DataSheet_1_Development of Specific DNA Barcodes for the Dinophyceae Family Kareniaceae and Their Application in the South China Sea.docx

Species from the family Kareniaceae (Dinophyceae) frequently cause harmful algal blooms (HABs), with serious ecological impacts and risks to human safety and aquaculture activities in coastal waters worldwide. However, due to their small size, lack of morphological divergence, and low abundance during non-bloom periods, the diversity within this family is not well understood. By comparing the commonly used molecular markers, the Internal Transcribed Spacer (ITS) region was found to have an appropriate mutation rate to distinguish three of the most common genera (Karenia, Karlodinium, and Takayama) within the Kareniaceae family and different geographical strains of Kareniaceae. Specific primers targeting the ITS region of Karenia, and the other primers specific to the genera Karlodinium and Takayama, were designed. Specificity of the primers was tested using 17 strains of Kareniaceae species and 15 non-target species. Representative Kareniaceae species could be successfully detected even at low concentrations of target DNA template with a limit of detection of 3.2 pg. The primers were also assessed using high-throughput sequencing with two environmental samples from the South China Sea (SCS). Analysis of the reads identified as Kareniaceae species revealed a high diversity and the existence of unreported Kareniaceae species in the SCS. In conclusion, the newly developed molecular barcodes specifically detected Kareniaceae species in the field and will provide technical support for the effective warning and monitoring of Kareniaceae HABs.来自(家庭)卡伦尼科科(Kareniaceae)的物种频繁引发有害藻华(HABs)，对全球沿海水域的生态系统造成严重破坏，并对人类安全和水产养殖活动构成风险。然而，鉴于其体型微小、形态分化不足以及非繁殖期低丰度，该科物种的多样性尚未得到充分理解。通过比较常用的分子标记，发现转录间隔区(ITS)具有适宜的突变率，可用于区分卡伦尼科科家族中三种最常见的属（卡伦尼亚(Karenia)、卡尔洛狄尼乌姆(Karlodinium)和池田(Takayama)）及其不同地理种群的菌株。针对卡伦尼亚属ITS区域的特异性引物，以及其他针对卡尔洛狄尼乌姆属和池田属的特异性引物已被设计。通过使用17个卡伦尼科科属物种和15个非目标物种进行特异性测试，验证了引物的特异性。即使在目标DNA模板的低浓度下，代表性卡伦尼科科物种也可成功检测，检测限为3.2 pg。此外，引物还通过高通量测序评估，使用了来自南海的两个环境样本。对被识别为卡伦尼科科物种的序列分析揭示了高多样性，并发现南海存在未报道的卡伦尼科科物种。综上所述，新开发的分子条形码在野外成功检测了卡伦尼科科物种，将为卡伦尼科科有害藻华的有效预警和监测提供技术支持。