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藻液的不同处理方法对石珊瑚共生虫黄藻密度测定准确性的影响

Influences of different treatment methods of algae suspensing on the accurate determination of zooxanthellae density

  • 摘要: 珊瑚和虫黄藻形成的共生关系是珊瑚礁生态系统的营养和结构基础。共生虫黄藻密度的测定成为了研究珊瑚和虫黄藻间共生关系的关键技术。由于虫黄藻共生于珊瑚虫的内胚层细胞中,传统的虫黄藻液的处理方法存在藻细胞分离不彻底和藻细胞聚团等问题,严重影响到虫黄藻密度测定的准确性。本研究使用曲拉通(TritonX-100)、十二烷基硫酸钠(SDS)和氨苄西林(AMP)3种化学试剂并结合物理振荡,采用两两组合的方式,处理盾形杜沙珊瑚(Duncanopsammia peltata)的虫黄藻液。通过比较处理后测得的虫黄藻密度差异,探究藻液的不同处理方法对虫黄藻密度测定准确性的影响,筛选出最佳的藻液处理方法。实验结果显示,虫黄藻液经过化学处理和物理处理后,测得的虫黄藻密度均有显著提升。在化学处理中,经过SDS处理后的虫黄藻液测得的虫黄藻密度最高。SDS处理既能显著减少上清液中的藻细胞浓度,又能显著减少藻液中藻团的数量。振荡处理能够显著减少藻液中藻团数量和藻团面积,减轻藻细胞聚团程度。研究结果表明,经过SDS和振荡处理后,虫黄藻液测得的虫黄藻密度最高,是最佳的藻液处理组合。本研究有助于提高珊瑚生理和生态等研究中共生虫黄藻密度测定的准确性。

     

    Abstract: The symbiotic association between corals and zooxanthellae forms the nutritional and structural foundation of coral reef ecosystems. Accurate determination of zooxanthellae density is critical for studying this symbiotic relationship. However, conventional processing methods for algal liquid often face challenges such as incomplete separation of algal cells and algal cell aggregation, significantly compromising measurement accuracy, as zooxanthellae live in the endoderm cells of polyps. In this study, three chemical reagents-TritonX-100 (TRI), ampicillin (AMP), and sodium dodecyl sulfate (SDS) were applied in pairwise combinations alongside physical oscillation to obtain algal suspension from Duncanopsammia peltata. By comparing the differences in zooxanthellae density following the treatments, we investigated the effects of different processing methods on measurement accuracy of zooxanthellae and identified the optimal protocol. Results demonstrated that both chemical and physical treatments significantly increased measured zooxanthellae densities. Among the chemical treatments, SDS treatment yielded the highest zooxanthellae density while markedly reducing algal cell concentrations in supernatants and minimizing algal clumping in the final algal liquid and physical oscillation significantly reduced both the number and area of algal clumps, effectively alleviating algal cell aggregation. The combination of SDS treatment and physical oscillation achieved the highest zooxanthellae density and was identified as the optimal processing protocol. Thus, it enhances the precison of zooxanthellae density for coral physiological and ecological studies.

     

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