长心卡帕藻的形态结构观察与分子鉴定

通过形态构造观察和分子生物学相结合的方法,对栽培于我国海南的长心卡帕藻(Kap-paphycus alvarezii)株系XC-03(陵水)和麒麟菜族未知种株系LD-02(三亚)和LD-06(三亚)进行了鉴定,并与细齿麒麟菜(Eucheuma denticulatum)株系TSNY-05进行比较分析,结果表明:长心卡帕藻株系XC-03与麒麟菜族未知种株系LD-02和LD-06之间在分枝形状、长短、分布以及突起数量等藻体外部形态特征方面具有明显差异;它们的表皮、皮层和髓部细胞大小、皮层细胞层数和胶质层厚度等显微结构特征亦不相同,但分子鉴定表明该3个株系为同一个种,即长心卡帕藻。它们的ITS序列相似度为99.69%~99.85%,株系XC-03与LD-02之间仅有1个碱基转换和1个碱基颠换,株系XC-03与LD-06和株系LD-02与LD-06之间均分别仅有1个碱基转换和1个碱基颠换。长心卡帕藻与细齿麒麟菜的ITS序列相似度只有41.28%,两者差异较大。用RAPD,ISSR和ITS标记分析表明长心卡帕藻3个株系的两两间遗传距离分别介于0.122 9~0.153 5,0.068 8~0.112 4和0.002~0.005。用neighbor-joining和UPGMA聚类分析皆显示长心卡帕藻株系XC-03与LD-06的亲缘关系较近,与株系LD-02相对较远,而细齿麒麟菜单独列为一支。该研究为长心卡帕藻的种质鉴定和遗传育种研究提供了依据。     

4株形态不同的长心卡帕藻（Kappaphycus alvarezii）的初步ISSR分析

为了研究高产胶量品系的遗传学特征,建立了长心卡帕藻(Kappaphycus alvarezii)物种的ISSR分析方法.用ISSR技术对4株同一来源,形态各异的长心卡帕藻进行了遗传学分析,与11条ISSR引物共扩增得到60务带,其中1条带具有多态性并推测与稳定的经济性状连锁.说明ISSR技术可应用于长心卡帕藻品系筛选,结合其他手段,为分子标记辅助育种提供了可能.     

盐度降低对长心卡帕藻原初光化学反应的影响

为探讨大量降雨如何导致热带产胶海藻长心卡帕藻死亡,通过快速叶绿素荧光诱导动力学曲线分析,研究了盐度降低对热带产胶海藻长心卡帕藻光化学反应的影响,结果表明,盐度降低对PSⅡ反应中心、供体和受体侧的光化学过程产生不同程度的影响。盐度为28时,虽然反应中心开放程度下降11%,受体侧PQ库减小,QA被还原次数下降,反应中心吸收光能出现下降同时热耗散能量出现增加,但是单位面积反应中心RC/CS0,RC/CSm数量显著增加,单位反应中心(或面积)用于电子传递能量都增加,因此最大光能转化效率未降低,光合性能指数甚至增加。上述结果意味着虽然盐度下降到28改变了长心卡帕藻光化学过程,但并没有严重影响到光合作用效率,属于可逆胁变。盐度降低到22时,长心卡帕藻反应中心吸收、捕获和用于电子传递的能量均下降,但是热耗散能上升,反应中心受体侧PQ库和QA被还原次数均显著下降,向反应中心PSⅠ传递的电子受阻,即盐度低于22时长心卡帕藻未有效转化所吸收光能,过剩激发能对光合机构产生损伤,藻体光合开始发生不可逆胁变。当盐度为18时,该藻PSⅡ开放的反应中心数量大大减少,单位面积内电子传递能量也减少,其最大光能转化效率和光合性能指数均明显下降,因此盐度低于18时藻体不可避免地死亡。     

海带愈伤组织的高效率诱导

于１９９４年１月和１１月在青岛太平角和荣成海带育苗场分别采集海带成熟孢子体和幼孢子体,采用培养基诱导方法,以４种培养基诱导海带愈伤组织的效果,成熟海带孢子体的不同部位在ＭＳ固体增富２基中形成愈 组织的能力进行了比较;研究了添另不同成分的ＭＳ固体增富２基对海带愈伤组织的影响,并比较了４种除菌方法的效果。结果表明,ＰＥＳＩ固体２基和ＭＳ固体在是诱导海带愈伤组织的理想培养基,其诱导率分别为７５．５％和６     

长心卡帕藻应对海洋酸化和升温的环境适应性研究

长心卡帕藻(Kappaphycusalvarezii)是主要生长于热带海域的大型经济红藻,在营养物生物提取方面具有重要的经济和生态价值,作为提取κ-卡拉胶的重要原料,广泛应用于食品和医学等多种行业。由于人类活动导致大气中CO₂浓度持续升高,引发海洋表层海水逐渐酸化和升温,对大型海藻产生耦合效应。因此,本文设置了两种CO₂浓度梯度(450×10^–6和1 200×10^–6)和三种温度梯度(26℃、29℃和32℃),从光合作用、细胞代谢产物、碳氮积累和酶活性方面探讨长心卡帕藻响应海洋酸化和海水升温的环境适应性变化。结果表明, CO₂浓度与温度变化对长心卡帕藻的F_v/F_m影响显著(P<0.05),在两种CO₂浓度条件下,Fv/Fm均随温度升高而增加;酸化条件与非酸化条件下可溶性糖浓度变化趋势相反,海水升温对游离氨基酸浓度影响极显著(P<0.01),在两种CO₂水平下,升温均会降低藻体游离...     

长心卡帕藻（Kappaphycus alvarezii）表达序列标记分析

分别采用BlastX和Blast2go软件对453条非冗余的长心卡帕藻EST序列进行匹配分析和GO注释。结果表明,有281条序列与蛋白数据库中序列匹配,有132条序列被归属为3个子本体：分子功能、生物学过程和细胞组成。同时筛选到24个与逆境适应相关的基因,包括应对胁迫反应和抗氧化反应的基因。采用GCUA软件,进行长心卡...     

光照强度和盐度对长心卡帕藻生长的影响

在室内采用静置培养法研究了光照强度和盐度对红色和绿色品系长心卡帕藻Kappaphycus alvarezii生长的影响,主要测定了不同光照强度(30、65、110、150和200μmol·(m2·s)-1)和不同盐度(23、28、32、35和41)对这2种藻的生长率、成活率、色素含量及其吸收光谱的影响.红色品系长心卡帕藻适宜的光照强度范围为30-65μmol·(m2·s)-1,光照强度为30μmol·(m2·s)-1时培养,其成活率和日生长率都最高,分别达100%和1.25%,强光下培养生长率和成活率明显下降;红色品系长心卡帕藻适宜的盐度范围为28-32,最适为32.绿色品系长心卡帕藻适宜150-200μmol·(m2·s)-1的强光,150μmol·(m2·s)-1光照强度下培养日生长率最高,达1.05%,各种光照强度下培养其成活率变化不大;绿色品系长心卡帕藻适宜的盐度范围为23-32,最适为23.不同光照强度和盐度条件下培养,长心卡帕藻体内的色素含量会发生变化,而色素含量与其生长率有或没有明显的相关关系.不同光照强度、盐度条件下培养,藻体内的色素吸收光谱特性不变.以上研究结果对我国长心卡帕藻的栽培生产具有一定的指导意义.     

长心卡帕藻原生质体分离、培养研究

In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30°C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL-1, 65.20×104 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20°C, light intensity of 1 500–2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated:(1) callus-like cell mass and regenerated plantlet occurred on protoplast;(2) young shoots and calluslike cell mass occurred in tissue blocks after enzymolysis.     

篮子鱼内脏粗提液制备长心卡帕藻和细齿麒麟菜原生质体的初步研究

于2011年9月至2013年1月,在海南陵水热带海藻实验基地,利用篮子鱼(Siganus fuscessens)内脏研磨制备消化酶,开展了酶解制备大型产胶红藻长心卡帕藻(Kappaphycus alvarezii)和细齿麒麟菜(Eucheuma denticulatum)的原生质体研究,观察了不同酶解条件对两种产胶海藻原生质体产量和产率的影响。结果表明,细齿麒麟菜原生质体得率高于长心卡帕藻;在藻泥与酶浓度配比中,长心卡帕藻或细齿麒麟菜藻泥越多,所制备出的原生质体产量越高,而其制备效率却越低;在温度(20~30℃)梯度中,有最高原生质体产量和效率的酶解温度为25℃;在pH 6.0~8.0的实验酶解梯度中,酶解pH 6.0时的酶解效果最好;在12~48 h实验酶解时间梯度中,酶解时间为48 h时原生质体产量和得率最高;酶解时间越长,长心卡帕藻或细齿麒麟菜原生质体制备效率越高。为此建议在25℃和pH6.0条件下,适当增加海藻藻泥用量和延长酶解时间,以更有效获得原生质体。     

温度、氮浓度和氮磷比对长心卡帕藻(Kappaphycus alvarezii)吸收氮速率的影响

分别在室内培养箱、海滨室外跑道池和不同自然海区,通过一次性和半连续添加营养、以及检测海区水质和藻体生长的方法,研究了不同氮浓度、温度和氮磷比条件下,长心卡帕藻氮吸收速率的变化和氮吸收速率随时间变化,以及栽培该藻的环境生态贡献。小型实验、中试放大和海区规模栽培结果表明:(1)在10—50μmol/L范围内,该藻吸收氮速率随氮浓度增加而增大;(2)当氮浓度一定时,氮磷比在1—50范围内对该藻吸收氮速率没有产生显著影响(P>0.05);(3)温度对该藻吸收氮速率有显著影响(P<0.05),其中温度在28℃时氮的吸收速率最高;(4)尽管一次性添加营养实验中长心卡帕藻吸收氮速率随时间变化表现出先快后慢的趋势,但是进一步的半连续添加营养实验证实,导致吸收速率下降系底物氮浓度限制,而不是藻本身吸收能力下降,结果还显示卡帕藻具有连续吸收同化无机氮能力;在自然光温度变化和不受底物浓度限制条件下,该藻藻体去除无机氮效率最大维持在0.3μmol/(gFW·h);(5)海南陵水黎安海湾水质数据显示,栽培该藻去除海水富营养化和净化水质作用显著,其去除海水富营养化的年贡献为33t氮素。     

Assessment of introduced Kappaphycus (Solieriaceae, Rhodophyta) species relationships in China with molecular markers

Due to morphological plasticity and paucity of diagnostic morphological characters, the taxonomy of Kappaphycus gets more and more confused with the expanding of commercial cultivation. In this study, the phylogenetic relationship of 13 strains of introduced Kappaphycus species in China was defined using DNA molecular markers, such as 18S rDNA, rbcL and cox2-cox3 spacer region. The resolutions obtained by three different molecular markers were compared: both cox2-cox3 spacer region and rbcL sequences are eligible in interspecies identification of Kappaphycus, whereas cox2-cox3 spacer region is more variable than rbcL sequence. There is several basepairs’ discrepancy among 18S rDNA sequences, while it is 100% identical among both cox2-cox3 spacer region and rbcL sequences of the ten strains of K. alvarezii. We suppose that 18S rDNA sequence can provide more information in biogeography study of Kappaphycus than other two DNA sequences.     

泥蚶、缢蛏和僧帽牡蛎呼吸与排泄的周年变化

于2005年5月至2006年3月对浙江沿岸重要的3种经济贝类泥蚶、缢蛏和僧帽牡蛎的呼吸率与排泄率进行了周年研究。结果表明:泥蚶、缢蛏和僧帽牡蛎耗氧率的变化范围分别为0.11~5.05 mg/（g·h）、0.77~5.97 mg/（g·h）和0.33~5.47 mg/（g·h）,排氨率的变化范围分别为21.51~1 078.63 μg/（g·h）,26.97~990.73 μg/（g·h）和32.12~1 378.67 μg/（g·h）;耗氧率与排氨率9月份最高,1月份最低。经单因素方差分析发现,3种实验贝类月际间的耗氧率与排氨率均存在极显著差异（p<0.01）,经配对T检验发现,缢蛏（SC）和僧帽牡蛎（SCA）之间的耗氧率存在着显著差异T_（SC-SCA）=3.184,p=0.024）,泥蚶（TG）与缢蛏、僧帽牡蛎的耗氧率之间都不存在差异（T_（TG-SC）=1.887,p=0.118;T_（TG-SCA）=0.246,p=0.815）,泥蚶、缢蛏和僧帽牡蛎之间的排氨率的差异均不显著（T_（TG-SC）=0.977,p=0.373;T_（TG-SCA）=2.369,p=0.064;T_（SC-SCA）=1.002,p=0.362）。3种贝类的耗氧率与排氨率均随温度的升高而升高,经回归分析发现,耗氧率和排氨率与温度的变化均呈较显著的幂指数性相关lnY=lna+blnT（Y=aebT）。泥蚶呼吸排泄O:N变化范围为4.69~28.55,缢蛏和僧帽牡蛎的O:N变化范围分别为:4.68~37.89和3.04~10.27。     

Spatial spring distribution of the copepod Eurytemora affinis (Copepoda,Calanoida) in a restoring estuary,the Scheldt (Belgium)

The spatial spring distribution of Eurytemora affinis (adults and C5) in the Scheldt estuary (Belgium) brackish and freshwater reaches was studied in between 1996 and 2007. The bulk of the E. affinis population being generally situated in the brackish water reach (salinity > 0.5); we studied which environmental factors are responsible for its recent sporadic occurrence in the freshwater estuarine reach. Using PLS analysis, it is shown that its presence upstream is limited by a sufficient oxygen concentration (>4 mg l⁻¹) that is associated with temperature. Not only are the environmental conditions in the upstream zone important, but also the frequent presence of an O₂ minimum zone in the mid-estuary (O₂ min < 1.3 mg l⁻¹) seems to block the movement of the downstream E. affinis population in an upstream direction. Occasionally, the bulk of the population is however situated upstream. During these periods, high E. affinis abundance was also observed in the Durme tributary. Our findings suggest the possibility to use E. affinis as an “indicator” species of water quality, but also lead us to stress the necessity to consider conditions over the entire estuary when studying restoration effects, not exclusively in the zone of interest.     

Distribution of amphipods Ischyrocerus on the red king crab, Paralithodes camtschaticus: Possible interactions with the host in the Barents Sea

The determination of costs and benefits experienced by crustaceans as a result of occupation by their symbionts has received increased attention from marine ecologists. However, the interactions between some important species and their associates remain unclear. We examined the distribution of amphipods in the genus Ischyrocerus on the red king crab Paralithodes camtschaticus, a commercially important species, in two areas of the Barents Sea. Ischyrocerus commensalis was found on 30.5% of crabs in Dalnezelenetskaya Bay (DZB) with the mean number per crab being 55.1, in Dolgaya Bay (DLB) these rates were 28.6% with 19.3 specimens per crab. Sympatric species Ischyrocerus anguipes was found on 13.5% of crabs in DZB with a mean of 7.3 individuals per host, in DLB it had much lower occurrence (1.3% and 1.5 specimens per host). There were no significant differences between proportions of male and female crabs infested by amphipods in both areas examined. Prevalence of amphipods was similar among years examined except for I. commensalis on small crabs (carapace length CL <90 mm) and I. anguipes on large crabs (CL >90 mm) in DZB. We found that I. commensalis and I. anguipes are not egg predators of P. camtschaticus in the Barents Sea, at least in summer. High numbers of I. commensalis occur in crab gills, and both mean intensity of the amphipods and their empty tubes increased with crab size. In the gills, I. commensalis predominated in the section nearest the mouth parts. Possible negative impacts for the hosts due to gills infestation are discussed. In contrast, I. anguipes were predominately found on the carapace and limbs of crabs and appears to be a less specific symbiont of P. camtschaticus. Both amphipod species seem to be commensals, however possible negative impacts for the host could not be excluded.     

Negative effects of blue mussel (Mytilus edulis) presence in eelgrass (Zostera marina) beds in Flensborg fjord,Denmark

The effect of blue mussel (Mytilus edulis) presence in eelgrass (Zostera marina) beds was studied from June 2004 to July 2005 in Flensborg fjord, Denmark. The field experiments were conducted at two stations, one with only Z. marina (Eelgrass station) present and one where M. edulis were present in the Z. marina beds (Mixed station). Zostera marina parameters were measured (growth of leaves, shoot density, leaf length, and nutrient content) in combination with epiphyte cover and sediment parameters (sulphate reduction rates, sediment nutrient fluxes, organic content, C, N and P content) to examine possible positive and negative effects of the mussels on eelgrass performance. The fluxes of ammonium from the sediments were stimulated at all sampling dates at the Mixed station, and possibly stimulated epiphyte growth at this station. Further ¹⁵N signals in epiphytes from the Mixed station suggested that excretion products from the mussels were important nitrogen sources at this station. Sulphate reduction rates were enhanced at the Mixed station and also sediment sulphide concentrations increased under mussel influence, which may have resulted in sulphide toxicity and decreased growth of Z. marina at this station. The study indicates that for Z. marina beds in Flensborg Fjord the effects of M. edulis in seagrass beds are primarily negative, and raises the question whether this leads to negative effects on the stability and expansion of Z. marina beds.