摘要
传统鉴定藻种的方法主要是通过形态学观察的方法加以判断。蓝藻在自然条件和人工培养过程中 ,其形态、代谢能力等都可能发生变化 ,同时该过程需要的时间长 ,难以区分种或种以下的分类、单位 ,亦难以在水华暴发早期阶段准确鉴定。本文利用rDNA通用引物扩增 ,表明在 5 0 μL的反应体系中加入 2 0个鱼腥藻细胞能扩增出目的条带 ;对已知的鱼腥藻PC基因的分析设计引物 ,在BSA浓度为 0 2 %— 1% (w/v)下 ,全细胞扩增出实验室保存的四种鱼腥藻的部分PC以及PC IGS序列 ,序列分析结果表明PC
The traditional way to identify algae is usually depend on the observation of the morphology. This way is time consuming and usually difficult to identify the algae during the early algal growth stage in which the character is not obvious and can not detect the subtle differences between species or strains; also, the morphology and the ability to produce toxicants of cyanobacteria may change under different natural or cultural conditions. In this paper, we use the molecular method to identify Anabaena species, the results showed that BSA are necessary for the PCR amplification and the optimal concentration is 1%(w/v) in the reacion buffer. DMSO increased or decreased the quantity of the amplification products selectively, but the effect is not significant. The self designed primers after comparison of other Anabaena PC sequences amplified the part of pcb and pca gene along with PC-IGS sequence when using both extracted DNA and pretreated whole cells as template, the banding pattern is not uniform but all produced the target bands, cloning and sequencing analyses indicated that PC in line with PC-IGS was amplified, even though the nucleotide between two Anabaena strains collected from two different regions in China show that only 2 sites in a fragment about 400 bp are different, but the PC-IGS sequences are quite different from others and thus can be applied to identify the different species or strains. The universal primers to amplify part of 16S rDNA and 23S rDNA along with their ITS sequence using the extracted DNA and pretreated whole cell showed that the banding pattern of amplification products are uniform when the whole cells were at the concentration of 10 5/mL in the reaction buffer, the minimum amount of whole cell to amplify the target band is 20 cells in 50μL reaction buffer.
出处
《水生生物学报》
CAS
CSCD
北大核心
2004年第2期159-162,共4页
Acta Hydrobiologica Sinica
基金
中国科学院知识创新工程重大项目 (KZCX1 SW 12 )
973项目 (2 0 0 2CB412 3 0 0 )
国家重大环境课题"滇池蓝藻水华污染控制技术研究"(K99 0 5 3 5 0 1)
中国科学院领域前沿课题 (No.2 2 0 3 16)资助
