BIOCHEMICAL AND BIOTECHNOLOGICAL INVESTIGATIONS ON THE WATER—FERN SALVINIA MOLESTA MITCHELL. THESIS SUBMITTED TO THE COCHIN UNIVERSITY OF SCIENCE AND TECHNOLOGY IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in PLANT BIOCHEMISTRY AND BIOTECHNOLOGY UNDER THE FACULTY OF MARINE SCIENCES. By PETER K. MAN]. M.Sc Reg. No: l487 Department of Marine Biology, Microbiology & Biochemistry, School of Marine Sciences, Cochin University of Science and Technology. Cochin 682 016. 1998 Dedicated to the Almighty, My Parents, Teachers and Well Wishers DECLgARATls0N I hereby declare that the thesis entitled “Biochemical and Biotechnological investigations on the water-fern Salvinia molesta Mitchell” is an authentic record of research work carried out by me under the supervision and guidance of Prof. Babu Philip, in partial fulfilment of the requirements for the award of the Ph. D degree under the faculty of Marine Sciences, Cochin University of Science and Technology and that no part of it has "previously formed the basis for the award of any degree, diploma or associateship in any University. Kochi. - 16g Date: 5“ l [qqg Peter K. Mani CERTIFICATE This is to certify that the thesis entitled “Biochemical and Biotechnological investigations on the water-fern Salvinia molesta Mitchell” submitted herewith by Mr. Peter K. Mani is an authentic record of the research work carried out by him in the Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, Kochi-16, under my supervision and guidance in partial fulfilment of the requirements for the award of Ph.D degree of Cochin University of Science and Technology and that no part thereof has been presented before, for any other degree or diploma in any University. / 1/»3 »/ iz /1z&é1 <l/ Dr. Babu Philip,/ Prof. in Marine Biochemistry, Dept. of Marine Biology, Microbiology & Biochemistry School of Marine Sciences. Kochi- I6, /.‘/'\'5'\;ii~iiigiY "i Sfiiancg ,5 )­ Date: 5 I", [0[¢{<Z /é,<‘$"“ ¢:g,;/\ i -P our ms AVENUE ‘E’.¢_f\, E=?|'=PAfiuLA.M 5-, ' Cfifiril it-562916 . 3' 1' Q‘ \.\‘*~-b.” 3122/ . . E$$§i// /;,’i.€l"0i A(?i<\'0\vLEoGE@1,r.i~1Ts I wish to express my sincere thankfulness and profound indebtedness to my supervising teacher Dr. Babu Philip, Professor in Marine Biochemistry, Department of Marine Biology, Microbiology and Biochemistry, School of Marine Sciences, for his bounteous supervision, valuable guidance and loving encouragements throughout the tenure of my research work. I am thankful to Prof. (Dr.) N. R. Menon, Head, Department of Marine Biology, Microbiology and Biochemistry and Director, School of Marine Sciences for providing me with all the necessary facilities for the research work. I would like to express my eamest thanks to Prof. (Dr.) R. Damodaran, Dean, Faculty of Marine Sciences, for his valuable suggestions during the early period of research work. I would like to place on record my sincere gratitude to Dr. K. J. Joseph, Professor in Marine Botany for timely suggestions and scrutiny of the work. I wish to thank Dr. Rosamma Philip, and Dr. A.V. Saramma, Lecturers in Microbiology for their valuable suggestions and encouragements and all my colleagues in the Biochemistry laboratory and other Departments of Marine Sciences, especially to Mr. Vinu Chandran R, for all the help they have rendered­ I wish to thank the Directors of C M F R I Cochin, C I F T Cochin, Kerala Agricultural University Mannuthy, R A R S Kumarakom, I R R I Puthupally, Kottayam, T B G R I Palode, Thimvananthapuram and N I O Goa for extending library facilities and the Director of Agrivision, Kottayam and R A R S Kumarakom for extending laboratory facilities. I wish to record my indebtedness to all my teachers especially to Prof. K. S. Alexander, K. E. College Mannanam (Retd.), Dr. Thomas James, D. Sc. (Former Head of the Department of Botany, S. B. College, Changanassery) and Dr. J. G. Ray who inspired me much into the enchanting fields of plant sciences and directed me on research line. I am grateful to the Patron, Manager, Pro-Manager, Principal and Head of the Department of Botany, B. C. M. College, Kottayam for permitting me to continue my research work on a part time schedule. I wish to express my gratefulness to all my colleagues in B. C. M. College, especially to the members ofthe Botany Department and also to the members of Kerala Botanical Society for their sincere co-operation. l wish to place on record my sincere thanks to Miss. Elizabeth Kozhi, Mrs. Monamma Kokkad, lecturer, B. C. M. College and Mr. Chandrasekharan, lecturer, Baseli us College, Kortayam for all the help they have rendered. I wish to thank the staff of B. C. M. Computer centre for typing services. I greatly acknowledge the financial assistance from C S I R for the first two years of research in the form, of Junior Research Fellowship and also the authorities of Cochin University of Science and Technology for providing me with all the facilities. Let me also avail of this opportunity to thank my family members and all those who have lent their whole-hearted co-operation and support in making this thesis a reality. Abbreviations used in the thesis. % Percentage 0 C Degree centigrade 2,4-D 2,4- Dichloro Phenoxy Acetic Acid ADF Acid Detergent Fibre AOAC Association of Ofiicial Analytical Chemists BOD Biological Oxygen Demand CEC Cation exchange capacity cm Centimetre CNT Control COD Chemical Oxygen Demand Conc. Concentration DNA Deoxy Ribo Nucleic acid DS Dried Salvinia EC Eichhomia crassipes, EDTA Ethylene Diamine Tetra Acetate Fig. Figure g & gm gram h Hour (s) ha Hectare IAA lndole 3- acetic acid ICBN Intemational Code For Botanical Nomenclature K cal Kilo calorie Ks Kilo gram LAR Leaf Area Ratio M Molar (Moles per litre) MCPA Methyl C hloro- Phenoxy Acetic acid meq / gm Milliequivalents per gram mg Milligram min Minutes ml Milli litre N Normal NAD Nicotinamide Adenine Dinucleotide NADP Nicotinamide Adenine Dinucleotide Phosphate NASA National Aeronautics & Space Administration nm Nano metre NPM Normal potting mixture OC Organic carbon PC Pleurorus cilrinopileatus PF Pleurotusflorida pH Potentia hydrogenii Ppm Parts per million PS Paddy straw PS Pleurotus sajor-caju RBD Randomized block design RNA Ribo Nucleic Acid rpm Revolutions per minute SB Salvinia before mushroom cultivation SEM Standard Error of Mean SH Sulfhydril group SM Salvinia moiesta TCA Tricarboxylic acid cycle WT weight CONTENTS GENERAL INTRODUCTION PAGE CHAPTER I. NO AQUATIC WEEDS AND THEIR IMPACT ON I ECOSYSTEM EXPERIMENTAL MATERIAL, AFRICAN WEED OR 7 THE WATER- FERN SALVINIA MOLLISTA MITCHELL METHODS OF CONTROL OF AQUATIC WEEDS WITH 12 SPECIAL REFERENCE TO UTILIZATION. SCOPE OF THE PRESENT STUDY 21 OBJECTIVES 24 CHAPTER II ANALYSIS OF THE CHEMICAL CONSTITUENTS OF THE WATER-FERN SALVINL4 MOLESTA MITCHELL INTRODUCTION 25 MATERIALS AND METHODS 26 RESULTS AND DISCUSSION 30 CHAPTER III POTENTIAL APPLICATION OF AFRICAN WEED (SALVINIA MOLESTA MITCHELL) FOR THE CULTIVATION OF OYSTER MUSHROOM. (PLEUROTUS SAJOR-CAJU (FR) SINGER). INTRODUCTION MATERIALS AND METHODS 35 RESULTS AND DISCUSSION. 36 CHAPTER IV COMPARATIVE EFFICIENCY FOR LlGNO­ CELLULOSE CONVERSION ON AQUATIC WEED SUBSTRATE BY DIFFERENT SPECIES OF OYSTER MUSHROOMS (PLEUROTUS SAJOR-CAJU, PLEUROTUS FLORIDA AND PLEUR 0 TUS CI TR INOPIL EA TUS). INTRODUCTION 38 MATERIALS AND METHODS 39 U) L4) 4.3 RESULTS AND DISCUSSION. 42 CHAPTER V ALTERATIONS IN THE NUTRITIVE VALUE OF MUSHROOMS IN RESPONSE TO SALVINIA AS SUBSTRATE. 5.1 INTRODUCTION 50 5.2 MATERIALS AND METHODS 51 5.3 RESULTS AND DISCUSSION 58 CHAPTER VI RESIDUAL SUBSTRATE AFTER MUSHROOM HARVEST (SPENT SUBSTRATE) :-A PROS-PECTIVE ORGANIC MANURE AND ITS IMPACTS ON (I) SOIL CHEMICAL STATUS (II) SOIL MICROBIAL POPULATION DYNAMICS & (III) PLANT GROWTH (ANTHURIUM ANDREANUM) 6. 1 SOIL CHEMICAL STATUS 6.1.] INTRODUCTION 67 6.1.2 MATERIALS AND METHODS 70 6.1.3 RESULTS AND DISCUSSION 80 6. 2 SOIL MICROBIAL POPULATION DYNAMICS 6.2.1 INTRODUCTION 87 6.2.2 MATERIALS AND METHODS 89 6.2.3 RESULTS AND DISCUSSION 91 6. 3 PLANT GROWTH (ANTHURIUM ANDREANUM) 6.3.1 INTRODUCTION 94 6.3.2 MATERIALS AND METHODS 96 6.3.3 RESULTS AND DISCUSSION. 98 CHAPTER VII SEED-BED PREPARED FROM AFRICAN WEED A RELIABLE SUBSTRATE FOR ANTHURIUM SEED GERMINATION 7.1 INTRODUCTION I02 7.2 MATERIALS AN D METHODS 103