中国蚌螨属主要种类线粒体基因和食性分析及幼螨的分子分类
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摘要
本研究结合普通PCR技术和Long PCR技术测定了弯弓蚌螨(U. arcuata)线粒体基因组全序列,该序列全长14574bp,A+T含量明显高于G+C含量,高含量的A+T是螨类生物线粒体基因组序列中普遍存在的现象。弯弓蚌螨COI基因全序列长1539bp,该基因共编码512个氨基酸,其中起始密码子为ATG,终止密码子为TAA, TGA编码色氨酸Trp而不是终止密码子,亮氨酸Leu的含量最高,半胱氨酸Cys的含量最低。利用线粒体COI基因序列分析弯弓蚌螨与其它种类蚌螨间的系统发生关系,发现敏捷蚌螨可能是最早从祖先种分化出来的种,而弯弓蚌螨与佛氏蚌螨、丫纹蚌螨可能起源于蚌螨属的同一个祖先。
分析弯弓蚌螨群体间的线粒体12S rRNA基因序列差异和系统发生关系,发现相同宿主的弯弓蚌螨群体间的12S rRNA序列遗传差异值不超过0.5%,不同宿主的弯弓蚌螨群体间的12S rRNA序列遗传差异值超过了6.7%；相同宿主的弯弓蚌螨群体首先聚于一支,而不同宿主的弯弓蚌螨群体却相隔甚远。采自不同宿主内的弯弓蚌螨群体中可能存有蚌螨属内的隐蔽种(Cryptic species)。生物地理隔离对采自相同宿主内的弯弓蚌螨群体间的12S rRNA序列差异的影响微小。然而,宿主的选择与利用却导致不同淡水贝类内的弯弓蚌螨群体间产生了高度的遗传分化。
利用线粒体16S rRNA基因对弯弓蚌螨和丫纹蚌螨(U. ypsilophora)的食物组份进行分析,结果发现：以褶纹冠蚌DNA、弯弓蚌螨整个个体DNA和内脏组织DNA、丫纹蚌螨整个个体DNA和内脏组织DNA为模板扩增出的序列完全相同,序列长度均为587bp；弯弓蚌螨和丫纹蚌螨内脏组织中均含有未消化的褶纹冠蚌DNA。从褶纹冠蚌鳃部采集到的弯弓蚌螨和丫纹蚌螨均取食了褶纹冠蚌的组织,弯弓蚌螨和丫纹蚌螨可能在褶纹冠蚌内营寄生生活。
以线粒体12S rRNA基因部分序列作为分子标记,鉴定出了我国蚌螨属(Unionicola)4种蚌螨幼螨,即：丫纹蚌螨(U. ypsilophora)、弯弓蚌螨(U. arcuata)、螯爪蚌螨(U. chelata)和敏捷蚌螨(U. agilex)幼螨,并对这4种幼螨的形态进行了描述,整理出了我国蚌螨属4种幼螨的分类检索表,归纳了它们共同的形态特征：肩刚毛(Hu)是幼螨背面最为粗壮且长度最大的一根刚毛；泄殖孔板(Ep)和泄殖孔(Exp)均为椭圆形,且泄殖孔上无刚毛着生；在泄殖孔板四周分布的刚毛中,腹侧刚毛4(V4)的长度远超其它刚毛；幼螨的足Ⅰ最短,足Ⅲ最长,足Ⅱ的长度介于足Ⅰ和足Ⅲ之间；幼螨的足Ⅰ转节上均仅着生有一根刚毛Tr1,Tr1短而粗；跗节是幼螨足上最长的一节。
In this research, the complete sequence of mitochondrial genome was amplified by Standard PCR and Long PCR for Unionicola arcuata, and its whole sequence was14574bp in length. In the complete sequence of mitochondrial genome for U. arcuata, the percentages of A+T apparently higher than G+C, high content of A+T for mitochondrial genome from mites was commonly phenomenon. The complete sequence of mitochondrial COI gene for U. arcuata was1539bp in length, which totally encodes512amino acids with ATG as start codon, TAA as stop codon, TGA as encoding Trp instead of a termination codon, and the content of Leu is the highest, while Cys is the lowest. Using mitochondrial COI gene sequence to analyzed phylogenetic relationships between U. arcuata and other unionicolid mites, we found that the U. agilex might be the first separated from ancestral species, and U. arcuata, U. foili and U. ypsilophora might originate in the same ancestor.
Be analyzed sequence divergences and phylogenetic relationships of the mitochondrial12S rRNA gene among groups of U. arcuata, we found that the12S genetic divergence among groups of U. arcuata from the same host species never exceeded0.5%, whereas the divergence among groups of U. arcuata from different host species was6.7%or higher. The groups of U. arcuata from different molluscan hosts may actually represent cryptic species of Unionicola. The biogeographic separation among groups of U. arcuata from the same species of host had little impact on divergence in12S. However, host selection and utilization resulted in a high degree of genetic differentiation among U. arcuata from different freshwater mussels.
Using mitochondrial16S rRNA gene partial sequence to analyzed alimental composition of U. arcuata and U. ypsilophora, the result shows that utilized DNA of Cristaria plicata, DNA of entire individual and splanchnic tissues for U. arcuata and U. ypsilophora as templates were amplified the same sequence, and the length was587bp; Containing undigested DNA of C. plicata in splanchnic tissues for U. arcuata and U. ypsilophora. U. arcuata and U. ypsilophora were collected from the gills and feeding on the tissues of C. plicata. U. arcuata and U. ypsilophora might be parasitic association with C. plicata.
Using mitochondrial12S rRNA gene partial sequence as a molecular marker to succeed on classified four species of unionicolid larval mites from China, these larval mites were U. ypsilophora, U. arcuata, U. chelata and U. agilex. Morphological description of these four species of unionicolid larval mites from China, and the key to the larval mites was listed. The larval mites show some the same morphological characters:humeral seta in dorsum of the larval mites was a widest and longest seta; excretory pore plate and excretory pore were oval in shape, and never seta grow in excretory pore; ventral seta4was a longest seta in excretory pore plate around; leg Ⅰ for larval mites was the shortest leg, while leg Ⅲ was the longest leg, the length of leg Ⅱ was between leg Ⅰ and leg Ⅲ; Only one seta Trl grow in trochanter of leg Ⅰ from the larval mites, this seta was short and thick; tarsus in leg of the larval mites was a longest segment.
分析弯弓蚌螨群体间的线粒体12S rRNA基因序列差异和系统发生关系,发现相同宿主的弯弓蚌螨群体间的12S rRNA序列遗传差异值不超过0.5%,不同宿主的弯弓蚌螨群体间的12S rRNA序列遗传差异值超过了6.7%；相同宿主的弯弓蚌螨群体首先聚于一支,而不同宿主的弯弓蚌螨群体却相隔甚远。采自不同宿主内的弯弓蚌螨群体中可能存有蚌螨属内的隐蔽种(Cryptic species)。生物地理隔离对采自相同宿主内的弯弓蚌螨群体间的12S rRNA序列差异的影响微小。然而,宿主的选择与利用却导致不同淡水贝类内的弯弓蚌螨群体间产生了高度的遗传分化。
利用线粒体16S rRNA基因对弯弓蚌螨和丫纹蚌螨(U. ypsilophora)的食物组份进行分析,结果发现：以褶纹冠蚌DNA、弯弓蚌螨整个个体DNA和内脏组织DNA、丫纹蚌螨整个个体DNA和内脏组织DNA为模板扩增出的序列完全相同,序列长度均为587bp；弯弓蚌螨和丫纹蚌螨内脏组织中均含有未消化的褶纹冠蚌DNA。从褶纹冠蚌鳃部采集到的弯弓蚌螨和丫纹蚌螨均取食了褶纹冠蚌的组织,弯弓蚌螨和丫纹蚌螨可能在褶纹冠蚌内营寄生生活。
以线粒体12S rRNA基因部分序列作为分子标记,鉴定出了我国蚌螨属(Unionicola)4种蚌螨幼螨,即：丫纹蚌螨(U. ypsilophora)、弯弓蚌螨(U. arcuata)、螯爪蚌螨(U. chelata)和敏捷蚌螨(U. agilex)幼螨,并对这4种幼螨的形态进行了描述,整理出了我国蚌螨属4种幼螨的分类检索表,归纳了它们共同的形态特征：肩刚毛(Hu)是幼螨背面最为粗壮且长度最大的一根刚毛；泄殖孔板(Ep)和泄殖孔(Exp)均为椭圆形,且泄殖孔上无刚毛着生；在泄殖孔板四周分布的刚毛中,腹侧刚毛4(V4)的长度远超其它刚毛；幼螨的足Ⅰ最短,足Ⅲ最长,足Ⅱ的长度介于足Ⅰ和足Ⅲ之间；幼螨的足Ⅰ转节上均仅着生有一根刚毛Tr1,Tr1短而粗；跗节是幼螨足上最长的一节。
In this research, the complete sequence of mitochondrial genome was amplified by Standard PCR and Long PCR for Unionicola arcuata, and its whole sequence was14574bp in length. In the complete sequence of mitochondrial genome for U. arcuata, the percentages of A+T apparently higher than G+C, high content of A+T for mitochondrial genome from mites was commonly phenomenon. The complete sequence of mitochondrial COI gene for U. arcuata was1539bp in length, which totally encodes512amino acids with ATG as start codon, TAA as stop codon, TGA as encoding Trp instead of a termination codon, and the content of Leu is the highest, while Cys is the lowest. Using mitochondrial COI gene sequence to analyzed phylogenetic relationships between U. arcuata and other unionicolid mites, we found that the U. agilex might be the first separated from ancestral species, and U. arcuata, U. foili and U. ypsilophora might originate in the same ancestor.
Be analyzed sequence divergences and phylogenetic relationships of the mitochondrial12S rRNA gene among groups of U. arcuata, we found that the12S genetic divergence among groups of U. arcuata from the same host species never exceeded0.5%, whereas the divergence among groups of U. arcuata from different host species was6.7%or higher. The groups of U. arcuata from different molluscan hosts may actually represent cryptic species of Unionicola. The biogeographic separation among groups of U. arcuata from the same species of host had little impact on divergence in12S. However, host selection and utilization resulted in a high degree of genetic differentiation among U. arcuata from different freshwater mussels.
Using mitochondrial16S rRNA gene partial sequence to analyzed alimental composition of U. arcuata and U. ypsilophora, the result shows that utilized DNA of Cristaria plicata, DNA of entire individual and splanchnic tissues for U. arcuata and U. ypsilophora as templates were amplified the same sequence, and the length was587bp; Containing undigested DNA of C. plicata in splanchnic tissues for U. arcuata and U. ypsilophora. U. arcuata and U. ypsilophora were collected from the gills and feeding on the tissues of C. plicata. U. arcuata and U. ypsilophora might be parasitic association with C. plicata.
Using mitochondrial12S rRNA gene partial sequence as a molecular marker to succeed on classified four species of unionicolid larval mites from China, these larval mites were U. ypsilophora, U. arcuata, U. chelata and U. agilex. Morphological description of these four species of unionicolid larval mites from China, and the key to the larval mites was listed. The larval mites show some the same morphological characters:humeral seta in dorsum of the larval mites was a widest and longest seta; excretory pore plate and excretory pore were oval in shape, and never seta grow in excretory pore; ventral seta4was a longest seta in excretory pore plate around; leg Ⅰ for larval mites was the shortest leg, while leg Ⅲ was the longest leg, the length of leg Ⅱ was between leg Ⅰ and leg Ⅲ; Only one seta Trl grow in trochanter of leg Ⅰ from the larval mites, this seta was short and thick; tarsus in leg of the larval mites was a longest segment.
引文
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