CN114793875B - 一种诱导细齿麒麟菜侧枝发生的方法 - Google Patents
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Abstract
本发明提供了一种诱导麒麟菜侧枝发生的方法,涉及海洋生物技术领域。本发明在200‑300μmol·m‑2·s‑1的光照条件下对麒麟菜进行通气培养。在高光强诱导期间,不断有新的侧枝长出,高光处理结束后,即可将麒麟菜重新置于原养殖环境。通过高光强诱导麒麟菜藻枝细胞脱分化成为分生组织,进而发育成侧枝,侧枝的增加将会提高麒麟菜的产量,同时高光处理会显著增加麒麟菜干物质含量,增加收益。
Description
技术领域
本发明属于海洋生物技术领域,具体涉及一种诱导细齿麒麟菜侧枝发生的方法。
背景技术
麒麟菜族海藻是世界范围内产量最大的藻类养殖品种,包括卡帕藻属(Kappaphycus)、麒麟菜属(Eucheuma)和琼枝藻属(Betaphycus),主要养殖于东南亚等热带-亚热带海域,是工业提取卡拉胶重要的原料藻。我国是世界上卡拉胶产量最大的国家,但麒麟菜养殖规模逐年萎缩,长期处于低水平,原料藻几乎全部依赖于进口。2020年我国麒麟菜的产量为3856吨(干重),仅占国内藻类养殖总产量的0.147%。
细齿麒麟菜(Eucheuma denticulatum)是麒麟菜属中重要的养殖品种,藻体直立,具多分支结构。海南省具有广袤的热带海域具有得天独厚的养殖条件,但目前细齿麒麟菜养殖模式单一,均以营养繁殖的方式进行,缺乏野生种质资源的补充,导致养殖群体遗传多样性低,环境适应性下降,使得产量逐年萎缩,亟需新的养殖模式改变这一情况。
分支作为农业生产中重要的性状,如禾本科作物分蘖,其与作物生物量及产量直接相关。同样,对于多侧枝的多细胞红藻,诱导侧枝的发生能够显著提升单位水体细齿麒麟菜的产量,但是目前并没有可促进多细胞红藻侧枝发生的方法。
发明内容
有鉴于此,本发明的目的在于提供一种诱导细齿麒麟菜侧枝发生的方法,高效诱导多细胞细齿麒麟菜产生侧枝,提高单位水体的养殖产量。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种诱导细齿麒麟菜(E. denticulatum)侧枝发生的方法,包括以下步骤:在200-300μmol·m-2·s-1的光照条件下对红藻进行通气培养。
优选的,所述通气培养在PES(Provasoli’s enrichment solution)培养基上进行。
优选的,所述光照条件包括每天连续的12h光照和连续12h黑暗。
优选的,所述通气培养的时间为至少10d。
有益效果:本发明提供了一种诱导细齿麒麟菜侧枝发生的方法,在200-300μmol·m-2·s-1的光照条件下对细齿麒麟菜进行通气培养。在高光强诱导期间,不断有新的侧枝长出,高光处理结束后,即可将细齿麒麟菜重新置于原养殖环境。通过高光强诱导细齿麒麟菜藻枝细胞脱分化成为分生组织,进而发育成侧枝,侧枝的增加将会提高细齿麒麟菜的产量,同时高光处理会显著增加细齿麒麟菜干物质含量,增加收益。
附图说明
图1为不同光强下诱导侧枝累积发生数;图中a、b、c表示在第10天,不同光照强度下单位厘米侧枝累积增加数的显著性差异;
图2为第10天干物质含量(%);图中a、b、c表示在第10天,不同光照强度下干物质含量的显著性差异;
图3为同一株细齿麒麟菜先后在不同光照强度下培养10d的状态,其中A的光照强度为50μmol·m-2·s-1,B为300μmol·m-2·s-1;
图4为同一株细齿麒麟菜先后在不同光照强度下培养10d的状态,其中A的光照强度为50μmol·m-2·s-1,B为300μmol·m-2·s-1;
图5为不同生物量的细齿麒麟菜经300μmol·m-2·s-1的高光处理15天后的图片。
具体实施方式
本发明提供了一种诱导细齿麒麟菜侧枝发生的方法,包括以下步骤:在200-300μmol·m-2·s-1的光照条件下对细齿麒麟菜进行通气培养。
本发明在高光强下对细齿麒麟菜进行侧枝诱导,所述诱导优选在PES培养基上进行,且培养至少10d。本发明所述PES培养基优选包括NaNO3 2.35g/L、Na2glycerophosphate·5H2O 0.35g/L、ES Fe Solution 162.5mL/L、P-II MetalSolution 162.5mL/L和HEPES buffer 3.25g/L,本发明对所述PES培养基的配制方法并没有特殊限定,优选包括将上述原料依次加入水中搅拌,调节pH至7.8,定容至1L后,于121℃0.1MPa灭菌20min,4℃保存即可。本发明所述ES Fe Solution优选包括以下浓度的组分:Fe(NH4)2(SO4)2·6H2O 0.7g/L和Na2EDTA·2H2O 0.6g/L;所述P-II Metal Solution 优选包括以下浓度的组分:Na2EDTA·2H2O 1g/L、H3BO3 1.14g/L、FeCl3·6H2O 0.049g/L、MnSO4·H2O 0.164g/L、ZnSO4·7H2O 0.022g/L和CoCl2·6H2O 0.0048 g/L。
本发明在进行所述诱导时,优选的包括每天连续的12h光照和连续12h黑暗,并且保持通气培养,本发明对所述通气培养的通气量并没有特殊限定。在本发明中,在高光强诱导期间,不断有新的侧枝长出,高光处理结束后,即可将细齿麒麟菜重新置于原养殖环境。通过高光强诱导细齿麒麟菜藻枝细胞脱分化成为分生组织,进而发育成侧枝,侧枝的增加将会提高细齿麒麟菜的产量,同时高光处理会显著增加细齿麒麟菜干物质含量,增加收益。
下面结合实施例对本发明提供的一种诱导细齿麒麟菜侧枝发生的方法进行详细的说明,但是不能把它们理解为对本发明保护范围的限定。
实施例1
细齿麒麟菜分别在50、100、200、300μmol·m-2·s-1的光照条件下进行通气培养10天,每种条件下做四个生物学重复。
测定初始的侧枝数,记作第0天,每两天对单位厘米侧枝发生数进行统计。
诱导侧枝发生数计算方式:第n天侧枝数/第n天长度-第0天侧枝数/第0天长度。
干物质含量测定:藻体用吸水纸擦干后的重量为鲜重m1,藻体置于60℃烘箱烘至重量不再变化,为干重m2,干物质含量为:(m2/m1)×100%。
不同光强下诱导侧枝发生数如表1和图1所示:第10天时,300μmol·m-2·s-1光强下的藻体单位厘米侧枝增加数达到7.62±1.16个,显著高于其他光强下的侧枝增加数(P<0.01)。说明300μmol·m-2·s-1的高光可以高效的诱导细齿麒麟菜侧枝的发生。
备注:a,b,c表示在第10天,不同光照强度下单位厘米侧枝累积增加数的显著性差异。
第10天干物质含量如表2和图2所示,随着光照强度的增加,藻体干物质含量呈上升趋势。300μmol·m-2·s-1光强下的藻体干物质含量达到16.95±0.34%,显著高于其他光强下的干物质含量(P<0.01)。说明300μmol·m-2·s-1的高光能有效提高藻体干物质含量。
对同一株细齿麒麟菜在50μmol·m-2·s-1的光照条件下培养10d后进行拍照,结果如图3所示;而后将该株细齿麒麟菜转至300μmol·m-2·s-1的光照条件下培养10d后进行拍照,结果如图4所示,在300μmol·m-2·s-1的光照条件下培养10d,可显著提高侧枝数量;在300μmol·m-2·s-1的光照条件下培养15d,进行拍照,如图5所示,侧枝数量更进一步提高。
表2 不同光强下第10天干物质含量
光强(μmol·m<sup>-2</sup>·s<sup>-1</sup>) | 50 | 100 | 200 | 300 |
干物质含量(%) | 12.75±0.12 | 13.85±0.23 | 14.13±0.28 | 16.95±0.34 |
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 (2)
1.一种诱导细齿麒麟菜(Eucheuma denticulatum)侧枝发生的方法,其特征在于,包括以下步骤:在300μmol·m-2·s-1的光照条件下对细齿麒麟菜进行通气培养;
所述通气培养在PES培养基上进行;
所述光照条件包括每天连续的12h光照和连续12h黑暗;
所述PES培养基由以下组分组成:NaNO3 2.35g/L、C3H7Na2O6P·5H2O 0.35g/L、ES Fe 溶液 162.5mL/L、P-II 金属溶液 162.5mL/L和HEPES 缓冲液 3.25g/L;
所述ES Fe 溶液由以下浓度的组分组成:Fe(NH4)2(SO4)2·6H2O 0.7g/L和Na2EDTA·2H2O 0.6g/L;
所述P-II 金属溶液由以下浓度的组分组成:Na2EDTA·2H2O 1g/L、H3BO3 1.14g/L、FeCl3·6H2O 0.049g/L、MnSO4·H2O 0.164g/L、ZnSO4·7H2O 0.022g/L和CoCl2·6H2O0.0048 g/L。
2.根据权利要求1所述方法,其特征在于,所述通气培养的时间为至少10d。
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