Supplanting resistance of the Mi gene by root-knot nematode in industrial tomato in the Cerrado in Goiás State of Brazil

ABSTRACT: Problems with root-knot nematodes, caused by genus Meloidogyne, can be controlled through the introduction of resistance genes in commercial varieties of tomatoes. However, plants of the hybrid ‘Heinz 9992’ for industrial processing, carrying the Mi gene, were reported with their roots severely infected in experimental cultivation in Goiás State of Brazil with symptoms typical of galls caused by Meloidogyne spp. After dissecting the galls present in the root samples, the females of the nematodes were removed and afterwards, performed the analysis of the perineal pattern and the biochemical characterization by electrophoresis of isoenzymes, besides the pathogenicity test. The pathogen analyzed was identified as Meloidogyne incognita. The pathogenicity test confirmed the supplanting resistance of the tomato hybrid with the Mi gene by this virulent population of M. incognita.

Responses of Tomato Genotypes to Avirulent and Mi-Virulent Meloidogyne javanica Isolates Occurring in Israel

The behavior of naturally virulent Meloidogyne isolates toward the tomato resistance gene Mi in major tomato-growing areas in Israel was studied for the first time. Virulence of seven selected isolates was confirmed over three successive generations on resistant (Mi-carrying) and susceptible (non-Mi-carrying) tomato cultivars. Diagnostic markers verified the predominance of Meloidogyne javanica among virulent isolates selected on resistant tomato cultivars or rootstocks. To better understand the determinants of nematode selection on Mi-carrying plants, reproduction of Mi-avirulent and virulent isolates Mjav1 and Mjv2, respectively, measured as eggs per gram of root, on non-Mi-carrying, heterozygous (Mi/mi) and homozygous (Mi/Mi) genotypes was evaluated. Although no reproduction of Mjav1 was observed on Mi/Mi genotypes, some reproduction was consistently observed on Mi/mi plants; reproduction of Mjv2 on the homozygous and heterozygous genotypes was similar to that on susceptible cultivars, suggesting a limited quantitative effect of the Mi gene. Histological examination of giant cells induced by Mi-virulent versus avirulent isolates confirmed the high virulence of Mjv2 on Mi/mi and Mi/Mi genotypes, allowing the formation of well-developed giant-cell systems despite the Mi gene. Analysis of the plant defense response in tomato Mi/Mi, Mi/mi, and mi/mi genotypes to both avirulent and virulent isolates was investigated by quantitative real-time polymerase chain reaction. Although the jasmonate (JA)-signaling pathway was clearly upregulated by avirulent and virulent isolates on the susceptible (not carrying Mi) and heterozygous (Mi/mi) plants, no change in signaling was observed in the homozygous (Mi/Mi) resistant line following incompatible interaction with the avirulent isolate. Thus, similar to infection promoted by the avirulent isolate on the susceptible genotype, the Mi-virulent isolate induced the JA-dependent pathway, which might promote tomato susceptibility during the compatible interaction with the homozygous (Mi/Mi) resistant line. These results have important consequences for the management of Mi resistance genes for ensuring sustainable tomato farming.

Meloidogyne Virulence Locus Molecular Marker for Characterization of Selected Mi-Virulent Populations of Meloidogyne spp. Is Correlated with Several Genera of Betaproteobacteria

Resistance to root-knot nematodes in tomato is conferred by the Mi resistance gene to the three most important species of Meloidogyne: M. arenaria, M. incognita, and M. javanica. Nevertheless, the Mi gene is unable to inhibit the reproduction of selected and naturally Mi-virulent populations of root-knot nematodes. As pathogenicity assays are time consuming, molecular markers were developed for the easy identification of Mi-virulent populations of Meloidogyne. The sequence characterized amplified region-Meloidogyne virulence locus (MVC) molecular marker is reported to differentiate Mi-avirulent and naturally Mi-virulent from selected Mi-virulent populations. This marker was used to compare acquired virulence in populations of M. javanica from Spain. The original populations used to develop the MVC marker were included as control for reference. Results showed that this marker did not amplify genomic DNA extracted from single juveniles or females of any of the populations tested either from Spain or Japan. In silico analyses performed with the recently published complete genome of M. incognita, indicated that the MVC marker is not correlated to a MVC or to any eukaryotic organism but to several betaproteobacteria genus from the family Comamonadaceae.

Effect of the Mi gene on reproduction of Meloidogyne hispanica on tomato genotypes

The root-knot nematode resistance (Mi) gene was screened in 25 tomato genotypes of Solanum lycopersicum, by amplification of REX-1 and Mi23 markers. Ten heterozygous tomato genotypes (Mimi), nine homozygous (MiMi) at the Mi locus and six lacking the Mi gene for resistance to root-knot nematode were identified using the marker REX-1. The results obtained with Mi23 marker confirmed the Mi gene status of the tomato genotypes, except for genotype Valouro RZ F1 that was homozygous (MiMi) and heterozygous (Mimi) at the Mi locus when using the REX-1 and Mi23 markers, respectively. The pathogenicity of Meloidogyne hispanica on the 25 tomato genotypes was assessed 60 days after inoculation with 5000 eggs on the basis of root gall index (GI) and reproduction factor (Rf). All the tomato genotypes were susceptible (excellent or good hosts), with GI > 4 and Rf > 2, except the genotype Rapit (Mimi), considered as resistant/hypersensitive (poor host). In this genotype, the nematode induced galls (GI = 4) on its roots and a small number of eggs were produced (Pf = 3085 ± 485). Significant differences in reproduction were detected between the Mi allelic conditions and genotypes within Mi allelic conditions. The increasing number of Mi alleles (0, 1 or 2) is associated with decreasing Rf, which suggests a possible dosage effect of the Mi gene. The variability observed in the Rf values for MiMi tomato genotypes may reflect an influence of the genetic background of the plants containing the Mi gene. Ten of the 25 tomato genotypes with Mi gene are commercially available. However, only Rapit can be used to control the three most common Meloidogyne spp. and inhibit the increasing of M. hispanica populations, and may have potential to be included in an integrated pest management programme. However, it is advisable to evaluate the pathogenicity of local populations of this nematode species associated with different environmental factors.