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1.
利用线粒体16SrRNA基因和线粒体细胞色素氧化酶亚基Ⅰ（cvtocllrome oxidase Ⅰ，COI）基因片段初步研究钦州湾牡蛎（Ostxea）的物种组成。以特异引物进行PCR扩增，对扩增产物进行纯化、测序，分析表明，16SrRNA基因部分长度为413bp，COI基因部分长度为535bp，2种牡蛎序列的碱基组成均显示出较高的A＋T比例：16SrRNA基因598％；COI基因605％。对位排序比较表明，16SrRNA片段种内个体间变异较小，存在7个变异位点，4种单倍型，其中包括5个转换位点突变，2个颠换位点突变；COI片段有30个碱基存在变异，7种单倍型，其中包括16个转换位点突变，14个颠换位点突变。运用MEGA4软件计算出不同个体间的遗传距离，并构建了NJ和UPGMA系统树。香港牡蛎（Crassostrea hongkongensis）16SrRNA和COI序列与白肉牡蛎的遗传距离均为0000，有明巨牡蛎（Crassostrea ariakensis）16SrRNA和COI序列和红肉牡蛎（redmeatostxea）的遗传距离分别为0000和0011，白肉牡蛎16SrRNA和COI序列和红肉牡蛎之间的遗传距离分别为0035和0146。结果表明，钦州湾牡蛎分为2个不同的种，线粒体16SrRNA和COI基因在种间存在明显的多态性，证实了16SrRNA和COI基因序列适用于牡蛎的系统学分析。 相似文献
2.
利用线粒体16S rRNA基因和线粒体细胞色素氧化酶亚基Ⅰ(cytochrome oxidaseⅠ,COⅠ)基因片段初步研究钦州湾牡蛎(Ostrea)的物种组成。以特异引物进行PCR扩增,对扩增产物进行纯化、测序,分析表明,16S rRNA基因部分长度为413bp,COⅠ基因部分长度为535bp,2种牡蛎序列的碱基组成均显示出较高的A+T比例:16S rRNA基因59.8%;COⅠ基因60.5%。对位排序比较表明,16S rRNA片段种内个体间变异较小,存在7个变异位点,4种单倍型,其中包括5个转换位点突变,2个颠换位点突变;COⅠ片段有30个碱基存在变异,7种单倍型,其中包括16个转换位点突变,14个颠换位点突变。运用MEGA4软件计算出不同个体间的遗传距离,并构建了NJ和UPGMA系统树。香港牡蛎(Crassostrea hongkongensis)16S rRNA和COⅠ序列与白肉牡蛎的遗传距离均为0.000,有明巨牡蛎(Crassostrea ariakensis)16S rRNA和COⅠ序列和红肉牡蛎(red meat ostrea)的遗传距离分别为0.000和0.011,白肉牡蛎16S rRNA和COⅠ序列和红肉牡蛎之间的遗传距离分别为0.035和0.146。结果表明,钦州湾牡蛎分为2个不同的种,线粒体16S rRNA和COⅠ基因在种间存在明显的多态性,证实了16S rRNA和COⅠ基因序列适用于牡蛎的系统学分析。 相似文献
3.
对裸体方格星虫(Sipunculus nudus)、可口革囊星虫(Phascolosoma esculenta)和澳洲管体星虫(Siphonosoma australe)的线粒体16S rRNA、COI和细胞色素b(Cytb)基因片段序列进行比较,并对其系统发生进行了初步探讨。采用PCR方法得到总长度分别为531～544bp(16S)、652～675bp(COI)和406～453bp(Cytb)的线粒体片段。片段碱基A+T比例较高(16S rRNA基因58.3%,COI基因56.9%,Cytb基因59.5%)。16S rRNA片段存在169个碱基变异位点(其中包括167个简约信息位点)和44个碱基插入/缺失,种内个体间变异较小;COI片段有512个碱基(333个简约信息位点)存在变异,79个碱基插入/缺失;Cytb片段存在347个碱基(318个简约信息位点)变异位点,16个碱基插入/缺失。数据分析结果支持3种星虫和环节动物的分类地位较近,与软体动物较远的分类观点。此外,裸体方格星虫与澳洲管体星虫之间亲缘关系较近(D=0.3159、0.3156、0.2361)。认为3种星虫线粒体16S rRNA、COI和Cytb基因在种间存在明显的多态性,证实了三种基因序列均普遍适用于星虫种及以上阶元的系统学分析。 相似文献
4.
《广东海洋大学学报》2017,(6)
运用PCR技术扩增吉富罗非鱼选育群体第20和21世代18S-ITS1-5.8S序列,分析二序列的遗传变异。结果表明,18S-ITS1-5.8S序列中,18S、内转录间隔区(ITS)1和5.8S序列分别为156、483~540、64 bp,主要变异位点位于ITS1中;基于ITS1序列的第20代吉富罗非鱼(17尾)群体内遗传距离为0.001,保守位点530个,变异位点10个,单倍型4个,单倍型多样性为0.331±0.143,核苷酸多样性为0.002,平均核苷酸差异为1.279;基于ITS1的第21代吉富罗非鱼(42尾)群体内遗传距离为0.015,6个个体存在严重碱基缺失现象,保守位点492个,变异位点49个,单倍型12个,单倍型多样性为0.638±0.083,核苷酸多样性为0.014,平均核苷酸差异为6.945。ITS序列在该两吉富罗非鱼群体中变异较小,基于ITS1序列分析的两代群体遗传多样性水平较低。 相似文献
5.
《广东海洋大学学报》2016,(4)
采用DNA条形码技术辅以传统形态分类方法,对雷州半岛马尾藻属(Sargassum)6常见种进行物种鉴定和系统进化分析。测得6种马尾藻线粒体COI基因序列,该序列与Gen Bank和BOLD数据库中马尾藻序列同源性均大于或等于99%,该结果与形态分类学鉴定结果一致。COI基因序列特征分析表明,6种马尾藻保守位点421个,变异位点58个,简约信息位点21个,单一突变位点37个,变异率7.7%,T、C、A、G平均含量分别为39.1%、18.9%、19.1%、22.9%,A+T含量(58.2%)高于C+G含量(41.8%),UUU所编码的苯丙氨酸(F)使用频率最高,达12.5%,最大似然法估算的转换颠换比值为2.42,种间遗传距离多在0.030±0.013~0.100±0.016之间。聚类分析结果与形态学鉴定、同源性分析结果一致。 相似文献
6.
近江牡蛎两个野生种群的遗传多样性分析 总被引：1，自引：0，他引：1
采用RAPD分子标记和rDNA-ITS1序列分析了近江牡蛎Crassostrea rivularis湛江官渡和阳江程村野生种群的遗传多样性。12个RAPD引物共扩增出8838条片段,157个位点,平均每个引物可产生13个位点,片段长度在200～2200bp之间。湛江种群和阳江种群的多态位点比例分别为89.62%和89.57%,遗传多样性指数分别为0.4170和0.4334。种群间平均遗传距离为0.0327,平均遗传相似性为0.9681,平均遗传分化系数为0.0437。得到近江牡蛎18S、5.8S部分序列和ITS1全部序列,其中ITS1序列片段长度为478～485bp,共有11个变异位点,两个为转换(A/G),其他为插入/缺失(A/-、T/-)。湛江和阳江种群各获得8个单倍型,其中有一个单倍型为两个种群共享。两个种群的CG碱基平均含量较高,分别为58.29%和58.41%。种群间的遗传分化系数0.0254。结果说明,近江牡蛎湛江种群和阳江种群间具有较高的遗传多样性和较低的遗传分化。 相似文献
7.
利用PCR技术分别扩增连云港及启东沿海蛤蜊科的西施舌(Coelomactra antiquata)、中国蛤蜊(Mactrachinensis)和四角蛤蜊(Mactra veneriformis)3种双壳贝的16S rRNA基因片段和ITS2核苷酸序列,测序后用DNA star软件分析了核苷酸差异。结果显示:三种贝类16S rRNA基因片段长度相同,均为306bp(去除引物),核苷酸存在多态性,共有45个变异位点,54个核苷酸发生了变异,全部为碱基置换。西施舌与中国蛤蜊此片段核苷酸的同源性为88.9%,与四角蛤蜊的同源性为88.6%,中国蛤蜊与四角蛤蜊的同源性为90.6%。三种蛤蜊ITS2序列分别为390 bp(西施舌)4、41 bp(四角蛤蜊)和466 bp(中国蛤蜊),存在长度多态性,ITS2核苷酸差异分析结果显示,西施舌与中国蛤蜊的同源性为70.9%-71.1%,西施舌与四角蛤蜊的为70.5%-71.0%,中国蛤蜊与四角蛤蜊的同源性为88.1%-88.8%。ITS2序列分析结果与16S rRNA基因片段分析结果一致,2种分子分析法均显示中国蛤蜊与四角蛤蜊的亲缘关系近。 相似文献
8.
采用聚合酶链式反应(PCR)技术对粤西镇海湾水域的近江牡蛎Crassostrea rivularis (Gould)群体27个个体的线粒体DNA16S rRNA基因序列片段进行扩增，获得大约500bp的扩增产物。PCR产物经纯化后进行序列测定，经Clustal X同源排序，除去引物及部分端部序列，得到414bp的核苷酸片段。27个个体共检测到2个变异位点，均为颠换位点，没发现碱基位点插入、缺失及转换位点，共3种单倍型，每个单倍型只有一个碱基的差异。运用DNASP软件计算得该群体的核苷酸多样性和平均核苷酸差异数分别为0．00036和0．14815。此结果提示镇海湾近江牡蛎群体遗传多样性已很低，很有必要从其他分布区引进近江牡蛎亲贝来扩大该种群的遗传多样性。 相似文献
9.
【目的】探究功能基因在海洋弧菌物种鉴定中的作用,为海洋弧菌多样性研究以及致病性弧菌的监测和防控提供参考。【方法】根据已有的弧菌分类学研究成果,阐述16S rRNA基因、管家基因和毒力基因等相关功能基因在海洋弧菌鉴定与分类研究中的应用进展。【结果】16S rRNA基因和功能基因可用于海洋弧菌的分类学及其快速检测鉴定的研究,对致病性弧菌的诊治具有重要的意义。【结论】功能基因和16S rRNA基因在海洋弧菌菌种鉴定中的广泛应用,提高了海洋弧菌种间分类的准确性,对海洋弧菌的多样性研究以及水产养殖和人类健康的风险评估有重要价值。探索新型功能基因在海洋弧菌分类学研究和快速鉴定中的应用有现实意义。 相似文献
10.
《广东海洋大学学报》2020,(2)
【目的】探讨矢耳石地标法在笛鲷种内及种间中的判别作用。【方法】利用2017年购自广西北海、海南文昌、广东阳江的87尾红鳍笛鲷(Lutjanus erythropterus)和76尾紫红笛鲷(Lutjanus argentimaculatus)成鱼的矢耳石样本,基于地标点法分析耳石的形态差异,运用判别分析检验耳石形态差异在2种笛鲷的种间和同种不同群体间的判别功效。【结果】位于听沟前中部交叉点的两个地标点10、11贡献较大,解释了耳石形态变异的64.88%～65.85%,表明两种笛鲷耳石形态的种间差异和种内群体差异主要集中于听沟前中部。基于耳石形态的地标点方法对2种笛鲷的种间判别成功率为97.4%和100.0%;红鳍笛鲷和紫红笛鲷的种内不同群体判别成功率分别为85.7%、83.3%、80.0%和94.1%、78.1%、81.5%。【结论】矢耳石地标点法可作为2种笛鲷种间和种内判别的有效工具。 相似文献
11.
This study is part of the endeavor to construct a comprehensive DNA barcoding database for common seaweeds in China. Identifications of red seaweeds, which have simple morphology and anatomy, are sometimes difficult solely depending on morphological characteristics. In recent years, DNA barcode technique has become a more and more effective tool to help solve some of the taxonomic difficulties. Some DNA markers such as COI (cytochrome oxidase subunitⅠ) are proposed as standardized DNA barcodes for all seaweed species. In this study, COI, UPA (universal plastid amplicon, domain V of 23S rRNA), and ITS (nuclear internal transcribed spacer) were employed to analyze common species of intertidal red seaweeds in Qingdao (119.3 -121 E, 35.35 -37.09 N). The applicability of using one or a few combined barcodes to identify red seaweed species was tested. The results indicated that COI is a sensitive marker at species level. However, not all the tested species gave PCR amplification products due to lack of the universal primers. The second barcode UPA had effective universal primers but needed to be tested for the effectiveness of resolving closely related species. More than one ITS sequence types were found in some species in this investigation, which might lead to confusion in further analysis. Therefore ITS sequence is not recommended as a universal barcode for seaweeds identification. 相似文献
12.
《中国海洋大学学报(英文版)》2021,(4)
The dried shellfish products with rich nutrients and low-calorie content are favorite food in China, especially in coastal areas. However, the species of dried shellfish products in the market are usually unknown, as the taxonomic features were removed during the production process. This study described the application of DNA barcoding technique to the identification of 100 dried shellfish(scallop, squid, octopus and cuttlefish) products in markets. Samples were authenticated by comparing mitochondrial cytochrome oxidase subunit I(COI) gene and 16 S ribosomal RNA(16 S rRNA) gene sequences with public reference taxonomic databases. The results showed that all the 100 products can be identified at species level. Sixty four scallop adductor products were processed using the bay scallop, Argopecten irradians, and one was from Portuguese oyster, Crassostrea angulata. All the 27 squid, 2 cuttlefish and 6 octopus products were produced by the Jumbo flying squid, Dosidicus gigas. The neighbour-joining tree is in agreement with the results of DNA barcoding analysis. The 64 scallop samples formed one A. irradians cluster, leaving Sca65 clustered with the reference oyster sequence C. angulata(MH997922). All the 35 cephalopod(squid, octopus and cuttlefish) samples formed a D. gigas cluster. This investigation revealed a low variety of dried shellfish products sold on the market, and highlighted the high rate of mislabeling and species substitution. Our present work provides a practical method for tracing and authenticating shellfish products. 相似文献
13.
《中国海洋大学学报(英文版)》2017,(2)
The effects of high pressure(HP) treatment on spoilage characteristic and shelf life extension of Pacific oysters(Crassostrea gigas) during refrigerated storage were studied.Results showed that HP treatment of 275 MPa for 3 min or 300 MPa for 2 min could achieve 100% full release of oyster adductor muscle,pressures higher than 350 MPa caused excessive release as the shells of oysters were broken,thus use of higher pressures should be cautious in oyster processing industry because of its adverse impact on the appearance of shells.HP treatment(300 MPa,2 min) was proper for the shucking of Pacific oyster(Crassostrea gigas) in China.This treatment caused no organoleptic disadvantage.Moreover,HP treatment resulted in obvious differences in biochemical spoilage indicators(p H,TVB-N and TBARS) changes and volatile compounds profile determined by electronic nose during storage.HP treatment(300 MPa,2 min) also led to a reduction of aerobic bacterial count(APC) by 1.27 log cycles.Furthermore,the APC values of oysters treated by HP were always lower than those of the control samples during storage.Based on the organoleptic,biochemical and microbiological indicators,shelf life of 6-8 d for control and 12 d for HP-treated oysters could be expected.HP treatment showed great potential in oyster processing and preservation. 相似文献
14.
DNA barcoding is a new method for biological taxonomy, offering the ability to identify species from fragments in any life-history stage. Pleuronichthys cornutus and P. japonicus are two morphologically similar species. Pleuronichthys japonicus has never been found previously in China. However, in this study, we identified both species using DNA barcoding (cytochrome c oxidase subunit I (COI)), the mtDNA control region and cytochrome b. The results reveal that: 1) intraspecific variation in the DNA barcode ... 相似文献
15.
A new identification method for five species of oysters in genus Crassostrea from China based on high-resolution melting analysis 总被引：2，自引：0，他引：2
The high phenotypic plasticity in the shell of oysters presents a challenge during taxonomic and phylogenetie studies of these economically important bivalves. However, because DNA can exhibit marked differences among morphologically similar species, DNA barcoding offers a potential means for oyster identification. We analyzed the complete sequences of the cytochrome oxidase subunit I （COI） of five common Crassostrea species in China （including Hong Kong oyster C. hongkongensis, Jinjiang oyster C. ariakensis, Portuguese oyster C. angulata, Kumamoto oyster C. sikamea, and Pacific oyster C. gigas） and screened for distinct fragments. Using these distinct fragments on a high-resolution melting analysis platform, we developed an identification method that does not rely on species-specific PCR or fragment length polymorphism and is efficient, reliable, and easy to visualize. Using a single pair of primers （Oyster- COI-1）, we were able to successfully distinguish among the five oyster species. This new method provides a simple and powerful tool for the identification of oyster species. 相似文献
16.
【目的】探明湛江市售鱼干中曲霉菌(Aspergillus spp.)分离株产AFB1的特性。【方法】采用马铃薯-葡萄糖琼脂(PDA)对鱼干中的曲霉菌进行分离纯化,根据菌落形态、显微形态观察和ITS序列分析对分离株进行鉴定,采用液相色谱-串联质谱(LC-MS/MS)法分析菌株的AFB1合成量。【结果】鱼干中曲霉菌的分离率高达47.66%,经形态学鉴定发现以黄曲霉(Aspergillus flavus)A03、烟曲霉(Aspergillus fumigatus)H32、杂色曲霉(Aspergillus versicolor)M23、欧式曲霉(Aspergillus europaeus)M11、棘孢曲霉(Aspergillus aculeatus)B21等为主,其中红笛鲷干(Lutjanus sanguinaus)中黄曲霉菌的含量高达1200 CFU/g。黄曲霉A03在红笛鲷干培养基中常温(27~30℃)培养21 d,产AFB1能力达到3.8 ng/mL。【结论】鱼干中存在产AFB1的黄曲霉菌,警示鱼干中存在AFB1污染的风险。 相似文献
17.
【目的】为开发用于害虫生物防治的细菌杀虫剂,对分离自红树林的一株沙雷氏菌进行鉴定并初步测试其杀虫活性。【方法】采用形态特征比较、细菌理化特性测定和基于16S rDNA序列构建系统发育树对从湛江红树林根际分离的菌株ZJ9进行综合鉴定,采用喂毒法和注射法进行杀虫活性测定。【结果】该菌为黏质沙雷氏菌Serratia marcescens,对草地贪夜蛾3龄幼虫7d校正死亡率为80.21%,且与苏云金芽孢杆菌和隐地杆菌无显著差异,菌体及菌液对大蜡螟老熟幼虫血腔注射24 h死亡率均达100%。【结论】菌株ZJ9为黏质沙雷氏菌,对草地贪夜蛾具有很好杀虫活性,可用作害虫的生物防治资源。 相似文献
18.
In this study, we integrated a DNA barcoding project with an ecological survey on intertidal polychaete communities and investigated
the utility of CO1 gene sequence as a DNA barcode for the classification of the intertidal polychaetes. Using 16S rDNA as
a complementary marker and combining morphological and ecological characterization, some of dominant and common polychaete
species from Chinese coasts were assessed for their taxonomic status. We obtained 22 haplotype gene sequences of 13 taxa,
including 10 CO1 sequences and 12 16S rDNA sequences. Based on intra- and inter-specific distances, we built phylogenetic
trees using the neighbor-joining method. Our study suggested that the mitochondrial CO1 gene was a valid DNA barcoding marker
for species identification in polychaetes, but other genes, such as 16S rDNA, could be used as a complementary genetic marker.
For more accurate species identification and effective testing of species hypothesis, DNA barcoding should be incorporated
with morphological, ecological, biogeographical, and phylogenetic information. The application of DNA barcoding and molecular
identification in the ecological survey on the intertidal polychaete communities demonstrated the feasibility of integrating
DNA taxonomy and ecology. 相似文献
19.
《中国海洋大学学报(英文版)》2021,(4)
Shell color is regarded as an economic trait in many breeding programs of bivalve mollusks, but the relationship between shell color and production traits remains controversial. In our breeding program of Crassostrea gigas, solid white, black, and orange shell lines were obtained, and second-generation(F2) and testcross families with segregating shell colors were constructed by crossing these three lines. These segregating families provided ideal samples for investigating the relationship between shell color and production traits in C. gigas. The growth and survival of 7-month-old oysters with different shell colors sampled within the same families were compared in seven F2 families and 13 testcrosses. In addition, the growth and survival of oysters from the three shell color lines were compared at 4 and 16 months of age. The growth and survival rates of the orange shell line were significantly lower than those of the white and black shell lines. However, no significant difference in growth between oysters with different shell colors was observed within segregating families, except the testcrosses produced by crossing orange-and white-shelled grandparents, and no significant difference in survival was observed in any family. Overall, no significant correlation was observed between shell color and production traits in C. gigas. These results suggest that shell color cannot be used as a marker to guide the selection of growth and survival traits. Thus shell color and production traits should be selected independently in oyster breeding programs. 相似文献
20.
The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae. 相似文献