Preparation and characterization of agar,agarose,and agaropectin from the red alga Ahnfeltia plicata

Agar,agarose,and agaropectin were extracted from the red alga Ahnfeltia plicata,and their properties and structures were characterized.Agar was extracted by a comparatively low alkaline consumption of 1.2%.It exhibited a gel strength of 1 152.50±74.25 g/cm^2 and a sulfate content of 0.55%±0.08%.The yield of agar from A.plicata was 24.53%,which is higher than those of other agarophytes commonly used in China.Three kinds of the method were compared for the purification of agarose,and the physicochemical properties of agarose that was prepared under the optimal condition were identical to those of commercially available agarose.Furthermore,agaropectin was purified from A.plicata and characterized by GC,HPLC,UV-spectrum,and FI-IR to understand its composition and structure.It was the first time to comprehensively study the agar and its fractions from the red alga of A.plicata.This research provided an eco-friendly agar extraction method from A.plicata and revealed its potential application for the production of agar,agarose,and agaropectin.     

相手蟹属两种蟹类线粒体16SrRNA基因序列的比较

测定了相手蟹属（Sesarma）红螯相手蟹（S．haematocheir）和褶痕相手蟹（S．plicata）线粒体16SrRNA基因部分片段的序列,二者的序列长度相同,均为533bp,且A、T、G、C的含量相似,分别为198bp（37．1％）,206bp（38．6％）,84bp（15．8％）,45bp（8．4％）和200bp（37．5％）,205bp（38．5％）,81bp（15．2％）,47bp（8．8％）;二者的序列有49处差异,其中21个位点为转换、22个位点为颠换和6个缺失／插入位点。进一步对20种相手蟹属蟹类的长度为361bp的16SrRNA基因同源序列进行分析,发现AT的含量为78．6％～82．9％,明显高于GC的含量,且存有91个变异位点。从NJ树和遗传距离来看,在分布于中国的3种相手蟹中,无齿相手型（S．dehaani）和红螯相手蟹的亲缘关系最近（d=0．0151）,而它们与褶痕相手蟹的亲缘关系则较远（d=0．0924／0．09231。分布于中国的相手蟹和分布于北美的相手蟹之间存在着较大的遗传距离（差异）,表明它们之间有着较远的亲缘关系,互为单系起源。     

Bioaccumulation of cadmium in the ascidian Styela clava (Herdman 1881)

The processes of bioaccumulation and elimination of cadmium by tissues of Styela clava (Herdman 1881) were investigated based on a semi-static two-compartment model. The kinetic parameters (accumulation rate constant k ₁, elimination rate constant k ₂, bioconcentration factor BCF, biological half-life t _1/2 and maximum equilibrium concentration C _Amax) were obtained by non-linear regression. The results showed that S. clava accumulates cadmium from its aquatic environment. BCF decreased with increasing metal concentration in the water, and when the accumulation achieved balance, C _Amax correlated positively with metal concentrations in the water. Concentrations of cadmium in different tissues of S. clava were: gonad > digestive gland ≈ other parts > tunic. The t _1/2 values for cadmium were 15.54-50.40 days in the accumulation process and 11.53-24.55 days in the elimination phase. The high rate of accumulation and elimination of cadmium from the body reveals the potential of the organism to be used as a biomonitor of short-term cadmium fluctuation in marine systems.     

柄海鞘共附生细菌的分离培养与系统发育多样性研究

通过纯培养和16SrRNA基因序列的相似性比对,并构建系统发育树对威海海域柄海鞘共附生细菌的多样性进行了研究。用2216E培养基从柄海鞘样品中分离到78株细菌,通过形态学特征和TCBS培养基选择性筛选后,选取30株具有代表性的菌株进行了16SrRNA基因测序和基于16SrRNA基因序列的系统发育多样性分析。结果表明,30株菌株分别属于细菌域的4个系统发育类群(Actinobacteria,Bacteroidetes,Firmicutes,Gro-teobacteria)的11个科,11个属。根据16SrRNA基因序列,大多数菌株与其系统发育关系最密切的模式菌株之间存在一定的遗传差异(16SrRNA基因序列相似性为91.82%～98.93%)。其中菌株HQW7,HQYD1,HQWD4,HQE1和HQE2与相关模式菌株的16SrRNA基因最高相似度仅分别为91.82%,92.51%,92.99%,93.52%,93.87%,这5株菌可能代表新的分类单元。威海海域柄海鞘共附生细菌存在着较为丰富的物种多样性和系统发育多样性,并可能存在新的物种。     

柄海鞘Styela clava化学成分的研究(Ⅱ)

从采自青岛海域的柄海鞘 Styela clava中分离得到两个化合物 (1)和 (2 )。经过 IR、MS、1HNMR和 13 CNMR等现代波谱学方法 ,确定二者的结构分别为 1- O-十六烷酰 -甘油酯和胸腺嘧啶。它们均为首次从该海域海鞘中得到。     

柄海鞘Styela clava中的甾醇化合物的分离和鉴定

应用GC-MS技术从青岛海域产柄海鞘Styela clava中分离鉴定出5种甾醇化合物,分别是(22Z)-26,27-二降-麦角甾-5,22-二烯-3β-醇(1),胆甾-5,22-二稀-3β-醇(2),胆甾醇(3),胆甾烷醇(4),(22E)-麦角甾-5,22-二烯-3β-醇(5)。     

冠瘤海鞘同种群聚的化学诱因初步研究

研究了不同浓度的冠瘤海鞘成体被囊萃取物和幼体萃取物及不同的幼体密度对冠瘤海鞘(Styela canopus)幼体附着和变态的影响.结果表明,冠瘤海鞘被囊萃取物和幼体萃取物在5%和10%的浓度下均促进冠瘤海鞘幼体的附着和变态,但被囊萃取物的诱导效果明显强于幼体萃取物;幼体密度高于25只/cm3时,促进冠瘤海鞘幼体的附着和变态;幼体密度低于10只/cm3时,幼体的附着和变态不受密度影响.这些结果提示海区中冠瘤海鞘的同种群聚可能与其被囊或幼体内含诱导幼体附着和变态的化学因子有关.     

皱瘤海鞘神经递质和性类固醇激素含量与性腺发育成熟的关系

运用免疫组织化学和化学发光免疫分析方法研究了皱瘤海鞘(Styela plicata)神经递质多巴胺(DA)、5-羟色胺(5-HT)及其受体以及性类固醇激素含量与性腺发育成熟的关系.结果显示,DA和5-HT免疫活性定位在脑神经节中大小2种类型的神经细胞,DA免疫活性还定位在不同发育时期神经腺上皮细胞.5-HT受体免疫活性存在于脑神经节小型神经细胞和大生长期神经腺上皮细胞.在小生长期和大生长期性腺中均检测到DA、5-HT及其受体,提示DA和5-HT可能参与调节皱瘤海鞘的性腺发育.皱瘤海鞘为雌雄同体,化学发光免疫分析证实,海鞘性腺能够合成和分泌性类固醇激素,大生长期性腺中雄激素、孕激素和雌激素的含量分别为25.8±0.0、38.9±0.2和14.3±0.6pg/mg,提示性腺既是生殖组织也是内分泌组织.上述发现为证明尾索类海鞘可能像头索类文昌鱼一样,具有原始的生殖内分泌调控轴(脑神经节-神经腺-性腺)提供科学依据.     

厦门皱瘤海鞘性腺发育的组织学研究

皱瘤海鞘雌性同体。性腺组织学观察结果表明，在厦门皱瘤海鞘卵巢和精巢发育和成熟可分为５人时期，在同一人体中，卵巢和精巢的发育不同步，因而不发生自体受精，另外，根据性腺发育的季节性观察，皱瘤海鞘常年都可繁殖，但生殖旺季在６月。     

冠瘤海鞘幼体附着和变态的离子控制

研究了人工海水中三种金属离子K+,Mg2+和Ca2+分别在不同浓度下对冠瘤海鞘(Styelaca nopusSavigny)幼体附着和变态的影响,同时采用K+通道阻滞剂TEA检验其可兴奋细胞膜上的K+通道特性,对冠瘤海鞘幼体附着和变态的离子控制机制进行初步探讨,结果表明:K+的浓度为20~40mmol/dm3时诱导幼体附着和变态的效果最佳,低于45 mmol/dm3和高于80mmol/dm3的K+浓度都表现出抑制效果;Mg2+浓度在0~32 mmol/dm3时显著促进幼体附着和变态,达88mmol/dm3时起抑制作用;在人工海水中无论添加或减少Ca2+,均有效抑制幼体变态.TEA不能阻抑增加K+的诱导作用,这表明冠瘤海鞘幼体可兴奋细胞膜上的K+通道对外部TEA不敏感.另外,人工海水中缺乏K+或Ca2+或K+浓度达80mmol/dm3都会对冠瘤海鞘幼体产生毒害作用.