Pseudomonas syringae coffee blight is associated with the horizontal transfer of plasmid-encoded type III effectors.

The emergence of new pathogens is an ongoing threat to human health and agriculture. While zoonotic spillovers received considerable attention, the emergence of crop diseases is less well studied. Here, we identify genomic factors associated with the emergence of Pseudomonas syringae bacterial blight of coffee. Fifty-three P. syringae strains from diseased Brazilian coffee plants were sequenced. Comparative and evolutionary analyses were used to identify loci associated with coffee blight. Growth and symptomology assays were performed to validate the findings. Coffee isolates clustered in three lineages, including primary phylogroups PG3 and PG4, and secondary phylogroup PG11. Genome-wide association study of the primary PG strains identified 37 loci, including five effectors, most of which were encoded on a plasmid unique to the PG3 and PG4 coffee strains. Evolutionary analyses support the emergence of coffee blight in PG4 when the coffee-associated plasmid and associated effectors derived from a divergent plasmid carried by strains associated with other hosts. This plasmid was only recently transferred into PG3. Natural diversity and CRISPR-Cas9 plasmid curing were used to show that strains with the coffee-associated plasmid grow to higher densities and cause more severe disease symptoms in coffee. This work identifies possible evolutionary mechanisms underlying the emergence of a new lineage of coffee pathogens.

Cross preferences and genetic diversity of Psidium interspecific hybrids through morphoagronomic traits and resistance to Meloidogyne enterolobii.

The introgression of M. enterolobii resistance-related genes in guava breeding programs can be compromised by incompatibility among Psidium species. This study aimed to evaluate the female parent preference and genetic diversity of Psidium interspecific hybrids using morphoagronomic traits and resistance to M. enterolobii. There were evaluated cross successes and germination from crosses between accesses of P. cattleyanum, P. guineense and P. guajava and the genetic diversity by Ward-MLM method of hybrids according to descriptors developed for the genus. Crosses were more successful when P. cattleyanum was the female parent. Germination was more successful in crosses involving P. cattleyanum and P. guajava. Four groups were formed. The group IV clustered the most resistant genotypes, composed by genotypes of P. cattleyanum x P. guineense, while the group II was the most susceptible. The groups I and III grouped some genotypes of P. cattleyanum x P. guajava with low levels of susceptibility. There are preferences of female parent species among crosses. Some individuals of groups I and III can be used as source of resistance genes for the breeding program, due the presence of favorable alleles inherited from guava parent. The high susceptibility leads to reduction in root development.

Meloidogyne enterolobii-induced Changes in Guava Root Exudates Are Associated With Root Rotting Caused by Neocosmospora falciformis.

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.

Characterizing the Interaction between Root-knot Nematodes and Soil-borne Fungi which are Pathogenic to Passion Fruit (Passiflora Edulis).

For decades there have been anecdotal claims of synergistic interactions between plant-parasitic nematodes and soil-borne fungi causing decline of productivity of passion fruit (Passiflora edulis) orchards. An empirical confirmation of these disease complexes would impact disease management and plant breeding for resistance. To test those claims, we subjected passion fruit plants to single or concomitant parasitism by Meloidogyne javanica or M. incognita and Fusarium nirenbergiae or Neocosmospora sp. under controlled conditions. Non-inoculated plants served as control for the assays. The severity of shoot symptoms and variables related to plant growth, the extent of fungal lesions, and nematode reproduction were assessed to characterize the interactions. The shoot symptoms and effect on plant growth induced by the pathogens varied, but no synergy between the pathogens was observed. Moreover, the volume of tissue lesioned by the fungi was not affected by co-parasitism of the nematodes. Conversely, plant resistance to the nematodes was not affected by co-parasitism of the fungi. The interactions M. incognita-F. nirenbergiae, M. incognita-Neocosmospora sp., M. javanica-F. nirenbergiae, and M. javanica-Neocosmospora sp. were not synergistic as previously claimed, but instead neutral.

Genetic structuring of segregating populations of Psidium spp resistant to the southern root-knot nematode by Bayesian approach as basis for the guava breeding program.

There are no guava cultivars resistant to the Meloidogyne enterolobii; for this reason, genetic breeding has been performed by introgressing genes into the current cultivars through interspecific hybridization. We used 33 microsatellite markers for the genetic-molecular characterization of segregating populations of Psidium resistant to M. enterolobii, aiming at selection within and between populations for generation advancement in the guava breeding program. The average number of alleles per locus ranged from 1.60 to 2.09. Populations 1 (P. guineense × P. cattleyanum) and 5 (P. guajava × P. cattleyanum) obtained the greatest genetic diversity, which can be confirmed by the higher observed-heterozygosity values (0.422 and 0.312, respectively). Bayesian analysis showed that the populations were subdivided into three groups, agreeing with the number of groups observed by Nei's genetic distance. The population obtained from the P. guineense × P. cattleyanum cross differed from the others with a clear structuring, whereas the P. guajava × P. cattleyanum and P. cattleyanum × P. guineense populations were the most similar between each other. The SSR markers were efficient in discriminating the populations, and individual 80 may be employed in future crosses with guava, allowing generation advancement in the guava breeding program aimed at resistance to M. enterolobii.

Nematode trophic structure in the phytotelma of Neoregelia cruenta (Bromeliaceae) in relation to microenvironmental and climate variables.

The term phytotelma (pl. phytotelmata) designates a plant-associated reservoir of fresh water and organic debris. Phytotelmata in tank bromeliads are abundant in tropical forests, and they provide shelter, food, and water for many metazoans. Among the invertebrates known to inhabit phytotelmata, nematodes are the least studied, despite their important role in nutrient and energy cycles in freshwater ecosystems. This study was conceived to characterize the nematode trophic structure in the phytotelma of the bromeliad N. cruenta, and to identify climate and microenvironmental variables that impact it. Nematode abundance (total and per trophic group), rainfall, air temperature, the amount of organic debris fallen into the phytotelma, and eight physico-chemical properties (PCPs) of the water retained in the bromeliad tank - volume; temperature; pH; dissolved organic carbon, nitrogen, oxygen, and solids; and electrical conductivity - were monitored during two years in a natural reserve in Brazil. Bacterial and hyphal feeder nematodes predominated over other trophic groups. Nematode abundance (total and per trophic group) was not impacted by fluctuations in rainfall or air temperature. The amount of organic debris fallen into the phytotelma correlated positively with nematode abundance (total and per trophic group). Regarding the PCPs of water, the only significant correlation - positive - was between the amount of dissolved oxygen and the abundance of hyphal feeder nematodes. These results bring a clearer understanding of the ecology of nematodes inhabiting phytotelmata, which are peculiar and understudied freshwater ecosystems.

Efficacy of Heterorhabdits indica LPP35 against Aedes aegypti in domiciliary oviposition sites.

Entomopathogenic nematodes have been evaluated for control of mosquito species for decades. Depending on the nematode and mosquito involved, mortality rates of larvae (L) may reach 100% in vitro. Nonetheless, nematode efficacy at oviposition sites has rarely been assessed. Heterorhabditis indica LPP35 has been shown to kill over 75% of Aedes aegypti L3/L4 in cups and bottles outdoors. To assess its efficacy in indoor oviposition sites, different types/sizes of floor drains and pot saucers, and 65 liter water barrels, were infested with L3/L4 and treated with two doses of infective juveniles (IJs). In floor drains, mortality rates varied from 45 to 82%, with better results in the smallest drains. The adjustable dose of 25 IJs/cm2 of the drain's bottom internal surface gave better results than the fixed dose of 100 IJs/larva. Mortality rates were only 28 to 53% and 0.1 to 1.7% in pot saucers and water barrels, respectively, probably because ridges and grooves that marked the bottom internal surface of these containers hindered the encounter of larvae and IJs.Entomopathogenic nematodes have been evaluated for control of mosquito species for decades. Depending on the nematode and mosquito involved, mortality rates of larvae (L) may reach 100% in vitro. Nonetheless, nematode efficacy at oviposition sites has rarely been assessed. Heterorhabditis indica LPP35 has been shown to kill over 75% of Aedes aegypti L3/L4 in cups and bottles outdoors. To assess its efficacy in indoor oviposition sites, different types/sizes of floor drains and pot saucers, and 65 liter water barrels, were infested with L3/L4 and treated with two doses of infective juveniles (IJs). In floor drains, mortality rates varied from 45 to 82%, with better results in the smallest drains. The adjustable dose of 25 IJs/cm2 of the drain's bottom internal surface gave better results than the fixed dose of 100 IJs/larva. Mortality rates were only 28 to 53% and 0.1 to 1.7% in pot saucers and water barrels, respectively, probably because ridges and grooves that marked the bottom internal surface of these containers hindered the encounter of larvae and IJs.

Bio-based products control black rot (Xanthomonas campestris pv. campestris) and increase the nutraceutical and antioxidant components in kale.

Black rot of crucifers, (Xanthomonas campestris pv. campestris) is the principal yield-limiting and destructive pathogen of cruciferous crop worldwide. In order to validate a bio-based control alternative for this disease, whey, lime sulfur, biofertilizer, Bordeaux mixture or raw milk were applied to kale (Brassica oleracea var. acephala) plants. The disease control was achieved by most of the tested products. Milk-based products (raw milk and whey) and biofertilizer reduced the severity by 44 and 56% in the field. Antioxidants, crude fibber, crude protein and lipid contents and kale yield were verified in the five treatments on the leaves with and without X. campestris pv. campestris inoculation. In the absence of the pathogen (non-inoculated), lime sulfur and Bordeaux mixture improved plant nutritional value compared to organic treatments, nevertheless milk-based products and biofertilizer improved the evaluated variables more than the control. However, on leaves inoculated with X. campestris pv. campestris raw milk increased antioxidant activity, crude protein and fiber contents, whereas biofertilizer increased kale yield, lipid and antioxidant contents. Milk-based products and biofertilizer were further evaluated in greenhouse trials to determinate the activity of defense-related enzymes and lignin content. Biofertilizer treatment resulted in increased phenylalanine ammonia lyase, catalase, peroxidase activities and lignin content. Hence, the application of milk-based products and biofertilizer are promising to control black rot of crucifers and also improves food quality by boosting nutritional values and antioxidant activity.

Strains of the Group I Lineage of Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch of Cucurbitaceous Crops, are Predominant in Brazil.

Bacterial fruit blotch (BFB), caused by the seedborne bacterium Acidovorax citrulli, is an economically important threat to cucurbitaceous crops worldwide. Since the first report of BFB in Brazil in 1990, outbreaks have occurred sporadically on watermelon and, more frequently, on melon, resulting in significant yield losses. At present, the genetic diversity and the population structure of A. citrulli strains in Brazil remain unclear. A collection of 74 A. citrulli strains isolated from naturally infected tissues of different cucurbit hosts in Brazil between 2000 and 2014 and 18 A. citrulli reference strains from other countries were compared by pulsed-field gel electrophoresis (PFGE), multilocus sequence analysis (MLSA) of housekeeping and virulence-associated genes, and pathogenicity tests on seedlings of different cucurbit species. The Brazilian population comprised predominantly group I strains (98%), regardless of the year of isolation, geographical region, or host. Whole-genome restriction digestion and PFGE analysis revealed that three unique and previously unreported A. citrulli haplotypes (assigned as haplotypes B22, B23, and B24) occurred in Brazil. The greatest diversity of A. citrulli (four haplotypes) was found among strains collected from the northeastern region of Brazil, which accounts for more than 90% of the country's melon production. MLSA clearly distinguished A. citrulli strains into two well-supported clades, in agreement with observations based on PFGE analysis. Five Brazilian A. citrulli strains, representing different group I haplotypes, were moderately aggressive on watermelon seedlings compared with four group II strains that were highly aggressive. In contrast, no significant differences in BFB severity were observed between group I and II A. citrulli strains on melon and squash seedlings. Finally, we observed a differential effect of temperature on in vitro growth of representative group I and II A. citrulli haplotypes. Specifically, of 18 group II strains tested, all grew at 40 and 41°C, whereas only 3 of 15 group I strains (haplotypes B8[P], B3[K], and B15) grew at 40°C. Three strains representing haplotype B8(P) were the only group I strains that grew at 41°C. These results contribute to a better understanding of the genetic diversity of A. citrulli associated with BFB outbreaks in Brazil, and reinforce the efficiency of MLSA and PFGE analysis for assessing population structure. This study also provides the first evidence to suggest that temperature might be a driver in the ecological adaptation of A. citrulli populations.

Características da marcha de mulheres com fibromialgia: um padrão prematuro de envelhecimento / Gait characteristics of women with fibromyalgia: a premature aging pattern

Introdução: Fibromialgia é uma condição que envolve dor crônica generalizada. Além disso, mulheres de meia idade com fibromialgia apresentam alterações no padrão de marcha, expondo-se prematuramente a um padrão de marcha semelhante ao encontrado na população idosa. Objetivo: Determinar os parâmetros espaciais (linear e angular) da marcha de mulheres com fibromialgia e compará-los com idosas sem essa condição. Métodos: 25 mulheres (10 no grupo com fibromialgia e 15 no grupo de idosas) se qualificaram como voluntárias para participar do estudo. A análise cinemática foi realizada por meio de um sistema optoeletrônico, e as variáveis lineares e angulares foram determinadas. Resultados: Ambos os grupos apresentaram similaridades na velocidade da marcha, tamanho da passada, cadência e amplitude de movimento do quadril, joelho e tornozelo (p > 0,05), exceto para a rotação da pelve, na qual o grupo com fibromialgia apresentou maior rotação de quadril (p < 0,05) quando comparado ao grupo de idosas. Além disso, houve correlação negativa no grupo com fibromialgia entre rotação do quadril e dor no glúteo (r = -0,69; p < 0,05), e entre obliquidade da pelve e dor na região do trocanter maior (r = -0,69; p < 0,05). Conclusão: Mulheres de meia idade com fibromialgia apresentaram um padrão de marcha similar ao de idosas, o qual é caracterizado por amplitude de movimento, tamanho da passada e velocidade da marcha reduzidos. .

Background: Fibromyalgia is a condition which involves chronic pain. Middle-aged individuals with fibromyalgia seem to exhibit changes in gait pattern, which may prematurely expose them to a gait pattern which resembles that found in the elderly population. Objective: To determine the 3 D spatial (linear and angular) gait parameters of middle-aged women with fibromyalgia and compare to elderly women without this condition. Methods: 25 women (10 in the fibromyalgia group and 15 in the elderly group) volunteered to participate in the study. Kinematics was performed using an optoelectronic system, and linear and angular kinematic variables were determined. Results: There was no difference in walking speed, stride length, cadence, hip, knee and ankle joints range of motion between groups, except the pelvic rotation, in which the fibromyalgia group showed greater rotation (P < 0.05) compared to the elderly group. Also, there was a negative correlation with pelvic rotation and gluteus pain (r = - 0.69; P < 0.05), and between pelvic obliquity and greater trochanter pain (r = - 0.69; P < 0.05) in the fibromyalgia group. Conclusion: Middle-aged women with fibromyalgia showed gait pattern resemblances to elderly, women, which is characterized by reduced lower limb ROM, stride length and walking speed. .

[Gait characteristics of women with fibromyalgia: a premature aging pattern]. / Características da marcha de mulheres com fibromialgia: um padrão prematuro de envelhecimento.

BACKGROUND: Fibromyalgia is a condition which involves chronic pain. Middle-aged individuals with fibromyalgia seem to exhibit changes in gait pattern, which may prematurely expose them to a gait pattern which resembles that found in the elderly population. OBJECTIVE: To determine the 3D spatial (linear and angular) gait parameters of middle-aged women with fibromyalgia and compare to elderly women without this condition. METHODS: 25 women (10 in the fibromyalgia group and 15 in the elderly group) volunteered to participate in the study. Kinematics was performed using an optoelectronic system, and linear and angular kinematic variables were determined. RESULTS: There was no difference in walking speed, stride length, cadence, hip, knee and ankle joints range of motion between groups, except the pelvic rotation, in which the fibromyalgia group showed greater rotation (P<0.05) compared to the elderly group. Also, there was a negative correlation with pelvic rotation and gluteus pain (r = -0.69; P<0.05), and between pelvic obliquity and greater trochanter pain (r = -0.69; P<0.05) in the fibromyalgia group. CONCLUSION: Middle-aged women with fibromyalgia showed gait pattern resemblances to elderly, women, which is characterized by reduced lower limb ROM, stride length and walking speed.

Transient effects of stretching exercises on gait parameters of elderly women.

This study aimed to analyse the effects of a single stretching exercise session on a number of gait parameters in elderly participants in an attempt to determine whether these exercises can influence the risk of fall. Fifteen healthy women living in the community volunteered to participate in the study. A kinematic gait analysis was performed immediately before and after a session of static stretching exercises applied on hip flexor/extensor muscles. Results showed a significant influence of stretching exercises on a number of gait parameters, which have previously been proposed as fall predictors. Participants showed increased gait velocity, greater step length and reduced double support time during stance after performing stretching exercises, suggesting improved stability and mobility. Changes around the pelvis (increased anterior-posterior tilt and rotation range of motion) resulting from the stretching exercises were suggested to influence the gait parameters (velocity, step length and double support time). Therefore, stretching exercises were shown to be a promising strategy to facilitate changes in gait parameters related to the risk of fall. Some other gait variables related to the risk of fall remained unaltered (e.g., toe clearance). The stable pattern of segmental angular velocities was proposed to explain the stability of these unchanged gait variables. The results indicate that stretching exercises, performed on a regular (daily) basis, result in gait adaptations which can be considered as indicative of reduced fall risk. Other studies to determine whether regular stretching routines are an effective strategy to reduce the risk of fall are required.