深圳湾水域微藻的分离及分子生物学鉴定

Isolation and identification of microalgae in Shenzhen Bay with molecular biotechnology

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 are widely used in many areas. Using microalgae for biofuel production is a hot topic in recent years. The location and climatic conditions of Shenzhen are very suitable for the growth of microalgae. In this article, water samples were collected from two depths （shallow and five meters deep） at three different sites in Shenzhen Bay area. Microalgae strains in water samples were isolated by Soil-extract media and Kuhl media. Genomic DNA of each strain was extracted after cultivation and 18S rRNA was amplified by PCR. Then restriction enzymes pst I and Taq I were applied to select different strains. After comparing the PCR sequences results with the data in NCBI, the species of microalgae were identified. Seventy-one microalgal strains which belonged to 16 genera were collected and the phylogenetic tree of the microalgae was built by software MEGA 5. Meanwhile, a preliminary collection of native microalgae in Shenzhen was established.