Effects of vegetation and sewage load on mangrove crab condition using experimental mesocosms

Constructed wetlands, especially mangroves, have been studied for their usefulness in sewage treatment but the effects of mangrove vegetation and a sewage load on mangrove macrofauna have been given little attention. Ocypodid crabs are important components of mangrove forests and constitute good bioindicators of the functioning of the ecosystem as a whole. In constructed mangrove mesocosms, three vegetation treatments (bare substratum, and Avicennia marina and Rhizophora mucronata seedlings) were subjected to 0, 20, 60 and 100% sewage loads from a nearby hotel. The physiological condition of introduced Uca annulipes and Uca inversa was evaluated in terms of their RNA/DNA ratio after one, five and twelve months, and used as an indicator of ecological function in the system. Crab condition in 0% sewage load was similar to that of wild crabs throughout, suggesting no significant effects of the mesocosms on their RNA/DNA ratio. Overall, both species coped well with the administered sewage loads, suggesting good ecological function in the system. Both species manifested similar patterns in RNA/DNA ratio, being more affected by seasonal fluctuations than by sewage load and vegetation presence and type. Higher RNA/DNA ratios were recorded in the long compared to the short rainy season. Sewage enhanced crab condition in the bare substratum and R. mucronata treatments, especially after one year, probably as a result of enhanced food availability. Uca inversa may be more sensitive to sewage pollution than U. annulipes. In A. marina, no difference in crab condition was observed between sewage loads, and this mangrove yielded the best reduction in sewage impacts. Our results support the usefulness of constructed mangrove areas in sewage treatment, especially if planted with A. marina and inhabited by physiologically healthy ocypodid crabs to enhance the system's performance.     

Molecular ecological study of <Emphasis Type="Italic">Siganus spinus</Emphasis> and <Emphasis Type="Italic">S. guttatus</Emphasis> from Okinawan waters based on mitochondrial DNA control region sequences

The mitochondrial DNA (mtDNA) control region was sequenced to examine the genetic variability and gene flow of juvenile Siganus spinus and S. guttatus. In total, 461 nucleotide sequences were obtained from 69 specimens of juvenile S. spinus from Okinawa Island and Ishigaki Island, in which 17 variable sites and 22 haplotypes were identified. Haplotype diversity (h) was high (0.9244 in Okinawa and 0.8984 in Ishigaki), whereas nucleotide diversity (π) was low (0.0063 in Okinawa and 0.0059 in Ishigaki). The two populations were not genetically distinct. Siganus guttatus, which do not form large schools at recruitment, in contrast S. spinus, were also analyzed by studying 152 individuals collected off Okinawa Island, Miyako Island, and Ishigaki Island. Of 476 nucleotide sequences, 50 were variable, and 42 haplotypes were identified. Genetic variability values were high (h = 0.8766 and π = 0.0151 in Okinawa; h = 0.9640 and π = 0.0192 in Miyako; h = 0.9161 and π = 0.0199 in Ishigaki). The Okinawa population was genetically isolated from the Miyako and Ishigaki populations. As a result, genetic diversity was high for each of these siganid populations despite their being target species for fisheries; however, the degree of inter-population gene flow was higher for S. spinus than S. guttatus, suggesting that these species exhibit different dispersal strategies.     

用于养殖环境调控的微生物制剂评价方法的研究

为尝试建立对不同微生物制剂产品的客观评价方法体系,选择5种商品化复合微生物制剂,进行了菌数和主要组成种类等测定,并利用天然或人工模拟的养殖污水进行了不同微生物制剂对氨氮、亚硝酸态氮、COD等降解效果的比较,以及微生物制剂活化增效后的效果比较等.结果表明,不同的商业化复合微生物制剂在有益菌含量和水质污染降解效果方面存在较大差异.另外,微生物制剂在海水和淡水中以及是否经过活化,效果也存在较大差异.     

虾类遗传多样性的研究现状

同工酶技术已经被广泛应用于虾类群体遗传学及标记辅助选择育种等研究方面.同工酶电泳显示对虾有相应的亚群差异,暖水性虾类的遗传多样性水平高于冷温性种类;连续分布的且在不同生活史阶段所处的栖息地环境变化幅度大的种类往往表现出较低的遗传多态性,虾类群体的多态位点比例和杂合度都较低,养殖群体比野生群体更低.但同工酶电泳技术往往检测不出一些"隐性"或"中性"变异而低估了遗传多样性水平.随着分子生物学技术的发展,从20世纪90年代开始,虾类遗传多样性的研究逐渐向线粒体DNA和核DNA的多态性方向发展.限制性片断长度多态性(RFLP)在虾类线粒体多倍型、物种标记以及比较野生和养殖群体线粒体COI 基因的多态性比较等方面应用普遍.随机扩增多态性(RAPD)技术主要应用于虾类不同地理群体遗传多态性调查,养殖群体与野生群体之间、养殖群体世代之间的遗传多态性比较,该技术得到的虾类遗传多样性水平高于同工酶电泳技术.扩增片断长度多态性(AFLP)技术在虾类中主要应用于遗传连锁图谱的构建,有关虾类的AFLP遗传多样性分析的报道不多.利用AFLP技术对连续选育群体的遗传多样性研究显示,随着选育世代的增加,选育群体的遗传多样性呈现下降趋势,但随着选育时间的延长,群体之间的分化逐渐降低,群体的遗传结构开始趋于稳定,成为一个品系.单一重复序列(SSR)技术主要用于标记种群遗传特异性、不同地理群体间以及野生和养殖群体间的遗传多样性差异.DNA序列分析技术特别是线粒体序列分析技术近年来在虾类遗传多样性研究中的应用逐渐增多.mtDNACOI、mtDNA12S rRNA和mtDNA16S rRNA的基因序列差异被应用于形态相近物种的鉴别、亚属分类、确定个体间亲缘关系和野生与养殖群体间的遗传多样性差异等.为了实现对虾类种群遗传差异的更深入了解,人们把种群遗传结构的研究和系统地理学相结合,即用分子系统地理学的方法来研究遗传谱系空间分布的历史特征,通过种群遗传结构的分析来探讨种内系统地理格局的形成机制、系统发育关系以及现有分布特征,并结合种群的地理分布状况来发现和验证与其相关的地质事件,追溯和揭示种群的进化历程.从育种、养殖以及资源保护的角度出发,人们开始研究野生群体的种质基因库,希望根据群体遗传结构确定合理的增殖策略,制定科学的保护措施,确保虾类养殖业的持续发展.     

高效非损伤性扇贝DN A提取方法的建立及效果评价

本研究建立了一种高效、经济的非损伤性扇贝DNA提取方法,在不影响扇贝个体生存状态的前提下,采用擦拭法和室温裂解相结合的方式在20 min内即可获得个体基因组DNA。以虾夷扇贝(Patinopecten yessoensis)、栉孔扇贝(Chlamys farreri)和海湾扇贝(Argopecten irradians)为实验对象,研究了不同擦拭材料(棉签、滤纸)和不同擦拭部位(外套膜、鳃丝、内脏团)的核酸提取效果,评估了本方法在贝类基因分型领域应用的可行性。研究表明,用棉签、滤纸2种材料擦拭扇贝的鳃丝、内脏团或外套膜组织均能获得主带清晰完整的基因组DNA,且不影响扇贝的存活状态。在3种扇贝中采用棉签擦拭法的DNA得率均高于滤纸法,以这2种方式获得的DNA为模板进行SSR和SNP标记分析,能获得均一稳定的扩增条带,SSR标记分型结果准确可靠。本研究建立了简便、快速进行扇贝基因型分析的非损伤性DNA提取方法,为贝类全基因组选育和珍稀贝类资源的种质保护开发提供了新的技术手段。     

基于COI序列的翡翠股贻贝Perna viridis线粒体遗传特性分析及其近缘种间的系统关系探讨

应用通用引物 COIL 1490和 COIH 2198对翡翠股贻贝Perna viridis的性腺和体细胞线粒体DNA进行PCR扩增,获得661bp长度的COI基因片段,经过比对性腺与体细胞的COI片段,发现雄性性腺与体细胞COI基因均为一个单倍型,即体内只有一种线粒体DNA类型,没有发现双单性遗传现象,雌、雄性腺的COI基因片段变异率很低（0.31%）。应用PAUP构建了NJ树、MP树以及贝叶斯法构建了贝叶斯树,对股贻贝属3种间的系统关系进行了分析,结果表明,翡翠股贻贝P. viridis与P. canaliculus 和 P. perna 之间的分化与分歧年代的估算是相吻合的。     

珠江口滨海湿地退化现状、原因及保护对策

2007年和2008年秋季对珠江口滨海湿地进行了环境现状综合调查,结果表明,珠江口滨海湿地退化主要表现在4个方面:天然湿地面积减少、湿地生产力不断下降、湿地环境状况持续恶化和湿地景观破碎化。珠江口滨海湿地退化的原因主要为滩涂开发与围填海、污染物排放、养殖污染、海平面上升、过度捕捞、海砂开采和港口码头建设以及水土资源开发等因素。滨海湿地保护与利用对策主要有:制定有关滨海湿地保护的法律法规和湿地保护与利用规划;加强人才队伍建设和滨海湿地有关技术研究;建立滨海湿地动态监测体系;加大滨海湿地保护区建设的力度;加强宣传和教育。     

基于微卫星标记的三疣梭子蟹家系系谱认证

用6个微卫星标记对三疣梭子蟹(Portunus trituberculatus)的6个家系进行系谱鉴别和遗传多样性研究。6个微卫星标记在6个家系中显示了高度的遗传差异,6个位点的多态信息含量(PIC)分别为Pot09位点0.6045,Pot14位点0.6072,Pot17位点0.8130,Pot18位点0.6870,Pot25位点0.7839,Pot42位点0.6330。6个多态性位点共发现了32个等位基因,每个位点的等位基因数在4～8个之间。不同位点共发现7个家系特异性等位基因,2#和3#家系中各发现2个,1#、5#、6#家系中各发现1个,4#家系未发现特异性等位基因。根据已知亲本及子代基因型,可推断出6个家系中全部亲本的基因型,据此鉴别各家系。在Pot18位点可将1#家系和6#家系与其他家系相区别;Pot14位点和Pot17位点可将3#家系与其他家系相区别;Pot42位点可将5#与其他家系相区别;Pot17位点和Pot25可将2#家系与其他家系相区别。因此,Pot18位点、Pot14位点和Pot17位点、Pot42位点、Pot17位点和Pot25位点可分别用于鉴别1#和6#家系、3#、5#、2#家系的特异性标记。研究表明,在选用的6个微卫星标记中,最少选用3个标记可鉴别6个三疣梭子蟹家系。     

基于线粒体D-loop区分析黄颡鱼(Pelteobagrus fulvidraco)五个淡水湖泊群体的遗传多样性

采用线粒体D-loop区测序技术,对巢湖、滆湖、洪泽湖、鄱阳湖和太湖5个地理群体的黄颡鱼(Pelteobagrus fulvidraco)遗传多样性及种群遗传结构进行了研究。结果表明,890个位点中变异位点有151个,简约信息位点24个。143个个体共检测到72个单倍型,平均单倍型多样性指数为0.951±0.011,核苷酸多样性为指数为0.00614±0.00087。鄱阳湖群体和其它4个湖泊群体的遗传距离相对比较大,滆湖和巢湖的遗传距离最小,总体遗传分化指数Fst为0.0896(P<0.01)。群体间基因流系数Nm=2.22,表明5个黄颡鱼种群间存在着一定的基因流动。     

红藻DNA条形码分析技术研究进展

DNA条形码是指利用一段相对较短的标准DNA片段,对物种进行识别和鉴定。目前该技术在动物、植物物种鉴定领域已经得到了广泛的研究和应用。在藻类的研究中,尚未确定一条统一的标准条形码基因,现阶段都是使用2条或2条以上基因序列来完成物种鉴定。对于形态多样、种类繁多的海洋红藻,常用的DNA条形码基因有COI基因(Partial cytochrome c oxidase I gene)、UPA基因(Partial 23SrRNA gene,universal plastid amplicon)、LSU基因(Partial 28SrRNA gene)和rbcL基因(The large subunit of ribulose-1,5-bisphosphate carboxylase)等,这些基因中2个或3个基因的互补运用准确有效地提高了红藻的鉴定准确率,尤其是COI基因的种间差异大足够区分相近物种。本文在概述条形码的原理及其标准的基础上,阐述了红藻DNA条形码鉴定研究的新进展以及常用几种基因片段的优缺点,并对条形码在红藻鉴定中存在的问题进行了分析,对其应用前景进行了展望。