Marine sponge Hymeniacidon perlevis possesses high diversity Marine sponge Hymeniacidon perlevis possesses high diversity

A total of 59 actinobacteria associated with an intertidal marine sponge Hymeniacidon perlevis collected from the Yellow Sea, China were isolated using eight different media. Species diversity and natural product diversity of isolates were analyzed. Twenty-seven isolates were selected on the basis of their morphology on different media, and assigned to the phylum Actinobacteria by a combination of 16S rRNA gene based restriction enzymes digestion and 16S rRNA gene sequence analysis (Zhang et al., 2008). The 16S rRNA genes of 27 isolates were digested by restriction enzymes Hha I and assigned to different groups according to their restriction enzyme pattern. The phylogenetic analysis based on 16S rRNA gene sequencing showed that the isolates belong to the genera Streptomyces, Nocardiopsis, Micromonospora, Cellulosimicrobium, Gordonia, Nocardia, Prauseria, Pseudonocardia, Saccharomonospora and Microbacterium. Previous studies have isolated strains of the genera Actinoalloteichus, Micromonospora, Nocardiopsis, Nocardia, Rhodococcus, Pseudonocardia, and Streptomyces from the same marine sponge (Zhang et al., 2006). Together, 12 culturable actinobacteria genera have been isolated and identified from the marine sponge H. perlevis. To our knowledge, this is the first report to recover such a high diversity of culturable actinobacteria from any marine sponges. In addition, this is also the first report of actinobacteria classified as Prauseria sp. and Saccharomonospora azurea that have been isolated from amarine sponge. All of the 27 isolates were screened for genes encoding polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS). PKS-I and PKS-II sequences were detected in more than half of the isolates and the different “PKS-I–PKS-II–NRPS” combinations in different isolates belonging to the same species are indicators of their potential natural product diversity and divergent genetic evolution (Pathom-Aree et al., 2006)

海绵中氯丙酸脱氯微生物的分离及其初步研究

以氯丙酸为单一碳源,通过富集培养从大连海域繁茂膜海绵中分离筛选得到139株氯丙酸降解菌。经过初步筛选得到35株降解能力较强的菌株。对这些菌株的耐盐试验表明,分离到的大部分菌株均具有较强的盐耐受能力,多数菌株为中度嗜盐菌。其中部分菌株可以在14.6%以上盐度培养基中生长,可能为极端嗜盐菌。多底物降解试验表明,大多数菌株均能降解多种底物,这些菌株可用于有机氯污染环境生物修复。同时也说明海绵对水体中有机氯污染物具有去除作用

一种海洋卤代酸脱卤微生物的分离筛选方法

本发明涉及一种海洋卤代酸脱卤微生物的分离筛选方法，其以海洋样品为分离源，通过卤代酸单一碳源液体培养基富集培养，卤代酸单一碳源显色平板筛选，菌株的分离纯化，菌体在卤代酸液体培养条件下催化脱卤情况检测四个步骤可以最终得到海洋卤代酸脱卤微生物。本发明适用于环境保护、污水处理、医药中间体、含氯农药等工业领域和实验研究领域。带填

海绵中氯丙酸脱氯微生物的分离及其初步研究

以氯丙酸为单一碳源,通过富集培养从大连海域繁茂膜海绵中分离筛选得到139株氯丙酸降解菌。经过初步筛选得到35株降解能力较强的菌株。对这些菌株的耐盐试验表明,分离到的大部分菌株均具有较强的盐耐受能力,多数菌株为中度嗜盐菌。其中部分菌株可以在14.6%以上盐度培养基中生长,可能为极端嗜盐菌。多底物降解试验表明,大多数菌株均能降解多种底物,这些菌株可用于有机氯污染环境生物修复。同时也说明海绵对水体中有机氯污染物具有去除作用

Culture-independent nested PCR method reveals high diversity of actinobacteria associated with the marine sponges Hymeniacidon perleve and Sponge sp.

A culture-independent nested polymerase chain reaction (PCR) technique was used to investigate the diversity of actinobacteria communities associated with the sponges Hymeniacidon perleve and Sponge sp. The phylogenetic affiliation of sponge-derived actinobacteria was then assessed by 16S rRNA sequencing of cloned DNA fragments. A total of 196 positive clones were screened by restriction fragment length polymorphism (RFLP) analysis; 48 unique operational taxonomic units (OTUs) were selected for sequencing. Rarefaction analysis indicated that the clone libraries represented 93% and 94% of the total estimated diversity for the two species, respectively. Phylogenetic analysis of sequence data revealed representatives of various phylogenetic divisions, which were related to the following ten actinobacterial genera: Acidimicrobium, Corynebacterium, Propionibacterium, Actinomyces, Micrococcus, Microbacterium, Streptomyces, Mycobacterium, Cellulosimicrobium, Sporichthya, and unidentified actinobacterial clones. A sponge-specific, previously uncultured actinobacteria community grouped within the subclass Acidimicrobidae was discovered from both H. perleve and Sponge sp. Sequences belonging to Acidimicrobium in the H. perleve and the Sponge sp. clone libraries represented 33% and 24% of the clones, respectively. In the Sponge sp. clone library Mycobacterium dominated, accounting for 70% of all clones. The presence of Acidimicrobium and mycobacteria within two sponges can lay the groundwork for attempts to culture these interesting bacteria for industrial applications

一株可降解石油烃的滕黄单胞菌及其应用

本发明公开了一株可降解石油烃的滕黄单胞菌及其应用。海连滕黄单胞菌（Luteimonas?dalianensis）OB44-3，保藏于中国微生物菌种保藏管理委员会普通微生物中心，保藏日期2013年1月18日，保藏编号CGMCC?N0.7165。本发明的海连滕黄单胞菌OB44-3是滕黄单胞菌属的新菌种，分离自石油污染的海洋环境，能够有效降解石油烃并利用石油作为生长的唯一碳源。在人工海水培养基中，7天内对石油烃（初始浓度为1000mg/L）的平均降解率为69.08%；同时该菌株的生长依赖于盐的存在，属于海洋细菌，对控制海洋环境中石油烃污染有独特的优势。本发明的海连滕黄单胞菌OB44-3可用于石油烃的降解

Comparative study on the phylogenetic diversity of culturable actinobacteria associated with marine sponges and ascidians in six isolation media

在相同的分离培养条件下,为比较地域差别较大的福建海域海绵动物(山海绵MyCAlE SP.和网架海绵STylISSA SP.)和海南海域海鞘动物(皱瘤海鞘STyElA PlICATA和乳突皮海鞘MOlgulA MAnHATTEnSIS)之间可培养放线菌多样性的差异,作者采用5种放线菌分离培养基和1种细菌通用培养基,对海绵和海鞘中的放线菌进行分离培养。采用16S rrnA基因限制性片段长度多态性(rESTrICTIOn frAgMEnT lEngTH POlyMOrPHISM,rflP)分析和序列分析,揭示其多样性。共获得可培养放线菌198株,其中从海绵中分离到87株放线菌,从海鞘中分离到111株放线菌。rflP分析表现为38种不同的图谱类型。16S rrnA基因序列分析表明,从海绵中分离到的放线菌包括6个放线菌属,其中有2株菌的16S rrnA基因序列与最相近的菌株相似性低于97%,可能是潜在的新菌株;从海鞘中分离到的放线菌包括7个放线菌属,有8株可能是潜在的新菌株。比较海绵和海鞘中可培养放线菌的多样性发现,从海绵中分离到的放线菌,除节细菌(ArTHrObACTEr)以外,均包括在海鞘分离的放线菌属中。海鞘相关放线菌多样性水平不容忽视,是除海绵之外另一获得新型放线菌资源以及药用天然活性产物的重要来源。The diversity of culturable actinobacteria was investigated and compared between two sponges(Mycale sp.and Stylissa sp.) collected from Fujian province and two ascidians(Styela plicata and Molgula manhattensis) collected from Hainan province under the same isolation condition.Five actinobacteria selective isolation media and one general bacteria isolation medium were selected to isolate and culture actinobacteria from both sponges and ascidians.The phylogenetic affiliation of the actinobacterial isolates was assessed using a combination of 16S rRNA gene amplification-restriction fragment length polymorphism(RFLP) analysis and 16S rRNA gene sequencing.A total of 198 purified actinobacteria were cultivated,including 87 from sponges and 111 from ascidians.There are 38 different RFLP patterns and the phylogenetic analysis based on sequencing results reveal a high and distinctive diversity of actinobacteria associated with these two sponge species and two ascidian species,including six and seven actinomycetes genera,respectively.The BLAST results show that two actinobacterial isolates derived from sponges are probably novel strains whose similarity are lower than 97%,whereas there are eight isolates from ascidians regarded as potential novel actinobacteria.Comparing the diversity of actinobacteria associated with sponges and ascidians,most genera from sponge are also found in ascidians except Arthrobacter.The diversity of ascidian-associated actinobacteria is high and cannot be underestimated.Together with sponges,ascidians are also excellent and promising sources for novel marine actinobacteria currently under exploited for the discovery of new bioactive marine natural products.海洋公益性行业科研专项经费资助项目(No.201305016