CN113999773B - 一种耐高光的扁藻及其应用 - Google Patents
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Abstract
本发明涉及微生物技术领域,特别涉及一株耐高光的扁藻及其应用。本发明从水样中分离获得一株扁藻(Tetraselmis sp.)MEM‑A‑412,于2021年8月25日保藏在中国典型培养物保藏中心,保藏编号为CCTCC NO:M20211080。该藻株可最高适应250μmol·m‑2·s‑1的高光环境,可广泛用于中国南方地区养殖,为南方地区微藻的利用提供了广阔的前景。
Description
技术领域
本发明涉及微生物技术领域,特别涉及一种耐高光扁藻及其应用。
背景技术
Tetraselmis是绿藻门绿枝藻纲团藻目衣藻科扁藻属的一种运动很快的单细胞绿藻,具有较高的脂质储存能力,具有生产生物柴油油脂的巨大潜力。此外,还产生长链w-3脂肪酸,即二十二碳六烯酸(DHA)和二十碳五烯酸(EPA)。能进行光合放氧作用,繁殖速度快,并含有丰富的营养物质,是鱼、虾幼体和贝类的优良饵料。因扁藻特殊的化学组成和系统位置,受到国内、外学者的普遍关注。目前,在水产养殖的饵料生产上,人工培养扁藻技术受到广泛重视,国外已有室内培养和大规模培养的报道,国内也有一些相应的报道。如何提高扁藻细胞生长速率,缩短培养周期,是当前需要致力解决的关键问题。
扁藻在90~180μmol·m-2·s-1光强度下均能生长繁殖,是一种适应光照范围广的品种。但在南方地区如海南、广东、福建等地,太阳高度角较大,获得的太阳辐射较高,夏季的平均光照强度达500μmol·m-2·s-1,远远超过了光合作用可利用的水平,过量的光照会对微藻的光系统造成伤害,抑制光合作用,降低光合效率。因此,针对于南方沿海地区,扁藻的培养一直是个难题,大大限制了扁藻的产业化生产。
发明内容
有鉴于此,本发明提供了一种耐高光的扁藻及其应用,该藻株可最高适应250μmol·m-2·s-1的高光环境。
本发明从海口市海域的海水中分离获得藻株MEM412,经形态学和ITS rRNA鉴定为扁藻(Tetraselmissp.),其保藏编号为CCTCC NO:M20211080。
本发明还提供了所述扁藻的培养方法,将保藏编号为CCTCC NO:M20211080的扁藻接种于F2培养基中培养。
其中,所述培养为24~26℃,50~250μmol·m-2·s-1光照强度下培养5-10天。
所述F2培养基的组成为:
每1L所述平板培养基的组成如下:
每200ml所述营养液母液的组成为:
硝酸钠 40g
一水合磷酸二氢钠 2.66g
水 补足至200mL。
每100ml所述维生素母液的组成为:
每1L所述微量元素母液的组成为:
每10ml所述抗生素溶液的组成为:
氨苄青霉素钠 1mg
头孢氨噻肟 1mg
水 补足至10mL。
本发明还提供了所述的扁藻在制备生物柴油或微生态制剂中的应用。本发明还提供一种微生态制剂,包括所述的扁藻MEM-A-412。
本发明还提供了所述的扁藻在制备水产动物饲料中的应用。
所述水产动物包括鱼、虾幼体和贝类。
本发明还提供一种水产动物饲料,包括本发明所述的扁藻MEM-A-412。
本发明还提供了一种饲喂水产动物的方法,其特征在于,向水产动物投喂所述的水产动物饲料。
本发明提供的扁藻MEM-A-412在高光条件下依然能够保持较高的生长速度,最高适应250μmol·m-2·s-1的高光环境,可广泛用于中国南方地区养殖,为南方地区微藻的利用提供了广阔的前景。
生物保藏说明
扁藻MEM-A-412(Tetraselmis sp.MEM-A-412),于2021年8月25日保藏在中国典型培养物保藏中心,地址为:中国,武汉,武汉大学,保藏编号为CCTCC NO:M20211080。
附图说明
图1为扁藻MEM-A-412与其他藻类的生长曲线对比结果;
图2为扁藻MEM-A-412与其他藻类的干重对比结果。
具体实施方式
本发明公开了一种耐高光的扁藻及其应用,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
下面结合实施例,进一步阐述本发明:
实施例1
1、从海口市周边海域的养殖水体中,采集微藻藻液,通过平板划线及涂布法,分离出较纯的单藻落(纯度80%以上),分离获得的微藻MEM-A-412经ITS rRNA鉴定为扁藻。
其中,平板培养基配置如下:
表1营养液母液配方(200mL)
表2维生素母液配方(100mL)
物料名称 | 分子式 | 级别 | 称量量(mg) |
维生素B12 | Vitamin B12 | AR | 1 |
生物素 | Biotin | AR | 1 |
维生素B1 | Vitamin B1 | AR | 20 |
表3微量元素溶液母液配方(1L)
物料名称 | 分子式 | 级别 | 称量量(g) |
乙二胺四乙酸二钠 | Na2EDTA | AR | 847 |
六水合氯化铁 | FeCl3.6H2O | AR | 730 |
五水硫酸铜 | CuSO4·5H2O | AR | 0.784 |
七水硫酸锌 | ZnSO4·7H2O | AR | 1.76 |
六水氯化钴 | CoCl2·6H2O | AR | 0.4368 |
四水氯化锰 | MnCl2·4H2O | AR | 14.4 |
二水钼酸钠 | Na2MoO4·2H2O | AR | 0.504 |
表4抗生素配方(10mL)
将15g琼脂溶入1L纯化水中,加入35gNaCl,加热溶解后放入灭菌锅中121℃,灭菌20min。灭菌完成后趁热在超净工作台上用移液器吸取5mL营养液母液,0.5mL维生素母液,1mL微量元素溶液,1mL抗生素加入琼脂液中,摇匀,封口,待冷却。(营养液母液及微量元素溶液均需要提前灭菌,121℃,20min,0.22膜过滤除菌)。
2、将平板中的MEM-A-412单藻落,挑选至30ml F/2液体培养基中进行培养,每个单藻落挑选入一个50ml锥形瓶中。培养所需培养基为液体,与平板培养基基本一致,不加琼脂。将挑选好的试管放置在光照培养箱中进行培养。试管培养温度;25(±1)℃,光照强度50μmol·m-2·s-1,培养5-10天。
3、将培养一段时间后的藻体离心收集,取藻泥接种于无菌F/2液体培养基,分别置于温度25℃、光强50μmol·m-2·s-1和25℃、光强250μmol·m-2·s-1在光照时间24h的培养箱中培养。
4、以微拟球藻IMET1作为对照,按照步骤1~3进行培养。
5、于2、4、6、8d测定两组(试验组和对照组)的D750nm;于0、1、3、6、12、24、48、96h测定各组的Fv/Fm,结果见图1,于第8d测定各组干重,结果见图2。
在正常培养光照强度50μmol·m-2·s-1和高光光强250μmol·m-2·s-1培养下MEM-A-412的生长比微拟球藻IMET1快,而且培养10天后的干重也比IMET1高。MEM-A-412在250μmol·m-2·s-1的光强下长得更快且生物量累积更高。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.保藏编号为CCTCC NO:M20211080的扁藻(Tetraselmis sp.)。
2.权利要求1所述的扁藻的培养方法,其特征在于,将保藏编号为CCTCC NO:M20211080的扁藻接种于F2培养基中培养。
3.根据权利要求2所述扁藻的培养方法,其特征在于,所述培养为:24~26℃,50~250μmol·m-2·s-1光照强度下培养5-10天。
4.权利要求1所述的扁藻在制备生物柴油或微生态制剂中的应用。
5.一种微生态制剂,其特征在于,包括权利要求1所述的扁藻。
6.权利要求1所述的扁藻在制备水产动物饲料中的应用。
7.根据权利要求6所述的应用,其特征在于,所述水产动物包括鱼、虾幼体和贝类。
8.一种水产动物饲料,其特征在于,包括权利要求1所述的扁藻。
9.一种饲喂水产动物的方法,其特征在于,向水产动物投喂权利要求8所述的水产动物饲料。
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Non-Patent Citations (3)
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产氢扁藻的培养优化;刘润国;中国优秀硕士学位论文 工程科技Ⅰ辑(第08期);第1-88页 * |
利用叶绿素荧光监测微藻的生长及生理状态;张文蕾 等;激光生物学报;第25卷(第1期);第48-55页 * |
单胞藻静置保种技术研究;王云鹏 等;齐鲁渔业;第14卷(第4期);第29-30页 * |
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