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TITLE
DEDICATION
CERTIFICATE
DECLARATION
ACKNOWLEDGEMENT
CONTENTS
1. INTRODUCTION
2. REVIEW OF LITERATURE
2.1 Biochemical changes
2.1.1 Changes in phenolic compounds
2.1.2 Changes in carbohydrates
2.1.3 Changes in nitrogenous compounds
2.1.3.1 Changes in amino nitrogen
2.1.3.2 Changes in total nitrogen
2.1.3.3 Changes in protein
2.1.4 Changes in ascorbic acid
2.1.5 Changes in oxidative enzymes
2.1.5.1 Ascorbic acid oxidase
2.1.5.2 Peroxidase
2.1.5.3 Polyphenol oxidase
2.1.6 Phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TALI
2.2 In vitro studies on the production of hydrolytic enzymes
2.2.1 Celluloly tic enzymes
2.2.2 Pectinolytic enzymes
2.2.3 Protease
2.3 Toxins
2.4 Changes in the composition of leaf leachates
2.5 Changes in phyllosphere microorganisms
2.6 Appressoria formation in the presence of phyllosphere microorganisms
3. MATERIALS AND METHODS
3.1 Establishment of piants and inoculation with C. cassikola
3.2 Blochemlcal changes
3.3 Oxidative Enzymes
3.4 Assay of phenylalanlne ammonldyase and tyroslne ammonla-lyase
3.5 Effect of nitrogen on the in vitro production of cellulase (C, and C, ) β-glucosidase, pectinolytic enzymes and protease of C. cassiicola
3.6 Toxin production in Corynespora cassiicola
3.7 Determination of nutrients on the leaf leachates
3.8 Enumeration of phyllosphere microflora of Hevea (Dickinson, 1965)
3.9 Effect of phyllosphere microorganlsms on the spore germination andappressoria formation in C. cassiicola
3.10 Statistical analysis of the experimental data
4. RESULTS
4.1 Disease incidence
Table1 Effect of different levels of nitrogen on Corynespora leaf spot disease intensity
Plate.1 Disease development in rubber seedlings applied with different levels of nitrogen (% of recommended dom)
4.2 Biochemical constituents
4.2.1 Changes in total phenols (Table 2)
Table 2 Changes in total phenols* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
4.2.2 Changes in ortho-dihydroxy phenols (Table 3)
4.2.3 Changes in phenolic acids
Table 3 Changes in ortho-dihydroxy phenols* in the healthy and C. cassiicola inoculated rubber seedhga at different levels of nitrogen
Fig.1 Changes in p. hydroxybenzoic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.2 Changes in ferulic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.3 Changes in p. coumarlc acid in the healthy and C.cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.4 Changes in vanillic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.5 Changes in caffeic acid in the healthy and C.cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.6 Changes in chlorogenic acid in the healthy and C cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.7 Changes in protocatechuic acid in the healthv and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.8 Changes in cinnamic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
4.2.4 Changes in reducing sugars (Table 4)
4.2.5 Changes in non-reducing sugar (Table 5)
Table 4 Changes in reducing sugars* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 5 Changea in non-reducing sugars* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
4.2.6 Changes in starch (Table 6)
4.2.7 Changes in amino nitrogen (Table 7)
4.2.8 Quantitative analysis of amino acids
Table 6 Changes in starch* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 7 Changes in amino nitrogen* in the healthy and C. cnssiicola inoculated rubber seedlings at different levels of nitrogen
Fig.9 Changes in asparagine in the healthy and C cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.10 Changes in phenylalanine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.11 Changes in glutamine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.12 Changes in methionine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.13 Changes in tyrosine in the healthy and C. canssiicola inoculated rubber seedlings at different levels of nitrogen
Fig.14 Changes in aspartic acid in the healthy and C.cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.15 Changes in glycine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.16 Changes in alanine in the healthy and C. cassiicola inoculatrd rubber seedlings at different levels of nitrogen
Fig.17 Changes in leucine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.18 Changes in cysteine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.19 Changes in glutamic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.20 Changes in arginine in the healthy and C.cassicola inoculated rubber seedlings at different levels of nitrogen
4.2.9 Changes in total nitrogen (Table 8)
Fig.21 Changes in histidine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.22 Changes in lysine in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 8 Changes in total nitrogen* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
4.2.10 Changes in protein (Table 9)
4.2.11 Changes in ascorbic acid (Table 10)
4.2.12 Changes in ascorbic acid oxidase (Table 11)
Table 9 Changes in protein in the healthy and C, cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 10 Changes in ascorbic acid in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 11 Changes in ascorbic acid oxidase* in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
4.2.13 Changes in peroxidase (Table 12)
4.2.14 Changes in polyphenol oxidase (Table 13)
4.2.15 Changes in phenylalanine ammonia-lyase (Table 14)
Table 12 Changes in peroxidase * in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 13 Changes in polyphenol oxidaeeSin the healthy and C. cassiicola inoculated rubber seedhgs at different levelsof nitrogen
Table 14 Changes in phenylalanine ammonia-lyase* in the healthy and C. cassiicola inoculated rubber seedlings atdifferent levels of nitrogen
4.2.16 Changes in tyrosine ammonia lyase (Table 15)
4.3 Effect of nitrogen on the in vitro production of hydrolytic enzymes
Table 15 Changes in tyrosine ammonia-lyase in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Table 16 Effect of nitrogen on biomass and production of cellulase (CI andC.) of C. cassiicola
Table 17 Effect of nitrogen on biomass and production of β-glucosidase of C. cassiicola
Table 18 Effect of nitrogen on biomass and production of pectionlytic enzymes of C. cassiicola
Table19 Effect of nitrogen on biomass and production of protease of C. cassiicola
4.4 In vitro studies on toxin production by C. cassiicola
Fig.23 Effect of nitrogen on toxin production by C.cassiicola in modified Czapeks medium
Fig.24 Effect of different levels of nitrogen application to rubber seedlings on toxin production in the leaf extracts byC. cassiicola
4.5 Changes in amino nitrogen, total sugars and total phenols in the leafleachates of healthy and C. cassiicola inoculated rubber seedlings atdifferent levels of nitrogen (Figure 25)
Table 20 Effect of phenolic acids on the inactivation of toxin (s) of C.cassiicola
Fig.25 Changs in amino nitrogen, total sugars and total phenols in the leaf leachates of healthy and C. cassiicola inoculated rubber seedlings at different levels (% of recommended dose) of nitrogen
4.6 Changes in the population of phyllosphere microorganisms in the healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Fig.26 Changes in the population of phyllosphere microorganisms in healthy and C. cassiicola inoculated rubber seedlings at different levels (% of recommended dose) of nitrogen
4.7 Percentage of phyllosphere bacteria and yeasts at different levels ofnitrogen inducing germination and appressoria formation inC. cassiicola
Table 21 Percentage of bacteria and yeasts inducing appressoria formation in Hevea leaves fertilized with different levels of nitrogen
5. DISCUSSION
Effect of graded levels of nitrogen on the incidence of Corynespma leafspot disease
Changes in Phenolic compounds
Fig. 27 Biosynthesis of phenolics
Carbohydrates
Nitrogenous compounds
Ascorbic acid and ascorbic acid oxidase
Peroxidases and polyphenol oxidases
Lyases
Hydrolytic enzymes
Proteolytic enzymes
Toxin production by the pathogen and inactivation by phenolic acids
Changes in phenols, amino nitrogen and total sugars in the leaf leachates of healthy and C. cassiicola inoculated rubber seedlings at different levels of nitrogen
Phyllosphere microflora
Interaction of phyllosphere microorganisms
6. SUMMARY
BIBLIOGRAPHY
ANNEXURE 1
ANNEXURE 2