Developing a high-throughput screen for the isolation of the model green alga Chlamydomonas reinhardtii from soil samples

藻类是一种多样性惊人的生物，是生态系统功能的组成部分，约占全球固碳量的一半。此外，藻类在应对气候变化方面发挥着关键作用，因为它们可以用来生产环境清洁的生物燃料和其他有价值的生物产品。尽管它们很重要，但我们对这些物种如何在自然环境中生活知之甚少。这包括几乎完全缺乏关于不同物种是否适应特定生态条件的知识，以及对物种的种群遗传学的了解很少（例如，如果物种内存在不同的遗传种群，种群之间存在什么迁移率等）。莱茵衣藻（Chlamydomonas reinhardtii）是藻类生物学研究的前沿物种。几十年来，细胞生物学、生物化学和遗传学研究都是利用这种物种进行的，这使它成为研究许多相关问题的理想模型。这些范围从探索藻类的生态学和种群遗传学，到了解生物燃料生产背后的生化途径。尽管它在多个领域得到了广泛的应用，并对该物种进行了大量的采样尝试，但它只被成功分离了36次，而且几乎所有的研究都是在单一的相关实验室菌株上进行的。虽然这严重限制了我们对种群遗传学的理解，但这也意味着生物技术研究忽略了物种中存在的遗传和表型多样性（例如，没有进行选择性育种以提高生物燃料产量）。在这份提交给英国-加拿大Globalink博士交流计划的提案中，我们的目标是开发一种高通量筛选方法来检测环境样本中莱茵假梭菌的存在，并随后从存在莱茵假梭菌的样本中分离出该物种。这将通过开发一种独特的莱茵梭菌遗传标记来实现，该标记可以使用标准分子生物学技术从环境样本中扩增和检测。一旦发展起来，这项技术将扩展到各种不同环境的地点，使我们能够对物种的生态偏好有一些初步的了解。此外，我们将开始建立一个可用于群体遗传学和生物技术应用的分离株集合。这项工作将与多伦多大学的Rob Ness教授及其研究小组一起进行，作为爱丁堡大学为期3个月的研究交流的一部分。样本将从安大略省和魁北克省周围收集，从以前分离过该物种的地方开始。如果成功，该方法也可以在未来扩展到其他藻类物种，这一点尤其重要，因为仅从单个培养的分离物中就知道了几种工业模式物种（例如佐芬吉色绿藻）。总之，我们的目标是开发一种实验室技术，该技术将有助于对生物学上重要的物种进行高通量采样，从而在藻类生物学的不同领域探索几个新问题。

BBSRC : Rory Craig : BB/M010996/1Algae are an amazingly diverse group of organisms that are integral to the function of ecosystems, performing approximately half of global carbon fixation. Furthermore, algae have a key role to play in the fight against climate change, as they can be used to produce environmentally clean biofuels and other valuable bioproducts. Despite their importance, we know very little about how these species actually live in natural environments. This includes almost a complete lack of knowledge concerning whether different species are adapted to specific ecological conditions, and little understanding of the population genetics of species (e.g. if there are distinct genetic populations within species, what rates of migration exist between populations, etc.).The green alga Chlamydomonas reinhardtii is the foremost research species in algal biology. Several decades of cell biology, biochemistry and genetics research have been performed using this species, making it the ideal model to investigate a number of pertinent questions. These range from exploring the ecology and population genetics of algae, to understanding the biochemical pathways underlying biofuel production. Despite its wide use across several fields and extensive attempts to sample the species, it has only been successfully isolated 36 times, and almost all research has been performed on a single line of related laboratory strains. While this severely limits our understanding of population genetics, it also means that the genetic and phenotypic diversity present in the species is ignored in biotechnology research (e.g. no selective breeding has been performed to increase biofuel yields). In this proposal to the UK-Canada Globalink Doctoral Exchange Scheme, we aim to develop a high-throughput screen to detect the presence of C. reinhardtii in environmental samples, and to subsequently isolate the species from samples where it is present. This would be achieved by developing a unique genetic marker for C. reinhardtii, that could be amplified and detected from environmental samples using standard molecular biology techniques. Once developed, this technique would be extended to a wide variety of sites differing in their environment, allowing us to gain some of the first insights into the ecological preferences of the species. Furthermore, we would begin to build a collection of isolates that could be used for both population genetics and biotechnology applications. This work would be performed with Prof. Rob Ness and his research group at the University of Toronto, as part of a 3-month research exchange from the University of Edinburgh. Samples would be collected from around Ontario and Quebec, starting in locations where the species has been isolated previously. If successful, the approach could also be extended to other algal species in the future, which is especially important given that several industrial model species are known from only a single cultured isolate (e.g. Chromochloris zofingiensis).In summary, we aim to develop a laboratory technique that will facilitate the high-throughput sampling of a biologically important species, enabling several new questions to be explored across the diverse field of algal biology.