RT Journal Article SR Electronic T1 A low-cost and open-source solution to automate imaging and analysis of cyst nematode infection assays for Arabidopsis thaliana JF bioRxiv FD Cold Spring Harbor Laboratory SP 2022.07.14.500020 DO 10.1101/2022.07.14.500020 A1 Olaf Prosper Kranse A1 Itsuhiro Ko A1 Roberta Healey A1 Unnati Sonawala A1 Siyuan Wei A1 Beatrice Senatori A1 Francesco De Batté A1 Ji Zhou A1 Sebastian Eves-van den Akker YR 2022 UL http://biorxiv.org/content/early/2022/07/15/2022.07.14.500020.abstract AB Background Cyst nematodes are one of the major groups of plant-parasitic nematode, responsible for considerable crop losses worldwide. Improving genetic resources, and therefore resistant cultivars, is an ongoing focus of many pest management strategies. One of the major bottlenecks in identifying the plant genes that impact the infection, and thus the yield, is phenotyping. The current available screening method is slow, has unidimensional quantification of infection limiting the range of scorable parameters, and does not account for phenotypic variation of the host. The ever-evolving field of computer vision may be the solution for both the above-mentioned issues. To utilise these tools, a specialised imaging platform is required to take consistent images of nematode infection in quick succession.Results Here, we describe an open-source, easy to adopt, imaging hardware and trait analysis software method based on a pre-existing nematode infection screening method in axenic culture. A cost-effective, easy-to-build and -use, 3D-printed imaging device was developed to acquire images of the root system of Arabidopsis thaliana infected with the cyst nematode Heterodera schachtii, replacing costly microscopy equipment. Coupling the output of this device to simple analysis scripts allowed the measurement of some key traits such as nematode number and size from collected images, in a semi-automated manner. Additionally, we used this combined solution to quantify an additional trait, root area before infection, and showed both the confounding relationship of this trait on nematode infection and a method to account for it.Conclusion Taken together, this manuscript provides a low-cost and open-source method for nematode phenotyping that includes the biologically relevant nematode size as a scorable parameter, and a method to account for phenotypic variation of the host. Together these tools highlight great potential in aiding our understanding of nematode parasitism.Competing Interest StatementThe authors have declared no competing interest.