Effects Of Cryopreservation On Main Developmental Genes Expression Of Early Embryo In Honeybee

Segmental development and morphological establishment are regulated by a series of genes in the early embryos of honeybee, including the pair-rule genes such as prd、poxn and eve, the segmental polarity genes such as ben, and the homeotic genes such as dfd and scr. These genes participate in the morphological development of segments at different developmental period by corresponding to molecular regulatory mechanism, so gene mutation will lead to segmental deficiency, abnormality, larval deformity, and even death before adulthood. At present, the deep frozen cryopreservative technique is a very effective method to keep honeybee eggs (early embryos), however, the damage of nuclear DNA of honeybee eggs (early embryos) treated by this method have been detected out, discovering a deficient segmental boundary which cannot tell the truth of genetic damage at a molecular level. Some probes used in this study such as prd, poxn, eve, ben, dfd and scrwere therefore synthetized through a transcriptional method and then were hybridized in situ with the transcripts in honeybee embryos at different hour ages in control and experimental groups, respectively. The results showed that:(1) it was only gene scr that had a different expression between two groups at stage4among these6genes;(2) These six genes had no any expression at stage1but had an obvious expression at stage9on every segments; gene prdhad a striped expression on the centre of embryo at stage5with an obvious segmental boundary; gene poxn, eve and ben had a punctuated expression at stage8with bilateral segmental boundary including head; gene eve had an obvious expression at stage7on segments; gene ben had no expression at stage4but had a linear expression from head to tail at stage9on bilateral segments; gene dfd had an obvious expression at stage4on the pre-torso of embryo; gene scr had a larger area of expression at stage3on the head of embryo in experimental group than in control group;(3) gene eve, dfd, scr and ben had an obvious expression at larval stage, gene eve and ben had a striped expression on segments horizontally and vertically, gene dfd and scr had a dotted expression on bilateral boundary of segments. It was concluded that no obvious adverse effect of cryopreservation on main developmental genes expression of early embryo in honeybee was found and the presently-used method to cryopreserve early embryo in honeybee was also feasible as a whole.