Research Methodology in Plant Science

Narayana, P. Suresh

Research Methodology in Plant Science - 2nd - Jodhpur Scientific Publisher 2021 - 457

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

978-93-89832-27-3

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