Development of molecular tools for the identification of powdery mildew-resistant Cannabis sativa strains through a genomics-based approach

OrganiGram是根据Marihuana for Medical Purposes获得许可的医用大麻生产商之一。 （MMPR）。与任何农作物一样，植物病害在大麻中起着主要的限制作用。 生产，为此OrganiGram正在寻找控制方法。使用农药防治植物病害 引发了严重的健康和环境问题，这决定了有机害虫管理的必要性 技术，如使用抗病菌株。白粉病是大麻的一种主要疾病，但 已知菌株之间存在抗性的变异。在大麦、西红柿或豌豆等多种作物中， 对白粉病的持久控制由Mlo基因中的功能丧失突变赋予。的可用性 Cannabis sativa基因组的鉴定和研究使得能够鉴定和研究Mlo直向同源物，并开发 筛选大麻品系中天然存在的功能丧失型MLO变体的测定。为了 为了有信心地鉴定所有潜在的Mlo直系同源物，大麻基因组将首先通过测序来改进， 使用长读序测序平台Pacific Biosciences的额外DNA文库。然后PCR引物将 被开发以允许特异性和可靠地扩增怀疑参与以下的Mlo直系同源物： 白粉病抗性，引物将针对不同菌株进行测试以确保敏感性。最后， 这些引物可用于筛选数百种大麻品系或育种材料， 编码具有功能丧失突变的Mlo直向同源物的基因组变体，例如抗白粉病的 植物

OrganiGram is one of ~15 licensed producers of medical marijuana under the Marihuana for Medical Purposes Regulations (MMPR). As for any agricultural crop, plant diseases play a major limiting role in Cannabis production, for which OrganiGram is looking for control methods. Control of plant diseases using pesticides raises serious health and environmental concerns, which have dictated the need for organic pest management techniques, such as the use of disease-resistant strains. Powdery mildew is a major disease of Cannabis, but variation in resistance is known to exist between strains. In crops as diverse as barley, tomatoes or peas, durable control of powdery mildew is conferred by loss-of-function mutations in the Mlo gene. The availability of the Cannabis sativa genome enables the identification and study of Mlo orthologues, and the development of screening assays for naturally occurring loss-of-function mlo variants in marijuana strains. In order to confidently identify all potential Mlo orthologues, the Cannabis genome will first be improved by sequencing additional DNA libraries using the long-read sequencing platform Pacific Biosciences. PCR primers will then be developed to allow specific and reliable amplification of Mlo orthologues suspected to be involved in powdery mildew resistance, and primers will be tested against different strains to ensure sensitivity. Ultimately, these primers could be used to screen hundreds of Cannabis sativa strains or breeding material to uncover genomic variants encoding Mlo orthologues with loss-of-function mutations, e.g. powdery mildew-resistant plants.