Eco-Physiological Studies on Cyanobacteria in the Sudan By: Eltigani Hamid Shihaba Ali B.Sc. (Agric.) honours (Agricultural Biotechnology) August 2002 A Thesis Submitted to the University of Khartoum in partial fulfillment for the degree of Master of Science (Agric.) Supervisor: Prof. Ahmed Ali Ahmed Mahdi Department of Botany and Agricultural Biotechnology Faculty of Agriculture University of Khartoum February 2010 ﺑﺴﻢ اﷲ اﻟﺮﲪﻦ اﻟﺮﺣﻴﻢ ۩واﷲُ ﺧَﻠﻖ آُﻞَ داﺑﺔٍ ﻣِﻦْ ﻣﺎءٍ ﻓﻤﻨْﻬﻢ ﻣﻦْ ﳝﺸِﻲ ﻋﻠﻲ ﺑﻄْﻨِﻪِ و ﻣﻨْﻬﻢ ﻣﻦ ﳝﺸﻲ ﻋﻠﻲ ِرﺟﻠِﲔ و ﻣﻨْﻬﻢ ﻣﻦْ ﳝﺸِﻲ ﻋﻠﻲ أرﺑﻊ ﻳﺨْﻠُﻖ اﷲُ ﻣﺎ ﻳﺸَﺎءُ إن اﷲَ ﻋﻠَﻲ آُﻞ ﺷﻲءٍ ﻗَﺪِﻳﺮ ۩ اﻟﻨﻮر ﺻﺪﻕ ﺍﷲ ﺍﻟﻌﻈﻴﻢ The locomotion type of cyanobacteria is the gliding!!!! TABLE OF CONTENTS Item Page No. Dedication …………………………………………………………...…i List of Tables ……………………….…………………………………ii List of Figures ………………………………………………………..iii List of plates…………………………………………………………..vi Acknowledgement …………………………………………..............vii Abstract (English) ………………………………………..…………….ix Abstract (Arabic) ………………………………………… ………….xi CHAPTER ONE: INTRODUCTION ……………..……….. ………1 CHAPTER TWO: LITERATURE REVIEW ………………..…….6 2. Cyanobacteria…………………………………………………………6 2.1- Definition and characterization ……………………………...……...6 2.2- Biological diversity of cyanobacteria………………………...……...9 2.3- Cyanobacteria and relation with other organisms ………...…..10 2.4- Classification of cyanobacteria………………………....................13 2.5- Factors affecting growth of Cyanobacteria ……………………....16 2.5.1- Light intensity ………………………………………………..…16 2.5.2-Temperature…………………………………......…......................19 2.5.3- Salinity………….……………………………………. ……...20 2.5.4- pH …………………………………………….……………….22 2.5.5- Phosphorous and Nitrogen ……………………………………22 2.6- Cyanobacteria and Crusting ability………………..…………….23 2.7 -Cyanobacteria and photosynthesis ……………….………………25 2.8 -Cyanobacteria and Nitrogen fixation……………….…………….25 2.9 - Cyanobacterial risks………………………………...…………...27 2.10- Population stability………………………………………........29 CHAPTER THREE: MATERIALS AND METHODS……….......30 3.1- Study area and sampling sites………………………………...……30 3.1- Samples collection, culturing and media …………………………..30 3.3- Isolation and purification ……………………………………...….34 3.4- Identification…………………………………………………….....34 3.5- Physiological studies………………………………………….…...34 3.6- Effects of temperature…………………………………...…………37 3.7- Effects of pH……………………………………….………………37 3.8- Effect of light intensity……………………………….…………….39 3.9- Effects of salinity………………………….………………………39 3. 10- Crusting ability……………………….……………….………….40 CHAPTER FOUR: RESULTS……………………………………….42 Temporal distribution of cyanobacteria……………………………........42 Temporal comparison of location……………………………………….42 Spatial distribution of cyanobacteria……………………………………47 Distribution of cyanobacteria on three types of samples………………..47 Frequency of the five cyanobacterial orders…………………………….47 Isolates of the genera of the order Oscillatoriales……….……………...47 Isolates of the genera of the order Nostocales…………………………..53 Isolates of the genera of the order Chroococcales………………………53 Temporal distribution for prevalent orders………………………...........53 The most frequency cyanobacteria species……………………………..57 Effects of Salinity on cyanobacterial growth……………………….......57 Effects of Light on cyanobacterial growth ……………………………..57 Effects of pH on cyanobacterial growth ……………………………...61 Effects of temperature on cyanobacterial growth ………………….......61 Effects of cyanobacteria on sand soils crusting…………………….......61 Effects of cyanobacteria on clay soils crusting…………………………61 CHAPTER FIVE: DISCUSSION ……………………………….....66 CHAPTER SIX CONCLUSIONS AND RECOMMENDATIONS …………………………………………………………………………..82 REFERENCES ……………………………………………………....84 Appendixes.………………………………………………………….114 Dedication To MY FAMILY: TO MOTHER AND WIFE, TO BROTHERS AND SISTERS, TO THE NEWBORN WHO RESEMPLES HIS FATHER THE MOMENT HEWAS BORN: MY SON i LIST OF TABLES Table No. Page No. 1- The principal groups of cyanobacteria …………………………….15 2- Isolates used in the physiological studies………………….……….37 3- The total number of isolates in the two location…………………....43 ii LIST OF FIGURES Fig. No. Page No. 1- Total number of cyanobacterial isolates in the 10 sampling points during the period January-November 2008…..………………….43 2- Number of cyanobacterial isolates in each of the 10 sampling points for the months Jan, Mar, May, July Sep and Nov 2008…….………44 3- Total number of cyanobacterial isolates in each sampling point during the period January-November 2008……..………………….……....48 4- Temporal distribution of total isolates of the water samples, dry and wet soil samples……………………………………………………..49 5- Frequency of isolation of members of each of the five cyanobacterial orders in all sampling sites for the months of January, March, May, July, September and November 2008…………………………….......50 6- Number of isolates of the genera of the order Oscillatoriales in all 10 sampling sites for the period January-November 2008………………51 7- Number of isolates of the genera of the order Nostocales in all 10 sampling sites for the months January-November 2008...…………..54 iii 8- Number of isolates of the genera of the order Chroococcales in all 10 sampling sites for the months January-November 2008..……………55 9- Temporal distribution of the most prevalent cyanobacterial orders during the period January-November 2008…………………………56 10- Frequency of isolation of the most prevalent cyanobacterial spp. in the sampled areas…………………………………..………………58 11- Effect of salinity on growth of the six selected cynobacterial isolates……………………………………………..………………59 12- Effect of light intensities on the growth of the six chosen cyanobacterial isolates..……………………………………………..60 13- Effect of pH on the growth of the six selected cyanobacterial isolates……………………………………………………………..62 14- Effect of temperature on the growth of the six selected cyanobacterial isolates……………………………………… ……………………..63 15- Sand aggregating activity of the growth of the six selected cyanobacterial isolates...………………………………………...….64 iv 16- Clay aggregating activity of the growth of the six selected cyanobacterial isolates……………………………………………..65 v LIST OF PLATES Plate No. Page No. 1- Map of the Sudan………………….…………………...…………5 2- El-Rahad dry sand site……………………………………….….. 32 3- El-Rahad Turda site………………………………………………33 4- Cultures of cyanobacteria ………………………………………..35 5- Micrographs of hormogona………………………………..…..…36 6- Micrographs of Microcystis sp., Synechocystis sp. and Dermocarpa sp……………………………….………………...……….………38 7- The oscillator apparatus……………………………….…………… 41 8- Micrographs of Lyngbya sp. And Spirulina sp…….………………..46 9- Micrographs of Anabaena sp., Calothrix sp. and Cylindrospermopsis sp……………………………………………….……………….……….52 vi APPENDIX 1- Mean sequent for the effect of salinity on chosen strains and their interactions on dry weight …………………………………...……..…112 2- Mean sequent for the effect of light on chosen strains and their interactions on dry weight……………… …………………………..112 3- Mean sequent for the effect of pH on chosen strains and their interactions on dry weight ……………………………………...….112 4- Mean sequent for the effect of temperature on chosen strains and their interactions on dry weight ………………………………………….....113 5- Mean sequent for the effect of selected isolates on sand soil particle aggregation….……………..……………………………...……….…114 6- Mean sequent for the effect of selected isolates on clay soil particle aggregation ……………….…………………………………….…...115 vii Acknowledgements I would like particularly to express my gratitude to: Prof. Ahmed Ali Mahdi, my supervisor: first, for accepting me as an M.Sc. student at the Dept. of Botany and Agricultural Biotechnology. Second, for his unlimited patience on many challenges providing, many nice discussions and so many ideas. He definitely has taught me a great deal of useful things, ranging from Science to Systems dynamics, and even the simple, but hard mission of smiling at the most rough times. Thank you for all the support and I hope that the time of thanking you would not come soon. A special gratitude to my friends who like brothers to me; Aboalgasim Mohmmed Ahmed, Eltyeb Abd allatif (Hulfawi) and Ismail Ahmed Mohmmed who, played an important role in my work. All the members of the Department of Botany and Agricultural Biotechnology; Thank you all for the good moments and teamwork spirit, while sharing knowledge with so many colleagues for their precious technical help during my time as lab assistants: always prepared, always very organized, always with a big smile! Thank you! Thanks extend to Biotechnology Commission at the National Center For Research, particularly to Hesien (Hennery), Mu'taz, Eihab, Aiman, Hajo, and all who used smile when we met. Appreciation is expressed to my “better-half”: Ashraf Musa for showing peculiar interest in helping me, for unconditional support from the very beginning, remarkable sense of patience, trust and hope. To my family: Mother, Wife, Brothers, Sisters, and my eldest son Mohmmed Alfatih, who lived without father's kindness during the period of the study. Unending gratefulness goes to all who helped and supported me but are not mentioned here. viii ABSTRACT This study was carried out in the Biofertilizer Laboratory of the Department of Botany and Agricultural Biotechnology, Faculty of Agriculture, University of Khartoum at Shambat during 2008 - 2009 to investigate the abundance, spatial and temporal distribution of cyanobacteria in Shambat and Omdurman in Khartoum State and El Rahad town in North Kordofan State (Sudan) and to determine the optimal growth conditions and role of some factors affecting cyanobacterial growth. This