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微藻应用广泛,微藻生物燃料是近年研究热点。深圳的地理和气候条件都非常适宜微藻的生长,具有丰富的微藻资源。从深圳湾水域3个不同位置分别采集了表层和深层(5 m)的水样,利用Soil extrat培养基和Kuhl培养基对水样中的微藻进行分离。将分离得到的菌株扩大培养后提取其基因组DNA,经聚合酶链式反应(PCR)放大18S rRNA,利用内切酶pstⅠ和TaqⅠ对PCR产物进行双酶切反应以筛选不同的菌株。将不同菌株的PCR产物测序后通过与美国国家生物技术信息中心数据库比对,确定微藻品种。共得到微藻菌株71个,分属于16个不同的属,利用MEGA 5软件建立了系统发育树。同时,初步建立了深圳本地微藻种质资源库。
Microalgae is widely used, microalgae biofuel is a hot research in recent years. Shenzhen’s geographical and climatic conditions are very suitable for the growth of microalgae, with rich microalgae resources. Water samples from the surface and the deep layer (5 m) were collected from three different locations in the waters of Shenzhen Bay. Soil micro-algae were separated using soil extrat medium and Kuhl medium. The isolated strains were expanded and cultured to extract the genomic DNA. The 18S rRNA was amplified by polymerase chain reaction (PCR) and double digested with pstⅠand TaqⅠ to screen out different strains. The PCR products of different strains were sequenced and compared with the database of the National Center for Biotechnology Information to determine the microalgal variety. A total of 71 microalgae strains were obtained, belonging to 16 different genera, and phylogenetic trees were established using MEGA 5 software. At the same time, the initial establishment of Shenzhen local microalgae germplasm resources.