摘要
通过形态学观察结合分子生物学方法,对东海浙江枸杞岛蜈蚣藻属的种类进行初步研究,确定该属在东海枸杞岛分布有7种,分别是Grateloupia filicina(蜈蚣藻)、Grateloupia catenata(链状蜈蚣藻)、Grateloupia ramossissima(繁枝蜈蚣藻)、Grateloupia imbricate(复瓦蜈蚣藻)、Grateloupia lanceolata(披针形蜈蚣藻)、Grateloupia turuturu(带形蜈蚣藻)和Grateloupia asiatica(亚洲蜈蚣藻)。通过对采集到的海藻样品的大小、形状、颜色、质地和分枝等形态学特征进行详细的观察,并与分子鉴定的结果比对,发现蜈蚣藻、链状蜈蚣藻、繁枝蜈蚣藻、复瓦蜈蚣藻、带形蜈蚣藻、亚洲蜈蚣藻等种类样品的形态学鉴定结果与分子生物学鉴定结果一致,而披针形蜈蚣藻样品中GQ-6和GQ-8形态差异较大,但通过分子生物学方法鉴定表明二者均为披针形蜈蚣藻。
Grateloupia is a species of high economic value. It is edible and could be used as industrial raw materials for agar, also as natural pigments and food additives. In addition, the medicinal values of Grateoupia were very high. Currently,the development and utilization of Grateoupia has drawn much attention at home and abroad. The traditional classification of Grateloupia was based on external morphology,reproductive organs and spore etc, but the morphology might change with the environment and the classification could be disturbed. The identification of nucleic acid sequences by molecular biology might help in morphological classification and improve the classification system. The rbc L gene sequence was used to identify Grateloupia from Gouqi Island of Zhejiang Province. Seven species of Grateloupia were determined. It might make contribution to the diversity research and the basic taxonomic research of Grateloupia on the island.Samples were collected from the intertidal zone and mussels raft of Gouqi Island during July 2013 –April 2014. The samples were carefully classified by morphological characteristics such as size,shape,color,texture and branch. Then they were rinsed by 30% hydrogen peroxide once and by ultrapure water for three times,dried and ground into powder by freezing liquid nitrogen. The genomic DNA was extracted according to the method of COLEMAN,the extracted genomic DNA was used as template,and the rbc L gene sequence of algae was amplified by PCR. Alkali basis degree was analyzed and the clustering tree for rbc L constructed.The neigh-bor-joining（ NJ） and UPGMA phylogenetic tree were constructed based on Kimura-2-parameter（ K2P） distance method,and bootstrap was 1 000. Through morphological observation and rcb L sequence analysis, seven species of Grateloupia were identified as Grateloupia turuturu, Grateloupia lanceolata,Grateloupia filicina,Grateloupia imbricate,Grateloupia ramossissima,Grateloupia asiatica,and Grateloupia catenata. The morphological identifications of six species（ except Grateloupia lanceolata） were consistent with molecular identification. For example,GQ-29 was frond cylindrical,with irregular forked branches,taper branches became hard after drying. GQ-29 was Grateloupia ramossissima by morphological identification.Using the method of molecular biology GQ-29 and Grateloupia ramossissima were on the same branch,bootstrap was 100%, sequence divergence value was 0. 2%. GQ-29 was Grateloupia ramossissima by molecular identification. The morphological identification of Grateloupia ramossissima was consistent with the molecular identification. GQ-31 was frond trust,flat and algae fanned. The morphological identification was Grateloupia imbricate. Using the method of molecular biology（ NJ and UPGMA trees） GQ-31 and Grateloupia imbricata were on the same branch,bootstrap was high,sequence divergence value was 0. 0%. GQ-31 was Grateloupia imbricate. The morphological identification of Grateloupia imbricata was consistent with the molecular identification. The morphological identifications of other four species were all consistent with their molecular identifications. These also proved that molecular biological method was reliable. But samples of Grateloupia lanceolata were different,GQ-6 and GQ-8 showed different morphological features,but both of them were identified as Grateloupia lanceolata by molecular biological method. GQ-8 was frond slimy,with leaves lanceolate and taper branches. The morphological identification was Grateloupia lanceolata. GQ-6 was frond flat,leaf margin smooth,with several pieces of lanceolate lobes,base tapering,algae surface slimy and leaves thick than Grateloupia turuturu,morphology of GQ-6 was similar to Grateloupia elliptica. But molecular identifications showed that GQ-6 and GQ-8 were on the same branch,they had high bootstrap values,their sequence divergence value with Grateloupia lanceolata was 0. 0%. The sequence divergence value of GQ-6and Grateloupia elliptica was 4. 6%. Therefore,GQ-6 and GQ-8 were Grateloupia lanceolata. Grateloupia morphology had a lot of changes due to different ecological environment. The Grateloupia lanceolata was difficult to identify by traditional morphological methods. The molecular biological method was easy to control and more objective. The rapid development of molecular biology was based on the inheritance and development of traditional taxonomy,but it couldn＇t completely replace the traditional taxonomic methods.The combination of those two methods could better complete the research of algae taxonomy.
出处
《海洋渔业》
CSCD
北大核心
2015年第3期215-222,共8页
Marine Fisheries
基金
国家海洋局海洋公益性行业科研专项"我国南方沿海大型海藻生态系统恢复技术集成与示范"(2011050088)
作者简介
李颖博（1989-），男，硕士研究生，研究方向为海洋环境藻类生物学研究。E-mail：shoulyb@163．com
通讯作者：马家海，教授。E-mail：jhma@shou．edu．cn
