王孝谦,黄翔鹄,李长玲,罗国玲,张宁,王新宇,张玉蕾

• 1:广东海洋大学水产学院
• 2:广东海洋大学广东省藻类养殖及应用工程技术研究中心

摘要(Abstract)：

【目的】探索波吉卵囊藻胞外滤液对铜绿微囊藻的抑制机理。【方法】采用Illumina平台,对使用BG11培养基(对照组)和波吉卵囊藻胞外滤液(实验组)培养的铜绿微囊藻(Microcystisaeruginosa)进行转录组测序,筛选差异表达基因并进行通路富集分析,并对部分差异表达基因进行q RT-PCR验证。【结果】与对照组相比,波吉卵囊藻胞外滤液处理后的铜绿微囊藻组中筛选1 483个表达差异显著的基因,其中上调表达基因788个,下调表达基因695个。GO功能分析将差异基因划分为35个子类别,包括催化活性、结合、代谢过程、细胞过程、膜、膜部分等。KEGG通路分析中,所有差异基因聚集在108个通路,其中84个差异基因注释在代谢功能类别中,而氧化磷酸化通路富集最显著。氧化磷酸化通路中,相关基因表达以上调为主(26个DEGs,20个上调,6个下调)。q RT-PCR验证结果与转录组测序结果的表达变化趋势一致。【结论】波吉卵囊藻滤液引起铜绿微囊藻氧化磷酸化途径上基因表达上调,ATP合成效率提高。

关键词(KeyWords)： 铜绿微囊藻;胞外滤液;转录组;波吉卵囊藻;化感作用;差异表达基因;基因功能

Abstract:

Keywords:

基金项目(Foundation): 广东省普通高校青年创新人才项目(2018KQNCX102,2018KQNCX110)

作者(Author): 王孝谦,黄翔鹄,李长玲,罗国玲,张宁,王新宇,张玉蕾

参考文献(References)：

• [1]SMITH V H,TILMAN G D,NEKOLA J C.Eutrophication:impacts of excess nutrient inputs on freshwater,marine,and terrestrial ecosystems[J].Environmental Pollution,1999,100(1/2/3):179-196.
• [2]LI M L,JIANG Y L,,CHUANG C Y,et al.Recovery of Alexandrium tamarense under chronic exposure of Ti O2nanoparticles and possible mechanisms[J].Aquatic Toxicology,2019,208:98-108.
• [3]TESTAI E,SCARDALA S,VICHI S,et al.Risk to human health associated with the environmental occurrence of cyanobacterial neurotoxic alkaloids anatoxins and saxitoxins[J].Critical Reviews in Toxicology,2016,46(5):385-419.
• [4]陈妍妍，李长玲，黄翔鹄.微囊藻毒素对凡纳滨对虾免疫相关酶活力的影响[J].广东海洋大学学报,2015,35(6):35-39.
• [5]ZI J M,PAN X F,MACISAAC H J,et al.Cyanobacteria blooms induce embryonic heart failure in an endangered fish species[J].Aquatic Toxicology,2018,194:78-85.
• [6]林森杰，姬南京，罗昊.海洋有害藻华研究进展[J].海洋与湖沼,2019,50(3):495-510.
• [7]EMILY,MYERS,TRACEY L,et al.PRINCE E K,MYERS T L,KUBANEK J.Effects of harmful algal blooms on competitors:Allelopathic mechanisms of the red tide dinoflagellate Karenia brevis[J].Limnology and Oceanography,2008,53(2):531-541.
• [8]ANDERSON D M.Approaches to monitoring,control and management of harmful algal blooms (HABs)[J].Ocean&Coastal Management,2009,52(7):342-347.
• [9]董文龙，闵水发，杨杰峰，等.湖泊富营养化防治对策研究[J].环境科学与管理,2014,39(11):82-85.
• [10]李由明，黄翔鹄，李晓梅.凡纳滨对虾养殖水体中浮游植物群落的组成分析[J].琼州学院学报,2012,19(5):42-45.
• [11]董龙香，程俊茗，胡鲲，等.波吉卵囊藻对重金属铬的耐受性研究[J].淡水渔业,2017,47(5):72-78.
• [12]黄翔鹄，魏少红，周美华，等.波吉卵囊藻对Cu2+、Zn2+耐受力和吸附作用的研究[J].上海海洋大学学报,2012,21(3):374-381.
• [13]任佳佳，洪廷，张宁，等.盐度对波吉卵囊藻叶绿素荧光参数的影响及Psb A基因的克隆与表达分析[J].广东海洋大学学报,2020,40(3):30-39.
• [14]李静红，黄翔鹄，李色东.波吉卵囊藻对弧菌生长的影响[J].广东海洋大学学报,2010,30(3):33-38.
• [15]刘梅，黄翔鹄，李长玲，等.不同条件下波吉卵囊藻对氨氮的吸收速率[J].广东海洋大学学报,2012,32(1):29-34.
• [16]李静红，黄翔鹄，李色东.波吉卵囊藻对弧菌生长的影响[J].广东海洋大学学报.2010,30(3):33-38.
• [17]蔡健，张玉蕾，黄翔鹄，等.波吉卵囊藻与蛋白核小球藻间的化感效应[J].海南热带海洋学院学报,2019,26(2):52-58.
• [18]WANG X Q,ZHANG Y L,LI C L,et al.Allelopathic effect of Oocystis borgei culture on Microcystis aeruginosa[J].Environmental Technology,2020,20:1-10.
• [19]李长玲，黄翔鹄，王孝谦.一种利用卵囊藻防控虾塘蓝藻水华的方法:CN110547237A[P].2019-12-10..
• [20]刘金豆，刘丽，黄元佳.稀杯盔形珊瑚转录组分析[J].广东海洋大学学报,2015,35(6):1-8.
• [21]王哲，王姿曼，郝瑞娟，等.马氏珠母贝金黄壳色选育群体植核后差异表达免疫基因筛选[J].广东海洋大学学报,2019,39(6):9-16.
• [22]成文龙，张永刚，张亚雷.转录组学在微藻研究中的应用[J].区域治理,2019(37):168-171.
• [23]张纯，唐承晨，王吉永，等.转录组学在植物应答逆境胁迫中的研究进展[J].生物学杂志,2017,34(2):86-90.
• [24]ZHAO P C,WANG Y M,LIN Z Y,et al.The alleviative effect of exogenous phytohormones on the growth,physiology and gene expression of Tetraselmis cordiformis under high ammonia-nitrogen stress[J].Bioresource Technology,2019,282:339-347.
• [25]DU C L,ZHANG B,HE Y L,et al.Biological effect of aqueous C60 aggregates on Scenedesmus obliquus revealed by transcriptomics and non-targeted metabolomics[J].Journal of Hazardous Materials,2017,324:221-229.
• [26]孙春蕾.铜绿微囊藻和水华鱼腥藻间化感作用的研究[D].南京:南京大学,2018.
• [27]CHEN L,MAO F J,KIRUMBA G C,et al.Changes in metabolites,antioxidant system,and gene expression in Microcystis aeruginosa under sodium chloride stress.[J].Ecotoxicology&Environmental Safety,2015,122:126-135.
• [28]金秋，董双林.海洋藻类化感作用研究进展[J].科技通报,2018,34(10):1-8.
• [29]JIN Q,WU H G,ZHANG X X,et al.Influence of non-nutrient environmental factors on Ulva pertusa's inhibitory effects on Heterosigma akashiwo growth[J].Journal of the Marine Biological Association of the United Kingdom,2016,96(5):1041-1052.
• [30]INDERJIT,DAKSHINI K M M.Algal allelopathy[J].The Botanical Review,1994,60(2):182-196.
• [31]孙春蕾.铜绿微囊藻和水华鱼腥藻间化感作用的研究[D].南京:南京大学,2018.
• [32]王蕊,黄文雯,吴佳俊,等.剧毒卡尔藻与东海原甲藻间的化感作用研究[J].海洋与湖沼,2019,50(4):858-863.
• [33]徐文静，王江涛.新月菱形藻和东海原甲藻间的化感作用研究[C]//中国第九届植物化感作用学术研讨会论文摘要集,杨凌:中国植物保护学会植物化感作用专业委员会,2019:35.
• [34]WANG T,HU Y D,ZHU M,et al.Integrated transcriptome and physiology analysis of Microcystis aeruginosa after exposure to copper sulfate[J].Journal of Oceanology and Limnology,2020,38(1):102-113.https://doi.org/10.1007/s00343-019-8357-9.
• [35]肖燕，姚珺玥，廖琼，等.拟南芥生态型Tor-1根系响应镉的转录组学与可变剪接分析[J].植物营养与肥料学报,2020,26(8):1513-1525.
• [36]翟秀明，唐敏，李解，等.基于RNA-Seq技术的茶树响应高温胁迫转录组差异性分析[J].分子植物育种,2020,18(17):5629-5637.
• [37]XIAO X,HUANG H M,GE Z W,et al.A pair of chiral flavonolignans as novel anti-cyanobacterial allelochemicals derived from barley straw (Hordeum vulgare):characterization and comparison of their anti-cyanobacterial activities[J].Environmental Microbiology,2014,16(5):1238-1251.
• [38]韩锡锡，于庆云，鞠莲，等.化感物质抑制短裸甲藻的机理研究[J].海洋开发与管理,2018,35(5):71-76.
• [39]KANEHISA M,GOTO S.KEGG:Kyoto encyclopedia of genes and genomes[J].Nucleic Acids Research,2000,28(1):27-30.
• [40]GUO R Y,LIM W A,KI J S.Genome-wide analysis of transcription and photosynthesis inhibition in the harmful dinoflagellate Prorocentrum minimum in response to the biocide copper sulfate[J].Harmful Algae,2016,57:27-38.
• [41]朱圣庚，徐长法，王镜岩.生物化学[M].3版.北京:高等教育出版社,2002.
• [42]KUNO S,BACHER A,SIMON H.Structure of enoate reductase from a Clostridium tyrobutyricum (C.spec.La1)[J].Biological Chemistry Hoppe-Seyler,1985,366(1):463-472.
• [43]WEST I C,MITCHELL P.The proton-translocating ATPase of Escherichia coli[J].FEBS Letters,1974,40(1):1-4.
• [44]CORTICEIRO S C,LIMA A I G,FIGUEIRA E M D A P.The importance of glutathione in oxidative status of Rhizobium leguminosarum biovar viciae under Cd exposure[J].Enzyme and Microbial Technology,2006,40(1):132-137.
• [45]LIN J.Stress responses of Acinetobacter strain Y during phenol degradation[J].Archives of Microbiology,2017,199(2):365-375.
• [46]SHAO J H,LIU D M,GONG D X,et al.Inhibitory effects of sanguinarine against the cyanobacterium Microcystis aeruginosa NIES-843 and possible mechanisms of action[J].Aquatic Toxicology,2013,142/143:257-263.
• [47]张礼.纳米银对典型水生生物的生物效应及毒性机理研究[D].上海:上海交通大学,2018.
• [48]MIWA S,ST-PIERRE J,PARTRIDGE L,et al.Superoxide and hydrogen peroxide production by Drosophila mitochondria[J].Free Radical Biology and Medicine,2003,35(8):938-948.
• [49]黄德玉.T-2毒素诱导的细胞线粒体毒性作用研究[D].武汉：华中农业大学，2019.
• [50]尹康.解偶联蛋白2在血管紧张素II致线粒体活性氧生成中的作用[D].南昌:南昌大学,2007.
• [51]FUENTES D,MENESES M,NUNES-NESI A,et al.Adeficiency in the flavoprotein of Arabidopsis mitochondrial complex II results in elevated photosynthesis and better growth in nitrogen-limiting conditions[J].Plant Physiology,2011,157(3):1114-1127.
• [52]BELT K,S B,THATCHER L F,et al.Salicylic acid-dependent plant stress signaling via mitochondrial succinate dehydrogenase[J].Plant Physiology,2017,173(4):2029-2040.
• [53]王盼，史文军，万夕和，等.低氧-复氧对脊尾白虾呼吸代谢和抗氧化酶活力的影响[J/OL].渔业科学进展,2020,:1-12.https://doi.org/10.19663/j.issn2095-9869.20200424002.
• [54]赵文鹏.氟损伤线粒体呼吸链致雌性小鼠生殖障碍的分子机制[D].洛阳：河南科技大学,2019.
• [55]ZUO S P,ZHOU S B,YE L T,et al.Synergistic and antagonistic interactions among five allelochemicals with antialgal effects on bloom-forming Microcystis aeruginosa[J].Ecological Engineering,2016,97:486-492.
• [56]ZHANG S L,ZHANG B,DAI W,et al.Oxidative damage and antioxidant responses in Microcystis aeruginosa exposed to the allelochemical berberine isolated from golden thread[J].Journal of Plant Physiology,2011,168(7):639-643.