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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Mycology and Nematology Genetic Diversity and Biology Laboratory » Research » Publications at this Location » Publication #313123
Title: A Taqman real-time PCR assay for detection of Meloidogyne hapla in root galls and in soil

Author
item SAPKOTA, RUMAKANTA - Aarhus University
item Skantar, Andrea
item NICOLAISEN, MOGENS - Aarhus University

Submitted to: Nematology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/27/2015
Publication Date: 11/24/2015
Publication URL: https://handle.nal.usda.gov/10113/62017
Citation: Sapkota, R., Skantar, A.M., Nicolaisen, M. 2015. A Taqman real-time PCR assay for detection of Meloidogyne hapla in root galls and in soil. Nematology. 18(1):147-154.

Interpretive Summary: Nematodes are microscopic worms that cause billions of dollars in crop losses worldwide each year. Root-knot nematodes are an important group of nematodes that damage many kinds of plants, including carrot, by invading the roots and interfering with nutrient uptake. Identifying these nematodes from soil can be very labor intensive and anatomical features of root knot nematodes are very similar in some species. In the present study, scientists from Denmark worked with an ARS scientist from Beltsville, MD to develop a new molecular test to detect the northern root knot nematode. The results are significant because the new test was sensitive, highly specific, and able to detect this species directly from infested soil. This research will be used by researchers and diagnosticians to accurately detect and quantify northern root knot nematodes from infested fields and for directing management decisions in carrot cultivation.

Technical Abstract: Meloidogyne hapla is one of the most widespread and serious soil-borne nematodes causing root knot diseases in various crops. Early and accurate detection and quantification of M. hapla in soil is essential for effective disease management. The purpose of this study was to develop an assay for detection of M. hapla in soil samples. Based on the alignment of available Meloidogyne internal transcribed spacer (ITS) ribosomal DNA sequences in Genbank, a primer pair and a probe were designed for M. hapla detection in a Taqman real-time PCR assay. The designed assay was able to specifically detect M. hapla and showed no amplification of DNA from other plant infesting nematode species including seven other Meloidogyne species. Moreover, the assay was able to detect M. hapla in a background of soil DNA and directly from infested soil. To our knowledge this is the first report describing a real-time PCR based tool for detection of M. hapla in carrot and infested soil. The assay will be useful for large scale survey studies to detect and quantify M. hapla in cultivated fields and possibly for directing management decisions in carrot cultivation.