Preliminary Study On Artemia Incubation And Development Of Genetic Markers On Artemia In Tibetan Plateau

As a kind of important experimental material and economic animal resource, the study on Artemia is of great significance. In this research,we studied the experimental conditions for incubation of Artemia eggs and developped six pairs of microsatellite primers. First, the study on the incubation conditions was mainly focused on the identifications of the optimal temperature and salinity concentration, Second,To isolate a set of available microsatellite primers, the (AC)n and (AG)n microsatellite enriched libraries of artemia were constructed using FIASCO method, and six polymorphic primers were finally discovered, which could be applied in future studies about resource assessments and genetic differentiations. The main results were as follows:1. The ideal salinity for Artemia cysts incubation and larvae development was 25‰.Within the salinity range tested, the mean incubation rate of Artemia cysts collected from Keke salt lake decreased with the rising of salinity. And Artemina larvae could not develop into viable nauplius though the egg membranes could be ruptured when salinity was below 25%o. Artemia couldn’t incubate when the salinity was 80%o.2.The optimal incubation temperature of Artemia cysts from Keke salt lake was 26℃ under the 25%o salinity. The incubation rate went down with the temperature increasing or decreasing; When the temperature went beyond the range 13℃-39℃, the incubation rate fell to zero.3.120 positive clones from (AC)n libraries and 200 from (AG)n libraries were randomly picked and cultured separately. Germ solution amplificatoions were carried out using universal primers M13 and corresponding pribe sequences not labelled by biotin. Finally,80 and 120 positive clones containing short tandem repeats were discovered,with a 66.7% and 60.0% positive clones rate respectively.4.63 microsatellite primers were designed and systhesised using Primer Premier 5.0. We tested these primers in 18 Artemia DNA samples from different geographical units in Qinghai and Xizang and got six polymorphic microsatellite primers succesfully.