浙江沿海鱼类DNA条形码及其在鱼卵仔鱼鉴定中的应用

发布时间：2018-05-24 23:24

本文选题：鱼卵 + 仔稚鱼　；参考：《浙江海洋大学》2017年硕士论文

【摘要】：浙江沿海渔业资源丰富,是多种海洋经济鱼类的产卵场和其仔稚鱼索饵场。众所周知鱼卵和仔鱼是重要的渔业补充资源,它们的数量和保护情况以及所处的生态环境直接影响了成鱼的资源量。本研究探讨应用DNA条形码技术对鱼卵和仔鱼进行分类鉴定,了解浙江沿海水域产卵区的物种组成,为渔业管理者提供帮助。主要研究内容如下:1.成鱼样品采集了来自浙江舟山、登步、桃花、六横、虾峙、朱家尖、岱山、小长涂、大长涂、衢山、泗礁和枸杞岛等岛屿以及苍南沿海海域的样本。采集时间为2011年-2014夏天,捕捞方法为近海捕鱼船和近海定点渔网。一共鉴定出71种海洋鱼类,隶属于8目25科。基于COI序列与Genbak数据库比对,相似度达到98%以上的有65种,可以鉴定到种的水平。相似度在92%-98%的有3种,可以鉴定到属的水平。剩余3种相似度低于92%。2.鱼卵仔鱼样品收集于2014年5月-2015年11月,在浙江沿海水域租用渔船采用大型浮游生物网(网长280cm、网口内径80cm、网口面积0.5m2网目0.5mm、拖速2海里/h、拖网时间10分钟)。通过形态学初步鉴定,可以鉴定到科的占样品总数的16.9%,鉴定到种的占1.4%。通过Genbank数据库比对,结果显示从73个样本序列中一共鉴定出21个鱼类物种,分属于9个科。其中相似度达到98%以上的有67个样品19种,可以鉴定到种的比例达到91.8%。相似度达到83%到92%的有6个样品2种,可以鉴定到科的比例为100%。DNA条形码技术是传统形态学强有力的补充,可以很好的辅助形态学,并验证形态学鉴定结果。它是一种准确的,规范化的,有效的物种鉴定工具,在鱼类生物多样性研究中具有广阔的应用前景。3.通过分析COI DNA序列种内平均K-2-P遗传距离0.56%,属内种间平均遗传距离9.19%,科内属间平均遗传距离21.72%,目内科间平均遗传距离25.25%,表明种内距离远小于种间距离,与绝大多数条形码研究结果一致。基于鱼卵仔鱼COI序列构建的系统发育树显示结果与形态学方法构建的成鱼系统进化关系一致,说明DNA条形码技术可以应用于鱼卵仔鱼的分类鉴定。
[Abstract]:Zhejiang is rich in fishery resources, which is the oviposition of many kinds of marine economic fish and the bait field of their larvae and juveniles. It is well known that fish eggs and larvae are important supplementary fishery resources. Their quantity and protection and ecological environment have a direct impact on the amount of adult fish resources. In this study, DNA barcode technique was used to classify and identify fish eggs and larvae, to understand the species composition of spawning areas in Zhejiang coastal waters, and to provide assistance to fishery managers. The main research contents are as follows: 1. Adult fish samples were collected from Zhoushan, Zhejiang Province, Dengbu, Taohua, Liuheng, Xianzhi, Zhu Jiajian, Daishan, Xiaochangtu, Dachang Tuo, Ququ Mountain, Sijiao and Lycium Island, as well as the coastal waters of Cangnan. The collection time is 2011-2014 summer, the fishing method is the offshore fishing vessel and the offshore fixed point fishing net. A total of 71 species of marine fish belonging to 25 families of 8 orders were identified. Based on the alignment of COI sequence and Genbak database, there are 65 species whose similarity is above 98%, which can be identified at the level of species. There are 3 species with similarity between 92% and 98%, which can be identified to the level of genus. The remaining 3 kinds of similarity are less than 92. 2. Fish eggs were collected from May 2014 to November 2015. Large plankton nets (net length 280 cm, net diameter 80 cm, net area 0.5m2 mesh 0.5 mm, drag speed 2 nautical miles / h, trawl time 10 minutes) were used to hire fishing vessels in coastal waters of Zhejiang Province (net length 280 cm, net diameter 80 cm, net area 0.5 mm, drag speed 2 nautical miles / h, trawl time 10 minutes). According to the preliminary morphological identification, 16.9% of the total samples of the family and 1.4% of the species can be identified. The results of Genbank database alignment showed that 21 fish species belonging to 9 families were identified from 73 sample sequences. Among them, there are 67 samples with more than 98% similarity, and the proportion of identified species is 91.8%. There are 6 samples with similarity of 83% to 92%. The proportion of 100%.DNA bar code can be identified as a powerful supplement to traditional morphology, which can be used to assist morphology and verify the results of morphological identification. It is an accurate, standardized and effective tool for species identification, and has a broad application prospect in the study of fish biodiversity. The average genetic distance of K-2-P in COI DNA sequence was 0.56, the average genetic distance among intra species was 9.19, the average genetic distance among Kone genera was 21.72, and the average genetic distance among orders was 25.25. The results showed that the average genetic distance within species was much smaller than that between species. The results are consistent with the results of most bar code studies. The results of phylogenetic tree constructed based on COI sequence were consistent with the phylogenetic relationship of adult fish constructed by morphological method, which indicated that DNA barcode technique could be applied to the classification and identification of larvae.
【学位授予单位】：浙江海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

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