Pfitzer René, Varrelmann Mark, Hesse Georgia, Eini Omid
Institute of Sugar Beet Research, Holtenser Landstraße 77, 37079 Göttingen, Germany.
Agricultural Entomology, Department of Crop Sciences, Faculty of Agricultural Sciences, University of Göttingen, Grisebachstrasse 6, 37077 Göttingen, Germany.
Insects. 2022 Oct 28;13(11):992. doi: 10.3390/insects13110992.
Monitoring of (Hemiptera: Auchenorrhyncha: Cixiidae), representing the main vector of the syndrome 'basses richesses' (SBR) disease in sugar beet is based on morphological identification. However, two other cixiid species, and with similar external characters are known to appear in sugar beet fields and are challenging to be distinguished from . We present a PCR-based method for species-specific detection of both male and female , directly after sweep net collection or after up to 18 months long term storage on sticky traps. Two methods of DNA template preparation, based on a commercial extraction kit or on simple grinding in phosphate-buffered saline (PBS) were compared. The latter method was also established for eggs and all five nymphal instars of from a rearing. Furthermore, in silico primer analysis showed that all Auchenorrhyncha species including far related species reported from sugar beet fields can be differentiated from . This was PCR-confirmed for the most common Auchenorrhyncha species from different German sugar beet fields. Sequence analysis of the mitochondrial cytochrome oxidase I gene () amplicon showed a close relationship to from but separated from the and species which are grouped into the same family Cixiidae. We present a sensitive, cost- and time-saving PCR-based method for reliable and specific detection of eggs and all nymphal instars, as well as male and female , after different methods of planthopper collection and template DNA template preparation that can be used in large scale monitoring assays.
对甜菜“basses richesses”（SBR）病主要传播媒介[半翅目：头喙亚目：蜡蝉科]的监测基于形态学鉴定。然而，已知另外两种具有相似外部特征的蜡蝉科物种也会出现在甜菜田中，且很难与[目标物种]区分开来。我们提出了一种基于PCR的方法，可在扫网采集后或在粘性诱捕器上长达18个月的长期保存后，直接对[目标物种]的雌雄个体进行物种特异性检测。比较了基于商业提取试剂盒或在磷酸盐缓冲盐水（PBS）中简单研磨的两种DNA模板制备方法。后一种方法也适用于从饲养中获得的[目标物种]的卵和所有五个若虫龄期。此外，电子引物分析表明，所有头喙亚目物种，包括从甜菜田报道的远缘物种，都可以与[目标物种]区分开来。这在德国不同甜菜田最常见的头喙亚目物种中得到了PCR验证。[目标物种]线粒体细胞色素氧化酶I基因（COI）扩增子的序列分析表明，它与[另一地区的目标物种]关系密切，但与属于同一蜡蝉科的[其他物种]分开。我们提出了一种灵敏、节省成本和时间的基于PCR的方法，可在不同的飞虱采集方法和模板DNA制备后，可靠且特异性地检测[目标物种]的卵、所有若虫龄期以及雌雄个体，该方法可用于大规模监测试验。
