viruses Article HTS-Based Diagnostics of Sugarcane Viruses: Seasonal Variation and Its Implications for Accurate Detection Martha Malapi-Wight 1,*,† , Bishwo Adhikari 1 , Jing Zhou 2, Leticia Hendrickson 1, Clarissa J. Maroon-Lango 3, Clint McFarland 4, Joseph A. Foster 1 and Oscar P. Hurtado-Gonzales 1,* 1 USDA-APHIS Plant Germplasm Quarantine Program, Beltsville, MD 20705, USA;
[email protected] (B.A.);
[email protected] (L.H.);
[email protected] (J.A.F.) 2 Department of Agriculture, Agribusiness, Environmental Sciences, Texas A&M University-Kingsville, Kingsville, TX 78363, USA;
[email protected] 3 USDA-APHIS-PPQ-Emergency and Domestic Program, Riverdale, MD 20737, USA;
[email protected] 4 USDA-APHIS-PPQ-Field Operations, Raleigh, NC 27606, USA;
[email protected] * Correspondence:
[email protected] (M.M.-W.);
[email protected] (O.P.H.-G.) † Current affiliation: USDA-APHIS BRS Biotechnology Risk Analysis Program, Riverdale, MD 20737, USA. Abstract: Rapid global germplasm trade has increased concern about the spread of plant pathogens and pests across borders that could become established, affecting agriculture and environment systems. Viral pathogens are of particular concern due to their difficulty to control once established. A comprehensive diagnostic platform that accurately detects both known and unknown virus species, as well as unreported variants, is playing a pivotal role across plant germplasm quarantine programs. Here we propose the addition of high-throughput sequencing (HTS) from total RNA to the routine Citation: Malapi-Wight, M.; quarantine diagnostic workflow of sugarcane viruses. We evaluated the impact of sequencing depth Adhikari, B.; Zhou, J.; Hendrickson, needed for the HTS-based identification of seven regulated sugarcane RNA/DNA viruses across L.; Maroon-Lango, C.J.; McFarland, two different growing seasons (spring and fall).