蟾蜍壶菌病病原遗传分化研究

Genetic Differentiation of the Pathogen of Batrachochytrium dendrobatidis in Toads

壶菌病为近年发现的两栖类动物重要传染病,对野生和养殖种群危害极大,为确定药用经济动物蟾蜍历史上壶菌病感染情况,提高养殖蟾蜍疾病防治水平,选取某博物馆馆藏采集于四川的蟾蜍标本32只,利用Taqman-MGB荧光探针定量PCR技术进行壶菌检测,并对定量PCR产物克隆、测序,通过序列比对和系统发育分析判定其来源。最终得到定量PCR标准曲线:Y=-3.0X+32.39;相关系数R2=0.999 6;检测结果为阳性样本12只,检出率37.5%。同时系统发育分析表明,我国的壶菌存在一定程度的分化,一类与北美洲、南美洲、欧洲菌株呈现高度的亲缘关系;另一类则表现出与世界其他地区分布的壶菌有明显的不同,显示其独特性。这提示我国蟾蜍壶菌病的防制应更具针对性,不可盲目照搬国外经验。

The chytridiomycosis is a significant emerging disease of amphibians in recent years which seriuosly does harm to the wild and breeding population. To ascertain the condition of the Batrachochytrium dendrobatidis infection history in the population of toad, the medicinal economic animal, and enhance the prevention for this disease, 32 toad specimen collected from Sichwan keeping in a museum were assayed by Taqman-MGB fluorescence probe quantitative polymerase chain reaction to detect Batrachochytrium dendrobatidis. Besides, the products of QPCR were cloned and sequenced to identify the origin of the pathogen by sequence alignment and phylogenetie analysis. Finally we got the standard curve： Y = - 3.0X ＋ 32.39 and the related coefficient： R2 = 0.999 6. Total 12 positive samples reported the detec- tion rate as 37.5 %. Meanwhile the phylogenetic analysis indicated that a certain extent differentiation of the chytrid fungi in our country exists. One kind of the tested strain showed highly genetic relationship with the strains from the North America, South America and Europe. Another one is obviously different from the strains found in the other areas of the world with special characteristics. It suggest that the pre- vention and treatment for chytridiomycosis of toad should be more targeted and should not copy foreign experience aimlessly.