ABSTRACT
CONTENTS List of Abbreviations .................................................................352 10.1 Introduction ....................................................................353
10.1.1 Taxonomy and Importance of Philaenus spp ....353 10.1.2 Classical Cytogenetics of Philaenus spp .......... 356
10.1.2.1 Standard Karyotypes ........................ 356 10.1.2.2 Nucleolus Organizer Region
Banding .............................................358 10.1.2.3 C-Banding .........................................359 10.1.2.4 Meiosis.............................................. 360 10.1.2.5 Karyotype Transformations in
Evolution ............................................361 10.1.3 Complete Mitochondrial Genome Sequence
of Philaenus spumarius .....................................361 10.1.4 Molecular Cytogenetics of Philaenus spp ........ 363
10.2 Fluorescence In Situ Hybridization Protocols ............... 363 10.2.1 Materials and Supplies ...................................... 363 10.2.2 Equipment ......................................................... 365 10.2.3 Chromosome Preparation ................................. 366 10.2.4 Probe Preparation ............................................. 366
10.2.4.1 Labeling of Ribosomal 18S rDNA Probe................................................. 366
10.2.4.2 Labeling of Telomeric (TTAGG)n Probe................................................. 366
10.2.5 Hybridization and Detection ............................. 367 10.2.5.1 Postlabeling Treatment of the
Probe................................................. 367 10.2.5.2 Slide Pretreatment and
Denaturation ..................................... 367 10.2.5.3 Fluorescence In Situ Hybridization .. 368 10.2.5.4 Slide Washing and Mounting ........... 368
10.2.6 Visualization and Mapping ............................... 369
10.2.7 Troubleshooting ................................................ 369 10.2.8 Representative Results ...................................... 370
10.2.8.1 Detection of a Tandem Telomere Repeat Sequence in Philaenus spp by Fluorescence In Situ Hybridization with the (TTAGG)n Probe................................................. 370
10.2.8.2 Detection of Ribosomal Sites in Philaenus spp by Fluorescence In Situ Hybridization with the 18S rDNA Probe ..................................... 370
10.3 Discussion ...................................................................... 373 10.3.1 Chromosome Organization and Evolution in
Philaenus .......................................................... 373 10.3.2 Molecular Organization of Telomeres in
Philaenus .......................................................... 373 10.3.3 Mapping a Ribosomal 18S rDNA Cluster to
Philaenus spp Chromosomes .............................374 10.4 Conclusions .....................................................................376 Acknowledgments .................................................................... 377 References ................................................................................ 377
LIST OF ABBREVIATIONS AgNOR, silver-binding nucleolus organizer region AT, adenine-thymine Bp, base pairs C-banding, staining of heterochromatic regions CCD-camera, charge-coupled device camera CGH, comparative genomic hybridization CMA3, chromomycin A3 COI, cytochrome c oxidase subunit I CytB, cytochrome b DAPI, 4′,6-diamidino-2-phenylindole dATP, deoxyadenosine triphosphate dCTP, deoxycytidine triphosphate 2D, two-dimensional ddH2O, double-distilled water dGTP, deoxyguanosine triphosphate DNA, deoxyribonucleic acid dNTP, deoxyribonucleotide triphosphate dTTP, deoxythymidine triphosphate EDTA, ethylene diaminetetraacetic acid FISH, uorescence in situ hybridization FITC, uorescein isothiocyanate GISH, genomic in situ hybridization
HCl, hydrochloric acid ITS2, internal transcribed spacer mt-genome, mitochondrial genome NORs, nucleolus organizing regions PBS, phosphate-buffered saline PCR, polymerase chain reaction rDNA, ribosomal DNA RNase A, ribonuclease A SAT-chromosome, satellite chromosome SSC, saline sodium citrate TE buffer, tris-ethylene diaminetetraacetic acid (EDTA) buffer TTAGG, thymine-thymine-adenine-guanine-guanine
The hemipteran (homopteran) suborder Auchenorrhyncha involves ve superfamilies including the Fulgoroidea (planthoppers), Cicadoidea (cicadas), Membracoidea (leafhoppers and treehoppers), Myerslopioidea stat. nov. (ground-dwelling leafhoppers), and Cercopoidea (spittlebugs or froghoppers). The object of the present study is the spittlebug genus Philaenus Stål, 1864 belonging to the cercopoid family Aphrophoridae.
