ABSTRACT
LIST OF ABBREVIATIONS AFRI, Agricultural Food and Research Initiative BSA, bovine serum albumin DAPI, 4 ,ʹ6-diamidino-2-phenylindole dNTP, the four deoxyribonucleotide triphosphates: dATP,
dCTP, dGTP and dTTP
CONTENTS List of Abbreviations .................................................................. 63 2.1 Introduction ....................................................................... 64 2.2 Hessian Fly Genome Organization .................................... 66 2.3 Hessian Fly as an Experimental Model ............................. 67 2.4 Review of Genetic and Physical Maps ............................... 68 2.5 Chromosome Preparations ................................................. 68
2.5.1 Mitotic S Chromosomes ......................................... 69 2.5.2 Mitotic S + E Chromosomes .................................. 69 2.5.3 Salivary Gland Polytene (S) Chromosomes ............ 69 2.5.4 Spermatogonial Preparations .................................. 71
2.6 Fluorescence In Situ Hybridization ................................... 71 2.6.1 Preparing Slides for Hybridization ......................... 71 2.6.2 Probe Preparation Using Nick Translation ............. 72 2.6.3 Hybridization .......................................................... 72 2.6.4 Detection ................................................................. 73 2.6.5 Solutions ................................................................. 73
2.7 Fiber-Fish ........................................................................... 74 2.7.1 Isolation of Hessian Fly Nuclei ............................... 74 2.7.2 Preparation of Extended DNA Fibers ..................... 75 2.7.3 Probe Labeling and Hybridization .......................... 75 2.7.4 Probe Detection ...................................................... 76
Acknowledgments ...................................................................... 76 References .................................................................................. 76
EDTA, ethylenediaminetetraacetic acid FISH, uorescence in situ hybridization NIB, nuclei isolation buffer NIFA, National Institute of Food and Agriculture PBS, phosphate buffered saline R, resistance SDS, sodium dodecyl sulfate SSC, saline-sodium citrate buffer STE, sodium chloride-Tris-EDTA buffer TE, tris EDTA buffer TMN1, tris magnesium chloride sodium chloride buffer 1
2.1 INTRODUCTION The Hessian y (Mayetiola destructor) is an important insect pest of wheat (Triticum spp.) (Harris et al. 2003). It has been a persistent problem in the United States since it was rst discovered along the Atlantic coast just after the American Revolutionary War (Pauly 2002). It is now present nearly everywhere where wheat is grown, but poses its greatest threat to agriculture in northern Africa, Southwest Asia, and the United States (Harris et al. 2003).
