Complete genome sequence of Bacillus velezensis strain Ag109, a biocontrol agent against plant-parasitic nematodes and Sclerotinia sclerotiorum.

Soybean is the main oilseed cultivated worldwide. Even though Brazil is the world's largest producer and exporter of soybean, its production is severely limited by biotic factors. Soil borne diseases are the most damaging biotic stressors since they significantly reduce yield and are challenging to manage. In this context, the present study aimed to evaluate the potential of a bacterial strain (Ag109) as a biocontrol agent for different soil pathogens (nematodes and fungi) of soybean. In addition, the genome of Ag109 was wholly sequenced and genes related to secondary metabolite production and plant growth promotion were mined. Ag109 showed nematode control in soybean and controlled 69 and 45% of the populations of Meloidogyne javanica and Pratylenchus brachyurus, respectively. Regarding antifungal activity, these strains showed activity against Macrophomia phaseolina, Rhizoctonia solani, and Sclerotinia sclerotiorum. For S. sclerotiorum, this strain increased the number of healthy plants and root dry mass compared to the control (with inoculation). Based on the average nucleotide identity and digital DNA-DNA hybridization, this strain was identified as Bacillus velezensis. Diverse clusters of specific genes related to secondary metabolite biosynthesis and root growth promotion were identified, highlighting the potential of this strain to be used as a multifunctional microbial inoculant that acts as a biological control agent while promoting plant growth in soybean.

Abscisic Acid Is Involved in the Resistance Response of Arabidopsis thaliana Against Meloidogyne paranaensis.

Abscisic acid (ABA) is a classical hormone involved in the plant defense against abiotic stresses, especially drought. However, its role in the defense response against biotic stresses is controversial: it can induce resistance to some pathogens but can also increase the susceptibility to other pathogens. Information regarding the effect of ABA on the relationship between plants and sedentary phytonematodes, such as Meloidogyne paranaensis, is scarce. In this study, we found that ABA changed the susceptibility level of Arabidopsis thaliana against M. paranaensis. The population of M. paranaensis was reduced by 58.3% with the exogenous application of ABA 24 h before the nematode inoculation, which demonstrated that ABA plays an important role in the preinfectional defense of A. thaliana against M. paranaensis. The increase in the nematode population density in the ABA biosynthesis mutant, aba2-1, corroborated the results observed with the exogenous application of ABA. The phytohormone did not show nematicide or nematostatic effects on M. paranaensis juveniles in in vitro tests, indicating that the response is linked to intrinsic plant factors, which was corroborated by the decrease in the number of nematodes in the abi4-1 mutant. This reduction indicates that the gene expression regulation by transcript factors is possibly related to regulatory cascades mediated by ABA in the response of A. thaliana against M. paranaensis.

Virulence and Reproductive Fitness of Meloidogyne paranaensis Field Populations in Coffee Genotypes.

The aim of this study was to characterize Meloidogyne paranaensis populations collected from infested coffee crops. Methodologies used to characterize the 11 studied populations from municipalities in Paraná and Minas Gerais States involved the morphological analysis of perineal patterns, biochemical analysis by isozyme electrophoresis, sequencing of internal transcribes spacer 1 (ITS-1) and D2/D3 ribosomal DNA (rDNA) regions, reproductive fitness, and virulence characterization in coffee genotypes. Morphological evaluations showed the existence of variation between populations, although the majority of them showed typical perineal patterns. The biochemical identification was based on α-esterase isozyme analyses and resulted in the appearance of three distinct profiles: P1 (typical), P2 (atypical), and a nondescribed profile, P2b. BLAST of the ITS-1 and D2/D3 rDNA regions indicated homology (>95%) with other sequences deposited in GenBank. For reproductive fitness and virulence characterization, 13 coffee genotypes (5 Coffea arabica and 8 C. canephora) were inoculated with 11 M. paranaensis populations. Variation in the reproductive fitness of populations was observed for cultivar Mundo Novo, a genotype without resistance genes, and variation in the virulence of populations was observed in genotypes carrying resistance genes. Three populations exhibited virulence combined with high reproductive fitness, while one showed virulence with low reproductive fitness. Some hosts were resistant to 11 populations, while one of the hosts was resistant to only one population, indicating the presence of different resistance genes. Nevertheless, no relationship was observed between the origin of population and their variations in perineal patterns, esterase profiles, phylogeny, or reproductive fitness in coffee genotypes, or between the different characterizations, although differences were observed within each characteristic.

Morus alba: Host reaction for Meloidogyne javanica, biological nematicides assessment and study of these relationships with yield and quality of leaves, cocoon and health of the silkworm.

Root-knot nematodes cause damage to several crops and the importance of each species can vary according with the crop and the agricultural region. In Brazil, Meloidogyne javanica is one of the most important nematode species parasitizing mulberry. To define management strategies, it is important to know if the crop species is damaged by the parasitism of the nematode and the best choices for control, as the use of nematicides. Biological nematicides have been extensively used in Brazil, but no information regarding its efficiency to control M. javanica in mulberry is available. Besides, it is not known if biological nematicides could improve the quality of leaves or if they alter the nutrient composition of leaves, which could interfere in the development of the silkworms that are feed with these leaves or in the quality of the silk produced. With the aim to address these questions, we propose a study that will start in the phenotyping of the main Brazilian mulberry cultivars to Meloidogyne species, passing through the test of efficiency of biological nematicides in the control of M. javanica in mulberry cultivar Miura, evaluation of the amount and quality of leaves produced and, using these leaves to feed silkworms, in the analyzes of the impact of these diet in the health of silkworms, and in the production and quality of the silk.

Aphelenchoides besseyi Parasitizing Common Bean in Brazil.

Aphelenchoides besseyi is the causal agent of soybean green stem and foliar retention syndrome known as "Soja Louca II." This nematode has recently been reported parasitizing cotton in Brazil. In Costa Rica, it causes the symptoms known as "amachamiento" and false angular spots in common bean (Phaseolus vulgaris). Due to the great importance of beans to Brazilian agriculture, the objective of this research was to study the pathogenicity of A. besseyi in common bean under greenhouse conditions, including its endoparasitic relationships by staining root and shoot system tissues with fuchsin acid. In addition, A. besseyi was collected and quantified from shoot systems 30 days after inoculation by washing the tissue in water and blender centrifugal flotation. We observed the symptoms of amachamiento, leaf and vein deformation in the expanded trifoliate leaves, and also leaves with necrotic, brown to reddish and angular lesions, characteristics from false angular spot, and deformed stems characterized by enlargement of nodes, retortions, and necrotic lesions. High numbers of nematodes were found inside common bean plants. This is the first report of the pathogenicity and symptoms caused by A. besseyi in common bean in Brazil. These findings are important for development of management strategies to avoid losses on bean cropped in infested areas.

Two novel potential pathogens for soybean.

Nematode diseases have a worldwide importance for several economic agricultural crops, such as soybean. Frequently, new or secondary pathogens arise as emergent diseases due to the intensified use of agricultural lands, causing extensive yield losses. Helicotylenchus dihystera (Hd) and Scutellonema brachyurus (Sb) appear as potential pathogens for soybean in Brazil, since their spread and population densities have been increased on soybean growing areas. Aiming to evaluate the quantitative distribution of Hd and Sb in soybean fields in South Brazil, a survey was conducted during the growing seasons of 2014-2015 and 2015-2016 in which 1,088 soil samples and 1,043 root samples were analyzed. Besides, two greenhouse experiments were conducted to evaluate the pathogenicity of both nematodes to soybean plants, in comparison with P. brachyurus (Pb), a known pathogen of the crop. The survey demonstrated that Pb, Hd and Sb are widely distributed in the States of Paraná, Santa Catarina and Mato Grosso do Sul. Besides, we proved that Hd and Sb multiply and cause root lesions in soybean cv. Potência, since they were found inside roots, and can be considered as potential pathogens for soybean plants.

Meloidogyne and Pratylenchus species in sugarcane fields in the state of Alagoas, Brazil / Espécies de Meloidogyne e de Pratylenchus em áreas cultivadas com cana-de-açúcar no estado de Alagoas

ABSTRACT: The objective of this study was to accomplish a survey on populations of Meloidogyne and Pratylenchus species in sugarcane farming areas in the state of Alagoas, Brazil. Twenty samples of soil and roots were processed to extract and quantify nematodes; however, the identification of Meloidogyne species was performed using only 12 samples. Pratylenchus spp. were reported at moderate population levels of 68-1556 specimens 50g-1 of roots and 2-298 specimens 100cm-3 of soil in twenty analyzed samples. For Meloidogyne spp., these values were of 12-487 specimens 50g-1 of roots and 0-140 specimens 100cm-3 of soil. Based on electrophoresis of esterase isozymes, M. incognita was reported to be the most frequent species, followed by M. javanica and M. arenaria. Pratylenchus species identified through morphometrical and morphological characteristics were P. zeae and P. brachyurus , with predominance for the first species. No significant correlation (P≤0.05) were reported between nematode populations and sugarcane cropping systems.

RESUMO: O objetivo deste trabalho foi realizar um levantamento de espécies de Meloidogyne e de Pratylenchus existentes em áreas de cultivo de cana-de-açúcar, no estado de Alagoas. Vinte amostras de solo e raízes foram processadas para a extração e quantificação, sendo a identificação das espécies de Meloidogyne realizada em 12 amostras. Detectou-se a presença de Pratylenchus spp. em níveis populacionais médios de 68-1556 espécimes 50g-1 de raízes e 2-298 100cm-3 de solo, em todas as amostras. Para Meloidogyne spp., esses valores foram de 12-487 50g-1 de raízes e 0-140 100cm-3 de solo. Com base na eletroforese da isoenzima esterase, M. incognita foi a espécie mais frequente, seguida por M. javanica e M. arenaria . As espécies de Pratylenchus identificadas pelas características morfológicas e morfométricas, foram P. zeae e P. brachyurus , com predomínio da primeira espécie. Não houve correlações significativas (P≤0,05) entre as populações de nematoides e as características dos sistemas de cultivo de cana-de-açúcar.

Root-Knot Nematodes (Meloidogyne spp.) Parasitizing Resistant Tobacco Cultivars in Southern Brazil.

Our study surveyed Meloidogyne spp. from tobacco cultivars in Brazil carrying the Rk gene, which confers resistance to Meloidogyne incognita races 1 and 3. Identification of Meloidogyne populations from 39 tobacco fields was based on the analysis of esterase phenotypes, perineal patterns, and 18S-internal transcribed spacer (ITS)1-5.8S ribosomal RNA sequences. Biometric characterization and differential host tests of isolates were determined as well. We detected M. incognita, M. javanica, M. enterolobii, M. arenaria, and M. inornata in 18 (46.2%), 16 (41%), 10 (25.6%), 2 (5.1%), and 1 (2.6%) samples, respectively. Mixtures of species were found in 25.6% of the samples. This is the first report of M. inornata parasitizing tobacco in Brazil's southern region. Two morphological and biochemically unusual populations had host ranges and ITS1 sequences nearly identical to M. enterolobii and M. incognita, respectively. Pathogenic and biometric intraspecific variations were observed. Based on our results and considering the limited efficiency of the Rk gene, it is indispensable not only to search for resistance sources to M. incognita virulent strains but also to consider resistance to M. enterolobii, M. inornata, M. arenaria, and M. javanica in tobacco breeding programs. Changes in integrated management procedures should be considered to avoid increased crop damage in the future.