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AN INVESTIGATION of FACTORS WHICH AFFECT COLONY FORM and GROWTH in GONIUM PECTORALE DISSERTATION Presented in Partial Fulfillmen

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AN INVESTIGATION of FACTORS WHICH AFFECT COLONY FORM and GROWTH in GONIUM PECTORALE DISSERTATION Presented in Partial Fulfillmen AN INVESTIGATION OF FACTORS WHICH AFFECT COLONY FORM AND GROWTH IN GONIUM PECTORALE DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By lynn Boyd Graves, Jr., B.A., M.Sc. The Ohio State University 1959 Approved by 0 4 ^ d . Adviser Department of Zoology and Entomology ACKNOWLEDGMENTS The author wishes to express his sincere appreciation to Professor W* J* Kostir for suggesting this problem and for his helpful advice and criticism throughout the course of this study, and to Dr* A. C* Broad for his help in the design and construction of the constant temperature illuminated water bath used in this investigation* ii TABLE OF CONTENTS Page INTRODUCTION............................................. 1 The Organism Studied...................................... I The Form of the Colony in Culture. ............... 3 Past Investigations of Changes in Form in Gonlun pectorale . • it The Scope of the Present Investigation ..................... 5 MATERIALS AND METHODS........... 7 Source of the Organism Studied ........................... 7 General Methods of Culture •••.•• ........ ....... 7 The Constant Temperature Water Bath. ...... ........... 12 Method of Sampling the Culture ....♦......... 13 Statistical Method ••••............... 15 A DESCRIPTION OF THE CHANGES IN THE FORM AND IN THE RATE OF GROWTH OF THE COLONY THAT ACCOMPANY AGING OF CULTURES IN 0.05 PER CENT KNOP SOLUTION.................................... 17 INVESTIGATIONS OF THE POSSIBILITY THAT A DEFICIENCY OF SOME SUBSTANCE OR SUBSTANCES IS RESPONSIBLE FOR THE FORMATION OF ABNORMAL COLONIES AND FOR THE RETARDATION OF GROWTH............19 The Influence of Different Coneaitratione of Carbonate on the Form of the Colony and on the Rate of Growth. • • • • 19 The Influence of Vitamin B12 on the Form of the Colony and on the Rate of Growth. ••••••.•• ............. 22 The Influence of Iron on the Form of the Colony and on the Rate of Growth 2h The Influence of the Calcium Ion Concentration on the Form of the Colony and on the Rate of Growth .......... 26 The Influence of the Concentration of Knop Solution on the Form of the Colony and on the Rate of G r o w t h ............ 29 The Influence of the Depletion of the Inorganic Carbon on the Form of the Colony and on the Rate of Growth. • • • • 30 INVESTIGATIONS OF THE POSSIBILITY THAT GONIUM PECTORALE PRODUCES AUTOTOXIC SUBSTANCES, WHICH INFLUENCE"HW15tSffi tftfOCRM OF THE COLONY AND THE RATE OF GROWTH........... 32 The Influence of Inocula Containing Fluid from Aged Cultures on the Form of the Colony and on the Rate of Growth. ••••.••••••••••••• .......... 32 iii iv Page The Influence of the Dilution of an Aged Culture on the Form of the Colony and on the Rate of Growth•••••••• 3U The Influence of Filtrates from Aged Cultures on Young Cultures .............. 35 The Relation of the pH of theMedium to theForm of the Colony and to Motility........ * ................ .. 38 DISCUSSION....................................................... hO SUMMARY.........................................................Uli BIBLIOGRAPHY..................................................... 16 AUTOBIOGRAPHY................................................... 81 LIST OF TABLES t a b l e p a g e 1 The Percentage of the Total Number of Cells which is Represented by the Number of Cells in Each of the Differently-Celled Forms in an Eight Day Old Culture in 0.0!? Per Cent Knop Solution............... 50 2 Changes in the Form of the Colony and in the Growth that Accompany Aging of Cultures in 0*05 Per Cent Knop Solution* • • « ......... * * . • ............. 51 3 The Influence of Different Concentrations of Carbonate on the Form of the Colony and on the Rate of Growth. • • 52 I* The Influence of Vitamin on the Form of the Colony and on the Rate of Growth............................ 53 5 The Influence of Iron on the Form of the Colony and on the Rate of Growth ........................... 5ii 6 The Influence of the Calcium Ion Concentration on the Form of the Colony and on the Rate of Growth ..•••• 55 7 The Influence of the Concentration of Knop Solution on the Form of the Colony and on the Rate of Growth .... 57 8 The Influence of the Depletion of the Inorganic Carbon on the Form of the Colony and on the Rate of Growth* * * 58 9 The Influence of Inocula Containing Fluid from Aged Cultures on the Form of the Colony and on the Rate of Growth* * ............... 59 10 The Influence of Inocula Containing Fluid from Aged Cultures on the F o m of the Colony and on the Rate of Growth. A Repetition ....................... 60 11 The Influence of the Dilution of an Aged Culture on the Form of the Colony and on the Rate of Growth • • • • 61 12 The Influence of Filtrates from Aged Cultures on Young Cultures ............. * .................. 63 13 The Relation of the pH of the Medium to the Form of the Colony and to the Motility ................... 66 v U S T OF FIGURES PAGE FIGURE 1 Normal and Abnormal Colonies of Goniuin pectorale ...•••« 67 2 The Constant Temperature Illuminated Water Bath.............. 68 3 Changes in the Form of the Colony and in the Rate of Growth of an Aging Culture. ........... 69 U The Influence of Different Concentrations of Carbonate on the Rate of Growth.......................... 70 5 The Influence of Vitamin Bqjj on the Rate of Growth.......... 71 6 The Influence of Iron on the Rate of Growth. 72 7 The Influence of the Calcium Ion Concentration on the Rate of Growth ......... 73 8 The Influence of the Calcium Ion Concentration on the Form of the Colony ............. 7li 9 The Influence of the Concentration of Knop Solution on the Rate of Growth ........... 75 10 The Influence of the Rate of Depletion of the Inorganic Carbon on the Rate of Growth ........... *••••• 76 11 The Influence of Inocula Containing Fluid from Aged Cultures on the Rate of Growth ............... ...... 77 12 The Influence of Dilution of Aged Cultures on the Rate of Growth .................................. 73 13 The Influence of Filtrates from Aged Cultures on the Rate of Growth of YoungCultures 79 lit The Influence of Filtrates from Aged Cultures on the Form of the Colony of Young Cultures 80 vi INTRODUCTION The Organism Studied Gonlum pectorale is a colonial protozoan which is frequently en­ countered in many bodies of fresh water. I have found it in rivers, lakes, farm ponds, road-side ditches, and water-filled ruts in dirt roads. Pocock (1955), who has collected Gonlum over large areas of North America, believes that G. pectorale is the most common of all the colonial Fhytomonadida* She found it in temporary fresh water pools, ponds in pasture lands, lakes, and in cultures of soil collected as much as four years previously. Crow (1927) observed the species in a rain barrel and Kostir (personal conmunication) observed a rich growth of Gonlum in a discarded pan which had previously been used for storing frogs in an ice box. Gonium pectorale was originally described by 0. F. ifiller in 1773. The 16-celled colony forms a nearly square plate which is usually bent back slightly along the two diagonals. The center of the colony is bounded by four of the sixteen cells, while the remaining twelve cells are distributed, three to a side, around these central four. The colo­ nies studied ranged in size from approximately 2h to lOOyu in diameter, with each cell being about 7 ** 13^1 wide to 7 - 1 Iji long. The pellicle of each cell adheres to the pellicles of the adjacent cells. When -the colony is quite small the pellicle of each cell fits tightly against the 1 2 plasma membrane of the cell and shows no distortion where it adheres to other cells* But as the colony grows, the space between the plasma mem­ brane and the pellicle widens, and there is a distortion of the pellicle to form short protuberances in the area in which one cell adheres to another* Each cell of the colony produces a gelatinous material which unites with that produced by the other cells of the colony and forms a common matrix about the entire colony* Each cell contains one vase-shaped chloroplast which has one or two large pyrenoids in its base* There are also two flagella, two contrac­ tile vacuoles, and a large eye-spot to each cell* It has been reported that delicate cytoplasmic processes connect the cells to one another (Ehreriberg, 18385 Harper, 1912; Mast, 1916; Bock, 1926; Crow, 1927; Tilden, 193$; Prescott et al*, 19ii9) • Stein (19$8b) was unable to ob­ serve such processes in either stained or unstained living material or in fixed material using either the ordinary light microscope or the phase microscope* The observations made in the present study agree with those of Stein* Pascher (1927) lists two species of Gonlum which possess sixteen n cells, G, formosua Pascher, and G, pectorale 0* F* Muller* The main dif­ ference between these two species is that the cells in G* formosum are widely separated and pear-shaped, while the cells in 0. pectorale are close together and oval* Prescott (19U2) described a 32-celled species, G. dlacoidemn* that rarely produces l6 -celled forms* Its cells were ir­ regularly pyriform with two basal pyrenoids* Pocock (1955) described an 8-ceUed species, Q* octonariun, which always has two center cells and six outer cells, an arrangement never found in G, pectorale* She also described a species of 8, 16, or 32 cells, G* milticoccum, whose cells may contain many pyrenoids while the cells of G, pectorale contain only one pyrenoid which divides into two Just before the cell divides* She also described a new variety of G, pectorale, G* pectorale var. praecox. The mature colonies of this variety are comparatively small, usually rather pale green and slightly oblong in shape* The cells are somewhat elongated, ovoid in side view, with the apex usually wider than the base, and contain a single pyrenoid.
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