驯鹿皮蝇蛆病病原形态及分类学研究
摘要
本研究对内蒙古根河地区寄生于驯鹿皮下的皮蝇蛆从形态学和分子生物学两个方面进行了鉴定,准确确定了其种属和分类地位。另外,还利用相关基因资料,对多种家畜蝇蛆的系统发生和进化关系进行了研究和比较。
在形态学方面,利用光学显微镜和扫描电子显微镜,对所检获的驯鹿皮蝇第二、三期幼虫进行了详细的观察及描述。特别是在伪头、体节刺(尤其是第十腹节腹面刺)和气门板构造上给出了其重要特征。
在分子生物学方面,对内蒙古驯鹿体发现的皮蝇蛆线粒体COI基因序列进行了研究,其DNA核苷酸测序表明该种皮蝇蛆线粒体COI种属特异性基因UEA7到UEA10特殊目标编码区域片段长度约为689 bp;种系发生进化树和同源性分析显示其与鹿皮蝇法国株同源性非常接近。因此确定内蒙古地区驯鹿皮蝇蛆病病原为鹿皮蝇(Hypoderma diana)。
多种家畜蝇蛆种间的差异性分析显示:皮蝇科、胃蝇科和狂蝇科不同科间序列差异明显,差异性在18.7 %～31.9%之间;同科不同属间差异性在12.3%～27.2 %之间;同属不同种间差异性在0.9%～18.7%之间;寄生于鹿科动物体内不同科属的蝇蛆差异性在1.9%～25.5%之间;种内差异性为0.6%～1.2%,鹿皮蝇其种内差异性为6.0%。说明生物线粒体COI基因核苷酸序列在一定程度上,可反映出种属及株间在进化上的差异性。
In order to indentify of Hypodermatinae which were found in reindeer in Genhe of Inner Mongolia, and study the characteristics of morphology and molecularbiology. In addition, the phylogenesis and the relation of evolution of the many kinds of domestic fly larvae were studied and compared by the related gene sequence .
In the morphology,the study was mainly to describe and observe the morphological characteristics of the pseudocephalon,the spines (especially the spines on the tenth ventral segment)and the spiracular plates in 2 and 3stage larvae in the unkown larva by optical microscope and scanning electron microscope . Determination of species was generally accorded to the characteristic of larvae of in stage 3 and fly morphous,and especially the latter was very important, so morphology was initially identified as Hypoderma Diana.
In the molecularbiology,the species specificity sequence of mitochondrial cytochrome oxidaseI(COI)gene sequence of warble fly larvae of unknown species from reindeer in Inner Mongolia and other related fly larvae isolates were studied by molecular biology techniques. The result of DNA nucleotide sequence was confirmed that the size of species-specificity of COI gene (partial cds.) from UEA7 to UEA10 of unknown species of reindeer in Inner Mongolia Region of China was approximately 689 bp. The analysis of phylogenic tree and homology showed that the unknown species and the France isolate of Hypoderma diana were extraordinarily near to each other. Thus, the larvae of unknown species from reindeer in Inner Mongolia is Hypoderma diana.
The interspecific difference showed that the sequences between Hypodermatidae and Oestridae and Gastrophilidae were obviously different, ranged from 18.7% to 31.9%. Dieffrent genus of variation ranged from12.3% to 27.2%. Different species pairwise divergences of same genus ranged from 0.9% to 18.7%; Different species pairwise divergences of fly larvae parasited in different deer ranged from 1.9% to 25.5%. Dieffrent isolates of the intraspecific variation of ranged from 0.6% to 1.2%, and intraspecific variation of Hypoderma diana was 6.0%. This study indicated that the nucleotide sequence of the mitochondrial cytochrome oxidase I(COI)gene could reflect the phylogenetic relationships among genera, species and strains to some extent.Considering the analytic result in morphology and molecularbiology,the larva founded in reindeer was Hypoderma diana.
在形态学方面,利用光学显微镜和扫描电子显微镜,对所检获的驯鹿皮蝇第二、三期幼虫进行了详细的观察及描述。特别是在伪头、体节刺(尤其是第十腹节腹面刺)和气门板构造上给出了其重要特征。
在分子生物学方面,对内蒙古驯鹿体发现的皮蝇蛆线粒体COI基因序列进行了研究,其DNA核苷酸测序表明该种皮蝇蛆线粒体COI种属特异性基因UEA7到UEA10特殊目标编码区域片段长度约为689 bp;种系发生进化树和同源性分析显示其与鹿皮蝇法国株同源性非常接近。因此确定内蒙古地区驯鹿皮蝇蛆病病原为鹿皮蝇(Hypoderma diana)。
多种家畜蝇蛆种间的差异性分析显示:皮蝇科、胃蝇科和狂蝇科不同科间序列差异明显,差异性在18.7 %～31.9%之间;同科不同属间差异性在12.3%～27.2 %之间;同属不同种间差异性在0.9%～18.7%之间;寄生于鹿科动物体内不同科属的蝇蛆差异性在1.9%～25.5%之间;种内差异性为0.6%～1.2%,鹿皮蝇其种内差异性为6.0%。说明生物线粒体COI基因核苷酸序列在一定程度上,可反映出种属及株间在进化上的差异性。
In order to indentify of Hypodermatinae which were found in reindeer in Genhe of Inner Mongolia, and study the characteristics of morphology and molecularbiology. In addition, the phylogenesis and the relation of evolution of the many kinds of domestic fly larvae were studied and compared by the related gene sequence .
In the morphology,the study was mainly to describe and observe the morphological characteristics of the pseudocephalon,the spines (especially the spines on the tenth ventral segment)and the spiracular plates in 2 and 3stage larvae in the unkown larva by optical microscope and scanning electron microscope . Determination of species was generally accorded to the characteristic of larvae of in stage 3 and fly morphous,and especially the latter was very important, so morphology was initially identified as Hypoderma Diana.
In the molecularbiology,the species specificity sequence of mitochondrial cytochrome oxidaseI(COI)gene sequence of warble fly larvae of unknown species from reindeer in Inner Mongolia and other related fly larvae isolates were studied by molecular biology techniques. The result of DNA nucleotide sequence was confirmed that the size of species-specificity of COI gene (partial cds.) from UEA7 to UEA10 of unknown species of reindeer in Inner Mongolia Region of China was approximately 689 bp. The analysis of phylogenic tree and homology showed that the unknown species and the France isolate of Hypoderma diana were extraordinarily near to each other. Thus, the larvae of unknown species from reindeer in Inner Mongolia is Hypoderma diana.
The interspecific difference showed that the sequences between Hypodermatidae and Oestridae and Gastrophilidae were obviously different, ranged from 18.7% to 31.9%. Dieffrent genus of variation ranged from12.3% to 27.2%. Different species pairwise divergences of same genus ranged from 0.9% to 18.7%; Different species pairwise divergences of fly larvae parasited in different deer ranged from 1.9% to 25.5%. Dieffrent isolates of the intraspecific variation of ranged from 0.6% to 1.2%, and intraspecific variation of Hypoderma diana was 6.0%. This study indicated that the nucleotide sequence of the mitochondrial cytochrome oxidase I(COI)gene could reflect the phylogenetic relationships among genera, species and strains to some extent.Considering the analytic result in morphology and molecularbiology,the larva founded in reindeer was Hypoderma diana.
引文
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