北黄海藻类DNA病毒遗传多样性研究
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摘要
浮游病毒是指游离在水体中的各类病毒,被认为是水体微生物群落中的重要成分,它在微食物环、生物地球化学循环、群落结构及多样性变化等方面起着重要作用。研究浮游病毒在不同海域和不同环境的丰度和分子多样性,可以探索浮游病毒的分布规律及生态学地位,为更好的理解生物群落结构变化规律奠定基础,并为环境评价等提供数据参考及理论依据。
北黄海海区代表了北温带海洋生态环境的一种重要类型,在该海域仅有少量利用荧光显微镜技术研究浮游病毒丰度的相关报道。本文利用流式细胞仪对北黄海水域秋季浮游病毒丰度进行了检测,并分析浮游病毒丰度变化与水文参数的相关性。各站点浮游病毒丰度在2.76×106～3.97×107个mL-1之间,平均值为2.12×107个mL-1；水平分布上,浮游病毒高值区主要分布在调查区域的东南部海域。垂直分布上,各水层浮游病毒丰度无显著差异。昼夜变化方面,浮游病毒丰度最高值出现在30m层19：00时刻,最低值出现在10m层4：00时刻。浮游病毒丰度在1：00时刻出现峰值后,急剧下降,在4：00降至最低(底层除外),在7：00出现一个峰值后从10：00到19：00呈上升趋势。相关性分析显示,浮游病毒丰度与异养细菌、聚球藻、微微型真核浮游植物均呈显著正相关,与VBR无相关性。浮游病毒丰度与温度正相关,与盐度和深度显著负相关。
藻类DNA病毒在海洋中的多样性极其丰富,而在国内尚未见相关报道。本文首次运用DGGE技术调查北黄海和青岛近海藻类DNA病毒群落多样性。在北黄海A604站、B107站和C803站分别得到10个、8个、8个OTUs (operational taxonomic units),在青岛近海ZQ站得到6个OTUs,说明不同海域藻类DNA病毒多样性有一定的差异,在北黄海较丰富,在青岛近海多样性稍低。北黄海3个站间相比,OTUs数量和位置均存在差异,这表明同一海域不同站位藻类DNA病毒多样性也存在差异。在所得OTUs中,有2个OTUs在4个站中均存在,这说明在北黄海和青岛近海存在着一些共同的藻类DNA病毒。
本文依据DGGE带谱从直观上对北黄海和青岛近海藻类DNA病毒多样性进行了初步的研究,而最终确定某一环境中的病毒组成及分类地位需要与基因测序、系统树的构建结合起来。作者运用PCR-RFLP技术,通过构建藻类DNA病毒系统进化树对北黄海藻类DNA病毒科遗传多样性进行了研究。本文建立了北黄海藻类DNA病毒DNA聚合酶文库,随机挑取329个阳性克隆进行RFLP分析。获得的18个OTUs分别属于3簇：簇Ⅰ与已分离的针胞藻病毒属的赤潮异湾藻病毒HaV01亲缘关系较近,推测可能同属于针胞藻病毒属；簇Ⅱ与褐藻病毒属FsV亲缘关系较近,推测可能同属于褐藻病毒属；簇Ⅲ为未获得培养的类群,其分类地位有待于进一步研究。本文在北黄海海域没有检测到寄生藻病毒属、绿藻病毒属、颗石藻病毒属和金藻病毒属的成员。
Virioplankton, viruses in aquaic ecosystem, are now considered to be one of the most abundant and active components of microbial communities. Viroplankton play an important role in the microbial loop, biogeochemical cycles, structure and diversity of microbial community. Studying of the abundance and diversity of viroplankton in different locations and environment will help in understanding of the rules of the distribution and the ecological status of viroplankton. And it will also help to understand population structure of biology and provide reference numerical dates and theoretical basis for environmental assessment.
The North Yellow Sea represents an important type of marine ecosystem of the north temperate zone. There were just a few reports about the abundance of virioplankton in the North Yellow Sea using epiflororence microscopy(EFM). In this paper, we studied the abundance of virioplankton by flow cytometry(FCM), and analyzed the correlation between virioplankton and environmental parameters in the North Yellow sea. Viral abundance ranged from 2.76×106 to 3.97×107 particles mL-1(2.12×107 particles mL-1 on average). In horizontal distribution, the area of the high viral abundance existed the southeast area of investigation area. In vertical distribution, there was no significant difference of virioplankton abundance in each water layer. In daily variations of virioplankton, its highest virioplankton abundance existed at the 30m water layer at 19:00, while the lowest abundance existed at the 10m water layer at 4:00. The abundance of virioplankton appeared uptrend from 10:00 to 19:00 in all layers. Virioplankton abundance has obvious positive correlation with heterotrophic bacteria, Synechococcus(Syn) and picoeukaryotes(Euk), however, there was no correlation between virioplankton abundance and VBR. The virioplankton abundance was positively correlated with temperature, while it has negative correlation with depth and salty.
The diversity of algae DNA viruses are extremely rich in the oceans, but there was no any reports about the study of viroplankton diversity in China. The diversity of algae DNA viruses in the North Yellow Sea and Qingdao coastal waters was analysed by denaturing gradient gel electrophoresis (DGGE). The amount of operational taxonomic units (OTUs) in the North Yellow Sea was 10,8,8 at stations of A604, B107 and C803, respectively, and it was 6 in ZQ station. These results implicated that the diversity of algal viruses was not the same in different waters, which was higher in the North Yellow Sea and lower in ZQ station. Furthermore, the diversity of algal viruses was also different in the three stations of the North Yellow Sea. And 2 similar OTUs were found throughout the whole four stations, suggesting that some kinds of genetically identical algal viruses have a ubiquitous distribution in the seas。
DGGE map indicated the diversity of algae DNA viruses in the North Yellow Sea and Qingdao coastal waters, but it was preliminary. In order to define the members and classification of virus, the target segments were sequenced and were used in sequence analysis. In this paper, we studied the genetic diversity of algae DNA viruses in the North Yellow Sea by PCR-RFLP.329 positive clones were digested by HaeⅢand MspⅠ, and we obtained 18 operational taxonomic units(OTUs). The 18 OTUs were attributed to three cluster. Cluster I was most closely related to DNA pol sequences of HaV01 which belongs to Raphidovirus. Cluster II was most closely related to DNA pol sequences of FsV which belongs to Phaeovirus. Cluster III was attributed to uncultured viruses, and its definite position in classification need further research. No member of Chlorovirus, Prasinovirus, Prymnesiovirus and Coccolithovirus was discovered in this study.
北黄海海区代表了北温带海洋生态环境的一种重要类型,在该海域仅有少量利用荧光显微镜技术研究浮游病毒丰度的相关报道。本文利用流式细胞仪对北黄海水域秋季浮游病毒丰度进行了检测,并分析浮游病毒丰度变化与水文参数的相关性。各站点浮游病毒丰度在2.76×106～3.97×107个mL-1之间,平均值为2.12×107个mL-1；水平分布上,浮游病毒高值区主要分布在调查区域的东南部海域。垂直分布上,各水层浮游病毒丰度无显著差异。昼夜变化方面,浮游病毒丰度最高值出现在30m层19：00时刻,最低值出现在10m层4：00时刻。浮游病毒丰度在1：00时刻出现峰值后,急剧下降,在4：00降至最低(底层除外),在7：00出现一个峰值后从10：00到19：00呈上升趋势。相关性分析显示,浮游病毒丰度与异养细菌、聚球藻、微微型真核浮游植物均呈显著正相关,与VBR无相关性。浮游病毒丰度与温度正相关,与盐度和深度显著负相关。
藻类DNA病毒在海洋中的多样性极其丰富,而在国内尚未见相关报道。本文首次运用DGGE技术调查北黄海和青岛近海藻类DNA病毒群落多样性。在北黄海A604站、B107站和C803站分别得到10个、8个、8个OTUs (operational taxonomic units),在青岛近海ZQ站得到6个OTUs,说明不同海域藻类DNA病毒多样性有一定的差异,在北黄海较丰富,在青岛近海多样性稍低。北黄海3个站间相比,OTUs数量和位置均存在差异,这表明同一海域不同站位藻类DNA病毒多样性也存在差异。在所得OTUs中,有2个OTUs在4个站中均存在,这说明在北黄海和青岛近海存在着一些共同的藻类DNA病毒。
本文依据DGGE带谱从直观上对北黄海和青岛近海藻类DNA病毒多样性进行了初步的研究,而最终确定某一环境中的病毒组成及分类地位需要与基因测序、系统树的构建结合起来。作者运用PCR-RFLP技术,通过构建藻类DNA病毒系统进化树对北黄海藻类DNA病毒科遗传多样性进行了研究。本文建立了北黄海藻类DNA病毒DNA聚合酶文库,随机挑取329个阳性克隆进行RFLP分析。获得的18个OTUs分别属于3簇：簇Ⅰ与已分离的针胞藻病毒属的赤潮异湾藻病毒HaV01亲缘关系较近,推测可能同属于针胞藻病毒属；簇Ⅱ与褐藻病毒属FsV亲缘关系较近,推测可能同属于褐藻病毒属；簇Ⅲ为未获得培养的类群,其分类地位有待于进一步研究。本文在北黄海海域没有检测到寄生藻病毒属、绿藻病毒属、颗石藻病毒属和金藻病毒属的成员。
Virioplankton, viruses in aquaic ecosystem, are now considered to be one of the most abundant and active components of microbial communities. Viroplankton play an important role in the microbial loop, biogeochemical cycles, structure and diversity of microbial community. Studying of the abundance and diversity of viroplankton in different locations and environment will help in understanding of the rules of the distribution and the ecological status of viroplankton. And it will also help to understand population structure of biology and provide reference numerical dates and theoretical basis for environmental assessment.
The North Yellow Sea represents an important type of marine ecosystem of the north temperate zone. There were just a few reports about the abundance of virioplankton in the North Yellow Sea using epiflororence microscopy(EFM). In this paper, we studied the abundance of virioplankton by flow cytometry(FCM), and analyzed the correlation between virioplankton and environmental parameters in the North Yellow sea. Viral abundance ranged from 2.76×106 to 3.97×107 particles mL-1(2.12×107 particles mL-1 on average). In horizontal distribution, the area of the high viral abundance existed the southeast area of investigation area. In vertical distribution, there was no significant difference of virioplankton abundance in each water layer. In daily variations of virioplankton, its highest virioplankton abundance existed at the 30m water layer at 19:00, while the lowest abundance existed at the 10m water layer at 4:00. The abundance of virioplankton appeared uptrend from 10:00 to 19:00 in all layers. Virioplankton abundance has obvious positive correlation with heterotrophic bacteria, Synechococcus(Syn) and picoeukaryotes(Euk), however, there was no correlation between virioplankton abundance and VBR. The virioplankton abundance was positively correlated with temperature, while it has negative correlation with depth and salty.
The diversity of algae DNA viruses are extremely rich in the oceans, but there was no any reports about the study of viroplankton diversity in China. The diversity of algae DNA viruses in the North Yellow Sea and Qingdao coastal waters was analysed by denaturing gradient gel electrophoresis (DGGE). The amount of operational taxonomic units (OTUs) in the North Yellow Sea was 10,8,8 at stations of A604, B107 and C803, respectively, and it was 6 in ZQ station. These results implicated that the diversity of algal viruses was not the same in different waters, which was higher in the North Yellow Sea and lower in ZQ station. Furthermore, the diversity of algal viruses was also different in the three stations of the North Yellow Sea. And 2 similar OTUs were found throughout the whole four stations, suggesting that some kinds of genetically identical algal viruses have a ubiquitous distribution in the seas。
DGGE map indicated the diversity of algae DNA viruses in the North Yellow Sea and Qingdao coastal waters, but it was preliminary. In order to define the members and classification of virus, the target segments were sequenced and were used in sequence analysis. In this paper, we studied the genetic diversity of algae DNA viruses in the North Yellow Sea by PCR-RFLP.329 positive clones were digested by HaeⅢand MspⅠ, and we obtained 18 operational taxonomic units(OTUs). The 18 OTUs were attributed to three cluster. Cluster I was most closely related to DNA pol sequences of HaV01 which belongs to Raphidovirus. Cluster II was most closely related to DNA pol sequences of FsV which belongs to Phaeovirus. Cluster III was attributed to uncultured viruses, and its definite position in classification need further research. No member of Chlorovirus, Prasinovirus, Prymnesiovirus and Coccolithovirus was discovered in this study.
引文
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