RESUMO

Infective second-stage juveniles (J2) of Meloidogyne spp. migrate towards host roots, which depends on several factors, including root exudates and soil temperature. Although Meloidogyne enterolobii is a highly virulent nematode that affects major agricultural crops worldwide, there is limited ecological data about it. The objective of this study was to determine the J2 migration pattern vertically in 14-cm long segmented soil columns towards tomato (Solanum lycopersicum) and marigold (Tagetes patula) roots, each grown at two soil temperatures (20 or 26ºC). Bottomless cups with tomatoes or marigolds were attached to the top of each column; cups with no plants were used as untreated controls. Juveniles (1,000/column) were injected into a hole located 1 cm from the bottom of each column. The apparatuses were placed in growth chambers at 20 or 26ºC, and J2 were allowed to migrate for 3, 6, 9, or 12 days after injection (DAI). At each harvest, J2 were extracted from each ring of the columns and counted to compare their distribution, and root systems were stained to observe root penetration. M. enterolobii migrated over 13 cm vertically 3 DAI regardless of temperature, even without plant stimuli. The vertical migration was greater at 26ºC, where 60% of active J2 were found at distances >13 cm at 12 DAI. Temperature did not affect root penetration. Overall, a greater number of J2 was observed in tomato roots, and root penetration increased over time.

RESUMO

Amazon chicory (Eryngium foetidum L. [Apiaceae]), also known as culantro, is native to Tropical America and the West Indies. It belongs to the unconventional food plants (UFPs) group, and in addition to be consumed as a spice herb, it possesses a wide range of ethnomedicinal uses (Paul et al. 2011). In 2019, in the eastern Amazon region of Brazil, state of Pará, producers of E. foetidum in the municipality of Castanhal (01°15'363" S 047°10'232" W) reported the occurrence of underdeveloped plants with leaf yellowing and a large number of galls in the root system, which are typical symptoms of root-knotting nematode. Soil and root samples were collected and sent to the Nematology Laboratory (LabNema) located at the Faculty of Agrarian and Veterinary Sciences, UNESP, Jaboticabal, São Paulo, Brazil. A total of 46 second-stage juveniles (J2s) were extracted per 100 cm3 of soil, and a total of 460 eggs and J2s Meloidogyne spp. were found per gram of root. Morphological and molecular techniques were used to identify the species. The analysis of the perineal patter of ten females revealed thin striations in an oval shape with a high and semi-trapezoidal dorsal arch. No striations were observed in the perivulvar region. The labial region of the ten males analyzed exhibited a non-prominent labial disc, fused and slightly recessed submedian lips, with no apparent annulations. The morphological characteristics observed in the adults were consistent with those originally described for Meloidogyne enterolobii (Yang; Eisenback, 1983), confirming the species purity of the recovered population. Three individual nematodes had their 18S rDNA region sequenced (Holterman et al. 2006) which showed an average identity of 99.7% with other sequences of M. enterolobii available in the GenBank database. A Bayesian phylogenetic tree was constructed, providing insights into the specific relationship of M. enterolobii recovered from E. foetidum with other related nematodes. Each of the three sequenced nematodes represented a unique haplotype, resulting in their separation into distinct clades. Moreover, the obtained sequences presented polymorphisms that differed from the M. enterolobii sequences already available in the database, highlighting the genetic diversity of this species in relation to its original host (Silva et al. 2021). The species M. enterolobii was also confirmed using species-specific primers for M. incognita, M. javanica, and M. enterolobii (Zijlstra et al. 2000; Tigano et al. 2010). To confirm the pathogenicity of M. enterolobii on E. foetidum, a modified Koch Postulate was conducted. Six seedlings of E. foetidum were transplanted individually to 10-liter pots containing autoclaved soil. Each pot was then inoculated with 5 mL of a suspension containing 3,000 eggs and J2s from the original population of M. enterolobii obtained from E. foetidum. After 90 days, the inoculated plants exhibited root galls with a plentiful egg mass, in contrast to the healthy non-inoculated plants. The average number of M. enterolobii nematodes recovered from the roots of the inoculated plants was 42,040 eggs and J2s, resulting in a reproduction factor (RF) of 14.0. The importance of reporting the occurrence of M. enterolobii in E. foetidum is due to the fact that this plant species is cultivated in a crop rotation system with other vegetables such as lettuce and coriander, which are also hosts of M. enterolobii. Consequently, different crop rotation strategies and control alternatives need to be considered in areas where E. foetidum is grown. This is the first report of E. foetidum serving as a host for the root-knot nematode M. enterolobii worldwide.

RESUMO

Despite the worldwide importance of disease complexes involving root-feeding nematodes and soilborne fungi, there have been few in-depth studies on how these organisms interact at the molecular level. Previous studies of guava decline have shown that root exudates from Meloidogyne enterolobii-parasitized guava plants (NP plants), but not from nematode-free plants (NF plants), enable the fungus Neocosmospora falciformis to rot guava roots, leading to plant death. To further characterize this interaction, NP and NF root exudates were lyophilized; extracted with distinct solvents; quantified regarding amino acids, soluble carbohydrates, sucrose, phenols, and alkaloids; and submitted to a bioassay to determine their ability to enable N. falciformis to rot the guava seedlings' roots. NP root exudates were richer than NF root exudates in amino acids, carbohydrates, and sucrose. Only the fractions NP-03 and NP-04 enabled fungal root rotting. NP-03 was then sequentially fractionated through chromatographic silica columns. At each step, the main fractions were reassessed in bioassay. The final fraction that enabled fungal root rotting was submitted to analysis using high performance liquid chromatography, nuclear magnetic resonance, mass spectrometry, energy-dispersive X-ray fluorescence, and computational calculations, leading to the identification of 1,5-dinitrobiuret as the predominant substance. In conclusion, parasitism by M. enterolobii causes an enrichment of guava root exudates that likely favors microorganisms capable of producing 1,5-dinitrobiuret in the rhizosphere. The accumulation of biuret, a known phytotoxic substance, possibly hampers root physiology and the innate immunity of guava to N. falciformis.

RESUMO

The sweetpotato (Ipomoea batatas L., Convolvulaceae family) originated in Latin America and is currently cultivated worldwide. The storage roots, rich in calories, have made this crop one of the main caloric sources for low-income populations, especially in developing countries. Brazil annually produces about 805,000 tons, with the Northeast region responsible for 34% of this production (Albuquerque et al. 2020). In October 2019, sweetpotato plants cv. Campina, from a field in the region of Touros, state of Rio Grande do Norte (RN), Brazil (5°12'31"S 35°34'42"W), presented deformed storage roots, with galls, typical of root-knot nematodes. The roots were sent to the Nematology Laboratory (LabNema) where 14,032 eggs and 3,312 second-stage juveniles (J2s) of Meloidogyne sp., in 10 g of roots, were recovered. The species of adults was identified through morphological, biochemical, and phylogenetic analysis. The perineal region of females (n = 10) presented an oval shape, with a high and semi-trapezoidal dorsal arch and streak-free perivulval region. The labial region of males (n=10) presented high and rounded head cap, labial region slightly set off from the body, without annulations. The morphological characters were compatible with the original description of Meloidogyne enterolobii (Yang and Eisenback 1983). The phenotype of esterase isoenzymes showed two major bands (VS1-S1) also characteristic of M. enterolobii (Esbenshade and Triantaphyllou 1985). Sequences of 18S rDNA (~1200bp) of individual females (Holterman et al. 2006) obtained from sweetpotatoes before (SPme1 and 2) and after inoculation (SPme3 and 6), and from guava, used as M. enterolobii species control, were submitted to Bayesian analysis. The sequences presented genetic diversity among them resulting from seven SNPs (Single Nucleotide Polymorphism) and 99.4 to 99.9% identity with M. enterolobii sequences deposited in the NCBI GenBank (accession numbers MW209034-MW209039). The pathogenicity test was carried out under greenhouse conditions, in which 3,000 eggs and J2s from the original population isolated of M. enterolobii were inoculated in sweetpotato seedlings cv. Campina (n = 6). After three months, the roots presented galls and deformations typical of root-knot nematodes, while non-inoculated plants did not present any symptoms. An average of 15,900 eggs and J2s of M. enterolobii (RF = 5.3) were recovered from the roots, proving that sweetpotatoes were a host of this species. Meloidogyne enterolobii is known to cause great damage to sweetpotato (Ye et al. 2020). In Brazil, Meloidogyne nematode had been reported once, isolated from a sweetpotato field in the Ceara state and the species suggested by the authors according to esterase electrophoresis was M. enterolobii. Nonetheless, the authors did not present taxonomic, isoenzyme phenotypes and molecular species identification integratively, nor included pathogenicity tests (Silva et al. 2016). Therefore, it is the first time that M. enterolobii, with reliable identification by different methods, including sequencing, was detected in commercial sweetpotato fields in the RN state and in Brazil. The local farmers reported that this nematode deforms the storage roots which make them useless for commercialization, resulting in minimal losses of 50% of production in the infested areas. Furthermore, as sweetpotatoes are vegetatively propagated, the spread of this nematode through planting material is favored. Considering the importance of this crop in Brazil, this report is essential for control measures of this pathogen to be taken in order to avoid its spread to other regions.

RESUMO

The Meloidogyne-based disease complexes (MDCs) are caused by the interaction of different root-knot nematode species and phytopathogenic fungi. These complexes are devastating several important crops worldwide including tomato and coffee. Despite their relevance, little is known about the role of the bacterial communities in the MDCs. In this study 16s rDNA gene sequencing was used to analyze the bacterial microbiome associated with healthy and infested roots, as well with females and eggs of Meloidogyne enterolobii and M. paranaensis, the causal agents of MDC in tomato and coffee, respectively. Each MDC pathosystems displayed a specific taxonomic diversity and relative abundances constituting a very complex system. The main bacterial drivers of the MDC infection process were identified for both crops at order level. While corky-root coffee samples presented an enrichment of Bacillales and Burkholderiales, the corcky-root tomato samples presented an enrichment on Saprospirales, Chthoniobacterales, Alteromonadales, and Xanthomonadales. At genus level, Nocardia was common to both systems, and it could be related to the development of tumor symptoms by altering both nematode and plant systems. Furthermore, we predicted the healthy metabolic profile of the roots microbiome and a shift that may result in an increment of activity of central metabolism and the presence of pathogenic genes in both crops.

RESUMO

Meloidogyne enterolobii has become an economically important plant parasitic nematode worldwide because of its high aggressiveness, increasing geographic distribution, wide host range, and pathogenicity in pepper (Capsicum annuum) cultivars carrying resistance genes to Meloidogyne incognita, Meloidogyne arenaria, and Meloidogyne javanica. The objectives of this study were to identify landraces of peppers resistant to M. enterolobii and analyze the relationship between resistance indicators and the phenotype parameters of plant height, stem width, leaf length, leaf width, relative chlorophyll, and number of flowers. Ninety landraces of C. annuum were collected from several states of Mexico and were inoculated with 2,000 eggs of M. enterolobii. Eleven resistant landraces were selected and confirmed with a second inoculation experiment. Seventy-five days after inoculation, in both experiments, the resistance of landraces UTC66, UTC90, UTC67, UTC88, and UTC81 to M. enterolobii was consistent. Although genotypes UTC24, UTC79, UTC65, UTC68, UTC69, and UTC25 were susceptible, these landraces had a significantly higher proportion of resistant plants, less root galling, and a lower reproductive index, in comparison with the rest of the 79 genotypes and the susceptible control, which were highly susceptible in both experiments. There was no correlation between resistant indicators and phenotypic parameters, although plant height, relative chlorophyll, and number of flowers were in general significantly affected compared with noninoculated controls, indicating that the nematodes reduce the growth and yield of peppers. Results indicate that all resistant plants from these landraces are promising sources of resistance for the development of pepper cultivars resistant to M. enterolobii.

Assuntos

RESUMO

The present study aimed to evaluate Capsicum accessions for resistance to Meloidogyne incognita race 3, Meloidogyne javanica and Meloidogyne enterolobii. Two experiments with different genotypes of hot and sweet peppers were carried out in a completely randomized design. The first experiment was conducted in a 31 x 3 factorial scheme with 27 genotypes of Capsicum annuum, two cultivars of hot pepper, one line of Capsicum frutescens and tomato 'Santa Cruz Kada', and three species of nematodes (M. incognita race 3, M. javanica and M. enterolobii). In the second experiment, we used a factorial scheme 39 x 3 with 36 accessions of C. annuum, two hot pepper cultivars and the 'Santa Cruz Kada' tomato and three nematodes species mentioned earlier. The total number of eggs and second-stage juveniles (TNEJ), number of eggs and second-stage juveniles per gram of root (NEJGR), reproduction index (RI) and reproduction factor (RF) were evaluated. Based on RI and RF, the genotypes CNPH 185, CNPH 187 and CNPH 680 were resistant and very resistant to M. incognita race 3 and M. javanica, simultaneously. The C. frutescens line presented resistance to the three root-knot nematode species.

O presente trabalho teve por objetivo avaliar acessos de Capsicum quanto à resistência a Meloidogyne incognita raça 3, Meloidogyne javanica e Meloidogyne enterolobii. Foram realizados dois experimentos, com diferentes genótipos de pimentas e pimentões, em delineamento inteiramente casualizado sendo o primeiro em esquema fatorial 31 x 3 com 27 genótipos de Capsicum annuum, duas cultivares de pimenta, uma linhagem de Capsicum frutescens, o tomateiro 'Santa Cruz Kada' e três espécies de nematoides (M. incognita raça 3, M. javanica e M. enterolobii). No segundo experimento foi utilizado esquema fatorial 39 x 3 com 36 acessos de C. annuum, duas cultivares de pimenta, o tomateiro 'Santa Cruz Kada' e três espécies de nematoides mencionadas anteriormente. Avaliou-se o número total de ovos e juvenis de segundo estádio (NTOJ), número de ovos e juvenis de segundo estádio por grama de raízes (NOJGR), índice de reprodução (IR) e fator de reprodução (FR). Com base no FR e IR os genótipos CNPH 185, CNPH 187 e CNPH 680 foram resistentes e muito resistentes a M. incognita raça 3 e M. javanica, simultaneamente. A linhagem de C. frutescens apresentou resistência às três espécies de nematoides de galha.

Assuntos

RESUMO

Meloidogyne enterolobii is one of the most important root-knot nematode in tropical regions, due to its ability to overcome resistance mechanisms of a number of host plants. The lack of new and safe active ingredients against this nematode has restricted control alternatives for growers. Egg-parasitic fungi have been considered as potential candidates for the development of bionematicides. In tissue culture plates, Pochonia chlamydosporia (var. catenulata and chlamydosporia) and Purpureocillium lilacinum strains were screened for their ability to infect eggs of the root-knot nematode M. enterolobii on water-agar surfaces. Reduction in the hatching of J2 varied from 13% to 84%, depending on strain. The more efficacious strains reduced hatchability of J2 by 57% to 84% when compared to untreated eggs, but average reductions were only 37% to 55% when the same strains were applied to egg masses. Combinations of fungal isolates (one of each species) did not increase the control efficacy in vitro. In experiments in which 10,000 nematode eggs were inoculated per plant, reductions in the number of eggs after 12 months were seen in three of four treatments in banana plants, reaching 34% for P. chlamydosporia var. catenulata. No significant reductions were seen in tomato plants after 3 mon. In another experiment with tomato plants using either P. chlamydosporia var. catenulata or P. lilacinum, the number of eggs was reduced by 34% and 44%, respectively, when initial infestation level was low (500 nematode eggs per plant), but tested strains were not effective under a moderate infestation level (5,000 eggs per plant). Under all infestation levels tested in this work, gall and egg mass indexes (MI) did not differ from the untreated controls, bringing concerns related to the practical adoption of this control strategy by farmers. In our opinion, if the fungi P. chlamydosporia and P. lilacinum are to be used as biocontrol tools toward M. entorolobii, they should focus on agricultural settings with low soil infestation levels and within an IPM approach.

RESUMO

The cassava crop (Manihot esculenta) has socioeconomic importance to Brazil, however, there is a paucity of studies regarding the plant parasitic nematodes associated with cassava and the host reaction of cassava cultivars to nematodes. The aims of this work were to report the geographic distributions and new records of plant parasitic nematodes collected from cassava areas from the States of Acre, Amapá, Pará and Rondônia, Brazil. Additionally, the host reaction of different cassava cultivars to Meloidogyne incognita race 3, Pratylenchus brachyurus and P. zeae was studied in a greenhouse experiment. Nematodes were extracted from either 10 g of roots or 250 cm³ of soil by centrifugation and identified by microscopy. The most prevalent species was P. brachyurus detected from 37.1% of samples tested (n=35), follow by M. incognita (14.2%). Among the identified nematode species, the following associations constitute new records for Brazil: Helicotylenchus erythrinae, M. enterolobii and Xiphinema longicaudatum. Cassava cultivars Amazonas, Colônia Matapi, Manteiga, Pretinha and Pó-da-China from Amapá State and Colonial and Caipora from Acre State were all susceptible to M. incognita race 3. In contrast, two cultivars, Caipora and Colonial, were resistant to P. brachyurus (reproduction factor = 0.5 and 0.9, respectively) but immune to P. zeae. Data generated in this study may be useful in the agronomic management of cassava production in areas infested with the root lesion nematodes P. brachyurus and/or P. zeae.

A cultura da mandioca (Manihot esculenta) tem importância socioeconômica no Brasil, no entanto há ainda uma escassez de informações sobre a associação de nematoides fitoparasitas, bem como ao comportamento de cultivares de mandioca ao parasitismo desses agentes. O objetivo do presente trabalho foi relatar a distribuição geográfica e novas ocorrências das principais espécies de nematoides encontradas associadas à mandioca em diferentes municípios dos estados do Acre, Amapá, Pará e Rondônia. Além disso, estudou-se, em casa de vegetação, a reação de algumas cultivares de mandioca ao parasitismo de Meloidogyne incognita raça 3, Pratylenchus brachyurus e P. zeae. Nematoides foram extraídos de 10 g de raízes ou 250 cm³ de solo e identificados sob microscópio de luz. A espécie mais prevalente foi P. brachyurus, presente em 37,1% das amostras analisadas (n=35), seguida de M. incognita, encontrada em de 14,2% das amostras. Constituíram novas ocorrências para mandioca no Brasil: Helicotylenchus erythrinae, M. enterolobii e Xiphinema longicaudatum. As cultivares de mandioca provenientes do Amapá (Amazonas, Colônia Matapi, Manteiga, Pretinha e Pó-da-China) e as cultivares de mandioca Colonial e Caipora, provenientes do Acre, comportaram-se como suscetíveis a M. incognita raça 3. Por outro lado, as cultivares Caipora e Colonial comportaram-se como resistentes a P. brachyurus (fator de reprodução = 0,5 e 0,9, respectivamente) e imunes a P. zeae. Informações geradas neste estudo podem ser úteis para o manejo de áreas cultivadas com mandioca e infestadas pelos nematoides das lesões, P. brachyurus e/ou P. zeae.

RESUMO

The cassava crop (Manihot esculenta) has socioeconomic importance to Brazil, however, there is a paucity of studies regarding the plant parasitic nematodes associated with cassava and the host reaction of cassava cultivars to nematodes. The aims of this work were to report the geographic distributions and new records of plant parasitic nematodes collected from cassava areas from the States of Acre, Amapá, Pará and Rondônia, Brazil. Additionally, the host reaction of different cassava cultivars to Meloidogyne incognita race 3, Pratylenchus brachyurus and P. zeae was studied in a greenhouse experiment. Nematodes were extracted from either 10 g of roots or 250 cm³ of soil by centrifugation and identified by microscopy. The most prevalent species was P. brachyurus detected from 37.1% of samples tested (n=35), follow by M. incognita (14.2%). Among the identified nematode species, the following associations constitute new records for Brazil: Helicotylenchus erythrinae, M. enterolobii and Xiphinema longicaudatum. Cassava cultivars Amazonas, Colônia Matapi, Manteiga, Pretinha and Pó-da-China from Amapá State and Colonial and Caipora from Acre State were all susceptible to M. incognita race 3. In contrast, two cultivars, Caipora and Colonial, were resistant to P. brachyurus (reproduction factor = 0.5 and 0.9, respectively) but immune to P. zeae. Data generated in this study may be useful in the agronomic management of cassava production in areas infested with the root lesion nematodes P. brachyurus and/or P. zeae.(AU)

A cultura da mandioca (Manihot esculenta) tem importância socioeconômica no Brasil, no entanto há ainda uma escassez de informações sobre a associação de nematoides fitoparasitas, bem como ao comportamento de cultivares de mandioca ao parasitismo desses agentes. O objetivo do presente trabalho foi relatar a distribuição geográfica e novas ocorrências das principais espécies de nematoides encontradas associadas à mandioca em diferentes municípios dos estados do Acre, Amapá, Pará e Rondônia. Além disso, estudou-se, em casa de vegetação, a reação de algumas cultivares de mandioca ao parasitismo de Meloidogyne incognita raça 3, Pratylenchus brachyurus e P. zeae. Nematoides foram extraídos de 10 g de raízes ou 250 cm³ de solo e identificados sob microscópio de luz. A espécie mais prevalente foi P. brachyurus, presente em 37,1% das amostras analisadas (n=35), seguida de M. incognita, encontrada em de 14,2% das amostras. Constituíram novas ocorrências para mandioca no Brasil: Helicotylenchus erythrinae, M. enterolobii e Xiphinema longicaudatum. As cultivares de mandioca provenientes do Amapá (Amazonas, Colônia Matapi, Manteiga, Pretinha e Pó-da-China) e as cultivares de mandioca Colonial e Caipora, provenientes do Acre, comportaram-se como suscetíveis a M. incognita raça 3. Por outro lado, as cultivares Caipora e Colonial comportaram-se como resistentes a P. brachyurus (fator de reprodução = 0,5 e 0,9, respectivamente) e imunes a P. zeae. Informações geradas neste estudo podem ser úteis para o manejo de áreas cultivadas com mandioca e infestadas pelos nematoides das lesões, P. brachyurus e/ou P. zeae.(AU)