Garcia-Benitez C, Melgarejo P, De Cal A
Department of Plant Protection, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), 28040 Madrid, Spain.
Plant Dis. 2017 Jun;101(6):1002-1008. doi: 10.1094/PDIS-11-16-1682-RE. Epub 2017 Mar 20.
Most stone fruit with a latent brown rot infection caused by Monilinia do not develop visible signs of disease until the arrival of fruit at the markets or the consumer's homes. The overnight freezing-incubation technique (ONFIT) is a well-established method for detecting latent brown rot infections, but it takes between 7 to 9 days. In this report, we inform on the advantages of applying a qPCR-based method to (i) detect a latent brown rot infection in the blossoms and fruit of nectarine trees (Prunus persica var. nucipersica) and (ii) distinguish between the Monilinia spp. in them. For applying this qPCR-based method, artificial latent infections were established in nectarine flowers and fruit using 10 Monilinia fructicola isolates, 8 M. fructigena isolates, and 10 M. laxa isolates. We detected greater amounts of M. fructicola DNA than M. laxa and M. fructigena DNA in latently infected flowers using qPCR. However, greater DNA amounts of M. laxa than M. fructicola were detected in the mesocarp of latently infected nectarines. We found that the qPCR-based method is more sensitive, reliable, and quicker than ONFIT for detecting a latent brown rot infection, and could be very useful in those countries where Monilinia spp. are classified as quarantine pathogens.
大多数受到链核盘菌引起的潜伏性褐腐病感染的核果,在果实到达市场或消费者家中之前不会出现明显的病害迹象。过夜冷冻培养技术(ONFIT)是一种成熟的检测潜伏性褐腐病感染的方法,但需要7到9天时间。在本报告中,我们介绍了应用基于qPCR的方法的优势,即(i)检测油桃花和果实中的潜伏性褐腐病感染,以及(ii)区分其中的链核盘菌种类。为了应用这种基于qPCR的方法,使用10株果生链核盘菌分离株、8株果产链核盘菌分离株和10株核果链核盘菌分离株在油桃花和果实中建立了人工潜伏感染。通过qPCR,我们在潜伏感染的花中检测到果生链核盘菌的DNA量比核果链核盘菌和果产链核盘菌的DNA量更多。然而,在潜伏感染的油桃中果皮中检测到核果链核盘菌的DNA量比果生链核盘菌更多。我们发现,基于qPCR的方法在检测潜伏性褐腐病感染方面比ONFIT更灵敏、可靠且快速,并且在那些将链核盘菌种类列为检疫性病原菌的国家可能非常有用。