TY - BOOK AU - Narayana, P. Suresh AU - Varalakshmi, D AU - Pullaiah, T. TI - Research Methodology in Plant Science SN - 978-93-89832-27-3 U1 - 574.028 NAR PY - 2021/// CY - Jodhpur PB - Scientific Publisher N1 - Description: The book comprises of different chapters associated with methodology in Plant Science (Botany), describing in a simple and comprehensive way. The importance of creativity and motivation in research, the planning and proposal of research project, the description of different scientific techniques involved in research are described in an elaborate way. It also includes the sources/collection of scientific information, method of scientific report/paper/thesis writing etc. The book is also a source of different aspects of research methodology tailored to the needs of postgraduate students/research scholars for easy understanding. The book is also profusely illustrated. Compared to the first edition, the entire text has been thoroughly revised and updated in addition to the inclusion of four new chapters (Recent Advances in Electrophoresis, Chromatography – Latest and Advanced Techniques, Polymerase Chain Reaction and Photon Counting and Photo-Multiplier Tubes). We hope this edition will be more useful to the readers and will continue to serve the purpose more effectively. Contents: 1. RESEACH METHODOLOGY Introduction Research - What it is? Aim and Purpose of Research Early History of Research Types of Research Possible Motives for Doing Research Methodology - What it is? Types of Research Methodology Research Design Research Plan Concepts and Context of Research Research Logistics Research Methods versus Research Methodology Research Proposal Different Stages of Research Criteria for Good Research Objectives of Research Why do people carry out Research? Importance of Research Significance of Research 2. CENTRIFUGATION Introduction Centrifugation - Definition Types of Centrifugation Preparative Ultracentrifugation - Separation Methods Centrifuge - The Instrument Centrifuge - Components and Parameters Centrifuge - Operation Common Centrifuges Used in Biological Sciences Centrifuges - Types Centrifuges - Classification Ultracentrifuge - Basic Principle Preparative Centrifugation - Types of Rotors Rotors - Selection Rotors and Accessories - Care Centrifuge - Containers Density Gradient Separation - Formation and Choice of Density Gradients Gradient Material - Nature and Use Suitable Density Gradient Medium - Selection Fractions - Collection Sample Application to the Gradient Recovery and Monitoring of Gradients Analysis of Sub-cellular Fractions Presentation of the Results Analytical Ultracentrifuge - Applications Centrifuges - Safety Aspects in Use Centrifugation - Applications in Biological Research Many Uses of Centrifuge 3. PLANT TISSUE CULTURE Introduction Plant Tissue Culture - Materials and Methods Explant Aseptic Environment Tissue Culture Media or Nutrient Media Nutritional Composition of Most Common Plant Cell/Tissue Culture Media (Angiosperms) Composition or Components of Tissue Culture Media General Technique of Plant Tissue Culture Plant Tissue Culture Protocol (General) Plant Tissue Culture - Laboratory and Equipment Methodology for the Establishment of Cultures Types of Plant Tissue Culture Applications of Plant Tissue Culture 4. ELECTROPHORESIS Introduction Definition How Separation Occurs in Electrophoresis? Requirements for Electrophoresis Basic Principles of Electrophoresis Electrophoresis - Method Factors that affect Electrophoresis Physical Background of Electrophoresis Instrumentation for Electrophoresis Electrophoresis – Apparatus Theory of Electrophoresis Types of Electrophoresis Specific Techniques of Electrophoresis Commonly Used in Biochemistry Detection Methods – Blotting Electrophoresis – Applications RECENT ADVANCES IN ELECTROPHORESIS Introduction Non-linear Frictiophoresis or Non-linear Electrofrictiophoresis ZIFE (Zero Integrated Field Electrophoresis) Pulsed - field Gel Electrophoresis (PFGE) Isotachophoresis (ITP) Transient Isotachophoresis (tITP) Multi-junction IEF System Dielectrophoresis (DEP) Optical Dielectrophoresis Affinity Electrophoresis Difference Gel Electrophoresis (DIGE) Discontinuous Electrophoresis (colloquially Disc Electrophoresis) Microfluidic Chip-based Electrophoresis Iontophoresis Temperature Gradient Gel Electrophoresis (TGGE) and Denaturing Gradient Gel Electrophoresis (DGGE) Free Flow Electrophoresis (FFE) or Carrier-Free Electrophoresis New Technique to Separate Microsatellite Alleles Conclusion 5. HERBARIUM TECHNIQUE Herbarium - What it is? Plant Collection Why Preserve Plant Specimens? Preservation of the Plant Specimens Equipment Needed for Collection of the Plant Specimens Method to keep Wet Plant Material in Adverse Conditions Methods of Drying the Plant Specimen What Makes a Good Specimen? The Data to be recorded in the Field Note Books Mounting of the Dry Plant Specimens Labeling the Plant Specimens Filling or Arrangement of the Plant Specimens Special Care of the Plant Specimens Safe Storage of the Sheets / Specimens Safe Preservation of the Specimens (Plants) Use and Significance of the Herbarium Role of Herbarium in Modern Plant Taxonomy 6. CHROMATOGRAPHY Introduction Definition Principle Chromatography - Early History Classification of Chromatographic Methods Special Techniques of Chromatography Hyphenated (Hybrid) Techniques and Alternative Methods of Detection in Chromatography Chromatography - Basic Principle Types of Chromatography Adsorption Chromatography Partition Chromatography Planar Chromatography General Procedure - Paper Chromatography (PC) Thin Layer Chromatography (TLC) High Pressure Liquid Chromatography (HPLC) or High Performance Liquid Chromatography General Procedure - High-Performance Liquid Chromatography (HPLC) Chromatography - Applications Chromatography - Applications in Biology and Chemistry Chromatography - Uses in Pharmacognosy Chromatography - Applications for the Analysis of Herbal Drugs Chromatography - Latest and Advanced Techniques Evolution of Chromatography Chromatography - Improvements - Multi-dimensional Analysis Dye-Ligand Chromatography Hydrophobic Interaction Chromatography (HIC) Pseudo-affinity Chromatography Immobilized Metal Affinity Chromatography (IMAC) Perfusion Chromatography (PC) Supercritical Fluid Chromatography (SFC) Expanded Bed Adsorption (EBA) Chromatography Nanoflow Liquid Chromatography (NLC) Micellar Liquid Chromatography (MLC) Electrochromatography Capillary Electrochromatography (CEC) Weak Affinity Chromatography (WAC) High Performance Liquid Chromatography (HPLC) Conclusion Hy-phe-nated Techniques or Hybrid Techniques Hy-phe-nated HPLC (High-performance Liquid Chromatography) Methods 7. BOTANICAL MICRO-TECHNIQUE Introduction Killing and Fixation Embedding the Material in Paraffin Stains and Staining Methods Procedure of Staining Dehydration of the Material Clearing the Sections Dewaxing the Sections Mounting the Material on to the Slide Adhering Sections to the Slides Ringing the Slide / Sealing the Cover slip Slide Label - Preparation Free Hand Sections or Sectioning by Hand Free Hand Sectioning - Procedure Epidermal Peeling - Preparation 8. MICROSCOPY Introduction Microscopy - Definition Types of Microscopes Optical (Light) Microscope Electron Microscope Scanning Electron Microscope (SEM) Transmission Electron Microscope (TEM) Scanning Probe Microscope (SPM) Optical or Light Microscopy The Compound Light Microscope Electron Microscope Cytophotometry or Microfluorometry Phase Contrast Microscopy (PCM) Fluorescence Microscopy (FM) Scanning Electron Microscopy (SEM) Transmission Electron Microscopy (TEM) Electron Microscopy (EM) 9. SMEAR AND SQUASH TECHNIQUE Introduction Smear and Squash Squash Technique for Root Tips - Protocol Preparation of the Onion Root Tip Squash Chromosome Squash Technique Preparation of the Anther Smear 10. SPECTROPHOTOMETRY Introduction History and Spectrophotometry Spectrophotometer - Definition Spectrophotometer - Components Spectrophotometer - Design of the Device Spectrophotometer - Basic Principle Using Spectronic-20 Spectrophotometer Spectrophotometers - Different Types Spectrophotometer - Source of Light Spectrophotometer - Methods of Measurement Spectrophotometer - Uses Spectrophotometer - Applications Colorimeter and Spectrophotometer - Differences 11. COLORIMETRY Introduction History and Colour/Colorimetry Colorimetry - Definition Colorimeter - The Instrument Principle of Colorimetry Chemistry of Colorimetry Colorimeter - Operational Procedure (Experimental Method) Colorimetry - Chemical Method Absorption Colorimeter Photo-Electric Colorimeter - Operation Colorimetric Assays Precautions to be taken in Colorimetry Factors that affect/influence Colorimetric Measurements Advantages and Disadvantages of a Colorimeter Uses of Colorimeter Colorimetry - Applications in Plant Sciences Colorimetry - Applications in Other Fields 12. HYDROGEN-ION CONCENTRATION (pH) Introduction Earliest pH Meter - Litmus Hydrogen Ion Concentration (pH) Measurement of pH pH Meter pH Meters - Types Accurate Measurement of pH pH Indicators Buffers and Buffer Solutions Determination of pH Care and Use of pH Meter pH Electrode / pH Meter pH Scale Uses of pH Meters pH Meters - Advantages and Disadvantages pH Controller Calibration of pH Meter - Why? Care of pH Electrodes pH Measurement - Applications pH in Nature pH and Life xiv Research Methodology in Plant Science 13. MICROTOMY AND MICROTOME Introduction History and Microtomy Parts of a Microtome Proposed Classification (Types) of Microtomes Microtome - Knife Design and Cut Types Sharpening of the Knives Method of Sectioning by a Microtome Maintenance of a Microtome Applications of a Microtome Conclusion 14. BIOSTATISTICS Introduction History and Biostatistics Scope of Biostatistics Principles of Experimental Design and Statistical Inference Tabular Design Tabular Presentation of the Raw Data Tabular Presentation of Data Types of Tables Best Practice of Presenting the Table Graphical Design Diagrammatic Representation of Data Statistical Method - Types Data (Information) - Types Central Tendencies - Measures Averages Standard Deviation (SD) Correlation Coefficient Standard Error of Mean (SEM) and its Applications Standard Deviation (SD) and Standard Error (SE) Confidence Interval (CI) or Fiducial Limits Distribution - Types Null Hypothesisμ Errors - Types Level of Significance Outliers One - tailed and Two - tailed Test Determination of Sample Size - Importance Power of Study Choosing an Appropriate Statistical Test Common Problems Faced by the Researcher Software for Biostatistics Statistical Applications in Biological Research Biostatistics - Applications Recent Development in Modern Biostatistics 15. COMPUTERS IN LIFE SCIENCES Introduction Computer - Basic Principles Computer - Parts Computers - Types according to Size and Power Brief History of Computing Computers - Applications in Life Sciences Computers in Agricultural Research Computers and Plant Sciences Importance of Computer Literacy and Useful Software for Students and Researchers in Life Sciences 16. APPLICATIONS OF RADIOISOTOPES IN BIOLOGY/PLANT SCIENCES Introduction Radioactive Isotopes Half-Life (t1/2) Half-life in Biology and Pharmacology Radioactive Tracers Tracer Studies / Tracer Techniques Instruments to Measure Ionizing Radiation Geiger-Muller Counter (GM Counter) Scintillation Counter Radioactivity - Applications in Life Sciences Radioisotopes - Applications in Plant Sciences Radioisotopes and Agriculture Radioisotopes - Applications in Agriculture Stable Isotopes - Use in Agriculture Radioisotopes - Uses in Molecular and Cell Biology Conclusion 17. BIOINFORMATICS Introduction Definition History and Bioinformatics Bioinformatics - Concept and Goal xvi Research Methodology in Plant Science Bioinformatics - Relevance to Biotechnology Bioinformatics - Important Thrust Areas Bioinformatics - Uses Goals of Bioinformatics Approaches of Bioinformatics Bioinformatics in relation to other fields Conclusion 18. POLYMERASE CHAIN REACTION (PCR) Introduction PCR - What it is? PCR - Principle PCR - General Protocol PCR - Different Schemes and Types Applications of PCR in Different Disciplines PCR - Benefits, Drawbacks and Limitations Conclusion 19. PHOTON COUNTING AND PHOTO-MULTIPLIER TUBES (PMTs) Introduction Photon Counting Photomultiplier Tube (PMT) Photomultipliers - Properties Photomultipliers - Modification Photon Counting: SPCMs, CPMs and CCDs: Which Technology Works Best? The 3 - Technologies SPCMs (Single Photon Counting Modules) CPMs (Channel Photo Multipliers) CCDs (Charged Coupled Devices) Photon Counting Mode (Analog and Digital) Photon Counting Methods Detectors for High Speed Photon Counting Photon Counting Techniques Detector Principles Photon Detection - Uses Photon Counting - Applications Photon Counting - Benefits Conclusion REFERENCES ER -