A LABORATORY COURSE IN PLANT PHYSIOLOGY
CONTENTS CHAPTER 1 - CARBOHYDRATES 1 1 ESTIMATION OF REDUCING SUGARS 1 2 DNS METHOD 1 3 ESTIMATION OF TOTAL SUGARS 1 4 PHENOL-SULPHURIC ACID METHOD 1 5 ESTIMATION OF NON-REDUCING SUGARS 1 6 ESTIMATION OF STARCH 1 7 ESTIMATION OF CELLULOSE 1 8 ESTIMATION OF HEMICELLULOSE 1 9 ESTIMATION OF CRUDE FIBER 1 10 ESTIMATION OF PECTIN SUBSTANCES - COLORIMETRIC METHOD CHAPTER 2- NITROGEN 2 1 ESTIMATION OF NITROGEN 2 2 ESTIMATION OF NON-PROTEIN NITROGEN 2 3 ESTIMATION OF AMINO ACIDS 2 4 ESTIMATION OF PROTEIN - BIURET METHOD 2 5 ESTIMATION OF PROTEIN - LOWRY'S METHOD 2 6 ESTIMATION OF SOLUBLE PROTEIN BY DYE-BINDING METHOD (BRADFORD'S METHOD) 2 7 POLYACRYLAMIDE-SODIUM DODECYL SULPHATE GEL ELECTROPHORESIS (SDS-PAGE) OF PROTEINS 2 8 ISOLATION OF PLANT DNA 2 9 EXTRACTION OF RNA FROM PLANT 2 10 ESTIMATION OF DNA 2 11 ESTIMATION OF RNA 2 12 ISOLATION OF PLASMIDS 2 13 ESTIMATION OF PROLINE 2 14 ESTIMATION OF METHIONINE 2 15 ESTIMATION OF LYSINE 2 16 ESTIMATION OF TRYPTOPHAN 2 17 PATHOGENESIS-RELATED PROTEINS (PR-PROTEINS) 2 18 ESTIMATION OF CHLOROPLAST PROTEIN 2 19 ESTIMATION OF HEAT SHOCK PROTEINS 2 20 NITROGEN FRACTIONS CHAPTER 3 - PHOSPHORUS 3 1 PHOSPHORUS FRACTIONS 3 2 PHYTIN PHOSPHORUS 3 3 ESTIMATION OF PHYTIC ACID CHAPTER 4 – FATS 4 1 EXTRACTION OF TOTAL LIPIDS 4 2 ESTIMATION OF OIL OR CRUDE FAT 4 3 ISOLATION AND ESTIMATION OF FREE FATTY ACIDS 4 4 ESTIMATION OF FREE FATTY ACIDS OR ACID VALUE OF OIL 4 5 ESTIMATION OF FREE FATTY ACIDS BY COLORIMETRY 4 6 ESTIMATION OF PEROXIDE VALUE OF AN OIL OR FAT 4 7 ESTIMATION OF IODINE VALUE OF OIL 4 8 ESTIMATION OF SAPONIFICATION VALUE / SAPONIFICATION NUMBER OF AN OIL OR FAT 4 9 ESTIMATION OF OIL BY NMR METHOD CHAPTER 5 - ENZYMES 5 1 EXTRACTION OF ENZYME 5 2 AMYLASES 5 3 INVERTASE (?-FRUCTOFURANOSIDASE) 5 4 DEOXYRIBONUCLEASE (DNASE) 5 5 UREASE 5 6 PHOSPHOENOLPYRUVATE CARBOXYLASE (PEP CARBOXYLASE) 5 7 GLUTAMINE SYNTHETASE (GS) 5 8 GLUTAMATE DEHYDROGENASE (GDH) 5 9 GLUTAMATE SYNTHASE (GOGAT) 5 10 NITRATE REDUCTASE (NR) 5 11 CATALASE 5 12 POLYGALACTOURONASE (PG) 5 13 PECTIN METHYL ESTERASE (PME) 5 14 L-PHENYLALANINE AMMONIA LYASE (PAL) 5 15 POLYPHENOL OXIDASE (PPO) OR DOPA OXIDASE 5 16 INDOLEACETIC ACID OXIDASE (IAA OXIDASE) 5 17 ASCORBIC ACID OXIDASE 5 18 PYRUVATE DECARBOXYLASE 5 19 LIPOXYGENASE (LIPOXIDASE) 5 20 NITRITE REDUCTASE (NIR) 5 21 PEROXIDASE (POD) 5 22 ACID PHOSPHATASE 5 23 SUCCINATE DEHYDROGENASE 5 24 ESTIMATION OF NITROGENASE ACTIVITY 5 25 ESTIMATION OF RUBISCO BY ELISA 5 26 ESTIMATION OF TOTAL DEHYDROGENASE ACTIVITY 5 27 ESTIMATION OF PEP – CARBOXYLASE 5 28 ESTIMATION OF RIBONUCLEASE ACTIVITY 5 29 ESTIMATION OF SUPER OXIDE DISMUTASE, CATALASE AND PEROXIDASE 5 30 ESTIMATION OF GLYCOLATE OXIDASE ACTIVITY 5 31 ESTIMATION OF LIPID PEROXIDATION 5 32 STARCH PHOSPHORYLASE 5 33 SUCROSE SYNTHASE (SUCROSE-6 PHOSPHATE ) 5 34 ISOZYMES IN PLANT SAMPLES CHAPTER 6 - MINERAL NUTRITION 6 1 ESTIMATION OF PHOSPHORUS 6 2 ESTIMATION OF POTASSIUM 6 3 ESTIMATION OF CALCIUM AND MAGNESIUM 6 4 ESTIMATION OF IRON AND MANGANESE 6 5 PLANT TISSUE TESTS 6 6 HYDROPONICS CHAPTER 7 - PLANT GROWTH REGULATORS 7 1 ESTIMATION OF AUXINS (INDOLE 3 ACETIC ACID OR IAA) 7 2 BIOASSAY OF IAA 7 3 ESTIMATION OF GIBBERELLINS BY CALORIMETRY 7 4 BIOASSAY OF GIBBERLLINS - AMYLASE RELEASE TEST FROM SEEDS 7 5 EXTRACTION AND ESTIMATION OF CYTOKININS BY CHROMOTOGRAPHY 7 6 BIOASSAY OF CYTOKININ - RADISH COTYLEDON TEST 7 7 ESTIMATION OF CYTOKININ BY ELISA 7 8 ABSCISIC ACID 7 9 BIOASSAY OF ABA - INHIBITION OF ?-AMYLASE SYNTHESIS IN BARLEY ENDOSPERM 7 10 ESTIMATION OF ABSCISSIC ACID BY ELISA 7 11 ESTIMATION OF ETHYLENE BY GAS CHROMOTOGRAPHY 7 12 ESTIMATION OF ETHYLENE BY COLORIMETRY 7 13 BIOASSAY OF ETHYLENE BY EPINASTIC RESPONSE 7 14 TRIPLE PEA TEST 7 15 PEA STEM SWELLING TEST 7 16 ANTHOCYANIN INHIBITION TEST 7 17 CHLOROPHYLL RETENTION TEST 7 18 CHLOROPHYLL FORMATION TEST 7 19 SPECIFIC PROTOCOLS FOR ABA AND CYTOKININ EXTRACTIONS CHAPTER 8 - SECONDARY METABOLITES 8 1 ESTIMATION OF CURCUMIN 8 2 ESTIMATION OF ANTHOCYANIN 8 3 ESTIMATION OF OXALIC ACID 8 4 ESTIMATION OF GOSSYPOL 8 5 IDENTIFICATION AND DETERMINATION OF POLYAMINES (BY TLC) 8 6 ANALYSIS OF POLYAMINES BY HPLC 8 7 ESTIMATION OF HCN 8 8 ESTIMATION OF TOTAL CAROTENOIDS 8 9 ESTIMATION OF LYCOPENE 8 10 ESTIMATION OF CHLOROPLAST PIGMENT COMPOSITION 8 11 ESTIMATION OF CHLOROPHYLL CONTENT WITHOUT HOMOGENISATION 8 12 ESTIMATION OF ANTHOCYANIN 8 13 ESTIMATION OF LEUCO-ANTHOCYANIN 8 14 ESTIMATION OF LIGNINS 8 15 ESTIMATION OF CAPSAICIN 8 16 ESTIMATION OF QUINONES 8 17 ESTIMATION OF TANNINS 8 18 ESTIMATION OF ASCORBIC ACID (VITAMIN C) 8 19 ESTIMATION OF ASCORBIC ACID BY COLORIMETRY 8 20 ESTIMATION OF TOTAL PHENOLS 8 21 ESTIMATION OF ORTHO-DIHYDRIC PHENOLS CHAPTER 9 - PHYSIOLOGY 9 1 MEASUREMENT OF HILL REACTION 9 2 ESTIMATION OF MITOCHONDRIA 9 3 ISOLATION OF CHLOROPLASTS 9 4 SULLIVAN?S HEAT TOLERANCE TEST 9 5 MEASUREMENT OF LOSS OF MEMBRANE PERMEABILITY 9 6 CHLOROPHYLL STABILITY INDEX (FOR DROUGHT TOLERANCE IN PLANTS) 9 7 PLANT TISSUE CULTURE 9 8 EQUIPMENT AND OTHER REQUIREMENTS FOR TISSUE CULTURE LABORATORY 9 9 PLANT TISSUE CULTURE TECHNIQUE: STEPS 9 10 CATION EXCHANGE CAPACITY (CEC) OF ROOTS 9 11 PHYSIOLOGICAL DISORDERS AND CORRECTIVE MEASURES 9 12 LEAF AREA DETERMINATION 9 13 PLANT GROWTH ANALYSIS 9 14 DETERMINATION OF LIGHT EXTINCTION COEFFICIENT 9 15 MEASUREMENT OF LIGHT INTERCEPTION 9 16 MEASUREMENT OF RATE OF GERMINATION 9 17 MEASUREMENT OF RESPIRATORY RATES OF FOLIAGE 9 18 MEASUREMENT OF AEROBIC RESPIRATION 9 19 ESTIMATION OF RELATIVE WATER CONTENT (RWC) 9 20 DETERMINATION OF LEAF EPICUTICULAR WAX 9 21 EFFECT OF OSMOTIC POTENTIAL UP ON IMBIBITION 9 22 DETERMINATION OF CHEMICAL OXYGEN DEMAND (COD) IN A WATER SAMPLE 9 23 TESTING FOR HEAT TOLERANCE IN PLANTS 9 24 DESICCATION TOLERANCE TEST FOR DROUGHT RESISTANCE 9 25 SEED GERMINATION TEST FOR DROUGHT TOLERANCE 9 26 CHLOROPHYLL STABILITY INDEX FOR DROUGHT TOLERANCE IN PLANTS 9 34 MEASURING PHOTOSYNTHESIS AND TRANSPIRATION IN WHOLE PLANT BY CONSTANT FLOW SYSTEM 9 35 MEASUREMENT OF LIGHT INTENSITY 9 36 ESTIMATION OF STOMATAL INDEX AND STOMATAL FRQEUENCY 9 37 TEST FOR POLLEN VIABILITY 9 38 SOIL ANALYSIS CHAPTER 10 - GENERAL PROCEDURES 10 1 CHROMATOGRAPHIC SEPARATION OF PLANT EXTRACTS 10 2 RADIO TRACER TECHNIQUES FOR 14CO2 STUDIES 10 3 CHROMATOGRAPHY 10 4 ATOMIC ABSORPTION SPECTROPHOTOMETRY 10 5 LOW VOLTAGE PAPER ELECTROPHORESIS 10 6 THE TRACER TECHNIQUE 10 7 POLY ACRYLAMIDE GEL ELECTROPHORESIS 10 8 INFRA RED GAS ANALYSIS FOR CO2 STUDIES (IRGA) 10 9 ENZYME - LINKED IMMUNOSORBANT ASSAY (ELISA) (GENERAL) 10 10 THE DOUBLE ANTIBODY SANDWICH TECHNIQUE 10 11 PREPARATION FOR LIGHT MICROSCOPY : WAX EMBEDDING TECHNIQUES 10 12 SCANNING ELECTRON MICROSCOPY TECHNIQUES