Investigation and integrated molecular diagnosis of root-knot nematodes in Panax notoginseng root in the field

Root-knot nematodes (RKNs, Meloidogyne spp.) are damaging pests that can infect thousands of plant species and cause enormous crop losses worldwide. Panax notoginseng is a common host of root-knot nematodes. In this study, we surveyed notoginseng gardens and determined the incidence of RKNs. Among the gardens surveyed, 71 % were infected with RKNs, and the RKN incidence index ranged from 8 % to 47 % in three randomly infected gardens. Meloidogyne hapla was identified as the pathogenic nematode based on 18S ribosomal RNA analysis by DNA barcoding. The results were qualitatively and quantitatively confirmed using a real-time PCR assay according to variations in the ITS1 and ITS2 regions. These results indicated that the combination of DNA barcoding and real-time PCR is a reliable and precise method for identifying parasitic nematodes from mixed-infected plant roots in the field. In addition, the abundance of ITS1 and ITS2 displayed a similar trend to the numbers of RKNs in the three gardens, which suggests that the results of real-time PCR can be used to determine the damage caused by M. hapla in the field. Our studies show that RKNs are common and can cause serious damage to notoginseng. We present an integrated method of detecting mixed nematode species in the field and confirm M. hapla as the target for parasitic nematode control in notoginseng gardens. Our results contribute to the improvement of RKN control in notoginseng and further promote the sustainable development of medicinal plants.