ICS 65.020 CCS B 16 湖 DB43 南 省 地 方 标 准 DB43/T 1 9 69—2020 穗状狐尾藻实验室培养技术规程 Myriophyllum Verticillatum L. Laboratory Culture Technology Procedures 2020-12-29发布 湖南省市场监督管理局 2021-03-29实施 发 布 DB43/T 1969—2020 目 次 前言························································································································ Ⅲ 1 范围 ····················································································································· 1 2 规范性引用文件 ······································································································ 1 3 术语和定义 ············································································································ 1 3.1 枝条 ··············································································································· 1 3.2 侧枝 ··············································································································· 1 3.3 节点 ··············································································································· 1 3.4 接种 ··············································································································· 1 4 环境要求 ··············································································································· 1 4.1 温度 ··············································································································· 1 4.2 光照 ··············································································································· 1 5 培养基用水 ············································································································ 2 6 设备与器具 ············································································································ 2 7 培养方法 ··············································································································· 2 7.1 取样 ··············································································································· 2 7.2 剪切 ··············································································································· 2 7.3 漂洗 ··············································································································· 2 7.4 培养 ··············································································································· 2 7.5 注意事项 ········································································································· 2 8 观察记录 ··············································································································· 2 附录 A（资料性） 狐尾藻的概述图 ················································································ 3 附录 B（资料性） 狐尾藻的接种效果图 ·········································································· 4 附录 C（资料性） ANDREWS 培养基配制方法 ····································································· 5 I DB43/T 1969—2020 II DB43/T 1969—2020 前 言 本文件按照 GB/T 1.1—2020 给出的规则起草。 本文件某些内容可能涉及专利。本文件发布机构不承担识别这些专利的责任。 本文件由湖南省农业农村厅提出。 本文件由湖南省农业标准化技术委员会归口。 本文件起草单位：湖南省植物保护研究所，长沙艾格里生物科技有限公司。 本文件主要起草人：陈昂、刘勇、张德咏、张战泓、蒋桂芳、罗杰、罗香文、陈武瑛、熊浩。 III DB43/T 1969—2020 IV DB43/T 1969—2020 穗状狐尾藻室内培养技术规程 1 范围 本文件规定了穗状狐尾藻实验室培养的环境要求、仪器设备、水质、培养方法等。 本文件适用于湖南省试验用穗状狐尾藻的室内培养。 2 规范性引用文件 下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中，注日期的引用文件， 仅该日期对应的版本适用于本文件；不注日期的引用文件，其最新版本（包括所有的修改单）适用于本 文件。 3 术语和定义 下列术语和定义适用于本文件。 3.1 枝条 Stem 维管植物地上部分的骨干，上面着生叶、花和果实。它具有输导营养物质和水分以及支持叶、花和 果实在一定空间的作用。 3.2 侧枝 Branch 从主枝或副主枝上生长出的小枝。又叫单位枝。由植物的芽原基发育而来。 3.3 节点 Node 植物茎上的节点，为植物茎中间的横隔。 3.4 接种 Inoculation 按无菌操作技术要求将目的生物进行一定处理之后移接到培养基质中的过程 4 环境要求 4.1 温度 18～22 ℃。 4.2 光照 冷荧光灯源。将光源固定在距水面 30～50 cm 的上方，可预防热损伤，且保证光合作用和生长充足 的光照。光照强度约 8000 Lux（水表面） 。每天光照时间 16h。 1 DB43/T 1969—2020 5 培养基用水 满足 pH 为 7.0～8.0。溶解氧＞3.0 mg/L。培养用水为去离子水，ANDREWS 培养基的配制方法参见 附录 C 的 C.1。 6 设备与器具 光源、培养架，标本瓶，超净工作台，酒精灯，镊子，剪刀等。 7 培养方法 7.1 取样 选取长势良好，未被异种侵染过的枝条。 7.2 剪切 将主枝放入无菌水中，用灭菌后的剪刀剪主枝上靠近顶端的枝条。于两个节点间剪切，即保留 2 个节点。 （如果是侧枝较长，也可剪切） 7.3 漂洗 使用塑料镊子辅助，在无菌水中轻轻漂洗狐尾藻新枝，漂洗 3 次。 7.4 培养 分装培养时，每个培养瓶约 1000 mL 培养基，液面高度维持在约 10-15 cm。将新剪切的狐尾藻枝 条约 20 根放入 1000mL 装有 ANDREWS 培养基的培养瓶，放在上述要求的环境下培养。穗状狐尾藻的保种 3～4 周接种一次。健康强壮没被侵染的藻株可用于试验。狐尾藻的接种效果图参见附录 B 的 B.1。 7.5 注意事项 野外采集的穗状狐尾藻，一定要进行无菌化处理后再用培养基培养，否则藻类的爆发会是依附在狐 尾藻上的藻类增多，增加无菌化的难度，可以先用采集的水暂养。 标本瓶等器皿使用前应进行清洗及杀菌消毒。 8 观察记录 记录培养过程中的环境条件监测数据，观察并记录狐尾藻生长的参数等数据。 2 DB43/T 1969—2020 附 录 A （资料性） 狐尾藻的概述图 图 A.1 穗状狐尾藻概述图 3 DB43/T 1969—2020 附 录 B （资料性） 狐尾藻的接种效果图 图 B.1 穗状狐尾藻的接种效果图 4 DB43/T