陈张翔,李淼淼,张达娟,程旭,姜曼,毕相东

• 1:天津农学院水产学院/天津市水产生态及养殖重点实验室
• 2:天津大来光电科技有限公司

摘要(Abstract)：

【目的】研究光质对雨生红球藻（Haematococcus pluvialis）的生长影响机制，为雨生红球藻的高效优质培养提供理论参考。【方法】将雨生红球藻置于LED白光（428～665 nm）（对照组）、LED红光（621～641 nm）和LED蓝光（452～459 nm）下培养8 d，测定并分析其藻细胞密度，叶绿素a、类胡萝卜素、虾青素质量浓度及蛋白表达等指标。【结果】培养至8 d时，蓝光组和红光组藻细胞密度分别为15.87×10~5 mL~(-1)和15.67×10~5 mL~(-1)，均极显著高于对照组（11.37×10~5 mL~(-1)）(P <0.01)；蓝光组雨生红球藻叶绿素a、类胡萝卜素和虾青素质量浓度分别为5.25、2.78和2.67μg·mL~(-1)，均显著高于红光组和对照组（P <0.05）。雨生红球藻细胞的定量蛋白质组学分析表明，红光组和对照组组间共鉴定出6 437个差异表达的蛋白，其中67个上调，126个下调，差异显著通路为卟啉与叶绿素代谢、糖酵解/糖异生；蓝光组和对照组间共鉴定出6 385个差异表达的蛋白，其中228个上调，649个下调，差异显著通路为氨酰-tRNA生物合成和光合作用-触角蛋白；蓝光组和红光组间共鉴定出6 433个差异表达的蛋白，但无差异显著的通路。亚细胞定位发现，在不同光质处理条件下，差异蛋白主要定位于叶绿体蛋白，红光与对照组之间的差异蛋白中有25.0%定位于叶绿体蛋白，蓝光与对照组间为26.9%，蓝光与红光组间为21.6%。【结论】红光与蓝光均可有效提高雨生红球藻的生物量，红光可下调藻细胞内的镁螯合酶亚基的表达，从而降低叶绿素a的质量浓度；8 d蓝光照射可对雨生红球藻造成更严重的光损伤，使其细胞内叶绿素a/b结合蛋白下调，从而促使雨生红球藻积累更多的虾青素。

关键词(KeyWords)： 雨生红球藻;光质;光合色素;虾青素;蛋白组学

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金（31170442）;; 天津市教委科研计划（2021KJ110,2020ZD06）;; 天津市淡水养殖产业技术体系创新团队-养殖水环境调控岗位项目（ITTFRS2021000-009）

作者(Author): 陈张翔,李淼淼,张达娟,程旭,姜曼,毕相东

参考文献(References)：

• [1]陈涛,沼泽彻,杨福梅,等.雨生红球藻中天然虾青素的应用研究[J].农业工程技术, 2023, 43(15):76-77.
• [2]田子豪,陈倩,刘乐丹,等.虾青素在水产养殖中的应用[J].饲料研究, 2023, 46(17):142-145.
• [3]李淼淼,陈张翔,王其鹏,等.响应面法优化雨生红球藻培养基[J].东北农业大学学报, 2023, 54(11):56-65.
• [4] CHEN Z, CHEN J, LIU J H, et al. Transcriptomic and metabolic analysis of an astaxanthin-hyperproducing Haematococcus pluvialis mutant obtained by low-temperature plasma(LTP)mutagenesis under high light irradiation[J].Algal Research, 2020, 45:101746.
• [5] SUN Y H, LIU J G, ZHANG X L, et al. Strain H2-419-4 of Haematococcus pluvialis induced by ethyl methanesulphonate and ultraviolet radiation[J]. Chinese Journal of Oceanology and Limnology, 2008, 26(2):152-156.
• [6] PANIS G, CARREON J R. Commercial astaxanthin production derived by green alga Haematococcus pluvialis:a microalgae process model and a techno-economic assessment all through production line[J]. Algal Research, 2016, 18:175-190.
• [7]孙双,张婷,方琰,等.雨生红球藻高产虾青素的培养条件优化[J].现代食品科技, 2021, 37(6):98-107.
• [8] WAN M X, ZHANG J K, HOU D M, et al. The effect of temperature on cell growth and astaxanthin accumulation of Haematococcus pluvialis during a light–dark cyclic cultivation[J]. Bioresource Technology, 2014, 167:276-283.
• [9] TRIPATHI U, SARADA R, RAVISHANKAR G A. Effect of culture conditions on growth of green alga:Haematococcus pluvialis and astaxanthin production[J]. Acta Physiologiae Plantarum, 2002, 24(3):323-329.
• [10] LIYANAARACHCHI V C, NISHSHANKA G K S H,PREMARATNE R G M M, et al. Astaxanthin accumulation in the green microalga Haematococcus pluvialis:Effect of initial phosphate concentration and stepwise/continuous light stress[J]. Biotechnology Reports, 2020, 28:e00538.
• [11]周泽鹏.促进雨生红球藻积累虾青素的方法研究[D].扬州:扬州大学, 2024.
• [12]戴颖,穆亮亮,黄旭雄.一株新分离雨生红球藻培养条件的优化[J].上海海洋大学学报, 2023, 32(1):68-78.
• [13] CHEN F, XU N J, LIU K, et al. Increasing production and bio-accessibility of natural astaxanthin in Haematococcus pluvialis by screening and culturing red motile cells under high light condition[J]. Bioresource Technology, 2022,364:128067.
• [14] LI L M, WU Y H, ACHEAMPONG A, et al. Red light promotes autotrophic growth of Haematococcus pluvialis with improved carbonic anhydrase activity and CO2utilization[J]. Aquaculture, 2023, 571:739462.
• [15] GAO X, WANG X Y, LI H J, et al. Parameterization of a light distribution model for green cell growth of microalgae:Haematococcus pluvialis cultured under red LED lights[J]. Algal Research, 2017, 23:20-27.
• [16]李腊梅.光照对雨生红球藻生长和虾青素积累的影响及机理研究[D].合肥:中国科学技术大学, 2023.
• [17] MA R J, THOMAS-HALL S R, CHUA E T, et al. Blue light enhances astaxanthin biosynthesis metabolism and extraction efficiency in Haematococcus pluvialis by inducing haematocyst germination[J]. Algal Research, 2018,35:215-222.
• [18] LI K, YE Q, LI Q Y, et al. Effects of the spatial and spectral distribution of red and blue light on Haematococcus pluvialis growth[J]. Algal Research, 2020, 51:102045.
• [19] PANG N, FU X, FERNANDEZ J S M, et al. Multilevel heuristic LED regime for stimulating lipid and bioproducts biosynthesis in Haematococcus pluvialis under mixotrophic conditions[J]. Bioresource Technology, 2019, 288:121525.
• [20]陈锋,胡朝阳,孙雪,等. microRNA介导雨生红球藻在高光胁迫下次级细胞壁形成和虾青素积累的分子机制[J].海洋与湖沼, 2023, 54(3):763-772.
• [21] MA R J, THOMAS-HALL S R, CHUA E T, et al. Gene expression profiling of astaxanthin and fatty acid pathways in Haematococcus pluvialis in response to different LED lighting conditions[J]. Bioresource Technology, 2018, 250:591-602.
• [22] WANG B B, PAN X S, WANG F, et al. Photoprotective carbon redistribution in mixotrophic Haematococcus pluvialis under high light stress[J]. Bioresource Technology,2022, 362:127761.
• [23] SHAH M M R, LIANG Y M, CHENG J J, et al. Astaxanthinproducing green microalga Haematococcus pluvialis:from single cell to high value commercial products[J]. Frontiers in Plant Science, 2016, 7:531.
• [24]李泽.雨生红球藻响应亚硒酸钠胁迫的蛋白组学研究[D].深圳:深圳大学, 2016.
• [25]戴立洲,成小英,俞珊,等.豆瓣菜有机提取物对铜绿微囊藻的抑制及成分初步分离[J].环境科学学报, 2015, 35(12):4159-4168.
• [26] WANG J, ZHAO F, CHEN B H, et al. Small water clusters stimulate microcystin biosynthesis in cyanobacterial Microcystis aeruginosa[J]. Journal of Applied Phycology, 2013, 25(1):329-336.
• [27] LEHMUSKERO A, SKOGEN CHAUTON M, BOSTR?M T. Light and photosynthetic microalgae:a review of cellular-and molecular-scale optical processes[J]. Progress in Oceanography, 2018, 168:43-56.
• [28]甘钰华,魏群,马湘蒙,等.光质对小球藻(Chlorella sorokiniana)生长和代谢机制的调控[J].中国环境科学,2022, 42(9):4296-4303.
• [29]张培书.不同单色光对雨生红球藻叶绿素荧光参数及ROS的影响[D].昆明:云南师范大学, 2021.
• [30] LI Y X, CAI X H, GU W H, et al. Transcriptome analysis of carotenoid biosynthesis in Dunaliella salina under red and blue light[J]. Journal of Oceanology and Limnology,2020, 38(1):177-185.
• [31]朱琴,张春辉,石颖,等.光质调控雨生红球藻虾青素合成及油脂积累的效应[J].植物生理学报, 2021, 57(9):1778-1788.
• [32] XIONG Q, FENG J, LI S T, et al. Integrated transcriptomic and proteomic analysis of the global response of Synechococcus to high light stress[J]. Molecular&Cellular Proteomics, 2015, 14(4):1038-1053.
• [33] WEI L, YOU W X, GONG Y H, et al. Transcriptomic and proteomic choreography in response to light quality variation reveals key adaption mechanisms in marine Nannochloropsis oceanica[J]. Science of The Total Environment, 2020,720:137667.
• [34] LI X W, ZHU Y L, CHEN C Y, et al. Cloning and characterization of two chlorophyll A/B binding protein genes and analysis of their gene family in Camellia sinensis[J].Scientific Reports, 2020, 10(1):4602.
• [35] LIU C, ZHANG Y J, CAO D R, et al. Structural and functional analysis of the antiparallel strands in the lumenal loop of the major light-harvesting chlorophyll a/b complex of photosystem II(LHCIIb)by site-directed mutagenesis[J].Journal of Biological Chemistry, 2008, 283(1):487-495.
• [36] TANAKA A, TANAKA R. Chlorophyll metabolism[J].Current Opinion in Plant Biology, 2006, 9(3):248-255.
• [37]刘子琦,汪佳.镁螯合酶的结构与功能[J].中国生物化学与分子生物学报, 2018, 34(1):18-24.