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This dissertation has been 65-9344 microfilmed exactly as received CJBULA, Adam Burt, 1934- THE RELATIONSHIP OF FREE AMINO ACIDS OF SOME SOLANACEOUS PLANTS TO GROWTH AND DEVELOPMENT OF LEPTINOTARSA DECEM- LINEATA (SAY). The Ohio State University, Ph.D., 1965 Zoology University Microfilms, Inc., Ann Arbor, Michigan THE RELATIONSHIP OF FREE AMINO ACIDS OF SOME SOLANACEOUS PLANTS TO GROWTH AND DEVELOPMENT OF LEPTINOTARSA DECEMLINEATA (SAY) DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Adam Burt Cibula, B.S. in Ed., M.A. ****** The Ohio State University 1965 Approved by: Adviser Department of Zoology and Entomology ACKNOWLEDGMENTS The author is indebted to Dr, Ralph H, Davidson, Dr, Frank W, Fisk, and Dr, Jules B, LaPidus for guidance, helpful suggestions, and loan of special apparatus during the course of this investigation. This study was supported in part by grant No, EF 00185 of the National Institutes of Health, U, S, Public Health Service, VITA June 4, 1934 Born - Salem, Ohio 1956 B.S. in Ed., Kent State University, Kent, Ohio 1956- 1958 Graduate Assistant, Department of Biology, Kent State University, Kent, Ohio 1958 M.A., Kent State University, Kent, Ohio 1958- 1960 Temporary Instructor, Kent State University, Kent, Ohio 1960- 1964 Graduate Assistant, Department of Zoology and Entomology, The Ohio State University, Columbus, Ohio 1964- Assistant Professor, Kent State University, Kent, Ohio FIELDS OF STUDY Major Field: Entomology Studies in Economic Entomology. Professor Ralph H. Davidson CONTENTS Page ACKNOWLEDGMENTS ii VITA iii TABLES V ILLUSTRATIONS vi INTRODUCTION 1 REVIEW OF LITERATURE 5 MATERIALS AND METHODS 13 Colorado potato beetle Maintenance of the beetle Plant varieties used Larval rearing Plant extraction methods Prepupal hemolymph extractions Chromatographic procedures OBSERVATIONS AND RESULTS 35 Free amino acids in plant foliage Larval growth and development Survival of first instar larvae to adults Weights of newly emerged adults Free amino acids in prepupal hemolymph Prepupal to adult survival DISCUSSION 55 SUMMARY AND CONCLUSIONS 61 LITERATURE CITED 63 IV TABLES Table Page 1. Relative amounts of free amino acids in alcoholic extracts of different solanaceous plants 36 2. Average weight gains in mg between molts of larvae fed on different solanaceous plants 43 3. Average number of days for each instar of larvae which had fed on different host plants 44 4. Percentage of individuals completing development 45 5. Range and average weights of all newly emerged adults 47 6. Range and average weights of newly emerged male adults 48 7. Range and average weights of newly emerged female adults 49 8. Relative amounts of free amino acids of prepupal hemolymph 50 9. Duration in days of prepupal and pupal periods and percentage of adult emergence 54 v ILLUSTRATIONS Figure Page 1 . Posterior abdominal sterna of adult beetles 15 2. Oviposition cage in which adults were reared 16 3. Humidity chamber improvised from a Boekel dessication chamber 19 4. Screened polystyrene crisperchest partially filled with sand 21 5. Hemolymph exuding from larva after aortic puncture 25 6. The chromatographic chambers employed in this study 28 7. A standard chromatographic map 29 8. One-way chromatogram of extracts of young and bloom stage plants 38 9. One-way chromatogram of extracts of senescent, young, and bloom stage plants 39 10. One-way chromatogram showing the free amino acids in prepupal hemolymph 51 vx INTRODUCTION The feeding habits of phytophagous insects axe varied. According to feeding habits, these insects may be categorized as monophagous, oligophagous, or polyphagous depending on whether they feed on only one, a few, or on many different species of plants. However, no matter what pattern is character­ istic for the insect the basic food requirements must be supplied by these plants since they are usually their only source of food. The physiological processes of these insects are adjusted to the chemical constituents of these plants, and the insects are dependent upon them as a means of maintaining their existence. Chin (1950) lists at least four requirements in the chemical properties of plants which determine whether they can be used as normal food plants. These include smell, taste, the lethal effects caused by groups of chemicals, and the nutritive value for promoting growth. However, an attractive smell, a preferred taste, the absence of poisons, and high nutritive contents may not always exist even in two closely related species of plants. Therefore, some insects may be attracted to some plants because of the odor and may feed on the plants and continue to feed since the taste is also agreeable. However, the nutritive value may be low which may cause adverse effects. Nutrition here refers to the proper utilization of food after ingestion and the adequacy of all the required nutrients and accessory factors such as vitamins and minerals. Even though a close relationship exists between most phytophagous insects and their host plants, it is not well known why it is possible for any particular species or race to survive on various food plants. Within the plant species there are varieties which differ in their susceptibility to injury. This insect resistance in crops may be morphological, phenologi- cal, or chemical (Thorsteinson, 1957). The absence of specific food materials in plant tissues upon which insects feed might lead to death. The lack of necessary vitamins, amino acids, or fatty acids in the part of the plant upon which the insect feeds offers a possible explanation of the effects of some resistant plants on insects (Painter, 1953). Besides differences in the death rate, insects fed on various food plants may exhibit differences in size and fecundity which may reflect the differing nutritional value of the plants. However, Fraenkel (1951, 1959a, 1959b) suggested that foliage from different plants differs relatively little in chemical composition as far as the nutri­ tional needs of insects for food substances are concerned. What attracts an insect to a particular plant is not well known. Studies by Beck (1956a, 1956b), Dadd (1960), Augustine (1962), and others have indicated that nutrients such as sugars in the plaint may play a role in some way. Numerous other chemicals such as glucosides, saponins, and essential oils have been described in the literature as possible attractants. It may be as Thorsteinson (1958, 1960) points out that other nutrients such as amino acids, amides, and vitamins may also be instrumen­ tal in attracting insects to plants, but very little work has been done along these lines. An excellent example of an oligophagous insect is the Colorado potato beetle, Leptinotarsa decemlineata (Say), which feeds on plants of the Solanaceae. Not all members of this plant family are equally attacked by this insect. Instead, there is a gradation from those readily accepted to those completely rejected. Also the growth and development of the beetle is either normally or partially completed on different members of this plant family. When the beetle was first discovered and described by Say in 1824, it was found feeding on sand-nettle, Solanum rostratum Dunal, a weed on the eastern slopes of the Rocky mountains. With the introduction of the cultivated potato, Solanum tuberosum L., the beetle deserted the weed for this new cultivated plant upon which it thrived very well and subsequently spread over the country and later into Europe. Accounts of its spread can be found in articles by Walsh auid Riley (1868) , Tower (1906), and others. From the time it became a pest of potatoes and spread over the United States and into Europe this insect has been 4 reported feeding on other solanaceous plants, some of which are also of economic importance. Recently it has been reported as a new pest of tomatoes in the Middle Atlantic states (Reed and Doolittle, 1961). Before this it was only occasionally reported on tomatoes. Along with these reports many studies have been carried on in an attempt to learn the relationships of the various species of solanaceous plants and the attraction and development of the beetle upon them. The studies described in this dissertation were made to determine whether there was a relationship between the free amino acid content of the solanaceous plants potato, tomato, Lycopersicum esculentum Mill., eggplant, Solanum melongena L,, and horse-nettie, Solanum carolinense L., in different stages of development, and the growth and development of the larvae of the Colorado potato beetle. This was accomplished by analyzing and recording: 1. the free amino acids of the different plants in various stages of development, 2. weight gains of larvae on the different plants, 3. survival of first instar to adult, 4. the weights of newly emerged adults, 5. the free amino acids in prepupal hemolymph of larvae which had fed on the different plants, 6. prepupal to adult survival. REVIEW OF LITERATURE Various studies have been made in an attempt to show the relationship between specific groups of phytophagous insects and the host plants upon which they feed. These have included studies of preference, effects upon survival and development, and attempts to link deficiencies of nutrient materials to resistance. The Colorado potato beetle is one of the phytophagous insects which has been studied extensively since it became an economic pest not <?nly in the United States, but also in many foreign countries. These studies have included ones of attraction, resistance, and development. However, very few studies have been made dealing with the identification of nutrients in plaints with an attempt to relate these findings to this insect host plant relationship. Even though the potato beetle was described in 1824, very little attention was paid to this insect until it became a pest some 40 yeairs later and then its actions were followed closely.

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