基于纳米孔测序技术的柑橘黄龙病检测方法建立

柑橘黄龙病(Huanglongbing)是柑橘生产上最具毁灭性的病害,及时快速地进行早期检测和诊断是防控黄龙病的关键措施之一.本文利用掌上纳米孔测序仪MinION对携带黄龙病菌Candidatus Liberibacter asiaticus的DNA样品进行测序,并利用Blast、GraphMap、minimap2以及两种bwa的比对方法将测序结果比对到黄龙病菌基因组上,比对结果均匀的比对到黄龙病菌基因组上,并未发现严重的偏倚现象,验证了其测序结果的可靠性.本技术可弥补因柑橘木虱Diaphorina citri(Kuwayama)虫体过小或损坏难以进行光学识别的不足,并可同时检测虫体是否携带有黄龙病菌,对有黄龙病发生风险但尚未有黄龙病实际发生的柑橘种植区提供实时实地的监控与预警.

Establishment of detection method of Huanglongbing based on nanopore sequencing technology

Huanglongbing is the most devastative disease in citrus production, early detection and diagnosis in a timely and rapid manner is one of the key measures to prevent and control huanglongbing. In this paper, the DNA sample carrying Candidatus Liberibacter asiaticus (CLas) was sequenced by MinION, and the sequencing results were compared to the genome of CLas by blast, graphmap, minimap2 and two BWA comparison methods. The comparison results were evenly compared to the genome of CLas, and no serious bias was found, which verified the reliability of the sequencing results. This sequencing technology could make up for the difficulty of optical identification due to the small or damaged Diaphorina citri (Kuwayama), and it could also detect whether the psyllids carry CLas at the same time, and provided real-time monitoring and early warning for citrus planting areas where there was a potential risk of huanglongbing.