【作者】 曹文浩； 严涛； 刘永宏； 黄日明； 杨斌； 胡煜峰； 董钰； 严文侠；

【Author】 CAO Wenhao,YAN Tao,LIU Yonghong, HUANG Riming,YANG Bin,HU Yufeng,DONG Yu,YAN Wenxia (South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301, Guangdong,China;)

【机构】 中国科学院南海海洋研究所；

【摘要】 海洋环境复杂多变,各物种间对空间的竞争十分激烈,任何大型动、植物都是营固着或附着生活种类的潜在附着污损对象,为避免其他生物附着可能产生的危害,海洋中一些生物在长期的进化过程中,形成了许多独特的自我保护机制来保持自身体表的洁净。其中,许多海洋生物可通过产生对环境无危害的、具有防污活性的次生代谢产物以防止其他生物污损附着。这些天然防污化合物是一类非常理想的防污剂,其通过驱赶而不是毒杀作用实现防污目的,不仅具有极好的防污效果,而且不会对环境产生危害。因此,开展海洋化学生态学研究,将其应用于海洋污损生物防除领域,不仅有利于弄清海洋污损生物的防除作用机制,还可以提供大量新型、无毒、环保、高效的天然防污化合物,在基础研究和生产实践中均具有重要意义。美丽属海绵(Callyspongia)在我国南方海域有较丰富的分布。自20世纪70年代开始研究该属海绵以来,已从中分离获得多炔、肽类、生物碱、脂肪酸、聚酮类及甾体等生物活性物质,且显示出较好的抗菌、抗肿瘤、细胞毒性及抑制HIV逆转录酶活性等作用。网纹藤壶(Balanus reticulatus)不仅是我国华南沿海污损生物群落中的绝对优势种,也是世界性热带和亚热带广布种。目前虽然已对美丽属海绵代谢产物的化学组成及其活性开展了研究,但尚未见这些化合物防污损研究的报道。从美丽海绵分离提取了2-去氧-1-氢-1,2,4,三唑、环(脯氨酸-甘氨酸)、尿嘧啶、环(脯氨酸-丙氨酸)、6-氨基嘌呤、4-(1-苯乙基)苯酚、1-氢-1,2,4,三唑和胆甾-5-烯-3β,7α-二醇等8种化合物,并研究了其对网纹藤壶金星幼虫的影响,其中6-氨基嘌呤和4-(1-苯乙基)苯酚对金星幼虫具有明显的毒杀作用。美丽海绵可能通过这些活性化合物的协同作用来防止它种生物附着污损。海洋生物化学防污作用十分复杂,海绵动物在长期进化过程中形成的天然化学防污损作用机制是现有防污涂料技术革新的理想研究模式。今后的工作重点除了进一步分离筛选新的天然活性防污化合物外,还需确定生物体表单位面积内相关活性化合物的含量及其之间的最佳比例,为研制经济、高效、环保的第二代污损生物防除涂料提供参考。更多还原

【Abstract】 Around the world,marine fouling can have serious impacts on human activities and other organisms.To survive in the highly competitive arena of the marine environment,many organisms have developed unique protection mechanisms against fouling,including tolerance,avoidance,low surface energy and the secretion of natural compounds.Some species of sponge and coral are rarely epiphytized because of the production of secondary metabolites against fouling.Unlike common man-made antifouling compounds,these natural chemicals are environmentally friendly in marine ecological systems. The majority of marine fouling organisms are algae,coelenterates,polychaetes,bivalves,bryozoans and barnacles.Of them,the acorn barnacle Balanus reticulatus is one of the most important dominant species in the fouling communities of tropical and subtropical waters,particularly in the East and South China Sea.Thus is can be considered as an appropriate test representative for antifouling bioassays. Barnacle nauplii were obtained by dissecting freshly collected adult barnacles from the aquaculture facilities at Daya Bay,Shenzhen,China.Barnacle larvae were reared in darkness,at 30℃and on a diet of the green algae Platymonas subcordiformis.Seawater was changed and more algae added as necessary.Cyprid larvae developed after around 5 days and were collected and stored at 4℃for subsequent use. Sponges of the genus Callyspongia are widely distributed in the southern waters of China（e.g.coastal water of Fujian,Guangdong,Guangxi and Hainan Provinces）.They are well known as sources of biologically active natural products,including polyacetylenes,peptides,alkaloids,fatty acids,polyketides,and sterols.Some of these compounds possess antimicrobial,anti-tumor,cytotoxicic,and HIV reverse transcriptase inhibition properties. Freshly collected samples of Callyspongia were minced and extracts taken by washing with dilute ethanol three times.The aqueous ethanol extracts were concentrated under vacuum.The combined extracts were partitioned into ethyl acetate and water.The ethyl acetate soluble portion was separated using column chromatography.A silica gel column was eluted with petroleum ether / ethyl acetate and then ethyl acetate / acetone.Similar isolates were identified using thin layer chromatography（TLC）.The structures of all isolates were determined using one dimensional and two dimensional nuclear resonance microscopy（NMR）,mass spectroscopy （MS） analyses,and by comparison of the spectroscopic data with those reported in the literature.A total of eight compounds were isolated and purified:Cyclo-（S-Pro-Gly）,Cyclo-Pro-Ala, 1-（2’-deoxy-β-D-erythro-pentofuranosyl）-1-H-1,2,4-triazole,uracil,4-（1-phenylethyl） phenol,6-NH₂-purine, 1-H-1,2,4,triazole and cholest-5-ene-3β,7α-diol. For antifouling tests,each compound was dissolved in lml methanol（or chloroform） and transferred into Petri dishes.Petri dishes were dried on a shaker prior to the addition of filtered seawater and larvae.Control dishes, using 1 ml of solvent（pre dried as above） and filtered seawater only were also prepared.The mean values of larval survival rate were compared among samples using least significant difference tests. Six compounds had no influence on the behavior and survival rate of larvae（P>0.05）: 1-（2’-deoxy-β-D-erythro-pentofuranosyl）-1-H-1,2,4-triazole,Cyclo-（S-Pro-Gly）,Cyclo-Pro-Ala,uracil, 1-H-1,2,4,triazole and cholest-5-ene-3β,7α-diol.The other two compounds 6-NH₂-purine（7.1μg/cm²） and 4-（1-phenylethyl） phenol（5.9μg/cm²） had significant toxic effects on the cyprid larvae of the acorn barnacle Balanus reticulatus（P<0.05）.The synergistic effect of these compounds may play an important role in the antifouling activities of the sponge.更多还原