Genetics of neotropical fish: from chromosomes to populations

The Neotropical freshwater fish fauna is very rich—according to the most recent catalogue 71 families and 4,475 species have been described. However, only a small amount of general information is available on the composition of Neotropical marine fishes. In Brazil, 1,298 marine species have been recorded. General analysis of available cytogenetic and population genetic data clearly indicates research has been mainly concentrated on freshwater fishes. Thus, today, cytogenetic information is available for 475 species of Characiformes, 318 species of Siluriformes, 48 species of Gymnotiformes, 199 freshwater species that do not belong to the superorder Ostariophysi, and only 109 species of marine fishes. For the species studied, only about 6% have sex chromosomes and about 5% have supernumerary or B chromosomes. A review of the cytogenetic studies shows that these data have provided valuable information about the relationships between fish groups, the occurrence of cryptic species and species complexes, the mechanism of sex determination and sex chromosome evolution, the distribution of nucleolus organizer regions, the existence supernumerary chromosomes, and the relationship between polyploidy and evolution. In relation to populations in Neotropical marine waters, the studies have shown the presence of cryptic species, which has important implications for fishery management. Different levels of genetic structuring can be found among Neotropical freshwater migratory fish species. This raises important implications for fish population genetic diversity and consequently its sustainable utilization in inland fisheries and aquaculture, specifically for conservation of ichthyo-diversity and survival.     

奥利亚罗非鱼（♀）、鳜（♂）及其子代间遗传关系的研究

罗非鱼生长快、味道好,是人们喜爱的鱼类,现已是世界性养殖鱼类,但它不能忍受低温,因此影响其养殖业的进一步发展。鳜则是名贵性鱼类,但其终生摄食活的鱼虾,这也限制了其养殖的发展。奥利亚罗非鱼(Oreochromisaurea)属鲈形目、鲡鱼科,鳜(Sinipercachuatsi)属鲈形目、脂科,它们间的杂交为科间杂交。鱼类科间杂交成功率很低,种间杂交有时可获得具有双亲性状的杂种鱼,有时也可发生雌核发育[1]。本研究中奥利亚罗非鱼雌鱼性染色体为ZW,其卵子如能在鳜精子刺激下行雌核发育,则可在短时间获得奥利亚罗非鱼纯系。在本文中,利用RAPD标记,对…     

Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Flavobacterium psychrophilum

Flavobacterium psychrophilum is the causative agent of bacterial cold-water disease and rainbow trout fry syndrome of salmonids. The pathogen has been reported from all regions in the world involved in salmonid aquaculture, but also from natural fresh-water environments. We established a quantitative loop-mediated isothermal amplification of DNA (LAMP) method to estimate quantities of F. psychrophilum . LAMP primers were designed based on the sequence of the DNA topoisomerase IV subunit B gene, parE , of F. psychrophilum. parE LAMP exhibited a high specificity for the parE gene of F. psychrophilum but not for other related species. parE LAMP detected the gene in a wide range of concentrations from 2.0 × 10¹ to 2.0 × 10⁹ copies/reaction within 70 min and revealed a good correlation between threshold times and gene copy number.     

栉孔扇贝、虾夷扇贝及其杂交子代的MSAP分析

运用甲基化敏感扩增多态性(methylation-sensitive amplification polymorphism,MSAP)技术对栉孔扇贝(♀)、虾夷扇贝(♂)及其杂交子一代、子二代基因组DNA胞嘧啶甲基化水平进行了研究,并分析了DNA甲基化与各性状的相关性及其与杂种优势的关系。结果表明,(1)DNA甲基化率与壳宽、总重等表型值呈正相关的关系,而与壳长、壳高、软体重和闭壳肌重4个性状表型值呈负相关的关系,其中闭壳肌重与甲基化率的相关性达到极显著水平(P<0.01);(2)虾夷扇贝、栉孔扇贝、F1代、F2代的总甲基化率分别为32.79%、24.13%、19.98%、20.18%,杂交种F1代的甲基化水平低于双亲,是两种扇贝杂交的结果;F1代的甲基化模式经过了重新调整,其变化相对其亲本主要有4种类型:甲基化水平相同、去甲基化、超甲基化、次甲基化,且去甲基化位点多于超甲基化位点。结果证实杂种优势的产生与杂交种F1代基因组DNA甲基化模式的改变和重新调整有关,丰富了杂种优势机理研究内容。     

改进的DNA指纹技术在玉米杂交种纯度鉴定上的应用研究

本试验以幼苗为对照,利用CTAB法建立了一套简单、快速的玉米干种子DNA提取程序,该程序提取的DNA分子量大小与幼苗法相当,能够利用SSR引物进行扩增,可应用于玉米杂交种纯度检测。     

发酵乳中沙门氏菌依赖解旋酶恒温基因扩增快速检测方法的建立

建立依赖解旋酶恒温基因扩增（helicase-dependent isothermal DNA amplification,HDA）快速检测发酵乳中沙门氏菌方法。以沙门氏菌invA基因序列为目的基因,设计特异性引物,优化反应体系中UvrD解旋酶及T4 gp32添加量,建立最优反应体系。通过HDA方法直接检测发酵乳中沙门氏菌,扩增其产物后进行电泳检测,验证方法特异性。结果表明：采用HDA快速检测法检测发酵乳中沙门氏菌特异性良好,优化后反应体系体积50 μL时,UvrD解旋酶添加量为0.10 μg,T4 gp32添加量为5.0 μg,得到与设计序列长度（304 bp）一致的扩增产物,检出限为2.6×102 CFU/g;该方法用于快速检测发酵乳中沙门氏菌能够满足检测需求,具有较高的灵敏度、易操作,可作为一种基础且快速的方法检测发酵乳中沙门氏菌。     

基于COI基因构建西北太平洋常见鱼类DNA条形码参考数据库

西北太平洋以其独特的地理位置和复杂的海洋环境,孕育了极为丰富的渔业资源,成为全球海洋生物多样性保护和渔业资源管理的热点区域。为了更有效地进行鱼类种类识别和多样性的调查,本研究建立了西北太平洋常见鱼类DNA条形码本地数据库。基于线粒体COI基因,对2023年6-8月在西北太平洋海域所采集的307份样品进行扩增和测序,共获得7目13科20属25种鱼类的COI基因序列。77.96%的COI序列在公共数据库中都能比对到高相似度序列。种间平均遗传距离为0.233,种内平均遗传距离为0.003,种间遗传距离是种内遗传距离的77.67倍,且能够形成明显的条形码间隙,不存在物种区分困难的现象。基于COI基因序列构建的系统发育树显示,同一属的鱼类首先聚为一支,随后与同一科的鱼类聚为一支,最后,同目不同科的鱼类聚为一支。综上所述,COI基因具有物种特异性,能够有效的区分西北太平洋常见鱼类物种,本数据库的初步建立,有利于后期利用环境DNA技术进行西北太平洋鱼类多样性的监测和调查,为西北太平洋海域生物多样性保护、资源管理和种群动态监测提供了技术支持。     

Isolation and molecular characterization of entomopathogenic nematode,Heterorhabditis sp. from an arable land in Nigeria

The occurrence of entomopathogenic nematodes(EPNs) in arable soil samples from Nigeria was investigated using Baermann extraction tray and insect-bait(White's trap) techniques. Isolates were tested for infectivity using the larvae of Galleria mellonella(greater moth) and Tenebrio molitor(mealworm). The study revealed a new species of Heterorhabditis(MT371593) in soil samples that were randomly collected from an arable farmland cultivated with cassava TMS-30572 at the Teaching and Research Farm of Landmark University, Nigeria. Amplification of the internal transcribed spacer region(ITS) of the ribosomal DNA produced a nucleotide sequence of 933 base pairs(bp). A BLASTN search of Gen Bank showed that the sequence of the Nigerian isolate is identical at 99% similarity to that of Heterorhabditis sp. from Thailand. Infectivity test of the isolate showed 100% mortality against T. molitor larvae within 48 h of exposure while only 80% mortality was recorded for G. mellonella after 1 week of exposure. This is the first account of Heterorhabditis sp. in Nigeria. The varying degrees of infectivity against mealworm and greater moth observed in this study proved that the Nigerian isolate of Heterorhabditis sp. could potentially be an attractive option in the management of insect pests of cash crops.     

BPA不影响卵母细胞减数分裂相关基因Dazl的甲基化

为了探讨环境雌激素BPA对小鼠卵母细胞减数分裂相关基因Dazl甲基化的影响,本研究通过给孕鼠饮用含有BPA的水方式使胎鼠在发育过程中接触BPA,利用重亚硫酸盐测序法,分析了胎鼠生殖嵴卵母细胞不同发育时期Dazl甲基化水平的变化。结果显示:Dazl在减数分裂期间处于低甲基化水平,无论对照组或处理组均低于10%,说明Dazl的低甲基化对维持减数分裂的正常进行有重要作用;对照组与处理组的甲基化水平相当,差异不显著,说明本研究的BPA浓度不影响卵母细胞Dazl的甲基化水平。     

Immunogenicity of multiple-epitope antigen gene of HCV carried by novel biodegradable polymers

In order to develop a promising vaccine candidate utilizing a combined approach to induce both antibody production and T-cell activity, the DNA fragment containing MA of HCV with five conserved epitopes was synthesized. Two types of HCV vaccine candidates (the DNA type and DNA/polymers) were constructed using MA. PLA-PEG-PLA and PLGA-PEG-PLGA were synthesized and used as micelles with encapsulated plasmid pcDNA3.1(+)-MA. The preparation of copolymers, the cloning and analysis of recombinant plasmid DNA, in vitro expression, and immunogenicity in transgenic mice were evaluated in detail. The results indicated that even single immunization and oral immunization with DNA/polymers achieved satisfying immune responses in vivo tests. As biodegradable and nontoxic triblock copolymers, the novel copolymers demonstrated a great advantage, as they made long-term and single-immunizing vaccines possible; in addition, the copolymers showed a better adjuvant effect and scarcely any side effects.