life cycle of all multicellular organisms. In higher plants however, relatively little is known about the cascade of genes and signalling molecules responsible for the initiation and execution of PCD. To aid with the discovery and analysis of plant PCD regulators, we have designed a novel cell death assay based on low cytosolic pH as a marker of PCD. 相似文献7.
Background
Salicylic acid (SA) is an important signalling molecule in plant defenses against biotrophic pathogens. It is also involved in several other processes such as heat production, flowering, and germination. SA exists in the plant as free SA and as an inert glucose conjugate (salicylic acid 2-O-β-D-glucoside or SAG). Recently, Huang et al. developed a bacterial biosensor that responds to free SA but not SAG, designated as Acinetobacter sp. ADPWH_lux. In this paper we describe an improved methodology for Acinetobacter sp. ADPWH_lux-based free SA quantification, enabling high-throughput analysis, and present an approach for the quantification of SAG from crude plant extracts. 相似文献8.
Sandra Dèrozier Franck Samson Jean-Philippe Tamby Cécile Guichard Véronique Brunaud Philippe Grevet Séverine Gagnot Philippe Label Jean-Charles Leplé Alain Lecharny Sébastien Aubourg 《Plant methods》2011,7(1):1-10
Background
Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.Results
To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.Conclusion
The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes. 相似文献9.
Background
The progress and completion of various plant genome sequencing projects has paved the way for diverse functional genomic studies that involve cloning, modification and subsequent expression of target genes. This requires flexible and efficient procedures for generating binary vectors containing: gene fusions, variants from site-directed mutagenesis, addition of protein tags together with domain swaps and deletions. Furthermore, efficient cloning procedures, ideally high throughput, are essential for pyramiding of multiple gene constructs. 相似文献10.
A decline in the hot-water-soluble pectin and an increase in the hot-water-insoluble pectin were observed when cucumber fruits were subjected to chilling temperatures. Infrared absorption spectra revealed the presence of highly esterified carboxyl groups in the soluble pectin, and of free carboxyl groups in the insoluble pectin. An increase of the insoluble pectin during chilling was also found in other chilling-sensitive plants. From these results it is suggested that a de-esterification of pectin and the concomitant increase of polymeric pectin takes place during chilling, making cell walls firmer, and that these pectic changes may be a characteristic common to a number of chilling-sensitive plants. 相似文献
11.
Jing Wang Chongnan Wang Yan Long Clare Hopkins Smita Kurup Kede Liu Graham J King Jinling Meng 《Plant methods》2011,7(1):1-7
Background
Protein extraction is a frequent procedure in biological research. For preparation of plant cell extracts, plant materials usually have to be ground and homogenized to physically break the robust cell wall, but this step is laborious and time-consuming when a large number of samples are handled at once.Results
We developed a chemical method for lysing Arabidopsis cells without grinding. In this method, plants are boiled for just 10 minutes in a solution containing a Ca2+ chelator and detergent. Cell extracts prepared by this method were suitable for SDS-PAGE and immunoblot analysis. This method was also applicable to genomic DNA extraction for PCR analysis. Our method was applied to many other plant species, and worked well for some of them.Conclusions
Our method is rapid and economical, and allows many samples to be prepared simultaneously for protein analysis. Our method is useful not only for Arabidopsis research but also research on certain other species. 相似文献12.
Background
Microsatellites are popular molecular markers in many plant species due to their stable and highly polymorphic nature. A number of analysis methods have been described but analyses of these markers are typically performed on cumbersome polyacrylamide gels or more conveniently by capillary electrophoresis on automated sequencers. However post-PCR handling steps are still required. High resolution melting can now combine detailed sequence analysis with the closed-tube benefits of real-time PCR and is described here as a novel way to verify the identity of plant varieties such as grapevine and olive. 相似文献13.
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15.
Thomas H Hansen Kristian H Laursen Daniel P Persson Pai Pedas Søren Husted Jan K Schjoerring 《Plant methods》2009,5(1):12-11
Background
Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. 相似文献16.
17.
Kim H Hebelstrup Michael W Christiansen Massimiliano Carciofi Birgitte Tauris Henrik Brinch-Pedersen Preben B Holm 《Plant methods》2010,6(1):15
Background
Cloning of gene casettes and other DNA sequences into the conventional vectors for biolistic or Agrobacterium-mediated transformation is hampered by a limited amount of unique restriction sites and by the difficulties often encountered when ligating small single strand DNA overhangs. These problems are obviated by "The Uracil Specific Excision Reagent (USER™)" technology (New England Biolabs) which thus offers a new and very time-efficient method for engineering of big and complex plasmids. 相似文献18.
Background
Simultaneous analysis of multiple functional-related phytohormones and their metabolites will improve our understanding of interactions among different hormones in the same biologic process. 相似文献19.
根据地毯草黄单胞菌花叶万年青致病变种（Xanthomonas axonopodis pv. dieffenbachiae）的RAPD序列设计特异性引物,并对羧基化磁珠的结合性能进行检验,从而建立和优化了免疫磁性分离–PCR体系,对安祖花细菌性枯萎病进行早期检测。结果表明：1 mg磁珠对多克隆抗体最大吸附值为0.268 mg,进行免疫磁性捕获时免疫磁珠的最佳浓度是0.566 ~ 0.741 mg ?mL-1。免疫磁性分离–PCR可以减少PCR反应中的抑制物质,对X. axonopodis pv. dieffenbachiae的检测灵敏度可以达到10 ~ 100 cfu ?mL-1,比常规PCR检测灵敏至少100倍。 相似文献
20.
Berendzen K Searle I Ravenscroft D Koncz C Batschauer A Coupland G Somssich IE Ulker B 《Plant methods》2005,1(1):4