Cowpea is one of the most important food and forage legumes in drier regions of the tropics and subtropics. However, cowpea yield worldwide is markedly below the known potential due to abiotic and biotic stresses, including parasitism by root-knot nematodes (Meloidogyne spp., RKN). Two resistance genes with dominant effect, Rk and Rk², have been reported to provide resistance against RKN in cowpea. Despite their description and use in breeding for resistance to RKN and particularly genetic mapping of the Rk locus, the exact genes conferring resistance to RKN remain unknown. In the present work, QTL mapping using recombinant inbred line (RIL) population 524B x IT84S-2049 segregating for a newly mapped locus and analysis of the transcriptome changes in two cowpea near-isogenic lines (NIL) were used to identify candidate genes for Rk and the newly mapped locus. A major QTL, designated QRk-vu9.1, associated with resistance to Meloidogyne javanica reproduction, was detected and mapped on linkage group LG9 at position 13.37 cM using egg production data. Transcriptome analysis on resistant and susceptible NILs 3 and 9 days after inoculation revealed up-regulation of 109 and 98 genes and down-regulation of 110 and 89 genes, respectively, out of 19,922 unique genes mapped to the common bean reference genome. Among the differentially expressed genes, four and nine genes were found within the QRk-vu9.1 and QRk-vu11.1 QTL intervals, respectively. Six of these genes belong to the TIR-NBS-LRR family of resistance genes and three were upregulated at one or more time-points. Quantitative RT-PCR validated gene expression to be positively correlated with RNA-seq expression pattern for eight genes. Future functional analysis of these cowpea genes will enhance our understanding of Rk-mediated resistance and identify the specific gene responsible for the resistance.

Nematodes are important pathogens in banana plants, and the lack of resistant genotypes is the biggest challenge of the banana breeding programs. Little is known on the behavior of banana triploids and tetraploids developed by Embrapa regarding parasitism by plant-parasitic nematodes in field conditions. Embrapa Mandioca e Fruticultura experimental areas, naturally infested in five Brazilian states (Embrapa Acre - Acre, Embrapa Semiárido - Pernambuco, Embrapa Cerrados - Distrito Federal, Palmital - São Paulo and Epagri - Santa Catarina) were evaluated for the distribution and population levels of plant-parasitic nematodes in commercial cultivars and triploid and tetraploid genotypes in the final breeding stage. The root-knot nematodes (Meloidogyne spp.) were the most frequent in roots (40 - 100%) and soil (85.71 - 100%), with a detectable number of juveniles (J2) varying between genotypes (4 - 148 J2.250g-1 roots, and 1 - 110 J2.100 cm-3soil). Four esterase phenotypes were characterized: M. incognita (Est I1 = Rm: 1.0), M. javanica (Est J3 = Rm: 1.0; 1.25 and 1.40 and Est J2 = Rm: 1.0 and 1.40) and M. arenaria (Est A2 = Rm: 1.20 and 1.35), M. javanica (Est J3) was predominant. Meloidogyne javanica and M. incognita were predominant, however mixed infestations between species were found. The occurrence of Meloidogyne spp. was: M. javanica (68.26%), M. incognita (64.73%) and M. arenaria (16.81%). Helicotylenchus multicinctus and Rotylenchulus reniformis was the second most frequent group. Radopholus similis, Scutellonema sp., Criconemoides sp. and Helicotylenchus sp. presented themselves in low frequency and population levels in banana plants.

Development and analysis of Multiparent Advanced Generation Inter-Cross (MAGIC) populations have been conducted with several crop plants to harness the potential for dissecting the genetic structure of traits and improving breeding populations. We developed a first MAGIC population for cowpea ( Vigna unguiculata L. Walp.) from eight founder parents which are genetically diverse and carry many abiotic and biotic stress resistance, seed quality and agronomic traits relevant to cowpea improvement in sub-Saharan Africa (SSA) where cowpea is vitally important in the human diet and in local economies. The eight parents were inter-crossed using structured matings to ensure the population would have balanced representation from each of the founder parents, followed by single-seed descent, resulting in 365 F8 recombinant inbred lines (RILs) each carrying a mosaic of genome blocks contributed from all founders. This was confirmed by SNP genotyping with the cowpea Illumina 60K iSelect BeadArray. Following filtering to eliminate duplicates, sister lines and accidental selfing events, a core set of 305 F8 RILs was chosen as the primary population. The F8 lines were on average 99.74% homozygous while also diverse in agronomic traits including flowering time, growth habit, maturity, yield potential and seed characteristics across environments. Trait-associated SNPs were identified for most of the parental traits. Loci with major effects on photoperiod sensitivity and seed size were also verified by genetic mapping in biparental RIL populations. The distribution of recombination frequency varied considerably between chromosomes, with recombination hotspots distributed mostly in the telomeric regions. Due to its broad genetic base, this cowpea MAGIC population promises breakthroughs in genetic gain and high-resolution genetic mapping for gene discovery, enhancement of breeding populations and, for some lines, direct releases as new varieties.

Summary: Meloidogyne enterolobii is an emerging pathogen in Brazil, considered highly destructive to tomato. Due to the lack of cultivars resistant to this nematode, the objective of this work was to evaluate the effect of Trichoderma spp. strains on the promotion of tomato growth and biological control of M. enterolobii populations. The use of eight strains from the CENARGEN collection and three formulated products at a concentration of 1.43 × 10⁶ conidia ml⁻¹ significantly increased the fresh weight of roots compared to control plants inoculated only with nematodes. These Trichoderma strains, except CEN316, also promoted a significant increase in the number of nematodes and in the reproduction factor (RF). With the increase of spore concentration (1 × 10⁸ conidia ml⁻¹) in the second assay, the inoculation of nematode or fungi plus nematode promoted an increase in the fresh weight of roots, except in treatments CEN162, CEN287, UFT 201, and CEN287 + nematodes, which maintained the fresh weight of roots equal to the control without nematodes. None of the spore concentrations (1.43 × 10⁶ and 1 × 10⁸ ml⁻¹) of different Trichoderma spp. strains influenced the plant height (45 and 90 days after nematode inoculation) or the dry plant weight 90 and 110 days after nematode inoculation, respectively. However, there was a positive and significant correlation between RF and fresh root weight, eggs (g root)⁻¹ and total eggs, in the two doses. Furthermore, the Trichoderma spp. strains did not result in control of M. enterolobii under glasshouse conditions after a complete life cycle of the tomato plant: high nematode infection was observed in both doses of the fungi, including in treatments with the commercial formulations.

The use of in vitro mass multiplication of nematodes under axenic conditions allows intensification of studies on taxonomy, biology, epidemiology and control of these pathogens. In the present study, two methods were compared for in vitro multiplication of two species of plant-parasitic nematodes, Radopholus similis and Pratylenchus brachyurus in 20, 40 and 60 day-periods. Both techniques involved the use of carrot cylinders placed in jars with lids, with water-agar in the bottom of the flasks (WA+) or without (WA-). In WA- the nematodes were treated with a 1% solution of ampicillin whilst for WA+ the axenization was carried out with a solution of mercuric chloride (0.01%), and streptomycin sulfate 0.02%. The WA+ condition was the most favorable for multiplication of both species of nematodes, resulting in a population increase of about 280 times for R. similis and 226 times for P. brachyurus in relation to the founding population of 25 individuals. For WA- a population increase of only five times after 60 days for R. similis and a duplication of the population after 60 days for P. brachyurus was obtained. Overall, the largest amount of nematodes occurred at 60 days after inoculation

The burrowing nematode Radopholus similis is among the most damaging pathogens. Resistant plants are one of the most promising approaches for nematode control, and knowledge of resistance and aggressiveness components is essential to understand resistance genetics and developing new cultivars. Therefore, this study aimed to assess the degree of resistance of eight banana accessions to populations of R. similis to verify differences in the reproduction capacity between nematode populations and to study the horizontal/vertical resistance and aggressiveness/ virulence components of Musa spp. and R. similis pathosystem. The accessions 4249-05, 4279-06, Yangambi Km5, 0323-03, 0337-02, 1304-06, Borneo and Grande Naine were inoculated with one of three R. similis populations from the Brazilian states of Pernambuco, Distrito Federal, and Santa Catarina and kept in a greenhouse. Accessions 4249-05, Yangambi Km5, 0323-03, and 4279-06 showed different resistance levels, and the Pernambuco population had the highest reproductive capacity. Using Griffing Model IV, evidence of vertical and horizontal resistance was observed, with accessions 4249-05 and Yangambi km5 showing the highest levels of horizontal resistance.