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Aspidiotus Nerii Bouchè (Insecta: Hemipthera: Diaspididae)

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University of Massachusetts Amherst ScholarWorks@UMass Amherst Masters Theses 1911 - February 2014 2003 Molecular systematics of a sexual and parthenogenetic species complex : Aspidiotus nerii Bouchè (Insecta: Hemipthera: Diaspididae). Lisa M. Provencher University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/theses Provencher, Lisa M., "Molecular systematics of a sexual and parthenogenetic species complex : Aspidiotus nerii Bouchè (Insecta: Hemipthera: Diaspididae)." (2003). Masters Theses 1911 - February 2014. 3090. Retrieved from https://scholarworks.umass.edu/theses/3090 This thesis is brought to you for free and open access by ScholarWorks@UMass Amherst. It has been accepted for inclusion in Masters Theses 1911 - February 2014 by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. MOLECULAR SYSTEMATICS OF A SEXUAL AND PARTHENOGENETIC SPECIES COMPLEX: Aspidiotus nerii BOUCHE (INSECTA: HEMIPTERA: DIASPIDIDAE). A Thesis Presented by LISA M. PROVENCHER Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE May 2003 Entomology MOLECULAR SYSTEMATICS OF A SEXUAL AND PARTHENOGENETIC SPECIES COMPLEX: Aspidiotus nerii BOUCHE (INSECTA: HEMIPTERA: DIASPIDIDAE). A Thesis Presented by Lisa M. Provencher Roy G. V^n Driesche, Department Head Department of Entomology What is the opposite of A. nerii? iuvf y :j3MSuy ACKNOWLEDGEMENTS I thank Edward and Mari, for their patience and understanding while I worked on this master’s thesis. And a thank you also goes to Michael Sacco for the A. nerii jokes. I would like to thank my advisor Benjamin Normark, and a special thank you to committee member Jason Cryan for all his generous guidance, assistance and time. I would like to recognize Stefano Colazza, Ezio Peri, and Paolo LoBue from the University of Palermo for their assistance with the sex pheromone analysis and Andrew Weeks at the University of Davis California for his work on the endosymbionts. I also thank the following for their help in collecting: J. Daneel (Nelspruit, South Africa), L. Erkilic (Adana-Gar, Turkey), A. Garonna (Napoli, Italy), U. Gerson (Rehovot, Israel), R. Gill (Sacramento, California), H. Glenn (Homestead, Florida), R. Henderson (New Zealand), F. Kozar (Budapest, Hungary), D. Miller (USDA), G. Stathas (Athens, Greece), G. M. Orphanides (Cyprus), P. Gullan (Davis, California). And J. Hare, P. Fanti, M. Hoddle and H. Schmutterer for their contacts. And special thanks to Ifedayo Nicholson for all her offerings to the PCR god. This work was supported by funds from the Massachusetts Agricultural Experiment Station (Hatch 839). IV ABSTRACT MOLECULAR SYSTEMATICS OF A SEXUAL AND PARTHENOGENETIC SPECIES COMPLEX: Aspidiotus nerii BOUCHE (INSECTA: HEMIPTERA: DIASPIDIDAE). MAY 2003 LISA M. PROVENCHER, B.S., UNIVERSITY OF MASSACHUSETTS AMHERST M.A., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Benjamin B. Normark Aspidiotus nerii Bouche, the oleander scale, is a worldwide pest of woody plants, including many of agricultural importance such as citrus, olive and ornamentals. There is a great amount of variation within this species with respect to genetic systems (a sexual system of haplodiploidy vs. apomictic parthenogenesis), host plant use (> 100 plant families), development time and fecundity. Aspidiotus nerii were obtained from 18 different world locations were tested for phylogenetic analysis using molecular markers. Using molecular analysis of a 774 bp fragment of mtDNA (including portions of the COI and COII genes; the Leucine tRNA was not found between the COI and COII as expected). Six 6 most parsimonious trees were recovered. The consensus tree was made up of three highly divergent clades with an uncorrected pairwise difference of 5.8-6.7%. Colleagues from Sicily tested sex pheromone emissions in sexual and parthenogenetic forms of A. nerii. They found the parthenogens did not emit sex pheromones. Here individuals were screened for endosymbiotic bacteria in the phylum Cytophaga- Flavobacterium-Bacteriods (CFB). These bacteria have been found to cause parthenogenesis in some mites. The findings revealed the parthenogens harbored the CFB v bacteria and the sexuals did not. Compiling all this evidence one clade was found to be made up of parthenogenetic lineages with two remaining clades made up of seven sexual haplotypes. This evidence suggests that A. nerii is a complex of at least two different species as Gerson and Hazan proposed in 1979 when they described a new parthenogenetic species, Aspidiotus paranerii Gerson. TABLE OF CONTENTS Page ACKNOWLEDGEMENTS...i\ ABSTRACT.....v LIST OF TABLES.viii LIST OF FIGURES.ix CHAPTER 1. INTRODUCTION.1 2. METHODS.5 2.1 Taxon Sampling ..5 2.2 Nucleotide Sampling and Laboratory Procedures.5 2.3 Sequence alignment and Phylogenetic Analysis.6 2.4 Bacterial Screen.7 2.5 Wolbachia Screen...7 3. RESULTS. 8 3.1 Sequence Analysis.8 3.2 Phylogenetic Analysis.8 3.3 CFB Analysis.8 3.4 Wolbachia Analysis.9 4. DISSCUSSION.10 4.1 Phylogenetic Analysis. 10 4.2 Biogeographical Analysis.12 4.3 CFB Analysis.12 4.4 Conclusion.13 BIBLIOGRAPHY.33 Vll LIST OF TABLES Table Page 1. Locations and information for the 18 specimens collected.15 2. Aspidiotus nerii haplotype sequences and out groups undet. Aspidiotini sp., Aodinella sp. and Hemiberlesia sp., showing the COI, intergenic space and COIL Note the Luecine tRNA is not between the COI and COII as normally found in other insect mtDNA.17 3. Primers used in polymerase chain reaction amplification and sequencing of the mitochondrial (CO I and CO II) DNA regions studied.23 4. Uncorrected Pairwise Distance matrix.25 5. Descriptive Statistics .. 27 vm LIST OF FIGURES Figure Page 1. Strict consensus tree of 11 haplotypes resolving into 3 main clades. Five haplotypes were placed in Clade 1, two haplotypes were placed in Clade 2, including Pomigliano Sicily Italy and San Juan California, four haplotypes were placed in Clade 3. Bootstrap (above) and Bremer support (below) are included...29 2. One of the six Maximum Parsimony analysis trees is shown here with character mapping. Mitochondrial DNA data results corroborate that the A. nerii complex is made up of sexual and parthenogenetic lineages. One clade is made up of parthenogenetic lineages. Confirmation for thelytoky in this clade is based on evidence that two of the tested populations were obtained from laboratory colonies that have produce no males in more than 10 years (Glenn 2001, Hare 2002 personal communication). CFB were found only in the parthenogenetic populations of A. nerii. The Sicilian individuals within the Group 1 tested negative for sex pheromone emissions and the Sicilian individuals within the Group 2 haplotype tested positive for sex pheromone emissions...31 IX CHAPTER 1 INTRODUCTION Why sex? One of the enduring questions in evolutionary biology is the significance of sexual vs. asexual reproduction in eukaryotes (Ghiselin, 1974; Williams, 1975; Smith, 1978; Bell, 1982; Stearns, 1987; Michod and Levin, 1988; Barton and Charlesworth, 1998; Hamilton, 2001; Kondrashov, 2001). In theory, a population with asexual reproduction is expected to increase at twice the rate of its sexual counterparts (Smith, 1971). Yet, sexual reproduction in eukaryotes predominates. Therefore a strong counterbalance for the cost of sex must exist. Many mechanisms (>20) (Kondrashov, 1993; Hurst and Peck, 1996; Barton and Charlesworth, 1998) have been proposed to explain the persistence of sexual reproduction over asexual reproduction. Some intriguing possibilities are resistance to parasites (Lively et al., 1990), the evasion of mutation accumulation (Muller, 1964; Kondrashov, 1988) and most recently a pluralist approach which considers the involvement of multiple mechanisms, environmental and mutational (West et al.,1999). A consensus, however, has not yet been established. An attempt to elucidate this question is to investigate the origin of thelytoky, the reproductive system in which females produce only daughters. Although, thelytoky is rare, it is one of the most widespread deviations from normal Mendelian sexual reproduction in eukaryotes (Normark, 2002). It has been found that some animal groups that exhibit low mobility such as terrestrial tube worms (Reynolds, 1974) soil mites (Norton et al., 1993) and scale insects (Nur, 1980) have high frequencies of thelytokous reproduction. This suggests a common evolutionary response in taxa that have limited dispersal and female philopatry 1 (Williams, 1975; Bell, 1982). Through extant thelytokes a direct comparison of the differences in the amounts of genetic recombination per generation and their current levels of genetic variation with ancestral sexual taxa can be examined. Insects have long played a critical role in the search for the significance of sex. There is a barrage of this type of research surrounding aphids (Hales et al., 1977; Weeks and Hoffmann, 1998; Spence and Blackman, 2000), walking sticks (Marescalchi, 1993; Law and Crespi, 2002), weevils (Normark et al., 1999) and scale insects (Nur, 1980), to mention a few. Additional advantages of using insects for investigating the hypotheses of sex include ease in rearing, short generation time, and a vast background in genetic manipulation. To further research in this direction of sexual vs. asexual reproduction, preliminary analysis was carried out on the armored scale insect Aspidiotus
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  • Baseline Study of Coconut Scale, Aspidiotus Destructor Signoret (Hemiptera: Diaspididae) and Its Management on Mango (Mangifera Indica L.)

    Baseline Study of Coconut Scale, Aspidiotus Destructor Signoret (Hemiptera: Diaspididae) and Its Management on Mango (Mangifera Indica L.)

    BASELINE STUDY OF COCONUT SCALE, ASPIDIOTUS DESTRUCTOR SIGNORET (HEMIPTERA: DIASPIDIDAE) AND ITS MANAGEMENT ON MANGO (MANGIFERA INDICA L.) By SALAHUDDIN A dissertation submitted for partial fulfillment of the requirement for the degree of DOCTOR OF PHILOSOPHY IN ENTOMOLOGY DEPARTMENT OF ENTOMOLOGY FACULTY OF AGRICULTURE GOMAL UNIVERSITY DERA ISMAIL KHAN 2016 © 2016 SALAHUDDIN DEDICATED TO MY PARENTS ACKNOWLEDGEMENT All glory be to Allah, the beneficial, the merciful. I would like to thank Almighty Allah who enabled me to complete this painstaking study of PhD well in time. I would like to express the deepest appreciation to my supervisor Dr. Habib ur Rahman, co- supervisors Dr. Inamullah Khan and Dr. Muhammad Daud Khan for their substantial guidance, continuous inspiration, invaluable assistance and suggestions from initial planning of experiments to the preparation of manuscripts in bringing this dissertation to its present form. I would like to thank my IRSIP program supervisor Dr. Steven P. Arthurs at Mid- Florida Research and Education Center, IFAS, University of Florida, USA for his cooperation to conduct research in his laboratory and proof reading and correction of my PhD dissertation. His deep knowledge and experience helped me to learn advance skills in Entomology and improve my scientific writing. I am grateful to my other supervisory committee members Professor Dr. Muhammad Ayaz Khan, Agha Shah Hussain and Dr. Muhammad Mamoon ur Rashid for their excellent guidance and suggestions on how to improve my work. Dr. Gillian W. Watson, Senior Insect Biosystematist, California Department of Food and Agriculture, Plant Pest Diagnostic Center, Sacramento California, U.S.A., is gratefully acknowledged for identification of Aspidiotus destructor.