Europe as a secondary distribution hub in the worldwide invasion of the potato cyst nematode Globodera rostochiensis.

The potato cyst nematode Globodera rostochiensis originates from the Andean Mountain region in South America and has unintentionally been introduced to all inhabited continents. Several studies have examined the population genetic structure of this pest in various countries by using microsatellite markers. However, merging microsatellite data produced from different laboratories is challenging and can introduce uncertainty when interpreting the results. To overcome this challenge and to explore invasion routes of this pest, we have genotyped 22 G. rostochiensis populations from all continents. Within populations, the highest genetic diversity was observed in the South American populations, the European populations showed an intermediate level of genetic diversity and the remaining populations were the less diverse. This confirmed pre-existing knowledge such as a first introduction event from South America to Europe, but the less diverse populations could originate either from South America or from Europe. At the continental scale, STRUCTURE genetic clustering output indicated that North America and Asia have experienced at least two introduction events. Comparing different evolutionary scenarios, the Approximate Bayesian Computation analysis showed that Europe served as a secondary distribution centre for the invasion of G. rostochiensis into all other continents (North America, Africa, Asia and Oceania).

Biology, pathotype, and virulence of Globodera rostochiensis populations from Kenya.

The potato cyst nematodes (PCN), Globodera rostochiensis (Woll.) and G. pallida (Stone), are important pests of potato globally. Due to their extensive damage potential and the challenge of managing them, these nematodes are under strict regulations in many countries; however, despite these regulations, PCN continue to spread into new areas and countries. In Kenya, G. rostochiensis was first reported in 2015 and G. pallida was reported three years later, both in Nyandarua County. Research was conducted to characterize the biology, pathotype, and virulence of G. rostochiensis populations from Kenya in glasshouse and laboratory studies. The development of G. rostochiensis was assessed in roots of susceptible potato 'Désirée' and resistant 'Laura' carrying the H1 resistance gene. The 'HAR1' population from Kenya and 'Ecosse' from Germany were not able to produce females in the roots of the resistant potato 'Laura'. The rate of root penetration by G. rostochiensis juveniles did not differ (p > 0.05) between populations and cultivars. However, in the resistant cultivar, juveniles developed into males only. A total of 736 cumulative degree-days at 6°C base temperature (DD6) were required by 'HAR1' to complete the life cycle on 'Désirée', whereas 'Ecosse' completed the life cycle within 645 DD6. The Kenyan populations lacked obligatory diapause and high numbers of juveniles hatched immediately after maturity. Consequently, the Kenyan populations had the potential to complete up to three reproduction cycles in less than a year. On selected potato cultivars, the populations from Kenya failed to reproduce on 10 out of 13 commercial cultivars tested. The 10 cultivars carried the H1 resistance gene, which suggests that the G. rostochiensis populations tested belong to the Ro1/4 pathotype group. The virulence of the G. rostochiensis populations from Kenya did not differ from that of the standard reference population 'Ecosse' and therefore can be effectively managed with the commercially available potato cultivars carrying the H1 resistance gene.

Author Correction: Genome assembly and annotation of Meloidogyne enterolobii, an emerging parthenogenetic root-knot nematode.

A Correction to this paper has been published: https://doi.org/10.1038/s41597-020-00747-0.

Genome assembly and annotation of Meloidogyne enterolobii, an emerging parthenogenetic root-knot nematode.

Root-knot nematodes (genus Meloidogyne) are plant parasites causing huge economic loss in the agricultural industry and affecting severely numerous developing countries. Control methods against these plant pests are sparse, the preferred one being the deployment of plant cultivars bearing resistance genes against Meloidogyne species. However, M. enterolobii is not controlled by the resistance genes deployed in the crop plants cultivated in Europe. The recent identification of this species in Europe is thus a major concern. Here, we sequenced the genome of M. enterolobii using short and long-read technologies. The genome assembly spans 240 Mbp with contig N50 size of 143 kbp, enabling high-quality annotations of 59,773 coding genes, 4,068 non-coding genes, and 10,944 transposable elements (spanning 8.7% of the genome). We validated the genome size by flow cytometry and the structure, quality and completeness by bioinformatics metrics. This ensemble of resources will fuel future projects aiming at pinpointing the genome singularities, the origin, diversity, and adaptive potential of this emerging plant pest.

Evaluation of fluopyram for the control of Ditylenchus dipsaci in sugar beet.

Fluopyram, a succinate dehydrogenase inhibitor fungicide, has shown potential in controlling Meloidogyne incognita and Rotylenchus reniformis in tomato. The effectiveness of this compound for the control of Ditylenchus dipsaci in sugar beet was evaluated. In this study, laboratory, growth chamber, glasshouse, and field experiments were conducted. In a motility bioassay, the EC50 value was determined with 3.00 µg/ml a.i. after 72 h exposure to fluopyram. The growth chamber experiment did not show any effects on D. dipsaci penetration rate; however, field experiments revealed a positive effect of fluopyram applied at planting in reducing D. dipsaci infectivity. The glasshouse experiment confirmed a limited effect of fluopyram on D. dipsaci population development. Under field conditions, despite a reduction of D. dipsaci penetration rates in spring, fluopyram was not effective in reducing the population development until harvest. Consequently, D. dipsaci densities in plant tissue and soil were high at harvest and not different among treatments. However, root-rot symptoms were significantly reduced at harvest. Fluopyram applied at planting showed good potential to reduce root-rot symptoms caused by D. dipsaci in sugar beet. However, for the long-term reduction of nematode populations in soil, further integrated control measures are needed to reduce the risks of substantial yield losses by D. dipsaci.Fluopyram, a succinate dehydrogenase inhibitor fungicide, has shown potential in controlling Meloidogyne incognita and Rotylenchus reniformis in tomato. The effectiveness of this compound for the control of Ditylenchus dipsaci in sugar beet was evaluated. In this study, laboratory, growth chamber, glasshouse, and field experiments were conducted. In a motility bioassay, the EC50 value was determined with 3.00 µg/ml a.i. after 72 h exposure to fluopyram. The growth chamber experiment did not show any effects on D. dipsaci penetration rate; however, field experiments revealed a positive effect of fluopyram applied at planting in reducing D. dipsaci infectivity. The glasshouse experiment confirmed a limited effect of fluopyram on D. dipsaci population development. Under field conditions, despite a reduction of D. dipsaci penetration rates in spring, fluopyram was not effective in reducing the population development until harvest. Consequently, D. dipsaci densities in plant tissue and soil were high at harvest and not different among treatments. However, root-rot symptoms were significantly reduced at harvest. Fluopyram applied at planting showed good potential to reduce root-rot symptoms caused by D. dipsaci in sugar beet. However, for the long-term reduction of nematode populations in soil, further integrated control measures are needed to reduce the risks of substantial yield losses by D. dipsaci.

Development and Validation of LNA-Based Quantitative Real-Time PCR Assays for Detection and Identification of the Root-Knot Nematode Meloidogyne enterolobii in Complex DNA Backgrounds.

Meloidogyne enterolobii is a quarantine root-knot nematode posing a major threat to agricultural production systems worldwide. It attacks many host plants, including important agricultural crops, ornamentals, and trees. M. enterolobii is a highly virulent and pathogenic root-knot nematode species, able to reproduce on plants resistant to other Meloidogyne spp. Significant crop damage has been reported in Asia, South America, Africa, the United States, France, and greenhouses in Switzerland. To identify potential introduction pathways and ensure appropriate phytosanitary measures and management strategies, accurate detection and identification tools are needed. Therefore, two real-time quantitative polymerase chain reaction (PCR) assays based on the second intergenic spacer region of the ribosomal DNA cistron and the cytochrome oxidase c subunit I (COI) gene using locked nucleic acid probes were developed and validated for fast and reliable detection and identification of M. enterolobii. Analytical specificity was confirmed with 16 M. enterolobii populations, 16 populations of eight closely related Meloidogyne spp., and four species from other nematode genera. Optimizing and testing the assays on two real-time PCR platforms revealed an analytical sensitivity of one juvenile in a background of 1,000 nematodes and the intended limit of detection of one juvenile per 100 ml of soil. Both assays performed equally well, with the COI-based assay showing a slightly better performance concerning detection of M. enterolobii target DNA in complex DNA backgrounds.

Effects of the Mi-1 and the N root-knot nematode-resistance gene on infection and reproduction of Meloidogyne enterolobii on tomato and pepper cultivars.

Meloidogyne enterolobii is widely considered to be an aggressive root-knot nematode species that is able to reproduce on root-knot nematode-resistant tomato and pepper cultivars. In greenhouse experiments, M. enterolobii isolates 1 and 2 from Switzerland were able to reproduce on tomato cultivars carrying the Mi-1 resistance gene as well as an N-carrying pepper cultivar. Reproduction factors (Rf) ranged between 12 and 109 depending on the plant cultivar, with M. enterolobii isolate 2 being more virulent when compared to isolate 1. In contrast, M. arenaria completely failed to reproduce on these resistant tomato and pepper cultivars. Although some variability in virulence and effectiveness of root-knot nematode-resistance genes was detected, none of the plant cultivars showed Rf values less than 1 or less than 10% of the reproduction observed on the susceptible cv. 'Moneymaker' (Rf = 23-44) used to characterize resistance. The ability of M. enterolobii to overcome the resistance of tomato and pepper carrying the Mi-1 and the N gene makes it difficult to manage this root-knot nematode species, particularly in organic farming systems where chemical control is not an option.

Integrated Control of Rhizoctonia Crown and Root Rot of Sugar Beet with Fungicides and Antagonistic Bacteria.

Rhizoctonia crown and root rot, caused by the fungus Rhizoctonia solani AG 2-2, is one of the most damaging sugar beet diseases worldwide and causes significant economic losses in more than 25% of the sugar beet production area in the United States. We report on field trials in the years 1996 to 1999 testing both experimental fungicides and antagonistic Bacillus sp. for their potential to reduce disease severity and increase sugar yield in trials inoculated with R. solani AG 2-2. Fungicides were applied as in-furrow sprays at planting or as band sprays directed at the crown at the four-leaf stage, or four- plus eight-leaf stage, while bacteria were applied at the four-leaf stage only. The fungicides azoxystrobin and tebuconazole reduced crown and root rot disease by 50 to 90% over 3 years when used at rates of 76 to 304 g a.i./ha and 250 g a.i./ha, respectively. The disease index at harvest was reduced and the root and sugar yield increased with azoxystrobin compared with tebuconazole. The combination of azoxystrobin applied at 76 g a.i./ha and the Bacillus isolate MSU-127 resulted in best disease reduction and greatest root and sucrose yield increase.