Over 90% of global commercial trade occurs between seaports, which are initial points-of-entry for nonnative, potentially invasive propagules. As such, there is a need to develop means to both rapidly intercept and identify propagules as they arrive. Here, we focus on plant propagules that are assumed to be non-native, in seed form. Because standard morphological techniques alone are laborious and require taxonomic expertise, we sought to address if identification through barcoding of the plastid DNA (rbcL + matK genes) of plant seeds could improve current processes in the early detection and rapid response to prevent entry/establishment of nonnative plant species. This research conducted a preliminary foray to evaluate the utility of widely accepted plant plastid DNA barcodes to identify plant propagules (seeds, hereafter) collected from the air-intake grilles of refrigerated shipping containers of a single agricultural commodity arriving at the Port of Savannah, Georgia, USA. We ask four questions: (1) Can DNA barcoding be used to detect seeds collected from shipping containers at the port? (2) What is the genetic composition of propagules entering the port? (3) How do morphological identifications compare to those based on genetic analysis? (4) Are nonnative invasive plant species present on shipping containers entering the Port of Savannah? This research collected 11,044 seeds from 628 refrigerated shipping containers between 2015 and 2017. Seeds were then morphologically sorted into Seed Types. Barcoding of the matK and rbcL gene regions of the plastid genomes directly isolated from seeds resulted in poor amplification. This is likely due to a host of potential confounding factors. Therefore, we germinated seeds and utilized leaf-tissues for sequencing of these two gene regions. From BLASTn analyses, results returned top hits for a variety of species, with up to 22 possible nonnative plant species and one definite Federal Noxious Weed. This work investigates the interception application of DNA barcoding to improve agro- and bio-security issues posed by nonnative and invasive species. Though this study required the germination of the seeds to obtain leaf-tissue suitable for our DNA barcoding method, we effectively demonstrated seed viability. Our seed identification process was lengthy and understandably not feasible for real-time application. Therefore, we seek to improve our methods for future applications by testing other approaches that may better complement morphological identification. Next reasonable steps include improved extraction protocols, metabarcoding to generate DNA barcode sequences directly from groups of seeds harvested from shipping containers and implementing other next-generation sequencing techniques.

全球超过90％的商业贸易发生在海港之间，这是非本地，潜在侵入性繁殖体的初始进入点。因此，需要开发一种手段，以在传播物到达时迅速拦截并识别它们。在这里，我们集中于假定为非本地种子形式的植物繁殖体。由于仅标准形态学技术很费力并且需要生物分类学专业知识，因此我们试图解决是否通过条形码识别质体DNA（rbcL  +  matK基因）可以改善早期检测和快速反应的当前过程，从而防止外来植物物种的进入/建立。这项研究进行了初步尝试，以评估广为接受的植物质体DNA条形码在识别从佐治亚州萨凡纳港的单一农业商品的冷藏运输集装箱的进气格栅中收集的植物繁殖体（种子，以下称为种子）的效用。 ， 美国。我们问四个问题：（1）DNA条码可以用于检测从港口的运输集装箱中收集的种子吗？（2）进入港口的繁殖体的遗传成分是什么？（3）形态学鉴定与基于遗传分析的形态学鉴定相比如何？（4）进入萨凡纳港口的运输集装箱上是否存在非本地入侵植物物种？这项研究在2015年至2017年之间，从628个冷藏运输集装箱中收集了11,044粒种子。然后按形态对种子进行分类。种子类型。该条码的的matK和rbcL基因直接从种子中分离出的质体基因组的基因区域导致扩增不良。这可能是由于许多潜在的混杂因素所致。因此，我们发芽了种子，并利用叶组织对这两个基因区域进行了测序。通过BLASTn分析，结果返回了多种物种的热门话题，其中包括多达22种可能的非本土植物物种和一种明确的联邦有害杂草。这项工作研究了DNA条形码的拦截应用，以改善外来物种和入侵物种带来的农业和生物安全问题。尽管这项研究要求种子发芽以获得适合我们DNA条形码方法的叶组织，但我们有效地证明了种子的生存能力。我们的种子识别过程很漫长，并且可以实时应用，这是可以理解的。因此，我们寻求通过测试其他方法来更好地完善我们的方法，这些方法可能会更好地补充形态学鉴定。接下来的合理步骤包括改进的提取方案，元条形码技术，以直接从运输集装箱中收获的种子组中生成DNA条形码序列，并实施其他下一代测序技术。