Studies On The Polarity Of Auricularia Auricula

The hybrid strain H2J3, obtained by mating of two protoplasted monokaryons H2 and J3 of Auricularia auricula, was cultivated. One hundred and twenty-six basidiospore isolates were collected by discharging spores from the fruit-body ofH2J3.The monokaryotic and dikaryotic mycelia of A.auricula were grown onto the surfaces of glass coverslips on CYM medium and were stained with fluorescence stains Calcofluor white M2R for septa, and Bisbenzimide(Hoechst 33258) and DAPI for nuclei respectively. The effects on fluorescence staining caused by various factors including buffer type, staining time, buffer concentration and mycelia age have been studied. The results showed that the optimal fluorescence staining condition for observation of nuclei and septa of A. auricula was as following: the mycelia were stained with 50ug/ml Bisbenzimide for 4 minutes firstly and followed with 2.5ug/ml Calcofluor white M2R for 1 minute.Sixty monospore cultures were divided into six groups and tested by pairing of 10X10. The method of double fluorescence staining for nuclei and septa were employed in observing the nuclei number of per cell of mated mycelia under fluorescence microscope. The clamp connections of mated mycelia were observed under light microscope, through the two approaches above, it was determined whether the dikaryotic mycelia were formed. The ratio of compatible and incompatible reactions was calculated and examined by modified x2 test.Furthermore, the mating type of all the sixty monokaryon cultures in mating test were determined by crosses between their parents or themselves each other.The result of fruiting test including twenty monokaryons and ten dikaryons indicated all the dikaryons could produce normal fruit-bodies, while only four monokaryons of tested twenty monokaryons could produce abnormal fruit-bodies.Comprehensively analyzing the results of mating test and fruiting test, the conclusion was drawn that the Auricularia auricula belongs to the bipolar heterothallic basidiomycete controlled by single factor.