微绿球藻(Nannochloropsis oculata)培养基的优化
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摘要
【目的】微绿球藻(Nannochloropsis oculata)是一种优良的单细胞饵料微藻,广泛应用于水产养殖。优化微绿球藻培养基,以期提高其生长速率,降低生产成本。【方法】以宁波大学3#配方为基础微藻培养液,以乙酸钠为碳源,硝酸钾、尿素和氯化铵为氮源,磷酸二氢钾和磷酸二氢钠为磷源,硫酸亚铁和柠檬酸铁为铁源,通过单因子和正交试验,研究了碳、氮、磷、铁、维生素B_1(VB_1)和B_(12)(VB_(12))等主要营养元素对微绿球藻生长、繁殖的影响。【结果】获得了以天然海水为基础的微绿球藻优化培养基:3 g/L CHCOONa、20 mg/L NH_4CL-N、2 mg/L KH_2PO_4-P、3 mg/L FeSO_4-Fe、0.05 mg/L VB_1和 0.005mg/L VB_(12);采用优化培养基与宁波大学3#培养基对比培养微绿球藻,结果表明,培养 2～6 d,优化培养基收获微绿球藻的生物量( 细胞密度)比宁波大学 3#培养基的分别提高了2.21、2.55、2.30、1.97、1.7倍;培养6 d,优化培养基中微绿球藻收获的生物量(细胞密度)达到1.74×10~7个/mL,是宁波大学3#培养基的1.7倍。【结论】优化培养基极显著地提高了微绿球藻的生物量,是微绿球藻的良好培养基。
【Objective】Nannochloropsis oculata is a kind of excellent single-celled diet microalgae, and widely used in aquaculture.The culture medium of Nannochloropsis oculata was optimized in order to improve its growth rate and reduce the production costs. 【Method】Taking Ningbo University 3# Formula as the basic microalgae culture medium,with CHCOONa as carbon source, KNO_3,(NH_2)_2 CO and NH_4 CL as nitrogen source, KH_2 PO_4 and NaH_2 PO_4 as phosphorus source, FeSO_4 and FeC_6 H_5 O_7 as iron source, the effects of different nutrient sources such as carbon, nitrogen, phosphorus,iron,Vitamin B_1 and Vitamin B_(12) on the growth and propagation of N. oculata were analyzed by one-factor experiment and orthogonal test. 【Result】The optimized culture medium based on nature sea water was formulated as follows: 3 g/L CHCOONa, 20 mg/L NH_4 Cl-N, 2 mg/L KH_2 PO_4-P, 3 mg/L FeSO_4-Fe, 0.05 mg/L Vitamin B_1 and 0.005 mg/L Vitamin B_(12). The N.oculata was cultured on optimized medium and Ningbo University 3# medium and the results showed that the biomass(cell density) of N. oculata cultured on optimized medium for 2-6 days was 2.21, 2.55, 2.30, 1.97 and 1.7 timeshigher than that of Ningbo University 3# Medium respectively. By the end of the sixth day, the biomass(cell density) of N. oculata on the optimized medium was up to 1.74×10~7 cell/mL, which was 1.7 times higher than that on Ningbo University 3# medium. 【Conclusion】The optimized medium extrenely enhamced the biomass of N. oculata and it was a good medium for N. oculata.
【Objective】Nannochloropsis oculata is a kind of excellent single-celled diet microalgae, and widely used in aquaculture.The culture medium of Nannochloropsis oculata was optimized in order to improve its growth rate and reduce the production costs. 【Method】Taking Ningbo University 3# Formula as the basic microalgae culture medium,with CHCOONa as carbon source, KNO_3,(NH_2)_2 CO and NH_4 CL as nitrogen source, KH_2 PO_4 and NaH_2 PO_4 as phosphorus source, FeSO_4 and FeC_6 H_5 O_7 as iron source, the effects of different nutrient sources such as carbon, nitrogen, phosphorus,iron,Vitamin B_1 and Vitamin B_(12) on the growth and propagation of N. oculata were analyzed by one-factor experiment and orthogonal test. 【Result】The optimized culture medium based on nature sea water was formulated as follows: 3 g/L CHCOONa, 20 mg/L NH_4 Cl-N, 2 mg/L KH_2 PO_4-P, 3 mg/L FeSO_4-Fe, 0.05 mg/L Vitamin B_1 and 0.005 mg/L Vitamin B_(12). The N.oculata was cultured on optimized medium and Ningbo University 3# medium and the results showed that the biomass(cell density) of N. oculata cultured on optimized medium for 2-6 days was 2.21, 2.55, 2.30, 1.97 and 1.7 timeshigher than that of Ningbo University 3# Medium respectively. By the end of the sixth day, the biomass(cell density) of N. oculata on the optimized medium was up to 1.74×10~7 cell/mL, which was 1.7 times higher than that on Ningbo University 3# medium. 【Conclusion】The optimized medium extrenely enhamced the biomass of N. oculata and it was a good medium for N. oculata.
引文
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[2]陈书秀,刘学迁,王青岩,李晓捷,曲善村.微绿球藻作为刺参幼体饵料的可行性研究[J].河北渔业,2014(7):53-55,66.doi:10.3969/j.issn.1004-6755.2014.07.020.CHEN S X,LIU X Q,WANG Q Y,LI X J,QU S C.Study on the feasibility of Nannochloris oculata as a feed for larvae sea cucumber Stichopus japonicus[J].Hebei Fisheries,2014(7):53-55,66.doi:10.3969/j.issn.1004-6755.2014.07.020.
[3]邓正华,陈明强,王雨,黄桂菊,于刚,吴开畅,李有宁.合浦珠母贝幼虫培育的饵料优化研究[J].水产科学,2018,37(6):721-727.doi:10.16378/j.cnki.1003-1111.2018.06.001.DENG Z H,CHEN M Q,WANG Y,HUANG G J,YU G,WU K C,LI YN.Optimization of species,density and combination of microalgae as food for larval pearl oyster Pinctada fucata[J].Fisheries Science,2018,37(6):721-727.doi:10.16378/j.cnki.1003-1111.2018.06.001.
[4]成文靖,蔡春芳.微绿球藻在罗氏沼虾育苗中的应用研究[J].水利渔业,2002,22(1):15-16.doi:10.15928/j.1674-3075.2002.01.010.CHENG W J,CAI C F.Application of Nannochloris oculata in the seedling of Macrobrachium rosenbergii[J].Journal of Hydroecology,20 02,22(1):15-16.doi:10.15928/j.1674-3075.2002.01.010.
[5]高腾,陈建酬.微绿球藻对罗氏沼虾幼体发育的影响[J].江西农业学报,2017,29(6):90-93.doi:10.19386/j.cnki.jxnyxb.2017.06.20.GAO T,CHEN J C.Influence of Nannochloris oculata on development of Macrobrachium rosenbergii larvae[J].Acta Agriculturae Jiangxi,2017,29(6):90-93.doi:10.19386/j.cnki.jxnyxb.2017.06.20.
[6]沈和定,黄旭雄.三种藻类对中华绒螯蟹I期溞状幼体培育效果的比较[J].上海水产大学学报,1999,8(3):202-209.SHEN H D,HUANG X X.Study on Metamorphosis and survival rate of zoea I of Eriocheir sinensis fed on Nannochloropsis oculata,Phaeodactylum tricornutum and powder of Spirulina platensis[J].Journal of Shanghai Fisheries University,1999,8(3):202-209.
[7]刘新富,雷霁霖,刘忠强,柳学周.真鲷饵料生物褶皱臂尾轮虫和眼点拟微绿球藻的大量培养[J].海洋科学,2002,24(5):47-51.LIU X F,LEI J L,LIU Z Q,LIU X Zl.Mass culture of Nannochloropsis oculata and rotifer Branchionus plicatilis as living diets for red sea bream[J].Marine Sciences,2002,24(5):47-51.
[8]LUBZENS E,GIBSON O,ZNIRA O,SUKENIK A.Potential advantages of frozen algae(Nannochloropsis sp.)for rotifer(Brachionus plicatilis)culture[J].Aquaculture,1995,133:295-309.
[9]CHBIL L,YAMASAKI S.Improvement of a rortifer ecosystem culture to promote recycling marine microalga,Nannochloropsis sp[J].Aquacultural Eng,1998,17:1-10.
[10]陈洁,蒋霞敏,段舜山.眼点拟微绿球藻生长的生态因子分析[J].生态科学,2002(1):50-52.CHEN J,JIANG X M,DUAN S S.Analysis of growth factor of Nannochloris oculata[J].Ecologic Science,2002(1):50-52.
[11]季祥,乔岩,成杰,刘鲜艳,赵昕宇,郑添慧,徐迪华,蔡禄.拟微绿球藻生长条件优化[J].江苏农业科学,2017,45(2):154-156.doi:10.15889/j.issn.1002-1302.2017.02.046.JI X,QIAO Y,CHENG J,LIU X Y,ZHAO X Y,ZHENG T H,XU D H,CAIL.Optimization of cultural conditions of Nannochloris oculata[J].Jiangsu Agricultural Sciences,2017,45(2):154-156.doi:10.15889/j.issn.1002-1302.2017.02.046.
[12]张海琪,蒋霞敏,潘双叶,赵明忠.光照、温度、碳源及接种密度对微绿球藻生长的影响[J].河北渔业,2001(1):9-13.ZHANG H Q,JIANG X M,PAN S Y,ZHAO M Z.Effect of light intensity,temperature,carbon source and vaccination density on the growth of Nannochloris oculata[J].Hebei Fisheries,2001(1):9-13.
[13]黄翔鹄,李长玲,刘楚吾,王中铎,陈建军.微绿球藻对氮和磷营养盐需求的研究[J].海洋科学,2002,26(8):13-17.HUANG X H,LI C L,LIU C W,WANG Z D,CHEN J J.Studies on the N and P nutrient demand of Nannochloris oculata[J].Marine Sciences,2002,26(8):13-17.
[14]丁丽欢,王珺,严虹羽,符新欢.热带小球藻培养模式的筛选及其培养基的优化[J].微生物学通报,2018,45(5):981-989.doi:10.13344/j.microbiol.china.170596.DING L H,WANG J,YAN H Y,FU X H.Optimization of tropical Chlorella vulgaris culture method and its medium formulation[J].Microbiology China tongbao,2018,45(5):981-989.doi:10.13344/j.microbiol.china.170596.
[15]王珺,王永强,陈国华,王军建,严虹羽,陈鸿林.生态和营养条件对日本星杆藻生长的影响[J].海洋渔业,2014,36(4):329-334.doi:10.13233/j.cnki.mar.fish.2014.04.001.WANG J,WANG Y Q,CHEN G H,WANG J J,YAN H Y,CHEN H Ll.Effects of ecological andnutritional factors on growth of Asterionella japonica[J].Marine Fisheries,2014,36(4):329-334.doi:10.13233/j.cnki.mar.fish.2014.04.001.
[16]胡章喜,安民,段舜山,徐宁,孙凯峰,刘晓娟,李爱芬,张成武.不同氮源对布朗葡萄藻生长、总脂和总烃含量的影响[J].生态学报,2009,29(6):3288-3294.HU Z X,AN M,DUAN S S,XU N,SUN K F,LIU X J,LI A F,ZHANGC W.Effects of nitrogen sources on the growth,contents of total lipids and total hydrocarbons of Botryococcus braunii[J].Acta Ecologica Sinica,2009,29(6):3288-3294.
[17]潘庭双,胡贤江,侯冠军,蒋业林,李海洋.氮对微绿球藻生长的影响[J].安徽农业科学,2001,29(4):548-555.doi:10.13989/j.cnki.0517-6611.2001.04.060.PAN T S,HU X J,HOU G J,JIANG Y L,LI H Y.Effect of Nitrogen Application on the growth of Nannochloropsis oculata[J].Journal of Anhui Agricultural Sciences,2001,29(4):548-555.doi:10.13989/j.cnki.0517-6611.2001.04.060.
[18]王丽卿,黄旭雄.不同营养盐浓度下微绿球藻的生长及水体中氮磷的变化[J].上海水产大学学报,2002,11(3):215-218.WANG L Q,HUANG X X.The growth of Nannochloropsis oculata cultured in different nutrient concentrations and the variations of nitrogen and phosphorus in culture tanks[J].Journal of Shanghai Fisheries University,2002,11(3):215-218.
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