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Standardized protocols for absolute quantification of potato virus Y (PVY) from potato tissue is critical for host-virus dynamic studies. Here, we developed a standardized protocol using a cloned viral sequence as standards to detect and quantify PVY. Starting with total RNA, concentrated via column-based kit, this protocol is able to detect approximately 50 viral copies/reaction from multiple PVY strains. Validation of this protocol confirmed linearity across 8 orders of magnitude with high repeatability, reproducibility and statistical robustness across three independent runs. This protocol offers reliable PVY quantification to manage potato crop health and enables comparative studies with other viral systems. 

The use of true potato seed (TPS) is fundamental to potato breeding and research, but can be hindered by poor germination and seed dormancy. TPS germination studies had focused mainly on seed treatment methods after seed extraction and not in combination with the seed extraction methods used. In potato, TPS extraction using water, using yeast fermentation or using sodium bicarbonate are common, but TPS extraction using dilute aqueous hydrochloric acid (HCl) followed by bleach treatment (3% sodium hypochlorite) had never been tested in potato even though this is standard practice for tomato seeds. Therefore, three seed extraction methods (water, 0.1 M HCl, and 0.8% yeast fermentation) in combination with three seed treatment methods (water, 1500 ppm GA3, and 3% sodium hypochlorite) were tested on diploid TPS at 1 week and 1 month after seed extraction. TPS treated with GA3 improved germination for both 1 week- and 1 month-old seeds, while TPS treated with 3% sodium hypochlorite only improved germination for 1 month-old seeds. This study shows that TPS extraction using water, yeast or HCl had no effect on germination, but supports the use of GA3 or bleach to promote TPS germination.