ISBN-13: 9783642832123 / Angielski / Miękka / 2011 / 304 str.
ISBN-13: 9783642832123 / Angielski / Miękka / 2011 / 304 str.
Modern Methods of Plant Analysis When the handbook Modern Methods of Plant Analysis was first introduced in 1954 the considerations were: 1. the dependence of scientific progress in biology on the improvement of existing and the introduction of new methods; 2. the difficulty in finding many new analytical methods in specialized journals which are normally not accessible to experimental plant biologists; 3. the fact that in the methods sections of papers the description of methods is frequently so compact, or even sometimes so incomplete that it is difficult to reproduce experiments. These considerations still stand today. The series was highly successful, seven volumes appearing between 1956 and 1964. Since there is still today a demand for the old series, the publisher has decided to resume publication of Modern Methods of Plant Analysis. It is hoped that the New Series will be just as acceptable to those working in plant sciences and related fields as the early volumes undoubtedly were. It is difficult to single out the major reasons for success of any publication, but we believe that the methods published in the first series were up-to-date at the time and presented in a way that made description, as applied to plant material, complete in itself with little need to consult other publications. Contributirig authors have attempted to follow these guidelines in this New Series of volumes.
Raw Material.- Barley and Malt Analysis.- 1 General Introduction.- 2 Barley Analysis.- 2.1 Visual Inspection.- 2.2 Mechanical Analysis.- 2.3 Germination Tests.- 2.4 Chemical Analysis.- 2.5 Varietal Identification.- 2.6 Micro-Malting.- 3 Malt Analysis.- 3.1 Wort Production and Analysis.- 3.2 Modification Analysis.- 3.3 Chemical Analysis.- 3.4 Enzyme Analysis.- 4 Further Reading.- References.- Reversed Phase HPLC as an Aid for the Identification of Barley Cultivars.- 1 Introduction.- 1.1 Reversed-Phase High Performance Liquid Chromatography.- 1.1.1 The RP-HPLC System.- 1.1.2 Sample Preparation.- 1.1.3 Hordeins.- 1.1.4 Albumins and Globulins.- 1.1.5 Glutelins.- 1.2 Hordein Identification.- 1.3 Repeatability of Hordein Elution.- 1.3.1 RP-HPLC Separations of Hordeins.- 1.3.2 RP-HPLC Separation of Albumins and Globulins.- 1.3.3 RP-HPLC Separation of Glutelins.- References.- The Application of Immunofluorescence for Detecting Specific Proteins in Barley Seeds.- 1 Introduction.- 2 Immunofluorescence.- 2.1 Fluorescent Dyes.- 2.2 Staining Methods.- 2.2.1 The Direct Method with Fluorochrome-Labelled Primary Antibodies.- 2.2.2 The Indirect Method with Fluorochrome-Labelled Secondary Antibodies.- 2.2.3 Use of Biotin-Labelled Primary Antibodies.- 2.2.4 Use of Biotin-Labelled Secondary Antibodies.- 3 Materials, Protocols and Comments.- 3.1 Production and Purification of Antibodies.- 3.2 Conjugation of Antibodies with Fluorochromes.- 3.2.1 Protocol for FITC Conjugation.- 3.2.2 Comments to the FITC Conjugation Procedure.- 3.2.3 Conjugation with Other Fluorochromes.- 3.3 Biotinylation.- 3.3.1 Protocol for Conjugation with Biotin.- 3.3.2 Comments to the Biotin Conjugation.- 3.4 Fixation.- 3.4.1 Protocol for PLP Fixation of Seeds to be Sectioned.- 3.4.2 Surface Fixation of Sanded Seeds for Retention of Germination Capacity.- 3.4.3 Comments to the Fixation.- 3.5 Preparation of Tissue.- 3.5.1 Thin Sections.- 3.5.2 Sanded Seeds.- 3.6 Staining Procedures.- 3.6.1 General Considerations.- 3.6.2 Direct Staining of Thin Sections.- 3.6.3 Indirect Staining of Thin Sections.- 3.6.4 Use of Biotin-Labelled Primary Antibodies.- 3.6.5 Use of Biotin-Labelled Secondary Antibodies.- 3.6.6 Two-Colour Staining of Thin Section.- 3.6.7 Direct Staining of Sanded Seeds.- 4 Concluding Remarks.- References.- Fluorimetric Methods for the Analysis of Malt Modification and Pre-Harvest Sprouting.- 1 Introduction.- 2 Modification and Homogeneity of Barley Malt.- 2.1 Field of Application.- 2.2 Sample Size.- 2.3 Reagents and Materials.- 2.4 Apparatus.- 2.5 Procedure.- 2.5.1 Fixation and Sanding.- 2.5.2 Staining.- 2.5.3 Examination.- 2.6 Expression of Results.- 2.7 Use of Malt Modification Analysis.- 3 Pre-Sprouting in Cereal Seeds.- 3.1 Field of Application.- 3.2 Sample Size.- 3.3 Reagents and Materials.- 3.4 Apparatus.- 3.5 Procedure.- 3.5.1 Fixation and Sanding.- 3.5.2 Staining.- 3.5.3 Examination.- 3.6 Expression of Results.- 3.7 Use of Pre-Sprouting Test.- 4 Further Applications of the Carlsberg Seed Fixation System.- References.- Hop Analysis.- 1 Introduction.- 2 Chemical Composition of Hops.- 3 Analysis.- 3.1 Resin Analysis of Hops.- 3.2 The Conductometric Lead Value of Hops.- 3.3 Verzele Conductometric Analysis of ?-Acids.- 3.4 Beer Bitterness Analysis.- 3.5 High Performance Liquid Chromatography (HPLC).- 4 Essential Oil of Hops.- 5 Polyphenols.- 6 Prospect.- References.- Determination of ?-Glucan in Barley, Malt, Wort and Beer.- 1 Introduction.- 2 The Nature of ?-Glucan.- 3 Quantification of Barley ?-Glucan.- 3.1 Methods Based on Extraction.- 3.1.1 Viscosity Methods.- 3.1.2 Difference Methods.- 3.1.3 Selective Precipitation Methods.- 3.2 Non-Extraction Methods.- 3.2.1 Near Infrared Reflectance.- 3.2.2 Calcofluor Staining Used in Flour Suspensions.- 3.3 Enzymatic Methods.- 3.3.1 Methods Using Bacterial ?-Glucanase.- 3.3.2 Methods Based on Fungal Cellulases.- 4 Quantification of ?-Glucan in Malt, Wort and Beer.- 4.1 Total ?-Glucan Content of Malt.- 4.1.1 Non-Enzymatic Methods.- 4.1.2 Enzymatic Methods.- 4.2 ?-Glucan Content of Wort and Beer.- 4.2.1 Non-Enzymatic Methods.- 4.2.2 Enzymatic Methods.- 5 An Automatic ?-Glucan Analyzer System.- 5.1 Principle.- 5.2 Apparatus and Conditions.- 5.3 Quantification of ?-Glucan in Barley, Malt, Wort and Beer.- 5.3.1 Sample Preparation for Wort and Beer.- 5.3.2 Sample Preparation for Barley and Malt.- 6 Evaluation of Methods.- 6.1 Specificity.- References.- Analysis of Proanthocyanidins and Phenolic Acids in Barley, Malt, Hops and Beer.- 1 Introduction.- 2 Choice of Analytical Methods.- 3 Chromatographic Methods.- 3.1 High Performance Liquid Chromatography (HPLC).- 3.1.1 Preparation of Barley and Malt Samples.- 3.1.2 Preparation of Hop Samples.- 3.1.3 Preparation of Beer Samples.- 3.1.4 Choice of HPLC Columns.- 3.1.5 Mobile Phase.- 3.1.6 Standards.- 3.1.7 Quantitation.- 3.2 Thin Layer Chromatography (TLC).- 3.3 Column Chromatography.- 4 Colourimetric Methods.- 4.1 Determination of Proanthocyanidins in Barley, Malt, Wort and Beer.- 4.2 Measurement of Total Polyphenols in Beer and Wort.- 4.3 Analysis of Proanthocyanidins and Catechins Using Acidified Vanillin.- 4.4 Determination of Proanthocyanidins and Catechins in Beer with p-Dimethylaminocinnamaldehyde as a Reagent.- 5 Vanillin Test of Barley Grains.- References.- Yeast.- Breeding and Characterization of Brewer’s Yeast Including Single Chromosome Transfer.- 1 Introduction.- 2 Sporulation, Isolation of Meiotic Progeny and Construction of Hybrids.- 2.1 Mass Spore Isolation.- 2.2 Isolation of Colonies and Spore Staining.- 2.3 Determination of Mating Type.- 2.4 Construction of Hybrids by Mating of Meiotic Segregants.- 2.5 Random Mating.- 3 Genetics of Diacetyl Formation.- 3.1 Isolation of Spontaneous Mutants Resistant to Sulfometuron Methyl.- 4 Transformation.- 5 Single Chromosome Transfer.- 5.1 Transfer of Chromosome V.- 5.2 Characterization of Chromosome V by Restriction Endonuclease Analysis.- References.- Genetic Analysis and Transformation of Distiller’s Yeast.- 1 Introduction.- 2 Genetic Analysis.- 2.1 Sporulation and Mating Ability.- 2.1.1 Sporulation Test.- 2.1.2 Spore Isolation.- 2.1.3 Mating Type Tests.- 2.1.3.1 Induction of Respiratory-Deficient Mutations.- 2.1.3.2 Determination of Mating Type.- 2.1.3.3 Rare Mating on Solid Medium.- 2.1.3.4 Rare Mating in Liquid Medium.- 2.1.4 Characterization of Auxotrophic Spore Clones.- 2.1.5 Sporulation and Mating Ability of Strain A.- 2.2 Mutagenesis.- 2.2.1 Procedure.- 2.2.2 Mutants Obtained from Spore Clones of Strain A.- 2.3 Crossing and Tetrad Analysis.- 3 Molecular Characterization.- 3.1 Procedure.- 3.2 Analysis of Chromosomes III of Strain A.- 4 Genetic Transformation of an Industrial Yeast Strain.- 4.1 Selection of Transformants by Acquired Prototrophy.- 4.1.1 Transformation Procedure.- 4.1.2 Screening for Amylase Activity.- 4.1.3 Stability Test of Transformants.- 4.1.4 Integration of a Self-Replicating Plasmid.- 4.1.5 Breeding Through Transformation of Spore-Derived Strains.- 4.2 Integration Vector with Drug Resistance as Selective Marker.- 4.2.1 Transformation Procedure Using a Non-Replicating Plasmid.- 4.2.2 Screening for ?-Galactosidase Activity.- References.- Automated Testing of Brewer’s Yeast Strains for Fermentation Characteristics (Multiferm, System Carlsberg).- 1 Introduction.- 2 Testing Techniques.- 2.1 Small-Scale Fermentations.- 2.2 MULTIFERM Fermentations.- 2.2.1 Description of the MULTIFERM System.- 2.2.2 Working Procedures.- 2.2.2.1 Propagation of Yeast.- 2.2.2.2 Wort Collection and Handling.- 2.2.2.3 Start of Fermentations.- 2.2.2.4 Sampling During Fermentations.- 2.2.2.5 End of Fermentations.- 2.2.2.6 Cleaning in Place of the MULTIFERM.- 2.2.2.7 Re-Use of Yeast.- 2.2.3 Analyses Performed.- 2.2.3.1 Yeast Cell Size and Yeast Cell Number.- 2.2.3.2 Pitching Rate.- 2.2.3.3 Extract Determination.- 2.2.3.4 Yeast Yield.- 2.2.3.5 Vicinal Diketones and Ester/Alcohol (Aroma Profile) Analyses.- 2.2.4 Data.- 2.2.4.1 Data Handling and Storage.- 2.2.4.2 Data Evaluation.- 3 Conclusion.- References.- The Use of Nucleotide Sequence Polymorphisms and DNA Karyotyping in the Identification of Brewer’s Yeast Strains and in Microbiological Control.- 1 Introduction.- 1.1 Restriction Fragment Length Polymorphisms.- 1.2 DNA Karyotypes.- 2 Storage and Propagation of Yeast Strains.- 3 Restriction Site Analysis of Yeast Genomic DNA.- 3.1 Isolation of Yeast Genomic DNA for Restriction Site Analysis.- 3.2 Analysis of Restriction Fragment Length Polymorphisms (RFLP).- 3.2.1 RFLP Level A.- 3.2.2 RFLP Level B.- 4 Southern Transfer and Molecular Hybridization of Radioactively Labelled Probes.- 4.1 Transfer of DNA Molecules from Agarose Gels onto Nitrocellulose Membranes by Southern Transfer.- 4.2 Molecular Hybridization.- 4.2.1 Hybridization Buffers.- 4.2.2 Nick Translation.- 4.2.3 Post Hybridization Wash.- 5 Isolation of Yeast Genomic DNA for Chromosome Separation.- 5.1 Orthogonal Field Alternation Gel Electrophoresis.- 5.2 Preparation of Chromosomal DNA for OFAGE Karyotypes.- References.- Screening and Testing New Distillers’ Yeasts for Their Potential in Molasses Ethanol Fermentations.- 1 Introduction.- 2 Screening and Testing Techniques.- 2.1 Small-Scale Batch Fermentations.- 2.1.1 Description of Apparatus.- 2.1.2 Propagation of Yeast.- 2.1.3 Fermentation and Analyses.- 2.2 Small-Scale Continous Fermentations.- 2.2.1 Propagation of Yeast.- 2.2.2 Fermentation and Analysis.- 2.3 Full-Scale-Fermentation.- 2.3.1 Propagation of Yeast.- 2.3.2 Fermentation Equipment.- 2.3.3 Analyses Performed.- 2.3.3.1 Oxidation Method of Ethanol Determination.- 2.3.3.2 Sugar Concentration Determined by Fermentation.- 2.3.4 Regulation of Fermentation.- 3 Evaluation of Results.- 3.1 Small-Scale Fermentations.- 3.2 Production-Scale Fermentations.- References.- Genetic Analysis and Manipulation of Mucor Species by DNA-Mediated Transformation.- 1 Introduction.- 1.1 Taxonomy of Mucor.- 1.2 Industrial Use of Mucor.- 1.3 Genetic Analysis and Manipulation.- 2 General Methods.- 2.1 Media.- 2.2 Growth Conditions.- 2.3 Harvesting, Storage and Germination of Sporangiospores.- 2.4 Replica Plating of Colonies.- 3 DNA-Mediated Transformation of Mucor.- 3.1 Plasmids pMCL006 and pMCL1302.- 3.2 Method for Transformation of Mucor with Plasmid DNA.- 3.3 Properties of Transformants.- 4 Isolation, Regeneration and Fusion of Protoplasts from Mucor.- 4.1 Formation of Protoplasts from Mucor.- 4.2 Preparation of a Lytic Enzyme from Streptomyces sp. no. 6 (Streptozyme).- 4.3 Regeneration of Protoplasts.- 4.4 Fusion of Protoplasts.- 5 Isolation of Marker Genes for Use in DNA-Mediated Transformation.- 5.1 Selectable Marker Genes Coding for Resistance to Antibiotics.- 5.2 Selectable Marker Genes Coding for Metabolic Enzymes.- 5.3 Isolation and Enrichment of Auxotrophic Mutants of Mucor.- 6 Analysis of Transformants.- 6.1 Isolation of RNA.- 6.2 Isolation of DNA.- 6.3 Recovery of Plasmid DNA.- 7 Genetic Manipulation of Mucor by DNA-Mediated Transformation. Concluding Remarks.- References.- Final Product.- Malt and Hop Flavanoids in Pilsner Beer.- 1 Importance.- 2 Identification of Flavanoids in Barley, Malt, Hops and Beer.- 2.1 Barley and Malt Flavanoids.- 2.2 Hop Flavanoids.- 2.3 Beer Flavanoids.- 3 Analysis of Beer Flavanoids.- 3.1 Analysis of Beer Anthocyanogens.- 3.1.1 Introduction.- 3.1.2 Method.- 3.2 Analysis of Total Polyphenolics.- 3.2.1 Introduction.- 3.2.2 Method.- 3.3 Analysis of Flavanoids.- 3.3.1 Introduction.- 3.3.2 Method.- 3.4 Comparison of the Methods.- 4 Significance of the Analytical Results.- 5 Analysis of Beer Flavanoids by High-Performance Liquid Chromatography.- 5.1 Gas Chromatography.- 5.2 High Performance Liquid Chromatography.- 5.2.1 Isolation of Proanthocyanidins from Beer.- 5.2.2 High Performance Liquid Chromatography.- References.- Analytical Measurement of Volatile Sulphur Compounds in Beer.- 1 Introduction.- 2 Volatile Sulphur Compounds in Beer.- 3 Analytical Methods for Determining Volatile Sulphur Compounds in Beer.- 3.1 General Remarks.- 3.2 Non-Chromatographic Methods.- 3.2.1 Sulphur Dioxide.- 3.2.1.1 The Monier-Williams Distillation/Titrimetric Procedure.- 3.2.1.2 The Para-Rosaniline Colorimetric Procedure.- 3.2.1.3 The DTNB Method.- 3.2.2 Hydrogen Sulphide.- 3.3 Chromatographic Methods.- 3.3.1 Analyte Detection.- 3.3.2 Chromatographic Separation.- 3.3.3 Sample Preparation/Introduction into Gas Chromatograph.- 3.3.3.1 Static Headspace Sampling.- 3.3.3.2 Dynamic Headspace Sampling (Purge and Trap Methods).- 3.3.3.3 Methods for Concentrating Sulphur Compounds of Lower Volatility.- 4 Conclusions.- References.- Short Introduction to Headspace Analysis.- 1 Introduction.- 2 The Static Headspace Method.- 3 The Dynamic Headspace Method.- 4 Sampling Methods.- 5 Factors Affecting Choice of Equilibrium Conditions.- 6 Quantitative Headspace Analysis.- 7 Final Remarks.- References.- The Use of Principal Components Analysis in Monitoring the Quality of Beer.- 1 Introduction.- 2 The Principles of PCA.- 2.1 The Objectives.- 2.2 Conceptualizing the Multivariate Problem.- 2.3 A Stepwise Explanation of PCA.- 3 The Use of PCA in Brewing Science.- 3.1 A Detailed Example of the Use of PCA in Brewing Science.- 3.2 Other Examples of the Use of PCA in Brewing Science.- 4 Conclusions.- References.- German Beer Purity Law and its Influences on the Properties and Analysis of Beer.- 1 The German Beer Purity Law.- 2 Complaint of the EEC Commission at the European Court of Law Against the Federal Republic of Germany.- 3 Arguments for Retaining the Beer Purity Law.- 4 Beers with Adjuncts.- 5 Analytical Analysis of Additives and Harmful Substances.- 5.1 Immunological and Immunochemical Methods.- 5.1.1 Qualitative and Half-Quantitative Indication Methods.- 5.1.1.1 Double Diffusion in Agar.- 5.1.1.2 Immunoelectrophoresis.- 5.1.1.3 Counter Current Electrophoresis.- 5.1.2 Quantitative Identification Methods.- 5.1.2.1 Basic Radial Immunodiffusion.- 5.1.2.2 Electroimmunodiffusion.- 5.1.2.3 Two-Dimensional Immunoelectrophoresis.- 5.1.3 Highly Sensitive Immunochemical Methods.- 5.1.3.1 Radio Immuno Assay.- 5.1.3.2 ELISA of EIA (Enzymelinked Immunosorbent Assay).- 5.2 Gas Chromatography and High Performance Liquid Chromatography (HPLC).- 5.3 Electrophoresis.- 5.4 Isotachophoresis.- 5.5 Atomic Absorption Spectrometry.- 6 Summary.- References.
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