Applications of three DNA barcodes in assorting intertidal red macroalgal flora in Qingdao, China

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.     

湛江海域6种常见经济海藻的营养成分分析

分析采自湛江海域的粗枝软骨藻Chondria crassicaulis、日本沙菜Hypnea japonica、鱼栖菜Acanthophora sp、芋根江蓠Gracilaria blodgettii、平卧松藻Codium repens、网地藻Dictyota dichotoma等6种经济海藻的营养成分。结果表明:碳水化合物是这些海藻的主要营养成分,占藻体干重的42.81%～66.11%,平均为55.93%;粗蛋白含量6.63%～11.13%;粗脂肪含量最低,除网地藻D.dichotoma(2.85%)外,其余5种皆不足藻体干重的1%;粗纤维和灰分含量在种间差别较大;矿物质含量丰富,其中磷、铁和锌分别在平卧松藻C.repens、网地藻D.dichotoma和鱼栖菜Acanthophora中占藻体干重的0.093%、0.13%和0.0055%;氨基酸含量较高,其中Thr、Val、Ile、Leu、Phe几种必需氨基酸的含量均高于FAO标准;不少种类中汞、镉、铅、砷等含量超出我国食品卫生标准。海藻体内的营养成分含量因海藻种类和生长海域的不同而有差异。     

EST-SSR标记的开发及其在南黄海浒苔研究中的应用

Ulva species can grow rapidly in nutrient-rich habitats causing green tides and marine fouling. A more complete understanding of the reasons behind these outbreaks is urgently required. Accordingly, this study attempts to use microsatellite markers based expressed sequence tag（EST） to analyze the genetic variation of several Ulva prolifera populations in the South Yellow Sea of China. Two hundred and thirty-eight SSRs were identified from 8 179 unique ESTs（6 203 newly sequenced and 1 976 downloaded from NCBI database） and 37 primer pairs were successfully designed according to the ESTs; 11 pairs were selected to detect the genetic diversity and relationship of 69 attached U. prolifera samples and 13 free-floating samples collected from coastal and off-coast areas of the South Yellow Sea. The results of cross-species transferability showed that six of the 11 EST-SSR primers could give good amplification in other five Ulva species and the average allele number was 4.67. Genetic variation analysis indicated that all 82 U. prolifera samples were clearly divided and most samples collected from the same site clustered together as a group in the dendrogram tree produced by unweighted pair-group mean analysis（UPGMA） method and the cluster results showed some consistency with the geographical origins. In addition, 13 free-floating samples（except HT-001-2） were grouped as a single clade separated from the attached samples.     

青岛岩礁海藻附植动物的区系组成和季节动态

于2007年11月～2008年10月对青岛太平角岩石潮间带鼠尾藻附植动物进行了连续12个月的逐月采样调查,并根据蜈蚣藻、角叉菜、扇形叉枝藻、海蒿子和叉节藻等海藻生长期的不同而在不同月份对其分别进行了采样,研究了附植动物的类群组成、丰度、生物量及其季节动态。所有海藻样品共鉴定出附植动物16个类群。鼠尾藻附植动物的年平均丰度为606ind/gdwtalgae,优势类群为线虫和桡足类,其次是腹足类和多毛类。鼠尾藻附植动物的平均丰度最高值出现在4月,最低值出现在7月。鼠尾藻附植动物的年平均生物量为282×103μg/g dwt algae,最高值出现在6月,最低值为2月。其他海藻附植动物的类群数、丰度、生物量均低于鼠尾藻。海藻附植动物的优势类群及其丰度和生物量在不同月份和不同海藻之间均显著不同。相对于海水理化因子季节变化的影响,不同海藻生长型形态的复杂性及同一种海藻随生长周期而发生的形态变化对附植动物的区系组成和季节动态的影响可能占居更主要地位。     

温度和盐度对几种大型海藻生长率和NH4-N吸收的影响

温度和盐度对细基江蓠繁枝变型(Gracilaria tenuistipitata var.liui)、孔石莼(Ulva pertusa)和蜈蚣藻(Grateloupia filicina)生长率及细基江蓠繁枝变型NH₄-N吸收的影响进行了实验研究.结果表明,温度和盐度对上述3种海藻生长率及NH₄-N吸收均有显著影响.3种海藻的生长适宜温度和盐度范围分别为细基江蓠繁枝变型20～30℃,15～30,孔石莼15～25℃,15～40,蜈蚣藻20～25℃,25～35;最大日特定生长率(SGR)分别为8.66%,12.28%,2.24%;N饥饿细基江蓠繁枝变型对NH₄-N的吸收存在短期的快吸收.细基江蓠繁枝变型NH₄-N吸收的最适温度和盐度范围分别为15～25℃,10～25.生长率及NH₄-N吸收速率与温度和盐度之间分别存在以下关系:SGR_G.t.=0.873(S-5.487)(T-9.007)e(-0.0708S-0.0745T);SGR_U.p.=0.222(S-2.665)e-0.047S(T-9.98)e-0.00057(T-9.98)2.7;SGR_G.f.=2.6×10-4(S-10.856)(T-11.704)e0.156T-3.36×10-6STe0.219T;V_upt=11.812(S-4.081)(T-9.221)e-0.1S-0.148T.     

中国沿海9种红藻hsp70基因序列及系统进化分析

本研究自山东青岛、浙江象山和江苏南通采集共9种红藻样品, 隶属于2纲、5目、6科、8属(据NCBI), 克隆各红藻hsp70 基因, 并对所获序列进行分析。利用特异性引物P1/P3扩增, 得到的目的条带约630 bp, 分析所推导的氨基酸序列发现:所获得片段均位于HSP70的ATPase结构域附近。9种红藻hsp70 序列之间的遗传距离在0.078～0.319之间, 序列相似度在73%～92%之间, 其保守性略低于HSP70蛋白;基因对A或T结尾的密码子表现出很高的偏好性, CGC与TGG这两种密码子在这9种红藻HSP70氨基酸密码子中未出现。上述表明hsp70 及HSP70密码子偏好性可应用于红藻分子系统学研究。基于多物种HSP70构建的进化树可见, Cyanidium caldarium与Cyanidioschyzon merolae strain 10D两种原始红藻的起源早于其他红藻, 紫菜次之, 本研究中9种红藻系统发生符合NCBI的描述。在真菌、藻类和植物中, 营养方式的差异可能是造成HSP70进化树分化的基本原因, 而相同形态类型的物种中, 环境适应是抗逆能力强、遗传结构稳定的物种生物分子进化的重要因素。     

Temperature and light tolerance of representative brown, green and red algae in tumble culture revealed by chlorophyll fluorescence measurements

Laminaria japonica, Undaria pinnatifida, Ulva lactuca, Grateloupia turuturu and Palmaria palmata are suitable species that fit the requirements of a seaweed-animal integrated aquaculture system in terms of their viable biomass, rapid growth and promising nutrient uptake rates. In this investigation, the responses of the optimal chlorophyll fluorescence yield of the five algal species in tumble culture were assessed at a temperature range of 10 - 30℃. The results revealed that Ulva lactuca was the most resistant species to high temperature, withstanding 30℃ for 4 h without apparent decline in the optimal chlorophyll fluorescence yield . While the arctic alga Palmaria palmata was the most vulnerable one, showing significant decline in the optimal chlorophyll fluorescence yield at 25℃ for 2 h. The cold-water species Laminaria japonica, however, demonstrated strong ability to cope with higher temperature （24 -26℃ ） for shorter time （within 24 h） without significant decline in the optimal chlorophyll fluorescence yield . Grateloupia turuturu showed a general decrease in the optimal chlorophyll fluorescence yield with the rising temperature from 23 to 30℃ , similar to the temperate kelp Undaria pinnatifida. Changes of chlorophyll fluorescence yields of these algae were characterized differently indicating the existence of species-unique strategy to cope with high light. Measurements of the optimal chlorophyll fluorescence yield after short exposure to direct solar irradiance revealed how long these exposures could be without significant photoinhibition or with promising recovery in photosynthetic activities. Seasonal pattern of alternation of algal species in tank culture in the Northern Hemisphere at the latitude of 36°N was proposed according to these basic measurements.     

龙须菜在鱼藻混养系统中的生态功能

近年来，随着海水养殖产业的迅速发展，养殖规模的不断扩大，养殖区的自身污染问题也日益显露，以人工投饵和网箱养殖方式为主的浅海鱼类养殖，残饵和鱼体的代谢产物往往导致水体的富营养化和底质的有机污染，既影响周围环境，又不利于鱼类的生长和健康，长远意义上还会制约整个鱼类养殖业的持续发展。怎样减轻海水养殖造成的自身污染和对生态环境的破坏，已成为人们日益关注的问题，国内外学者已提出多种解决这一问题的途径，其中一条重要途径是通过优化养殖结构，实施综合养殖，发展生态养殖技术(杨圣云等，1996；杨红生，2000；阎希柱等，2000)。目前国际上，主要的综合养殖方式包括鱼藻混养、鱼贝混养、鱼类与底栖沉积食性动物混养等。近年来的研究又发现，在富含营养盐的鱼类养殖水体中养殖大型海藻，不仅能显著降低养殖废水中的溶解营养盐，而且能提高海藻的产量(Troell，1999)，鱼藻的相互作用重新激起了人们的兴趣，但大型海藻与鱼类混养的基础理论和关键技术还刚刚起步，养殖容量和养殖配比等方面尚亟待研究。 江蓠属龙须菜(Gracilaria lemaneiformis)是一种大型红藻，具有适温范围较广(12-23℃)、生长快、适应环境能力强和经济价值高等优点，是改善生态环境、提高经济效益的理想品种。本项研究利用室内海水养殖系统，将龙须菜和我国北方海域的代表性养殖鱼类——黑鲪(Sebastodes fuscescens)进行混养，设计不同的养殖密度，测定若干水环境和生物因子指标，旨在查明江蓠在鱼类养殖系统中的生态效应，初步寻求合理的养殖配比，为今后大规模运用大型海藻改善养殖区环境提供科学依据。     

Seasonal variation in biomass and species composition of seaweeds stranded along Port Okha,northwest coast of India

Port Okha coast, which is known for its luxuriant growth of a diverse assemblage of seaweeds on Saurashtra coast, is found to have abundant quantities of seaweeds being drifted and washed ashore every year. Studies conducted for quantifying the stranded seaweeds from May 2004 to April 2005 showed an average biomass value of 3.10 kg fresh wt/m²/month with maximum being 6.60 kg fresh wt/m² in April. The stranded weeds constituted a total of 62 species during the entire study period. Of this, Rhodophyta ranked high with 26 species followed by Chlorophyta with 22 species and Phaeophyta with 14 species. The stranded seaweeds that were washed ashore provide valuable floristic information about the intertidal and near shore sub-tidal algae of the respective regions. Although natural senescence of seaweeds is one of the major factors, strong currents primarily forced by tides, also contribute to the uprooting and subsequent drifting of seaweeds on to the beach. This ultimately causes changes in floristic features of the existing algal beds.     

Biofilm: A crucial factor affecting the settlement of seaweed on intertidal rocky surfaces

We hypothesized that the presence of biofilm accelerated the settlement of dominant seaweed species and maintained high levels of species richness by regulating the blooming of particular species on an intertidal rocky shore. The coverage and species richness of macroalgae on sterile and cleared substrates (225 cm²) were measured to investigate the effects of biofilm on the settlement of macroalgae in intertidal zones at Noryang, Songmoon, and Sangnam in Gawngyang Bay on the southern coast of the Korean peninsula. Green algae coverage on cleared substrates was significantly higher than that on sterile substrates at both Noryang and Songmoon during the study period. This suggests that the presence of biofilm enhances the settlement of green algae by providing various habitat structures and, consequently, may lead to serious ‘green tide’ events. However, the coverage of algae other than green algae and algal species richness on cleared substrates remained high at Sangnam during the experimental period. Biofilm facilitated the settlement of macroalgae and inhibited the blooming of specific algae by inducing inter-specific space competition. Therefore, biofilm plays an important role on seaweed assemblages on intertidal rocky shores by accelerating the settlement of seaweed.