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United States Department of Agriculture

Agricultural Research Service

Title: Cryopreservation of Fruit Fly (Tephritidae) Embryos

Authors
item Wang, Wen - VIA CELL, WORCESTER, MA
item Leopold, Roger
item Handler, Alfred
item Mccombs, Susan - APHIS, HAWAII
item Freeman, Thomas - NORTH DAKOTA STATE UNIV

Submitted to: Society for Cryobiology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: January 2, 2002
Publication Date: N/A

Technical Abstract: Three species of tephritid fruit flies, the Caribbean (Anastrepha suspensa), Mexican (Anastrepha ludens), and Mediterranean (Ceratitis capitata) were used in this study. More than 73% of A. suspensa embryos withstand treatments of egg shell removal (dechorionation), permeabilization of embryonic membranes, loading with cryoprotectants, and dehydration in the vitrification solution. About 27% of A. suspensa larvae hatched from the embryos that had been vitrified in liquid nitrogen vapor, stored in liquid nitrogen and subsequently recovered by warming in vapor before placing in Schneider's cell culture medium. An average of 29% of these larvae developed into pupae and as high as 46% fertile adults emerged. The fertility and fecundity of the progeny of these adults was assessed and found to be at control levels. More than 80% of A. ludens embryos withstand treatments of dechorionation and permeabilization. Loading the embryos with cryoprotectant and dehydration in vitrification solution reduced the hatching to about 60%. Up to 50% of the A. ludens larvae hatched after storage in liquid nitrogen. Development of these larvae into adults was as high as 90%. Preliminary studies have also begun on the medfly using the caribfly protocol for embryo cryopreservation. More than 65% of the medfly embryos withstand dechorionation and permeabilization. Loading the embryos with cryoprotectant and the dehydration in vitrification solution further reduced the hatching rate. However, up to 31% of C. capitata embryos hatched and larval to adult emergence was observed after storage in liquid nitrogen has exceeded 54% (5 replications).

Last Modified: 10/24/2014
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