Comparative genomic analysis of Bradyrhizobium strains with natural variability in the efficiency of nitrogen fixation, competitiveness, and adaptation to stressful edaphoclimatic conditions.

Bradyrhizobium is known for fixing atmospheric nitrogen in symbiosis with agronomically important crops. This study focused on two groups of strains, each containing eight natural variants of the parental strains, Bradyrhizobium japonicum SEMIA 586 (=CNPSo 17) or Bradyrhizobium diazoefficiens SEMIA 566 (=CNPSo 10). CNPSo 17 and CNPSo 10 were used as commercial inoculants for soybean crops in Brazil at the beginning of the crop expansion in the southern region in the 1960s-1970s. Variants derived from these parental strains were obtained in the late 1980s through a strain selection program aimed at identifying elite strains adapted to a new cropping frontier in the central-western Cerrado region, with a higher capacity of biological nitrogen fixation (BNF) and competitiveness. Here, we aimed to detect genetic variations possibly related to BNF, competitiveness for nodule occupancy, and adaptation to the stressful conditions of the Brazilian Cerrado soils. High-quality genome assemblies were produced for all strains. The core genome phylogeny revealed that strains of each group are closely related, as confirmed by high average nucleotide identity values. However, variants accumulated divergences resulting from horizontal gene transfer, genomic rearrangements, and nucleotide polymorphisms. The B. japonicum group presented a larger pangenome and a higher number of nucleotide polymorphisms than the B. diazoefficiens group, possibly due to its longer adaptation time to the Cerrado soil. Interestingly, five strains of the B. japonicum group carry two plasmids. The genetic variability found in both groups is discussed considering the observed differences in their BNF capacity, competitiveness for nodule occupancy, and environmental adaptation.IMPORTANCEToday, Brazil is a global leader in the study and use of biological nitrogen fixation with soybean crops. As Brazilian soils are naturally void of soybean-compatible bradyrhizobia, strain selection programs were established, starting with foreign isolates. Selection searched for adaptation to the local edaphoclimatic conditions, higher efficiency of nitrogen fixation, and strong competitiveness for nodule occupancy. We analyzed the genomes of two parental strains of Bradyrhizobium japonicum and Bradyrhizobium diazoefficiens and eight variant strains derived from each parental strain. We detected two plasmids in five strains and several genetic differences that might be related to adaptation to the stressful conditions of the soils of the Brazilian Cerrado biome. We also detected genetic variations in specific regions that may impact symbiotic nitrogen fixation. Our analysis contributes to new insights into the evolution of Bradyrhizobium, and some of the identified differences may be applied as genetic markers to assist strain selection programs.

First Record of Ategmic Ovules in Orchidaceae Offers New Insights Into Mycoheterotrophic Plants.

The number of integuments found in angiosperm ovules is variable. In orchids, most species show bitegmic ovules, except for some mycoheterotrophic species that show ovules with only one integument. Analysis of ovules and the development of the seed coat provide important information regarding functional aspects such as dispersal and seed germination. This study aimed to analyze the origin and development of the seed coat of the mycoheterotrophic orchid Pogoniopsis schenckii and to compare this development with that of other photosynthetic species of the family. Flowers and fruits at different stages of development were collected, and the usual methodology for performing anatomical studies, scanning microscopy, and transmission microscopy following established protocols. P. schenckii have ategmic ovules, while the other species are bitegmic. No evidence of integument formation at any stage of development was found through anatomical studies. The reduction of integuments found in the ovules could facilitate fertilization in this species. The seeds of P. schenckii, Vanilla planifolia, and V. palmarum have hard seed coats, while the other species have seed coats formed by the testa alone, making them thin and transparent. P. schenckii, in contrast to the other species analyzed, has a seed coat that originates from the nucellar epidermis, while in other species, the seed coat originates from the outer integument.

Caracterización del crecimiento miceliar de cepas nativas de Neolentinus ponderosus y N. lepideus en diferentes pH y degradación de madera de Pinus spp / Mycelial growth of native strains of Neolentinus ponderosus and N. lepideus at different pH and their Pinus spp. wood degradation

Neolentinus ponderosus y N. lepideus son dos especies de hongos saprobios que poseen uso alimenticio tradicional en comunidades de los departamentos de Huehuetenango y Totonicapán, Guatemala. La capacidad degradadora de ambas especies les confiere potencial para el estudio de la producción de cuerpos fructíferos a nivel de sustrato, por lo que en esta investigación se evaluó el crecimiento de una cepa nativa de N. ponderosus y una de N. lepideus en el medio de cultivo agar extracto de malta (EMA) a diferente pH y también se estudió la degradación de la madera de dos especies de pino (Pinus tecunumanii y P. ayacahuite) en cámaras de podredumbre durante 12 meses. El mayor diámetro de crecimiento miceliar de la cepa N. ponderosus se observó a pH 7.0, en tanto que para N. lepideus se obtuvo a pH 5.0 y 5.6. Las colonias de ambas cepas evidenciaron textura afelpada, borde regular a irregular, color blanco, con o sin pigmento difusible, olor frutal, hifas de 1.0-5.0 µm de diámetro, con clamidosporas y fíbulas. La madera de P. tecunumanii degradada por la cepa de N. ponderosus y la madera de P. ayacahuite por la cepa N. lepideus, presentaron porcentajes de pérdida de peso de 8.76 (±5.58) y 12.07 (±5.66), respectivamente y en ambos casos se alcanzó solamente el estadio temprano de degradación. Los resultados de este estudio sientan las bases para investigaciones posteriores en los que se evalúe el cultivo en troncos de ambas especies para la producción de cuerpos fructíferos con fines alimenticios y comerciales

Neolentinus ponderosus and N. lepideus are two saprophytic fungi species used traditionally in Huehuetenango and Totonicapán, Guatemala. The degradative capacity of both species confers them potential for fruiting bodies production. This study evaluated the mycelial growth of two native strains of N. ponderosus and N. lepideus in malt extract agar (EMA) at different pH and the degradation of wood from two pine species in rot chambers during 12 months. pH 7.0 was the most appropriate for the mycelial growth of N. ponderosus and for N. lepideus were 5.0 and 5.6. The colonies of both strains showed fruity odor, velvety texture, regular to irregular edge, white color, with or without diffusible pigment, hyphae with 1-5 µm width, chlamydospores and clamp connections. Wood from Pinus tecunumanii and P. ayacahuite exhibit weight-loss percentages between 8.76 ± 5.58 and 12.07 ± 5.66, with N. ponderosus 145.2003 and N. lepideus 90.2002, respectively. In both cases reached the early stage of brown-rot decay. These results could be useful for future research that evaluate the fruiting bodies production in logs for food and commercial purposes

Larvicidal effects of endophytic and basidiomycete fungus extracts on Aedes and Anopheles larvae (Diptera, Culicidae)

Introduction In vitro bioassays were performed to access the larvicidal activity of crude extracts from the endophytic fungus Pestalotiopsis virgulata (Melanconiales, Amphisphaeriaceae) and the saprophytic fungus Pycnoporus sanguineus (Basidiomycetes, Polyporaceae) against the mosquitoes Aedes aegypti and Anopheles nuneztovari. Methods The extracts were tested at concentrations of 100, 200, 300, 400 and 500ppm. Ethyl acetate mycelia (EAM) extracts and liquid culture media (LCM) from Pe. virgulata and Py. sanguineus were tested against third instar larvae of Ae. aegypti and An. nuneztovari. Results The larvicidal activity of the EAM extracts from Pe. virgulata against Ae. aegypti had an LC50=101.8ppm, and the extract from the basidiomycete fungus Py. sanguineus had an LC50=156.8ppm against the Ae. aegypti larvae. The Pe. virgulata extract had an LC50=16.3ppm against the An. nuneztovari larvae, and the Py. sanguineus extract had an LC50=87.2ppm against these larvae. Conclusions These results highlight the larvicidal effect of EAM extracts from the endophyte Pe. virgulata against the two larval mosquitoes tested. Thus, Pe. virgulata and Py. sanguineus have the potential for the production of bioactive substances against larvae of these two tropical disease vectors, with An. nuneztovari being more susceptible to these extracts. .

Hydrolytic enzymes in Paracoccidioides brasiliensis--ecological aspects

Paracoccidioides brasiliensis is a thermally dimorphic fungus that causes paracoccidioidomycosis. The yeast form of this pathogen is found in the animal host whereas the mycelial form is recovered from living and non-living organic material. The sole carbon source available in these habitats is represented by polysaccharides from the plant cell wall. Hydrolytic enzymes are necessary to convert these polymers into simple sugars for fungal metabolism. We report on the presence of ortholog genes of hydrolytic enzymes identified in the P. brasiliensis transcriptome and on hydrolytic activities in supernatants of induced P. brasiliensis cultures of mycelium and yeast cells. Enzymatic assays have shown cellulase and xylanase activities, both being higher in mycelium than in the yeast form. Amylase and chitinase activities were detected only in mycelium. Data so far reinforce the idea that mycelial P. brasiliensis is a saprobe.