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Molecular approach (PCR-DGGE) to diet analysis in young Antarctic krill Euphausia superba

机译:分子方法(PCR-DGGE)对南极磷虾幼年磷虾的膳食进行分析

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Antarctic krill Euphausia superba Dana comprise a key component of the Southern Ocean food web, yet despite decades of research, questions concerning the regional, seasonal and ontogenetic differences in their diet remain. All current methods used to characterize krill diet have limitations for identifying the full complement of the diet. Using DNA as a marker molecule, our goal in this study has been to evaluate the efficacy of a PCR-DGGE (denaturing gradient gel electrophore-sis) approach targeting the 18S rDNA gene to discriminate among diet constituents in gut and fecal pellet samples from young Antarctic krill relative to their feeding environment—the seawater and sea ice microbial community. We conducted 2 laboratory-based feeding experiments with known food items and 3 field samplings of both the krill and their feeding environment. Sequenced PCR-DGGE phylotypes from laboratory trials clearly distinguished diatom and copepod prey, while in situ feeding analyses revealed that a broad diversity of taxa were ingested, including diatoms (Bacillario-phyta, the most prevalent group detected), dinoflagellates, cryptomonads, prasinophytes, ciliates, cercozoans, choanoflagellates, turbellarians and (possibly) sponge larvae. Band image analyses allowed environmental and diet phylotypes to be matched. On average, 32% of those from the environment were present in the diet; conversely, of the phylotypes detected in the diet, an average of 59% were in common with the environment. Changes in environmental phylotypes among sampling dates were reflected by similar changes in the krill diet as potential prey diversity (richness) decreased during a phytoplankton bloom.
机译:南极磷虾Euphausia superba Dana构成了南部海洋食物网的重要组成部分,尽管进行了数十年的研究,但有关其饮食的区域,季节和个体发育差异的问题仍然存在。当前用于表征磷虾饮食的所有方法都存在局限性,无法确定饮食的全部补充。使用DNA作为标记分子,我们在这项研究中的目标是评估针对18S rDNA基因的PCR-DGGE(变性梯度凝胶电泳)方法的功效,以区分年轻肠和粪便颗粒样品中的饮食成分相对于其摄食环境(海水和海冰微生物群落)的南极磷虾。我们对已知的食物进行了2次基于实验室的喂养实验,并对磷虾及其喂养环境进行了3次野外采样。来自实验室试验的测序PCR-DGGE系统型清楚地区分了硅藻和co足类的猎物,而就地喂养分析表明,摄入了各种各样的分类单元,包括硅藻(芽孢杆菌,检出的最普遍的群体),鞭毛藻,隐孢子虫,藻类,纤毛虫,天牛,鞭毛虫,turbellarians和(可能)海绵幼虫。带图像分析允许环境和饮食系统型匹配。饮食中平均有32%来自环境。相反,在饮食中检测到的系统型中,平均有59%与环境共有。磷虾饮食中的类似变化反映了采样日期之间环境系统型的变化,因为浮游植物开花期间潜在的猎物多样性(丰富度)降低。

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