Allelopathy of extracts of Ulva prolifera on green tides in the Yellow Sea and the identification of the allelochemicals
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摘要: 化感作用是浒苔绿潮暴发的重要原因之一,目前,对于浒苔产生化感物质的结构、种类以及产量的研究鲜有报道。本文针对黄海浒苔绿潮致灾藻种,利用毒性鉴别程序(TIE)对其体内的化感物质进行分离提取及结构鉴定,明确了浒苔产生化感物质的主要成分。首先用石油醚、乙酸乙酯和甲醇3种有机溶剂分别对新鲜浒苔进行提取,得到3种不同溶剂粗提物,其中乙酸乙酯粗提物的抑藻活性最强,对测试物种中肋骨条藻的半数有效浓度(96h-EC50)值为22.25 mg/L;其次对乙酸乙酯粗提物进行萃取分离得到初步提纯物,其对中肋骨条藻的96h-EC50值为21.12 mg/L;然后用硅胶层析柱对上述初步提纯物进行分离,共得到6种组分,其中抑藻活性最强的组分对中肋骨条藻的96h-EC50值为10.57 mg/L;最后,对新鲜浒苔提取物用红外光谱和气相色谱-质谱联用仪进行检测,实验结果表明主要化感物质为:4,7,10,13,16,19-二十二碳六烯酸、2-十六碳烯酸、棕榈酸、5,8,11,14,17-二十碳五烯酸、9,12-十八碳二烯酸、9,12,15-十六碳三烯酸、花生四烯酸、13-二十二碳烯酸等8种脂肪酸。研究结果对于系统阐释浒苔绿潮暴发机制具有重要意义。Abstract: Allelopathy is one of the important factors of green tide, while the studies of structure, category and yield of allelochemicals released by Ulva prolifera are lacking. Based on the procedure of toxicity identification evaluation(TIE), this study was carried to extract and identify the allelochemicals from the disaster-causing algaes on green tides in the yellow sea, so that we can confirm the main components of allelochemicals realeased by U. prolifer. Firstly, U. prolifera was extracted with three different organic solvents, which include petroleum ether, ethyl acetate and methanol. Three different solvents crude extracts were obtained and the ethyl acetate extract has the stongest effect on anti-algal activity, the 96h-EC50 value against S. costatum was 22.25 mg/L. Next, the primary extracts were obtained by isolating ethyl acetate extract, the 96h-EC50 value against S. costatum was 21.12 mg/L. Then, chromatography column was used to purify the above primary extracts and six components were certained, the 96h-EC50 value of the most toxic component against S. costatum was 10.57 mg/L. Finally, the possible allelochemicals of U. prolifera dry tissues were detected using Gas chromatography-mass spectrometry (GC-MS) and infrared spectroscopy (IR) analysis. Eight fatty acids were identified for both species:4,7,10,13,16,19-docosahexaenoic acid, 2-Hexadecenoic acid,Hexadecanoic acid, 5,8,11,14,17-eicosapentaenoate, 9,12-octadecadienoic acid, 9,12,15-octadecatrienoic acid, arachidonic acid, 13-docosenoic acid. The results of the study play a significant role in systematically illustrating the mechanism of the outbreak of green tides.
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Li Yan, Song Wei, Xiao Jie, et al. Tempo-spatial distribution and species diversity of green algae micro-propagules in the Yellow Sea during the large-scale green tide development[J]. Harmful Algae, 2014, 39:40-47. Zhang Cui, Lu Jian, Wu Jun, et al. Removal of phenanthrene from coastal waters by green tide algae Ulva prolifera[J]. Science of the Total Environment, 2017, 609:1322-1328. Liu Xiangqing, Wang Zongling, Zhang Xuelei. A review of the green tides in the Yellow Sea, China[J]. Marine Environmental Research, 2016, 119:189-196. Hiraoka M, Ohno M, Kawaguchi S, et al. Crossing test among floating Ulva thalli forming 'green tide' in Japan[J]. Hydrobiologia, 2004, 512(1/3):239-245. 李瑞香, 吴晓文, 韦钦胜, 等. 不同营养盐条件下浒苔的生长[J]. 海洋科学进展, 2009, 27(2):211-216. Li Ruixiang, Wu Xiaowen, Wei Qinsheng, et al. Growth of Enteromorpha prolifera under different uutrient conditions[J]. Advances in Marine Science, 2009, 27(2):211-216. Granéli E, Weberg M, Salomon P S. Harmful algal blooms of allelopathic microalgal species:the role of eutrophication[J]. Harmful Algae, 2008, 8(1):94-102. Zhang Jianheng, Huo Yuanzi, Yu Kefeng, et al. Growth characteristics and reproductive capability of green tide algae in Rudong coast, China[J]. Journal of Applied Phycology, 2013, 25(3):795-803. Zhang Xiaowen, Wang Hongxia, Mao Yuze, et al. Somatic cells serve as a potential propagule bank of Enteromorpha prolifera forming a green tide in the Yellow Sea, China[J]. Journal of Applied Phycology, 2010, 22(2):173-180. 范士亮, 傅明珠, 李艳, 等. 2009-2010年黄海绿潮起源与发生过程调查研究[J]. 海洋学报, 2012, 34(6):187-194. Fan Shiliang, Fu Mingzhu, Li Yan, et al. Origin and development of Huanghai (Yellow) Sea green-tides in 2009 and 2010[J]. Haiyang Xuebao, 2012, 34(6):187-194. 贾睿, 吴敏, 蔡春尔, 等. 浒苔对赤潮异湾藻的克生作用[J]. 水产学报, 2012, 36(4):562-567. Jia Rui, Wu Min, Cai Chuner, et al. Allelopathic effects of Ulva prolifera on Heterosigma akashiwo[J]. Journal of Fisheries of China, 2012, 36(4):562-567. 霍元子, 田千桃, 徐栅楠, 等. 浒苔对米氏凯伦藻生长的克生作用[J]. 海洋环境科学, 2010, 29(4):496-499, 508. Huo Yuanzi, Tian Qiantao, Xu Shannan, et al. Allelopathic effects of Ulva prolifera on growth of Karenia mikimotoi[J]. Marine Environmental Science, 2010, 29(4):496-499, 508. 王兰刚, 徐姗楠, 何文辉, 等. 海洋大型绿藻条浒苔与微藻三角褐指藻相生相克作用的研究[J]. 海洋渔业, 2007, 29(2):103-108. Wang Lan'gang, Xu Shannan, He Wenhui, et al. Studies on the antagonistic relationship between seaweed Enteromorpha clathrata and marine microalgae Phaeodactylum tricornutum[J]. Marine Fisheries, 2007, 29(2):103-108. Allen J L, Ten-hage L, Leflaive J. Allelopathic interactions involving benthic phototrophic microorganisms[J]. Environmental Microbiology Reports, 2016, 8(5):752-762. Granéli E, Salomon P S, Fistarol G O. The role of allelopathy for harmful algae bloom formation[M]//Evangelista V, Barsanti L, Frassanito A M, et al. Algal Toxins:Nature, Occurrence, Effect and Detection. Dordrecht:Springer, 2008:159-178. Belz R G, Hurle K. A novel laboratory screening bioassay for crop seedling allelopathy[J]. Journal of Chemical Ecology, 2004, 30(1):175-198. Gross E M, Meyer H, Schilling G. Release and ecological impact of algicidal hydrolysable polyphenols in Myriophyllum spicatum[J]. Phytochemistry, 1996, 41(1):133-138. Körner S, Nicklisch A. Allelopathic growth inhibition of selected phytoplankton species by submerged macrophytes[J]. Journal of Phycology, 2002, 38(5):862-871. Yang Cuiyun, Liu Sujing, Zhou Shiwei, et al. Allelochemical ethyl 2-methyl acetoacetate (EMA) induces oxidative damage and antioxidant responses in Phaeodactylum tricornutum[J]. Pesticide Biochemistry and Physiology, 2011, 100(1):93-103. 赵中华, 张路, 于鑫. 水体沉积物毒性的评价方法[J]. 环境与健康杂志, 2007, 24(5):360-363. Zhao Zhonghua, Zhang Lu, Yu Xin. Methods for sediment toxicity evaluation[J]. Journal of Environment and Health, 2007, 24(5):360-363. Wheelock C E, Phillips B M, Anderson B S, et al. Applications of carboxylesterase activity in environmental monitoring and toxicity identification evaluations (TIEs)[M]//Whitacre D M. Reviews of Environmental Contamination and Toxicology. New York:Springer, 2008:117-178. 狄兰兰. 孔石莼中防污活性物质的提取与分离[D]. 青岛:中国海洋大学, 2009. Di Lanlan. Extraction and separation of antifouling active substances in Ulva pertusa[D]. Qingdao:Ocean University of China, 2009. Tameishi M, Yamasaki Y, Nagasoe S, et al. Allelopathic effects of the dinophyte Prorocentrum minimum on the growth of the bacillariophyte Skeletonema costatum[J]. Harmful Algae, 2009, 8(3):421-429. Nan Chunrong, Zhang Haizhi, Zhao Guangqiang. Allelopathic interactions between the macroalga Ulva pertusa and eight microalgal species[J]. Journal of Sea Research, 2004, 52(4):259-268. Huang Xuguang, Lin Xiechang, Li Shunxing, et al. The influence of urea and nitrate nutrients on the bioavailability and toxicity of nickel to Prorocentrum donghaiense (Dinophyta) and Skeletonema costatum (Bacillariophyta)[J]. Aquatic Toxicology, 2016, 181:22-28. Wang Rui, Wang Jiangtao, Xue Qiaona, et al. Preliminary analysis of allelochemicals produced by the diatom Phaeodactylum tricornutum[J]. Chemosphere, 2016, 165:298-303. 高敏. 海洋微生物和微藻中防污活性物质的提取及其性能研究[D]. 青岛:中国海洋大学, 2014. Gao Min. Extraction and performance study of antifouling compounds produced by marine microorganism and microalgae[D]. Qingdao:Ocean University of China, 2014. Vanewijk P H, Hoekstra J A. Calculation of the EC50 and its confidence interval when subtoxic stimulus is present[J]. Ecotoxicology and Environmental Safety, 1993, 25(1):25-32. 江玉, 吴志宏, 韩秀荣, 等. 多环芳烃对海洋浮游植物的生物毒性研究[J]. 海洋科学, 2002, 26(1):46-50. Jiang Yu, Wu Zhihong, Han Xiurong, et al. Toxicity of polycyclic aromatic hydrocarbons (PAHs) to marine algae[J]. Marine Sciences, 2002, 26(1):46-50. 韩秀荣, 高嵩, 侯俊妮, 等. 浒苔干粉末提取物对东海原甲藻和中肋骨条藻的克生作用[J]. 生态学报, 2013, 33(23):7417-7429. Han Xiurong, Gao Song, Hou Junni, et al. Allelopathic effects of extracts from Ulva prolifera powders on the growth of Prorocentrum donghaiense and Skeletonema costatum[J]. Acta Ecologica Sinica, 2013, 33(23):7417-7429. An Zhen, Wang Zhenyu, Li Fengmin, et al. Allelopathic inhibition on red tide microalgae Skeletonema costatum by five macroalgal extracts[J]. Frontiers of Environmental Science & Engineering in China, 2008, 2(3):297-305. Gao Yunni, Liu Biyun, Xu Dong, et al. Phenolic compounds exuded from two submerged freshwater macrophytes and their allelopathic effects on microcystis aeruginosa[J]. Polish Journal of Environmental Studies, 2011, 20(5):1153-1159. 胡陈艳, 葛芳杰, 张胜花, 等. 马来眼子菜体内抑藻物质分离及常见脂肪酸抑藻效应[J]. 湖泊科学, 2010, 22(4):569-576. Hu Chenyan, Ge Fangjie, Zhang Shenghua, et al. Isolation of antialgal compounds from Potamogeton malaianus and algal inhibitory effects of common fatty acids[J]. Journal of Lake Sciences, 2010, 22(4):569-576. Gao Y N, Ge F J, Zhang L P, et al. Enhanced toxicity to the cyanobacterium Microcystis aeruginosa by low-dosage repeated exposure to the allelochemical N-phenyl-1-naphthylamine[J]. Chemosphere, 2017, 174:732-738. 赵妍. 大型海藻缘管浒苔(Enteromorpha linza)对两种海洋微藻克生效应的初步研究及克生物质的分离与鉴定[D]. 青岛:中国海洋大学, 2010. Zhao Yan. Allelopathic effect of Enteromorpha linza on two species of marine microalgae and the allelochemicals' isolation and characterization[D]. Qingdao:Ocean University of China, 2010. Nakai S, Yamada S, Hosomi M. Anti-cyanobacterial fatty acids released from Myriophyllum spicatum[J]. Hydrobiologia, 2005, 543(1):71-78. Alamsjah M A, Hirao S, Ishibashi F, et al. Algicidal activity of polyunsaturated fatty acids derived from Ulva fasciata and U. pertusa (Ulvaceae, Chlorophyta) on phytoplankton[J]. Journal of Applied Phycology, 2008, 20(5):713-720.
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