Validation of protocol to detect Candidatus Liberibacter solanacearum in carrot seeds II: Evaluation of subsample size
2018
Kagatsume, R. (Moji Plant Protection Station (Japan). Fukuoka Sub-station) | Oishi, M. | Kasukabe, R. | Motokura, Y.
It is commonly believed that Candidatus Liberibacter solanacearum (Lso) can infect carrots via psyllid vector transmission like Bactericera trigonica and Trioza apicalis. While the validity of seed-borne transmission of Lso via carrot seeds remains unclear, possible entry of Lso by way of carrot seeds is one phytosanitary concern since, for example, Bertolini et al. (2014) reported Lso was transmitted through carrot seeds. This study examined whether the subsample size to detect Lso can be increased from that in the protocol developed by Oishi et al. (2017). As the current protocol is time and labor consuming in the national plant protection scheme, a more efficient approach to detection is required. Based on the proposed subsample size of 2,850 in Oishi et al. (2017), it was concluded that 2,700 seeds were suitable as a subsample. The present study suggested an improvement to the protocol, in which six groups of 450 carrot seeds each would be ground and then mixed for nucleic acid extraction. This method reduces the cost and time for detection by approximately one-sixth.
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