Genetic diversity of populations and clones of Rhopilema esculentum in China based on AFLP analysis

Amplified fragment length polymorphisms(AFLP) markers were developed to assess the genetic variation of populations and clones of Rhopilema esculentum Kishinouye(Scyphozoa,Rhizostomatidae).One hundred and seventy-nine loci from 56 individuals of two hatchery populations and two wild populations were genotyped with five primer combinations.The polymorphic ratio,Shannon’s diversity index and average heterozygosity were 70.3%,0.346 and 0.228 for the white hatchery population,74.3%,0.313,and 0.201 for the red hatchery population,79.3%,0.349,and 0.224 for the Jiangsu wild population,and 74.9%,0.328 and 0.210 for the Penglai wild population,respectively.Thus,all populations had a relatively high level of genetic diversity.A specific band was identified that could separate the white from the red hatchery population.There was 84.85% genetic differentiation within populations.Individual cluster analysis using unweighted pair-group method with arithmetic mean(UPGMA) suggested that hatchery populations and wild populations could be divided.For the hatchery populations,the white and red populations clustered separately;however,for the wild populations,Penglai and Jiangsu populations clustered together.The genetic diversity at the clone level was also determined.Our data suggest that there are relatively high genetic diversities within populations but low genetic differentiation between populations,which may be related to the long-term use of germplasm resources from Jiangsu Province for artificial seeding and releasing.These findings will benefit the artificial seeding and conservation of the germplasm resources.     

海蜇(Rhopilema esculentum)浮游幼体对褐牙鲆(Paralichthys olivaceus)卵和初孵仔鱼捕食的比较研究

利用实验生态学方法研究不同个体大小的海蜇浮游幼体[平均伞径(2.5±0.1),(4.1±0.2),(11.7±0.5),(21.1±0.6)mm]对不同密度(10,30,50,80ind/L)的褐牙鲆卵[卵径(0.92±0.01)mm]和初孵仔鱼[全长(3.01±0.08)mm]的捕食率,解析海蜇浮游幼体对初孵仔鱼的捕食率随捕食时间(0.5,1,2,3,4,5h)的变化特征。结果表明,各个体组海蜇浮游幼体对卵的捕食率均显著低于对仔鱼的捕食率;它们对卵的捕食率与卵密度和海蜇个体大小的关系不显著,但对仔鱼的捕食率随海蜇个体大小及仔鱼密度的增大而显著升高;伞径21.1mm的个体对仔鱼的捕食率在开始捕食后1h时达到最大值[17.3ind/(predator.h)],此后随捕食时间的延长而逐渐下降。在自然水域中,如果二者发生时空上的匹配关系,海蜇浮游幼体对仔鱼的捕食可能影响褐牙鲆的早期存活及其资源补充量的变动。     

Genetic diversity and population structure of the golden cuttlefish,Sepia esculenta (Cephalopoda: Sepiidae) indicated by microsatellite DNA variations

The golden cuttlefish, Sepia esculenta Hoyle, 1885 (Cephalopoda: Sepiidae) is a valuable and important fishery resource for China, Japan and Korea. This fishery has experienced severe population decline largely due to overexploitation in past decades. To provide guidelines for fisheries management, we estimated genetic diversity and population structure across four locations along the coast of Japan and one location in China (a total 281 individuals) using nine microsatellite DNA loci. Sepia esculenta showed high genetic variability with mean allele richness ranging from 15.8 to 21.4, and mean heterozygosity from 0.80 to 0.90. Weak but significant genetic differentiation was present and the F_ST value was 0.020 across all five populations. The useful information obtained in this study will offer insights into how to fine‐tune conservation and fishery management measures for this species and resource in the future.     

Comparative analysis of methods for concentrating venom from jellyfish Rhopilema esculentum Kishinouye

In this study, several methods were compared for the efficiency to concentrate venom from the tentacles of jellyfish Rhopilema esculentum Kishinouye. The results show that the methods using either freezing-dry or gel absorption to remove water to concentrate venom are not applicable due to the low concentration of the compounds dissolved. Although the recovery efficiency and the total venom obtained using the dialysis dehydration method are high, some proteins can be lost during the concentrating process. Comparing to the lyophilization method, ultrafiltration is a simple way to concentrate the compounds at high percentage but the hemolytic activities of the proteins obtained by ultrafiltration appear to be lower. Our results suggest that overall lyophilization is the best and recommended method to concentrate venom from the tentacles of jellyfish. It shows not only the high recovery efficiency for the venoms but high hemolytic activities as well.     

可口革囊星虫(Phascoloma esculenta)铁结合蛋白基因的研究

根据已报道的铁结合蛋白基因的保守序列设计引物,用cDNA末端快速扩增(RACE)法,成功地从可口革囊星虫体液中获得铁结合蛋白基因的全长序列.结果表明,该基因cDNA全长1017bp,5'-端非编码区为15lbp,3'一非编码区为341bp,开放阅读框长度为525bp(括一个终止密码子),可编码175个氨基酸(GenBank:EU091352).该序列与加洲海兔、皱纹盘鲍、刺参、微小牛蜱、叉尾鲴、非洲爪蟾、小家鼠、人等的铁结合蛋白基因有67%-75%的同源性,而相应的氨基酸同源性为59%-76%.氨基酸相似性为74%*-89%%.分析结果表明,铁结合蛋白基因在动物进化中是高度保守的.     

闽江口海蜇生长的研究

分别采用高次方程和指数高次方程模拟闽江口海蜇伞径和体重的生长,结果表明海蜇生长明显分３个阶段：５月１０日以前为缓慢生长期;５月１１日至７月３０日为快速生长期;７月３０日以后为衰老期,个体收缩,体重减轻;伞径和体重的最大值分别出现在８月５日和７月３０日;伞径和体重生长速度最大值分别在６月２日和６月２５日。据此确定闽江口海蜇最佳捕捞期７月３０日,最佳捕捞伞径４９４．１ｍｍ;合理开捕期７月２０日,开捕伞     

海月水母与海蜇不同幼体阶段的种间短期竞争研究

近年来海蜇池塘养殖业的兴起,补充了传统海蜇渔业捕捞生产的不足,推动了海蜇产业的迅猛发展。海月水母常常在海蜇养殖池塘中泛滥成灾,对海蜇养殖业常常造成很大损失,但关于海蜇与海月水母的种间竞争关系尚缺乏研究。本文对海月水母与海蜇的螅状体、碟状体及水母体幼体相互之间的短期生存竞争关系进行了初步观察研究。结果表明:除海月水母碟状体可以被海蜇螅状体所捕食外,各种类型的海蜇幼体基本不能对海月水母幼体造成危害。但海月水母螅状体及其水母体幼体对海蜇幼体均存在一定的危害性,尤其是对海蜇碟状体的危害性最为严重,在饵料缺乏的情况下海蜇碟状体可以100%地被捕食。随着海蜇幼体的生长,其对于海月水母危害的抵抗力逐渐增强。本研究显示海月水母相对于海蜇而言,在幼体种间竞争关系中处于绝对优势,这是海月水母时常暴发成灾的一个重要因素。在海蜇池塘养殖生产及苗种培育过程中,应当严禁混入海月水母幼体,以避免对海蜇养殖生产造成损害。     

海蜇养殖群体及自然捕获群体ITS 序列遗传分析

为调查海蜇(Rhopilema esculentum)自然海区群体和养殖群体遗传多样性,对烟台莱州湾和江苏海州湾自然海区捕获群体及威海养殖群体24个个体的ITS序列进行了PCR扩增和序列分析,获得DNA片段长度在1 080~1 096 bp之间,包括完整的ITS1,5.8S和完整ITS2序列。结果表明,在所测碱基序列中共发现26个变异位点,4处多碱基插入位置,29个插入缺失位点。群体内平均核苷酸差异数K和平均核苷酸多样性指数Pi为2.179~2.750和0.002 02~0.002 54,群体间平均核苷酸差异数K值波动在2.797~3.031之间,遗传距离在0.002 30~0.002 81之间,遗传分化指数(Fst)值在0.032 50~0.730 8之间,群体内及群体间遗传多样性指标数值比较相近,群体间的遗传距离比较小,群体遗传结构相似,群体间没有明显的遗传分化。     

Isolation and characterization of venom from nematocysts of jellyfish Rhopilema esculentum Kishinouye

The present work is the first report of the biochemical characterization of the venom from nematocysts of the jellyfish Rhopilema esculentum Kishinouye. The nematocysts were isolated by autolysis and centrifugation and separated by flow cytometry. Four types of nematocysts were identified: mastigophores, euryteles, and atrichous and holotrichous isorhiza. SDS-PAGE and amino acid analyses demonstrated that most of the proteins in the nematocyst extract were between 10 kDa and 40 kDa, and that glutamic acid was the main amino acid. A hemolytic activity assay showed that the activity of the nematocyst venom (RNV) was strongest in Tris-HCl buffer (50 mmol/L, pH 7.8, 5% glycerol, 0.5 mmol/L EDTA, 0.1 mol/L NaCl). The hemolytic activity was related to protein concentration and the HU50 against chicken erythrocytes was 0.91 μg/mL.     

Comparative analysis of methods for concentrating venom from jellyfish Rhopilema esculentum Kishinouye

In this study, several methods were compared for the efficiency to concentrate venom from the tentacles of jellyfish Rhopilema esculentum Kishinouye. The results show that the methods using either freezing-dry or gel absorption to remove water to concentrate venom are not applicable due to the low concentration of the compounds dissolved. Although the recovery efficiency and the total venom obtained using the dialysis dehydration method are high, some proteins can be lost during the concentrating process. Comparing to the lyophilization method, ultrafiltration is a simple way to concentrate the compounds at high percentage but the hemolytic activities of the proteins obtained by ultrafiltration appear to be lower. Our results suggest that overall lyophilization is the best and recommended method to concentrate venom from the tentacles of jellyfish. It shows not only the high recovery efficiency for the venoms but high hemolytic activities as well. Supported by the Award Foundation of Scientific Research for Excellent Young and Middle-age Scientist of Shandong Province (No. 2006BS07003) and the Knowledge Innovation Project of Chinese Academy of Sciences (KZCX2-YW-R-104)