东海原甲藻cDNA文库构建及尝试性EST分析

东海原甲藻(Prorocentrumdonghaiense)是中国沿海频繁发生的大规模赤潮的原因种之一,大规模分离鉴定东海原甲藻功能基因是理解东海原甲藻赤潮形成过程和机理的重要基础。取对数生长期藻体,经微量总RNA抽提、cDNA合成、cDNA扩增和克隆等步骤,构建了东海原甲藻cDNA文库并进行了尝试性表达序列标签分析。从含有约5000个转化子的文库中随机取150个测序,获得126条EST序列。经网上BlastN及BlastX分析,共发现11个是已知功能的基因的标签。这些基因与东海原甲藻生长发育、物质转换和能量代谢相关。     

Isolation, cloning and sequencing of AFLP markers related to disease-resistance traits in Fenneropenaeus chinensis

Amplified fragment length polymorphism (AFLP) technique was used to analyze the fingerprinting of four successive generations ofFenneropenaeus chinensis to reveal their disease-resistance traits. Some loci showed quite different genetic frequencies due to artificial selection, which implied that these fragments were putative markers related to the disease-resistance trait. We developed a simple and effective method to further characterize these AFLP fragments. Specific AFLP bands were cut directly from polyacrylamide gels, re-amplified, cloned and sequenced. Eight putative genetic markers were sequenced and their sizes ranged from 63 to 209 bp. The sequences were submitted to dbGSS (database of Genome Sequence Survey); and the BLAST analysis showed low similarity to the function genes, indicating these markers were tightly linked to a disease-resistance trait but were not functional genes. This research was supported by special funds from the National Key Basic Research Program (G1999012007) and the National High-Tech Research and Development Program of China (863 Program. 2001AA620105).     

Study on gametophyte vegetative growth ofUndaria pinnatifida and its applications

When cultured under certain environmental conditions (25°C, light intensity 80 μmol/m²·s, LD 12/12, in enriched seawater medium with 7×10⁻⁴ mol/L NO₃−N, 1.56×10⁻⁴ mol/L, PO₄−P and supplements of other elements like Mn, Fe, I, etc.), male and female gametophytes ofU. pinnatifida kept growing vegetatively and propagated fast at average daily fresh weight increase rate of about 20%. The empirical formulaG _m=G _o·3^m was established to estimate the output of vegetative gametophytes. Vigorous vegetative gametophyte cells began to form reproductive structures (oogonium and spermatangium, when the temperature was lower than 25°C and other environmental factors were kept optimal. The sufficient supply of gametophyte cells provided enough seeds for raisingUndaria sporelings on production scale. Controlled cross-breeding experiments using selected male and female gametophyte clones which increase their cell number by mitosis instead of meiosis were also carried out in vitro. Juvenile sporophytes from the cross-breeding had almost the same length and width increase rates as those of the control. The fact that vegetative gametophytes can be purposely selected, propagated quickly, cross-bred, with the resulting sporophytes having almost the same characteristics leads to a new way to select desiredUndaria strains for long time use without losing the desired economic characteristics. Contribution No. 2696 from the Institute of Oceanology, Chinese Academy of Sciences. Supported by National Natural Science Foundation of China.     

南极欺骗岛土壤细菌群落结构和多样性分析

采用基于16S r DNA克隆文库的非培养方法和传统培养方法相结合的手段,对南极欺骗岛土壤细菌群落结构及多样性进行了初步分析。对来自16S r DNA克隆文库的118个阳性克隆进行测序和序列比对,结果显示细菌来自放线菌门(Actinobacteria)、变形杆菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)3大门的39个操作分离单元(OTU),其中优势菌群为放线菌门和变形杆菌门,分别占65.25%和28.81%。放线菌门的主要优势属为鱼孢菌属(Sporichthya)、类诺卡式氏菌属(Nocardioides)、束缚菌属(Conexibacter)、Gaiella和节杆菌属(Arthrobacter)。变形杆菌门的主要优势属为鞘氨醇单胞菌属(Sphingomonas)、硫杆菌属(Thiobacillus)、寡养单胞菌属(Stenotrophomonas)和热单胞菌属(Thermomonas)。采用6种培养基对细菌进行培养和分离,共得到57株菌,来自放线菌门、变形杆菌门、拟杆菌门和厚壁菌门4大门的13个OTU,其中优势菌群为放线菌门和变形杆菌门,分别占57.89%和31.58%。优势属是放线菌门的节杆菌属(Arthrobacter)、雷夫松氏菌属(Leifsonia)和变形杆菌门的假单胞菌属(Pseudomonas)。本论文为研究欺骗岛土壤细菌多样性以及有益菌种资源的开发和利用奠定了基础。     

GIS组件分类与检索技术的研究和应用

讨论了组件分类和匹配原理，并对签名匹配法进行了研究，把它作为从已知的组件中获取可重用组件的方法，最后提出了在国土信息管理领域中GIS组件的解决方案。     

基于资源三号卫星影像变化更新的应用浅析

选取哈尔滨市周边地区作为试验区,基于资源三号卫星影像和历史DLG信息,综合利用栅格计算方法和矢量套合比较方法进行变化发现,按照地物类别特性和指标构建变化检测知识库来消除大部分伪变化,辅助解译人员更新要素,为以后的常态性更新提供参考。     

Nucleotide sequence of β-tubulin gene from the soft coralscleronephthya gracillimum (kükenthal)

We cloned the complete cDNA of the β-tubulin from the soft coral,Scleronephthya gracillimum (Kükenthal) (Alcyonacea, Octocorallia, Anthozoa, Cnidaria), via the random sequencing of a cDNA library and the 5′-rapid amplification of cDNA end (RACE) technique. The full-length cDNA of theS. gracillimum β-tubulin comprised 1541 bp, not including the poly (A)⁺ stretch, also contained a complete open reading frame, which codes for a total of 445 amino acids. The amino acid residues 16-402 appeared to be in a state of conservation in a variety of animals. Northern blot analysis clearly demonstrated that the sequence we have obtained is, indeed, the full-length cDNA of the β-tubulin gene inS. gracillimum.     

三角褐指藻磷酸甘油酸变位酶基因可能侧翼序列的筛选、克隆以及序列测定

采用RT PCR的方法从酵母中成功地得到了磷酸甘油酸变位酶的cDNA基因 ,分别用32 P和地高辛 ddUTP标记以用作探针。以32 P标记的探针筛选三角褐指藻基因组文库 ,获得了 4kb的阳性DNA片段 ;进一步分析发现 ,该 4kb片段的真正阳性区域是位于片段端部的30 6bp的序列 ,因此认为该序列为三角褐指藻磷酸甘油酸变位酶基因的侧翼部分。克隆该30 6bp的DNA片段 ,并且测定其序列。结果表明 ,该 30 6bp的DNA片段包含两个同向重复序列 ,每个重复序列的大小为 1 1 6bp ,在每个重复序列中均含有GGTTCAATGT区域 ,这与一般常见的真核基因 5′端的CAATbox有相似之处。     

Complementary (secondary) metabolites in a soft coral: sex-specific variability, inter-clonal variability, and competition

Sex‐specific interactions involving competition for space between the dioecious alcyonacean soft coral Sarcophyton glaucum and the scleractinian coral Acropora robusta were assessed experimentally on Bald Rock, central region of the Great Barrier Reef. To examine this, plus inter‐clonal responses, one male colony of S. glaucum, known to produce sarcophytoxide as its predominant complementary (secondary) metabolite, was sectioned, producing 10 clones. The same was done for a female colony. These two sets of clones were then relocated to grids and placed in contact with Acropora clones. Relocated and non‐relocated controls were also monitored. High levels of tissue necrosis were observed in the hard coral under contact conditions with both the male and female clones after 20 days. The development of a protective polysaccharide layer in the alcyonacean was also observed. Differences observed in the concentrations of complementary metabolites within the two different S. glaucum colonies were related to sex. Both under competition and non‐competition conditions, females exhibited significantly higher concentrations of sarcophytoxide than males, and this increased with time. Fatty ester concentration was also higher in females than males, varying significantly through time, and falling dramatically just after spawning. Fatty ester concentrations decreased linearly through time in the male clones. When involved in competition for space, females possessed higher concentrations of fatty esters than males, both at the site of contact and in non‐contact sites, again, decreasing after spawning. No significant changes in sarcophytoxide levels were noted in the parental colonies, but such changes were observed in fatty esters, with the female producing higher concentrations until after spawning. The use of these two variates in the form of a ratio (sarcophytoxide concentration:fatty ester concentration) yielded a variable Rho (ρ) which was a more sensitive indicator of biochemical change than either of its components alone. These two sets of compounds appeared to have a negative association through time and varied highly significantly between sexes. The diterpene sarcophytoxide may be considered an allelopathic or stress metabolite, while the lipids act as energy storage metabolites.