A novel cost effective and high-throughput isolation and identification method for marine microalgae.
A novel cost effective and high-throughput isolation and identification method for marine microalgae.
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DOI：
10.1186/1746-4811-10-26
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发表时间：
2014
期刊：
影响因子：
5.1
通讯作者：
Mock T
中科院分区：
文献类型：
--
作者：
Jahn MT;Schmidt K;Mock T
海洋微藻具有重要的生态和新兴的经济重要性。微藻特定性状的生物技术筛选计划和实验室实验，以提高我们对藻类生物学和进化的知识，极大地受益于反映最大限度的自然种间和种内多样性的文化收藏。然而，菌株分离和鉴定的标准程序，即DNA提取、纯化、扩增、测序和分类鉴定仍然包括相当大的限制，增加了建立新培养物所需的时间。在这项研究中，我们报告了一个成本效益和高通量的海洋微藻分离和鉴定方法。通过在平板上应用菌株分离并通过系统发育标记基因的直接PCR（dPCR）结合基于新型测序电泳图的筛选方法进行分类鉴定以评估分离培养物的分类多样性和身份，来增加通量。为了验证这种方法的有效性，我们从北冰洋的自然浮游植物群落中分离并鉴定了一系列单藻培养物。这些培养物包括在几种不同的硅藻中分离出一种新的海洋绿藻菌株。我们提供了一个高效和有效的方法，导致从自然浮游植物群落分离和分类鉴定的藻类菌株在短短几周内。验证与敏感的北极浮游植物，这种方法克服了标准的分子表征和建立单藻文化的限制。
Marine microalgae are of major ecologic and emerging economic importance. Biotechnological screening schemes of microalgae for specific traits and laboratory experiments to advance our knowledge on algal biology and evolution strongly benefit from culture collections reflecting a maximum of the natural inter- and intraspecific diversity. However, standard procedures for strain isolation and identification, namely DNA extraction, purification, amplification, sequencing and taxonomic identification still include considerable constraints increasing the time required to establish new cultures. In this study, we report a cost effective and high-throughput isolation and identification method for marine microalgae. The throughput was increased by applying strain isolation on plates and taxonomic identification by direct PCR (dPCR) of phylogenetic marker genes in combination with a novel sequencing electropherogram based screening method to assess the taxonomic diversity and identity of the isolated cultures. For validation of the effectiveness of this approach, we isolated and identified a range of unialgal cultures from natural phytoplankton communities sampled in the Arctic Ocean. These cultures include the isolate of a novel marine Chlorophyceae strain among several different diatoms. We provide an efficient and effective approach leading from natural phytoplankton communities to isolated and taxonomically identified algal strains in only a few weeks. Validated with sensitive Arctic phytoplankton, this approach overcomes the constraints of standard molecular characterisation and establishment of unialgal cultures.
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影响因子：
2.2
作者：
Chen, Wei;Zhang, Chengwu;Hu, Qiang
通讯作者：
Hu, Qiang
影响因子：
3.3
作者：
Radha, Sudhakar;Fathima, Anwar Aliya;Ramya, Mohandass
通讯作者：
Ramya, Mohandass
DOI：
10.1073/pnas.1001665107
发表时间：
2010-08-17
影响因子：
11.1
作者：
Cuvelier, Marie L.;Allen, Andrew E.;Worden, Alexandra Z.
通讯作者：
Worden, Alexandra Z.
影响因子：
11.4
作者：
LEE, RF;VALKIRS, AO;SELIGMAN, PF
通讯作者：
SELIGMAN, PF
DOI：
10.1016/s0967-0645(97)00057-x
发表时间：
1997-01-01
影响因子：
3
作者：
Booth, BC;Horner, RA
通讯作者：
Horner, RA