84 JoURNAL oF THE ArranRrclrrl Moseurro CoNrnor, Assocrlrtor{ VoL.3, No. 1 GENETIC SEXING SYSTEM FOR THE PREFERENTIAL ELIMINATION OF FEMALES IN CULEX QUINQUEFASCIATUS N. J. SHETTYI Departntent of Zoology & Centre for Applied, Genetics, BangalnretJniuersity,Bangalnre India ABSTRACT. A genetic sexing strain T(1M;2)I of Cutex quinquefasciatus was synthesizedfor the preferential elimination of females during the larval stage.Translocations were induced which linked the-gene for resistance to malathion to the male-determining factor. Mitotic chromosomes were analyzed to determine the precisenature of the translocation. INTRODUCTION Culexquinquefascintus Say is the most impor- tant carrier of Bancroftianfilariasisin Asia. and the main urban nuisance mosquito. This species is found throughout the tropics and extendsinto the temperateregionsofthe northern and south- ern hemispheres (Mattingly 1951).Because of the seriousproblem of insecticide resistance in this species,alternative insect control metho- dologies based on genetic mechanisms are being explored. The success of genetic manipulation involving sterile-male releases can be enhanced by developing a method by which malescan be easily separated from females during mass pro- duction. Since the females of this species are potential vectors, and cause biting nuisance, they should be eliminated during the early de- velopmental stages by genetic methods. This will also help to lower the cost of massproduc- tion of males for releasepurposes.To achieve this objective, we haveused a male-linked trans- locationand a malathionresistance gene to syn- thesizea geneticsexing strain of Cx. quinquefas- clatus. MATERIALS AND METIIODS The following strains wereusedin the exper- iment. 1. MlNclLoRE (MNG)-This strain is ho- mozygous for malathion resistance and was orig- inally collectedfrom Mangalore,Karnataka, In- dia during April 1980. 2. MADRAS (MDS)-This strain is homozy- gousfor malathion susceptibility and was origi- nally collected from Madras,Tamil Nadu, India during May 1978. Our preliminary studies on the inheritance of malathion resistance in Cr. quinquefascintus confirmed the observation of Tadano (1969) that malathion resistanceis inherited as a par- tial dominant geneon linkage group 2. For the induction of translocations. two-to- I Present address: Insects Affecting Man and Ani- malsResearch Laboratory, U.S.D.A., P. O. Box 14565, Gainesville, FL 32604 three-day-old males from MNG strain were ex- posedto 4,500rads of gamma rays at the rate of 140 rads per minute from a 60Co sourceat the Kidwai Memorial Institute of Oncology, Ban- galore. The irradiated males were crossedto three-day-old, virgin MDS females. The result- ing F1males wereback crossed to MDS females, and after a blood meal. the females were indi- vidually isolated in vials for egglaying. The eggs were held for 72 hoursto ensure complete hatch- ing. The total number of eggs laid and the num- ber of larvae from each female was recorded. Egg batches with greater than 35% sterility, indicating the possible presence of a reciprocal translocation, were exposedas early fourth in- star larvae to 1.5 ppm malathion for 24 hours. The surviving larvae from each family were rearedto the adult stage. Families showing only males were saved and were mated with MDS femalesfor continuation of the line and further testing. Translocation breakpointswere ascer- tained from the mitotic chromosomes of pupal testes as described by French et al. (1962). RESULTS AND DISCUSSION Of the 119 backcross females that laid eggs, 87 families showing greater than 307osterility were treated with malathion and the survivors were reared to adults. Twenty-two families showed a significant sex distortion favoring males but only one family, later designatedas T(1M;2)1, contained no females. This clearly showed that in the T(1M;2)1 line a translocation was induced which linked the male-determining Iocus to the malathion resistance gene. Exami- nation of mitotic chromosomes from the testis of the T(1M;2)I line confirmed a reciprocal translocation involving the shortest chromo- some (sex chromosome) and one of the auto- somes. The translocation break points are near the distal end of the metacentric chromosome I and about the middle of the larger arm of chro- mosome 2 (Fig. 1B). In Cx. quinqu.efascintus,Iike other culicine mosquitoes, there is no hetero- morphic pair of sex chromosomes. Sex is deter- mined by a single pair of alleles, M and rn, for