海参核糖体基因间隔区2(ITS2)的克隆及序列分析

为探讨刺参科海参和海参科海参的系统进化关系,本研究通过PCR技术获取19种刺参科和海参科海参的ITS2序列,从NCBI上获取瓜参(C. salma)的ITS2序列。结果表明ITS2序列具有长度多态性,从318 bp (绿刺参)到591 bp (白尼参属)。海参属的ITS2序列长度多态性高,ITS2的GC含量从56.7%(糙海参)到70.6%(瓜参)。海参ITS2序列保守性不高,仅有48个保守位点,其余均为变异位点。基于ITS2的系统进化树结果显示进化树主要分成两支,一支包括海参科的4个属:海参属、白尼参属、辐肛参属和格皮氏海参属。辐肛参属和格皮氏海参属为姐妹关系,二者聚在一起后与白尼参属聚为一支,随后再与海参属聚在一起。白尼参属和辐肛参属为单系,海参属为复系。另一支为C. salma和刺参科。梅花参属与刺参属聚为一支后,再与仿刺参属聚在一起,3个属都是单系。在20种海参中,S. naso与B. argus的遗传距离最大(6.415)。刺参属中,S. monotuberculatus和S. horrens遗传距离最近(0.012),海参属中,糙海参与H. fuscopunctata的遗传距离最大(3.24)。本研究为从分子水平上研究海参科和刺参科之间的系统进化关系奠定了基础。     

重金属胁迫对刺参胚胎发育的影响简

正刺参(Apostichopus japonicus)又称仿刺参,隶属棘皮动物门海参纲楯手目刺参科,是一种营养和经济价值很高的海洋无脊椎动物[1,2]。主要分布于北起俄罗斯的海参崴,经日本海,朝鲜半岛南到我国黄、渤海以及江苏连云港外的西北太平洋沿岸[3]。近年来,随着人们对刺参高营养价值的认可,刺参的人工养殖规模不断扩     

海刺参i型溶菌酶基因的重组表达及抑菌谱分析

将本实验室已分离到的海刺参 (Stichopus japonicus) i型溶菌酶的基因(GenBank Accession No. EF036468)亚克隆进原核表达载体pET32a(+)中, 构建重组质粒pET32a(+)-SjLys, 转化至大肠杆菌BL21(DE3)pLysS。阳性克隆子经诱导表达、亲和纯化和透析复性, 得到了酶活力为19.2 U/mg的重组SjLys (rSjLys), 并对rSjLys进行了抑菌谱测定。结果表明, rSjLys对革兰氏阳性菌和阴性菌均有抑制作用, 尤其对常见的海洋致病菌副溶血弧菌和铜绿假单胞菌具有很强的抑菌活性。更为显著的结果是, rSjLys经100oC、40 min加热处理后, 失活的rSjLys对革兰氏阳性菌和阴性菌的抑菌能力高于rSjLys的9%~25%。上述结果表明, 海刺参溶菌酶是一种具有糖苷酶活性和非酶抑菌活性的特殊的i型溶菌酶, 且具有很广的抑菌活性, 在海刺参机体先天免疫系统中是一个重要的效应分子。     

刺参病害现状及其生物技术检测的研究进展

随着人工养殖规模的扩大,刺参养殖的病害问题已严重阻碍刺参养殖业的发展,如刺参腐皮综合症、化板病、病毒性疾病等.综述了近年来刺参的养殖工艺和模式,以及常见疾病的症状和防治手段.并针对刺参防御系统的特点,从免疫学和分子生物学方面做了相应的探讨和展望.     

刺参粉和江汉粉的结构功能和演化

剌参粉和江汉粉具有独特的萌发器和结构.依据结构形态,联系其出现的环境,分析了它们的结构和结构功能.从化石花粉与现生亲缘属花粉的差异,它们在上白垩统中出现的先后和形态结构的异、似情况,探讨了剌参粉的演化.概略地推测了刺参粉植物可能是从风媒植物到典型虫媒植物;由灌木到多年生草本植物;从生长于低山、丘陵到高山.描述了5新种.     

白花刺参中两个新三萜皂甙

从著名藏药白花刺参(Morina nepalensis var.alba Hand.-Mazz.)的水溶性部分分离到2个新三萜皂甙--刺参甙K(1)和刺参甙L(2),以及一个已知三萜皂甙mazusaponin I(3).应用波谱和化学方法,刺参甙K和刺参甙L的结构分别鉴定为3-O-α-L-arabinopyranosyl-(1→3)-β-D-xylopyranosyl siaresinolic acid(1)和3-O-β-D-glucopyranosyl-(1→3)-α-L-arabinopyranosyl siaresinolic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(2).     

白花刺参中两个新三萜皂甙

从著名藏药白花刺参（Morina nepalensis var.alba Hand.-Mazz)的水溶性部分分离到2个新三萜皂甙-刺参甙K（1）和刺参甙L（2）,以及一个已知三萜皂甙mazusaponinⅠ（3)。应用波谱和化学方法,刺参甙K和刺参甙L的结构分别鉴定为3-O-α-L-arabinopyranosyl-（1→）-β-D-xylopyranosyl siaresinolic acid(1)和3-O-β-D-glucopyranosyl-(1→3)-α-L-arabinopyranosyl siaresinolic acid28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(2)。     

刺参粉和江汉粉的结构功能和演化

剌参粉和江汉粉具有独特的萌发器和结构.依据结构形态,联系其出现的环境,分析了它们的结构和结构功能.从化石花粉与现生亲缘属花粉的差异,它们在上白垩统中出现的先后和形态结构的异、似情况,探讨了剌参粉的演化.概略地推测了刺参粉植物可能是从风媒植物到典型虫媒植物;由灌木到多年生草本植物;从生长于低山、丘陵到高山.描述了5新种.     

制作刺参标本的方法

刺参(Stichopus japonicus Selenka)又名沙噀,是一种名贵的食品,同时又是在教学上需要的一种实验和示教标本材料.它在我国北方沿岸浅海分布很广,从辽宁旅大沿海到山东半岛的南岸所产较多,尤以辽宁的旅大沿海和长海县各岛屿最多. 我们所制作的刺参标本,近年来多半是在烟台附近采到的,那里的产量很大,而且又有善于捕捞刺参的渔民.他们分春、秋两季进行捕捞:春季从4月初旬(清明前后)至5月初旬(立夏前后);捕捞秋参从10月中旬(霜降前后)至天寒水凉(大雪前后)不能作业时止.他们所捕捞的刺参是经过加工后供食品用的.我们在春季捕捞时期,(4月下旬至5月上旬)去进行采集的,制作过程大体分两个步骤,现在简单介绍如下:     

微生物方法降解刺参养殖池塘中氨态氮的研究进展

随着刺参养殖业的迅速发展,刺参池塘养殖暴露出诸多问题,如水体氨态氮含量高、病害严重、单位产量下降等.传统药物弊端较多,微生物方法成为降解水体中氨态氮含量和预防病害发生的新的有效途径.综合阐述了刺参池塘养殖过程中氨态氮对刺参的危害和微生物降解氨态氮的利弊,并介绍了水体中氨态氮高效降解菌株的筛选以及现有微生态制剂存在的问题,对微生物方法降解刺参养殖池塘氨态氮的前景进行了展望.     

Characterization of the gut bacterial community of the Japanese sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Comparative analysis of the composition and abundance of the gut bacterial community of the Japanese sea cucumber (trepang) Apostichopus japonicus and the sediment from its habitat was carried out. In spite of the presence of strictly aerobic bacteria in the sediments, gut microbiota of this holothurian was shown to be formed by facultative anaerobes from the environment. Irrespective of the geographical location of the habitat, the sea cucumber gut community had a constant composition, considered its normal microflora. Capacity of bacteria isolated from the Japanese sea cucumber for decomposition of starch, sodium alginate, chitin, chondroitin sulfate, Tweens, olive oil, casein, and gelatin was studied. Various degrees of enzymatic activity were shown for 33% of the studied bacterial strains, which indicates a considerable role of the trepang gut microflora in processing and assimilation of organic matter arriving with food.     

The Current Status of Populations of the Sea Cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> (Selenka, 1867) in the Far Eastern Marine Reserve

The spatial distribution of the sea cucumber Apostichopus japonicus (Selenka, 1867) in the Far Eastern Marine Reserve was studied using scuba. The abundance, size composition, population density, and distribution of A. japonicus were determined. The population density of this species at the mainland coast was higher than in waters around islands in the reserve. Seasonal movements of sea cucumber were observed: in autumn large individuals migrate to greater depths, but at the beginning of summer they move to shallow water. The average sizes of individuals and population density of sea cucumber in the reserve do not differ from those in unprotected areas of Peter the Great Bay. A comparative analysis of our own and literature data showed that the current estimates of sea cucumber population density in the reserve are 5?7 times lower than estimates for the period before the illegal fishing of sea cucumber began in Primorye.     

刺参海带饲料原料褐藻胶降解菌的分离与鉴定

从海带及刺参养殖环境中筛选有效降解褐藻胶,且对刺参无致病性的微生物,对海带饲料原料进行降解处理,以降低海带饲料中刺参难以消化的褐藻胶成分,显著提高饲料利用率,增加海带原料价值。以褐藻胶为唯一碳源选择培养基初筛;DNS法测定褐藻胶裂解酶酶活;16S r DNA测序及生理生化试验对菌种进行鉴定;高浓度腹腔攻毒试验考察筛选所得菌株对刺参的潜在致病性;分子排阻色谱及高效凝胶色谱法对微生物酶解褐藻胶的终产物进行分析。系统发育树分析表明,菌株WB1与Bacillus amyloliquifaciens有最高同源性,对刺参无潜在致病性;其褐藻胶裂解酶酶解褐藻胶的终产物主要为二糖和三糖,相对含量分别为74.1%和25.9%,平均分子量为516 Da。解淀粉芽胞杆菌WB1可作为一种安全的有益微生物用于刺参海带饲料原料中褐藻胶成分的降解。     

Screening and characterization of eicosapentaenoic acid-producing marine bacteria

To obtain eicosapentaenoic acid-producing bacteria, some 3,500 strains of bacteria were isolated from the intestine of marine animals from sea around the Korean peninsula. Forty two eicosapentaenoic acid-producing bacteria were obtained through screening with TLC and gas chromatography, and the strain with highest content of eicosapentaenoic acid (up to 33% of total fatty acid when grown at 5°C ) as the sole polyunsaturated fatty acid was identified as an Aeromonas sp. on the basis of physiological properties.     

A sulfated polysaccharide from the sarcoplasmic reticulum of sea cucumber smooth muscle is an endogenous inhibitor of the Ca(2+)-ATPase

Vesicles derived from the endoplasmic reticulum of sea cucumber smooth muscle retain a membrane bound Ca(2+)-ATPase that is able to transport Ca(2+) into the vesicles at the expense of ATP hydrolysis. In contrast with vesicles obtained from rabbit muscles, the activity of the Ca(2+)-dependent ATPase from sea cucumber is dependent on monovalent cations (K(+)>Na(+)>Li(+)). With the addition of highly sulfated polysaccharide to vesicle preparations from rabbit muscle, Ca(2+) uptake decreases sharply and becomes highly sensitive to monovalent cations, as observed with vesicles from sea cucumber muscle. These results led us to investigate the possible occurrence of a highly sulfated polysaccharide on vesicles from the endoplasmic reticulum of sea cucumber smooth muscle, acting as an "endogenous" Ca(2+)-ATPase inhibitor. In fact, vesicles derived from the invertebrate, but not from rabbit muscle, contain a highly sulfated polysaccharide. This compound inhibits Ca(2+) uptake in vesicles obtained from rabbit muscle and the inhibition is antagonized by monovalent cation. In addition, sea cucumber muscles contain high concentrations of another polysaccharide, which surrounds the muscle fibers, and was characterized as a fucosylated chondroitin sulfate. Possibly the occurrence of sulfated polysaccharides in the sea cucumber muscles is related with unique properties of the invertebrate body wall, which can rapidly and reversibly alter its mechanical properties, with change in length by more than 200%.     

Genetic structure of Japanese sea cucumbers (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis>) along the Sanriku coast supports the effect of earthquakes and related tsunamis

The Japanese common sea cucumber (Apostichopus japonicus) is a major marine product from Sanriku, Japan, but its populations were severely affected by the 2011 Tohoku earthquake, possibly decreasing its genetic diversity and increasing its extinction risk. In this study, we estimated the genetic structure and diversity of sea cucumbers from Touni and Yamada Bays of Sanriku over 4 years after the earthquake. The between-population genetic structure was estimated using two mitochondrial DNA regions (cytochrome c oxidase subunit I and 16S rDNA). Genetic differentiation (as measured by pairwise F_ST) was not significant between locations. Thus, even after the tsunami, gene flow and genetic diversity among the two sea cucumber populations were maintained. Our data also suggested that sea cucumbers in Sanriku experienced population expansion of about 0.20–0.24 million years ago, during the stable Mindel-Riss interglacial period. We conclude that A. japonicus populations in Sanriku could maintain their genetic structure throughout multiple disastrous tsunamis over the past several 1000 years. The high dispersal ability of planktonic larvae may enable the entrance of new recruits, thereby reducing risks associated with genetic structure and diversity changes stemming from mass die-offs in a given body of water from the past to the present.     

Screening of marine bacteria for fucoidan hydrolases

Twenty-five strains of epiphytic marine bacteria isolated from the brown algae Fucus evanescens and Chorda filum and fifty-three bacteria isolated from the sea cucumber Apostichopus japonicus were screened for fucoidanases using fucoidans prepared from the brown algae F. evanescens, Laminaria cichorioides, and L. japonica. Eighteen bacterial epiphytes and thirty-eight bacterial isolates from the sea cucumber were found to contain fucoidanases, which were able to hydrolyze either all of the fucoidans studied or some of them. Bacteria of the genera Cytophaga and Alteromonas/Pseudoalteromonas exhibited the highest fucoidanase activities, which, however, did not exceed the activity of fucoidanases from the already known sources.     

Auxin,biosynthesis of ethylene and sex expression in cucumber (Cucumis sativus)

Ethylene production, level of 1-aminocyclopropane-1-carboxylic acid (ACC) and activity of the ethylene forming enzyme (EFE) were higher in apices of gynoecious cucumber (Cucumis sativus cv. Alma) as compared to monoecious cucumber (C. sativus cv. Elem). Application of indole-3-acetic acid (IAA) enhanced ethylene and ACC production in both cultivars. The stimulatory effect of IAA was more pronounced in gynoecious apices. Induction of ethylene production and accumulation of ACC resulting from treatment with IAA were effectively blocked by aminoethoxyvinylglycine (AVG). Content of endogenous IAA, measured by an enzyme immunoassay, was lower in gynoecious cucumber as compared to monoecious one. Treatment of gynoecious plants with the antiauxins -(p-chlorophenoxy)isobutyric acid (PCIB) and -naphthaleneacetic acid (-NAA) did not inhibit female sex expression.It appears that although exogenous IAA enhances ACC and ethylene production, endogenous IAA might not have a major role in the control of sex expression in cucumber of the Beit-Alfa type.Prof. Rudich passed away in May 1986.     

The control of food mobilization in seeds of Cucumis sativus L.

An analysis of the in vitro activities of proteolytic enzymes from cotyledons of germinating cucumber seeds has been carried out and the effects of protein degradation products on such activities monitored. Aminopeptidase activity is substantially inhibited with either L-leucine or L-phenylalanine and trypsin activity with L-arginine. Aminopeptidase activity was also markedly reduced in the presence of individual di- and tripeptides. Of the peptides tested, however, only L-tryptophyl-L-phenylalanine inhibited the degradation of native cucumber seed protein by the endogenous cucumber seed protease(s) (autodigestive activity).Abbreviations TCA trichloroacetic acid - L-leuglygly L-leucylglycylglycine - L-pheglygly L-phenylalanylglycylglycine - L-phe-L-leu L-phenylalanyl-L-leucine - L-leu-L-phe L-leucyl-L-phenylalanine - L-tryp-L-phe L-tryptophyl-L-phenylalanine - LPA L-leucine-p nitroanilide - BAPNA -N-benzoyl-DL-arginine-p nitroanilide - ADA autodigestive activity