Abstract Introduction: A recent revision of the generic classification of the Trochilidae based on DNA sequences revealed many inconsistencies with the current generic classification, largely based on plumage characters subject to homoplasy, especially in the Trochilini, the largest tribe. A thorough generic reorganization brought the classification into accord with the phylogeny, but due to lack of genetic data, two species remained unclassified. One of these was the Mangrove Hummingbird, “Amazilia” boucardi, endemic to Costa Rica and included in the IUCN red list of threatened species. Objective: To obtain molecular evidence to clarify the generic relationships of “A.” boucardi. Methods: We isolated DNA from tissues of this species and amplified 4 nuclear and 4 mitochondrial fragments and compared these with homologous fragments from 56 species in the Trochilini, constructing phylogenetic trees with maximum likelihood and Bayesian methods. Results: Our phylogenetic analyses confirmed the placement of boucardi in the Trochilini and definitely excluded it from Amazilia but placed it with high confidence in the genus Chrysuronia Bonaparte, 1850, within which its closest relative is C. coeruleogularis, which also inhabits mangroves. Conclusions: Our genetic data based on nuclear and mitochondrial regions clearly indicate the relationship of A. boucardi and L. coeruleogularis. Moreover, it is also supported by their habitat distribution in the mangroves of the Pacific coast of Costa Rica and Western Panama. Therefore, we suggested to exclude A. boucardi as “incertae sedis”.

Resumen Introducción: Una revisión reciente de la clasificación de la familia Trochilidae con base en secuencias de ADN demostró muchas incongruencias con la clasificación genérica previa, que había sido hecho con base en caracteres del plumaje muy sujetos a homoplasia, especialmente en la tribu más grande, Trochillini. Una reorganización de los géneros logró llevar su clasificación genérica a la concordancia con la filogenia, pero debido a la ausencia de datos genéticos, dos especies permanecieron sin clasificar. Una de estas fue el colibrí de manglar Amazilia boucardi, una especie endémica de Costa Rica, considerada como amenazada en la lista roja de la UICN. Objetivo: Obtener evidencia molecular para esclarecer las relaciones genéricas de A. boucardi. Métodos: Se aisló ADN de tejidos de esta especie y se amplificaron 4 fragmentos de ADN del núcleo y 5 de la mitocondria, y se compararon con fragmentos homólogos de 56 especies en la tribu Trochillini, generando árboles filogenéticos con métodos de máxima verosimilitud y bayesiano. Resultados: Los análisis filogénticos obtenidos confirmaron la ubicación de boucardi en Trochilini y definitivamente la excluyó del género Amazilia, pero la ubicó con un alto grado de confianza en el género Chrysuronia Bonaparte, 1850, dentro los cuales su pariente más cercano es C. coeruleogularis, que también habita manglares. Conclusiones: Nuestros datos genéticos basados en regiones nucleares y mitocondriales indican claramente la relación entre A. boucardi and L. coeruleogularis. Es más, lo anterior se sustenta por su distribución en los manglares de la costa Pacífica de Costa Rica y oeste de Panamá. Por lo tanto, sugerimos excluir a A. boucardi como “incertae sedis”.

Abstract Background: The chloroplast genomes of Cactaceae exhibit boundary modifications in the inverted repeat regions (IRs), gene inversions, and deletions. Among nine Mammillaria species, three distinct chloroplast structures have been identified, although not all of these correspond to the morphology-based classification of the genus. Question: Is there a distinct chloroplast genome structure in the species of Mammillaria series Stylothelae? Studied species: Mammillaria bocasana and M. erythrosperma. Study site and dates: Mexico from 2019 to 2023. Methods: Chloroplast DNA was sequenced, and chloroplast genomes were de novo assembled using the Fast-Plast program. Complete plastome sequences were annotated and verified. The sequences were aligned in MAUVE program to detect possible structural changes. A maximum likelihood phylogeny was executed to evaluate the relationships of the studied species. Results: The plastomes ranged from 107,368 bp in Mammillaria bocasana to 108,069 bp in M. erythrosperma. Both presented a quadripartite structure and contained 108 genes. The IRs were ~ 1,600 bp long and included the genes rpl2, rpl23 (pseudo), and trnI-CAU. MAUVE identified a ~ 21 kb inversion in the large single copy containing a block of genes related to photosynthesis. The phylogenetic analysis placed both species in a single clade separated from the other species within Mammillaria subg. Mammillaria. Conclusions: The studied species of Mammillaria series Stylothelae exhibited a different and synapomorphic chloroplast genome structure. Other Mammillaria chloroplast genome structures have evolved independently in different lineages.

Resumen Antecedentes: Los genomas de cloroplasto de Cactaceae exhiben modificaciones en los límites de las regiones repetidas inversas (IRs), inversiones de genes y deleciones. En nueve especies de Mammillaria, se han encontrado tres estructuras de cloroplasto distintas, aunque no todas corresponden a las clasificaciones basadas en morfología. Pregunta: ¿Existe una estructura de genoma de cloroplasto distinta en las especies de Mammillaria serie Stylothelae? Especies estudiadas: Mammillaria bocasana y M. erythrosperma. Sitio y años de estudio: México, de 2019 a 2023. Métodos: ADN de cloroplasto fue secuenciado y los plastomas fueron ensamblados de novo usando el programa Fast-Plast. Las secuencias completas fueron anotadas y verificadas. Las secuencias fueron alineadas en el programa MAUVE para detectar posibles cambios estructurales. Un análisis filogenético con máxima verosimilitud evaluó las relaciones de las especies estudiadas. Resultados: Los plastomas variaron desde 107,368 pb de longitud en Mammillaria bocasana hasta 108,069 pb en M. erythrosperma. Ambos presentaron una estructura cuadripartita y contuvieron 108 genes. Las IRs tuvieron ~1,600 pb y contuvieron los genes rpl2, rpl23 (pseudo) y trnI-CAU. MAUVE identificó una inversión de ~21 kb en la región larga de copia única que contiene genes relacionados con la fotosíntesis. El análisis filogenético recuperó a ambas especies como grupos hermanos dentro de Mammillaria subg. Mammillaria. Conclusiones: Las dos especies estudiadas de Mammillaria serie Stylothelae presentaron una estructura de plastoma sinapomórfica. Otras estructuras de plastomas de Mammillaria han evolucionado independientemente en linajes diferentes.

Abstract Evolution has originated high levels of biodiversity that provide essential actual benefits to humans and future called option values. These resources are extracted and used to meet basic needs, such as traditional medicine, which is the result of human experience and its contact with nature over time. Traditional knowledge used as a guide for bioprospecting and ethnomedicinal research is the first step to protecting it and making it visible. The need to apply phylogenetic approaches and their importance for the development of ethnobotany in Mexico is discussed, as well as their implications for the discovery, use and conservation of medicinal resources for human well-being. This approaches, elucidating the evolutionary history of species, can improve bioprospecting practices since they point out related species which could produce useful chemical compounds, which is relevant to biotechnology and emphasizes the importance of preserving biocultural heritage.

Resumen La evolución ha producido una gran biodiversidad que proporciona al ser humano bienes esenciales actuales y opciones de valor de facturas. Estos recursos son extraídos y empleados para satisfacer necesidades básicas, como la medicina tradicional, resultado de la experiencia y el contacto humano con la naturaleza a través del tiempo. Los saberes tradicionales han sido aprovechados como guía para la bioprospección y los estudios etnobotánicos son el primer paso para visibilizarlos y protegerlos. Se plantea la necesidad de aplicar aproximaciones filogenéticas y su importancia para el desarrollo de la etnobotánica en México, así como su impacto en el descubrimiento, aprovechamiento y conservación de recursos medicinales para el bienestar humano. Estas aproximaciones, que revelan la historia evolutiva de las especies, pueden fortalecer la bioprospección, ya que señalan especies emparentadas que pudieran llegar a producir sustancias químicas útiles, lo que resulta relevante para la biotecnología y resalta la importancia de la conservación del patrimonio biocultural.

Abstract Aerenchyma is a characteristic tissue of aquatic plants, characterized by the presence of air lacunae commonly septated by diaphragms. These are formed by one or more layers of stellate cells, which allow for the passage of gases. Most species of Eleocharis grow in wet or flooded soils and have aerenchyma in their aerial stems. However, extensive studies on this structure, which could contribute to ecological and phylogenetic studies of the group, are lacking. This work describes the structure of the diaphragm in the stem of Eleocharis species and investigates the evolution of this characteristic in the genus. Fifty-three species were analyzed under light and scanning electron microscopy. We analyzed the evolution of the characteristics by reconstructing their ancestral states based on the previously published original phylogeny. The diaphragms in Eleocharis vary mainly in the number of layers, cell shape, and cell wall thickness. The typical diaphragm of the genus is composed of three to four layers of stellate cells, with microprojections and secretory cells. The diaphragm of the group’s ancestor had practically the same characteristics as the genus’s typical diaphragm. plants cells gases stems However group lacking Fiftythree Fifty microscopy phylogeny shape thickness groups s genuss

Resumo Aerênquima é um tecido comum nas plantas aquáticas, caracterizado pela presença de lacunas de ar comumente septadas por diafragmas. Esses, são formados por uma ou mais camadas de células, geralmente braciformes, que permitem a passagem de gases. A maioria das espécies de Eleocharis cresce em solos úmidos ou inundados e apresenta aerênquima no caule. No entanto, essa estrutura carece de estudos amplos que poderiam contribuir aos estudos ecológicos e filogenéticos do grupo. Este trabalho descreve a estrutura do diafragma no caule de espécies de Eleocharis e investiga a evolução desse caractere no gênero. 53 espécies foram analisadas em microscopia de luz e eletrônica de varredura através de secções transversais e longitudinais do caule. A análise da evolução dos caracteres foi feita pela reconstrução de estado ancestral, baseada na filogenia original anteriormente publicada. Os diafragmas em Eleocharis variam principalmente em número de camadas, formato das células e espessura da parede celular. O diafragma típico do gênero é composto de três a quatro camadas de células braciformes lobadas, com microprojeções e células secretoras. O diafragma do ancestral do grupo apresentava praticamente as mesmas características do diafragma típico para o gênero. aquáticas Esses gases entanto 5 publicada celular lobadas secretoras

Abstract We announce the mitochondrial genomes of seven species of the genus Sporophila (S. bouvreuil, S. iberaensis, S. melanogaster, S. minuta, S. nigrorufa, S. pileata, and S. ruficollis) which were validated by comparative genomic and phylogenetic analysis with related species. The mitochondrial genomes of seven passerines of the genus Sporophila were assembled (three complete and four nearly complete genomes) and were validated by reconstructing phylogenetic relations within Thraupidae. The complete mitogenomes ranged from 16,781 bp in S. ruficollis to 16,791 bp in S. minuta. We identified a conserved genome composition within all mitogenomes with 13 protein-coding genes, 22 tRNAs and two rRNAs. We observed a bias in the nucleotide composition and six mutational hotspots in Sporophila mitogenomes. Our mitogenome-based phylogenetic tree has S. minuta, S. maximiliani and S. nigricollis as sister species of the remaining species in the genus. We present new mitogenome sequences for seven Sporophila species, providing new genomic resources that may be useful for research on the evolution, comparative genetics, and conservation of this threatened group. S (S bouvreuil iberaensis melanogaster minuta nigrorufa pileata three Thraupidae 16781 16 781 16,78 16791 791 16,79 1 proteincoding protein coding genes 2 rRNAs mitogenomebased based evolution genetics group 1678 78 16,7 1679 79 167 7 16,

ABSTRACT Spodoptera cosmioides (Walker, 1858) is an economically relevant polyphagous moth, widely distributed in the Neotropics and part of the Spodoptera latifascia (Walker, 1856) species group. In this study, we used extensive sampling from different regions to describe the spatial distribution of S. cosmioides in Brazil and evaluate its variability both from morphological and molecular perspectives. Variable coloration and several morphological similarities were found among S. cosmioides and congeners of the S. latifascia complex, diverging from each 0.5 to 3.5% of mitochondrial DNA. The genetic divergence at the species level of S. cosmioides was 0.5% throughout Brazil, and a geographic structure was absent, including shared haplotypes with S. descoinsi Lalanne-Cassou & Silvain, 1994. Spodoptera cosmioides was found in all six biomes of Brazil, with the highest abundance recorded in the Cerrado, followed by the Amazon, Atlantic Rainforest, Caatinga, Pantanal, and Pampa. Walker, Walker (Walker 1858 moth 1856 group study S perspectives complex 05 0 5 0. 35 3 3.5 DNA absent LalanneCassou Lalanne Cassou Silvain 1994 Cerrado Amazon Rainforest Caatinga Pantanal Pampa 185 3. 199 18 19 1

ABSTRACT Changes in Neotropical bat taxonomy in the past two decades have caused significant shifts in our understanding of the diversity and distribution of the group. Taxonomic revisions have shown that the once widespread Platyrrhinus helleri (Peters, 1866) represented a species complex, which resulted in the elevation of Platyrrhinus helleri incarum (Thomas, 1912) to the species level for cis-Andean South America and the restriction of P. helleri to Central America and northern South America west of the Andes. Nevertheless, some studies still mention P. helleri for Brazil. Aiming to investigate if P. helleri occurs in Brazil, we collected specimens of small-sized Platyrrhinus in the Cerrado and Pantanal ecosystems, and revised vouchers in museum collections. Based on DNA sequence data and analysis of the pelage and craniodental morphology, we confirm that the only small Platyrrhinus species occurring in Cerrado and Pantanal of central Brazil corresponds to P. incarum. This is the first study to provide sequence data for Brazilian P. incarum. We also provide new records of P. incarum, extending its range by 390 km to southeast and 480 km to the east in Brazil. group Peters, Peters (Peters 1866 complex Thomas, Thomas (Thomas 1912 cisAndean cis Andean P Andes Nevertheless smallsized sized ecosystems collections morphology 39 48 186 191 3 4 18 19 1

ABSTRACT: Canine parvovirus 2 (CPV-2) is an important causative agent of segmental enteritis in young dogs and has globally distributed variants and subtypes. Viral mutations can alter the pathogenesis and clinical signs, making identifying the samples circulating in a given region relevant. This study described the epidemiological and clinical findings and the molecular characterization of CPV-2 samples circulating in the canine population of Uruguaiana, Rio Grande doSul (RS), Brazil. We analyzed 27 cases with a complete clinical history and at least one confirmatory etiologic diagnosis. In addition to clinical and epidemiological data, whole blood samples or tissues were tested by PCR for viral DNA detection. Amplified products were sequenced and analyzed, and phylogeny was generated with reference sequences. The disease was diagnosed especially in the summer months, and the most common clinical findings were diarrhea, anorexia, listlessness, and vomiting. Infection was predominant in young (< 6 months) unvaccinated or partially immunized dogs, with mortality exceeding 93%. It was possible to identify 15 CPV-2 samples, four of which were CPV-2a and 11 were new CPV-2a. It can be concluded that canine parvovirus is a disease with high mortality rates, with young unvaccinated dogs being more susceptible, with a predominance of the new CPV-2a variant in the western region of Rio Grande do Sul. ABSTRACT CPV2 CPV (CPV-2 subtypes signs relevant CPV- Uruguaiana RS, RS , (RS) Brazil diagnosis data detection sequences months diarrhea anorexia listlessness vomiting < ( 93 93% 1 CPV2a CPVa 2a CPV2a. 2a. rates susceptible Sul (CPV- (RS 9 (CPV

RESUMO: O parvovírus canino 2 (CPV-2) é um importante agente etiológico de enterite segmentar em cães jovens e apresenta variantes e subtipos distribuídos mundialmente. As mutações virais podem alterar a patogenia e os sinais clínicos, o que torna relevante identificar as amostras que circulam em determinada região. O objetivo do trabalho foi descrever os achados epidemiológicos, clínicos e a caracterização molecular das amostras de CPV-2 circulantes na população canina do município de Uruguaiana, Rio Grande do Sul (RS), Brasil. Foram analisados 27 casos clínicos que possuíam histórico clínico completo e pelo menos um diagnóstico etiológico confirmatório. Além dos dados clínicos e epidemiológicos, amostras de sangue total, suabes retais ou tecidos foram testados por PCR para detecção do DNA viral. Os produtos amplificados foram sequenciados, analisados e a filogenia gerada com sequências de referência. A doença foi diagnosticada especialmente nos meses de verão e os achados clínicos mais comuns foram diarreia, anorexia, apatia e vômito. A infecção foi predominante em cães jovens (< 6 meses) não vacinados ou parcialmente imunizados, com mortalidade superior a 93%. Foi possível identificar quinze amostras de CPV-2, sendo quatro CPV-2a e onze new CPV-2a. Pode-se concluir que a parvovirose canina é uma enfermidade com altos índices de mortalidade, sendo os cães jovens não vacinados mais susceptíveis, havendo predominância da variante new CPV-2a na região oeste do RS. RESUMO CPV2 CPV (CPV-2 mundialmente epidemiológicos CPV- Uruguaiana RS, RS , (RS) Brasil confirmatório total viral sequenciados referência diarreia anorexia vômito < ( imunizados 93 93% CPV2, 2, CPV2a CPVa 2a CPV2a. 2a. Podese Pode se susceptíveis (CPV- (RS 9 (CPV

ABSTRACT Noroviruses are highly infectious, genetically diverse viruses. Global outbreaks occur frequently, making molecular surveillance important for infection monitoring. This cross-sectional descriptive study aimed to monitor cases of norovirus gastroenteritis in the Brazilian Amazon. Fecal samples were tested by immunoenzymatic assay, RT-PCR and genetic sequencing for the ORF1/ORF2 and protease regions. Bayesian inference with a molecular clock was employed to construct the phylogeny. The norovirus prevalence was 25.8%, with a higher positivity rate among children aged 0-24 months. Genogroup GII accounted for 98.1% of the sequenced samples, while GI accounted for 1.9% of them. The GII.P16/GII.4 genotype was the most prevalent, with an evolution rate of 2.87x10−3 and TMRCA estimated in 2012. This study demonstrates that norovirus is a primary causative agent of gastroenteritis and provides data on viral genetic diversity that may facilitate infection surveillance and vaccine development. infectious viruses frequently monitoring crosssectional cross sectional Amazon assay RTPCR RT PCR ORF1ORF2 ORFORF ORF1 ORF2 ORF ORF1/ORF regions phylogeny 258 25 8 25.8% 024 0 24 0-2 months 981 98 1 98.1 19 9 1.9 them GIIP16GII4 GIIPGII P16 4 P GII.P16/GII. prevalent 287x103 x 2 87x10 3 2.87x10− 2012 development ORF1ORF 25.8 02 0- 98. 1. GIIP GIIP16GII P1 GII.P16/GII 287x10 87x1 2.87x10 201 25. 287x1 87x 2.87x1 20 287x 2.87x

Resumo Surtos frequentes do vírus da gripe aviária H9N2 no Paquistão revelaram que esse subtipo se tornou endêmico na avicultura e, além das perdas econômicas, representa uma ameaça à saúde pública. O presente estudo descreve a caracterização molecular e as alterações patológicas em frangos de corte naturalmente infectados com a atual cepa H9N2 e sua dinâmica filogenômica. Neste estudo, amostras de tecidos (traqueia, pulmões, rim e intestino) de 100 lotes comerciais de frangos foram coletadas de julho de 2018 a agosto de 2019. As amostras foram submetidas à detecção molecular, filogenia e posterior exame patológico. O comprimento completo do gene HA foi amplificado com sucesso em cinco amostras. O sequenciamento de nucleotídeos revelou amostras positivas colocadas em um clado pertencente à sublinhagem B2 do genótipo G1 e categorizado como LPAIV com base na sequência de aminoácidos do gene da hemaglutinina (HA) no local de clivagem (PAKSSR/G). A análise genética do gene da HA revelou: nt = 80,5%-99,5%; aa = 83,8%-98,9% de homologia com cepas de H9N2 relatadas anteriormente no Paquistão e em países vizinhos (A/Quail/Hong Kong/G1/97). As lesões macroscópicas incluíram aerossaculite leve, hemorragias leves, congestão difusa e exsudato purulento na mucosa traqueal, cilindro fibrinonecrótico no lúmen da traqueia e congestão pulmonar leve. As alterações histopatológicas incluíram descamação de células epiteliais, infiltração de células inflamatórias na traqueia, infiltração de células mononucleares (MNCs), congestão pulmonar e exsudato no lúmen dos parabrônquios, congestão peritubular nos rins com degeneração das células epiteliais tubulares, alterações degenerativas nas células epiteliais das vilosidades intestinais e hiperplasia de células caliciformes. A análise imunoistoquímica confirmou a presença do antígeno AIVH9N2 na traqueia, nos pulmões, no rim e no intestino. A microscopia eletrônica revelou alterações ultraestruturais na traqueia, incluindo cílios degenerados, inchaço mitocondrial e retículo endoplasmático aumentado. Com base em todas as análises, o presente estudo revelou a distribuição do vírus H9N2 do genótipo G1 em Punjab, Paquistão, com patogenicidade de leve a moderada.

Abstract Frequent outbreaks of avian influenza H9N2 virus in Pakistan revealed that this subtype has become endemic in the poultry industry and, besides economic losses, poses a threat to public health. The present study describes the molecular characterization and pathological alterations in naturally infected broiler chickens with the current H9N2 field strain and their phylogenomic dynamics. In this study, tissue samples (trachea, lung, kidney and intestine) from 100 commercial chicken flocks were collected from July 2018 to August 2019. Samples were subjected to molecular detection, phylogeny and subsequent pathological examination. The complete length of the HA gene was successfully amplified in five samples. Nucleotide sequencing revealed positive samples placed in a clade belonging to the B2 sub-lineage of the G1 genotype and categorized as LPAIV based on the amino acid sequence of the HA gene at the cleavage site (PAKSSR/G). Genetic analysis of the haemagglutinin (HA) gene revealed nt: 80.5%-99.5%; aa: 83.8%-98.9% homology to H9N2 strains reported previously from Pakistan, neighbouring countries, and (A/Quail/Hong Kong/G1/97). Gross lesions include a slight airsacculitis, mild hemorrhages, diffuse congestion and purulent exudate in tracheal mucosa, fibrinonecrotic cast in the trachea lumen and mild pulmonary congestion. Histopathological alterations include sloughing of epithelial cells and infiltration of inflammatory cells in the trachea, mononuclear cells (MNCs) infiltration, pulmonary congestion and exudate in the lumen of parabronchi, peritubular congestion in the kidneys with degeneration of tubular epithelial cells and degenerative changes in the intestinal villi epithelial cells and goblet cell hyperplasia. Immunohistochemistry analysis confirmed the presence of AIVH9N2 antigen in the trachea, lungs, kidney and intestine. Electron microscopy revealed ultrastructural changes in the trachea, including degenerated cilia, mitochondrial swelling and enlarged endoplasmic reticulum. Based on all essential analysis, the present study revealed the distribution of the H9N2 virus of G1 genotype in Punjab, Pakistan, with mild to moderate pathogenicity.

Abstract Endophytic microorganisms are those that inhabit the interior of plant tissues and organs without causing damage to the plant, triggering a mutualistic interaction. These endophytes are known to produce compounds with various applications in the areas of biotechnology, pharmaceutical industry and agriculture; therefore, molecular methods are used to investigate the phylogeny of these organisms. The Brazilian Cerrado is the second largest biome in Brazil and is considered a hotspot, with a high diversity of endemic fauna and flora. The objective of this work was to analyze, through in silico analysis, the phylogeny of cultivable endophytic fungi isolated from plants found in the Brazilian Cerrado biome. A search was carried out for articles that worked with endophytes in the period between 2012 and 2022. The dendrogram was generated using the Neighbor-joining method based on the ITS1-5.8S-ITS2 conserved region obtained by GenBank codes of isolated endophytes. The genera Xylaria, Diaporthe, and Colletotrichum were isolated in more than three plants in the seven articles found. Most of the articles found related to the biodiversity of endophytic fungi from the Brazilian Cerrado aim at the bioprospecting of bioactive compounds, through culture-dependent methods; as such, a part of endophytic diversity may be lost due to the inability of certain endophytes to grow in artificial media. In silico analysis can assist in the investigation of phylogenetic relationships between endophytic fungi and has the potential to guide future work aimed at prospecting for bioactive compounds, phylogenetic identification, or biodiversity of this group of endophytes. interaction biotechnology agriculture therefore organisms hotspot flora analyze 201 2022 Neighborjoining Neighbor joining ITS15.8SITS2 ITS158SITS2 ITSSITS ITS1 5.8S ITS2 5 8S ITS S ITS1-5.8S-ITS Xylaria Diaporthe culturedependent culture dependent such media identification 20 202 ITS15 8SITS2 SITS ITS15.8SITS ITS158SITS 58S 2 8SITS

Resumo Microrganismos endofíticos são aqueles que vivem no interior de tecidos e órgãos vegetais sem causar dano à planta, desencadeando uma interação mutualística. Esses endófitos são conhecidos por produzir compostos com diversas aplicações nas áreas de biotecnologia, indústria farmacêutica e agricultura; logo, métodos moleculares são utilizados para investigar a filogenia destes organismos. O Cerrado brasileiro é o segundo maior bioma do Brasil e é considerado um hotspot, com uma alta diversidade de fauna e flora endêmica. O objetivo deste trabalho foi analisar, por meio da análise in silico, a filogenia de fungos endofíticos cultiváveis isolados de plantas encontradas no bioma Cerrado brasileiro. Foi realizada uma busca por artigos que trabalharam com endófitos no período entre 2012 e 2022. O dendrograma foi gerado usando o método Neighbor-joining baseado na região conservada ITS1-5.8S-ITS2 obtida pelos códigos GenBank de endófitos isolados. Os gêneros Xylaria, Diaporthe e Colletotrichum foram isolados em mais de três plantas nos sete artigos encontrados. A maior parte dos artigos encontrados relacionados à biodiversidade de fungos endofíticos do Cerrado brasileiro visam à bioprospecção de compostos bioativos, através de métodos dependentes de cultura; como tal, uma parte da diversidade endofítica pode ser perdida devido à incapacidade de certos endófitos crescerem em meios artificiais. A análise in silico pode auxiliar na investigação das relações filogenéticas entre fungos endofíticos e tem potencial para orientar trabalhos futuros voltados à prospecção de compostos bioativos, identificação filogenética ou biodiversidade deste grupo de endófitos. planta mutualística biotecnologia agricultura logo organismos hotspot endêmica analisar 201 2022 Neighborjoining Neighbor joining ITS15.8SITS2 ITS158SITS2 ITSSITS ITS1 5.8S ITS2 5 8S ITS S ITS1-5.8S-ITS Xylaria bioativos cultura tal artificiais 20 202 ITS15 8SITS2 SITS ITS15.8SITS ITS158SITS 58S 2 8SITS

RESUMO Um dos gêneros mais importantes de Apocynaceae, Aspidosperma apresenta desafios na delimitação de espécies devido à sobreposição de caracteres morfológicos e o grande número de sinonímias. Assim, buscamos entender os caracteres morfológicos mais relevantes na delimitação de duas seções do gênero (Aspidosperma sect. Aspidosperma e A. sect. Pungentia), confrontando hipóteses de homologia primária com estudo filogenético molecular. Os dados dos caracteres foram obtidos através da literatura do gênero e confrontados em uma árvore filogenética para verificar a presença de características diagnósticas para as seções e espécies. Nossos resultados evidenciaram: (1) a confirmação de características diagnósticas para as duas seções; (2) a possibilidade de segregar a seção típica em três novas seções, com base nas características morfológicas; (3) a distinção de espécies tradicionalmente tratadas como sinônimos; (4) a importância da utilização de características da madeira para a delimitação de gêneros com ampla variação morfológica, como é o caso de Aspidosperma. Apocynaceae sinonímias Assim sect A Pungentia, Pungentia , Pungentia) molecular evidenciaram 1 (1 2 (2 morfológicas 3 (3 sinônimos 4 (4 morfológica (

ABSTRACT One of the most important genera of Apocynaceae, Aspidosperma presents challenges in species delimitation due to overlapping morphological characters and the large number of synonyms. Thus, we seek to understand the most relevant morphological characters in the delimitation of two sections of the genus (Aspidosperma sect. Aspidosperma and A. sect. Pungentia), confronting hypotheses of primary homology with a molecular phylogenetic study. Character data were obtained from the literature of the genus and confronted in a phylogenetic tree to verify the presence of diagnostic characteristics for sections and species. Our results showed: (1) confirmation of diagnostic characteristics for both sections; (2) the possibility of segregating the typical section into three new sections based on morphological characteristics; (3) the distinction of species traditionally treated as synonyms; (4) the importance of using wood characteristics for the delimitation of genera with wide morphological variation, as is the case of Aspidosperma. Apocynaceae synonyms Thus sect A Pungentia, Pungentia , Pungentia) study showed 1 (1 2 (2 3 (3 4 (4 variation (

Abstract The phylogenetic affinities of early dinosaurs are a controversial topic. There are disputing scenarios aiming to explain the evolutionary tree of these reptiles. A number of factors play a role on this issue. High levels of intraspecific variation experienced by the earliest forms are pointed by distinct authors as one of the main sources of phylogenetic biases. In the present study, we performed experimental analyses incorporating skeletally immature individuals as distinct operational taxonomic units in order to investigate the effect of ontogeny on the phylogeny and morphospace of early Late Triassic sauropodomorphs. The results support an “artificial” grouping of skeletally immature individuals of early dinosaurs in the phylogenetic trees, which is controlled by ontogenetic development. Such results are interpreted as the effect of ontogenetic control on the states of certain morphological characters, producing false synapomorphies. In addition, the morphological disparity analysis indicated a significant difference between the skeletally immature and mature individuals, corroborating previous assumptions. At this point, we suggest caution during the scoring process of early dinosaurs. Some approaches that avoid ontogenetically sensitive characters revealed a putative solution. Nevertheless, additional specimens and alternative tools are mandatory in order to properly deal with this issue. topic reptiles issue biases study sauropodomorphs artificial “artificial trees development synapomorphies addition assumptions point solution Nevertheless

Resumo Hypostomus está distribuído por bacias da América Central e do Sul, com grande diversidade de espécies com conflitos taxonômicos. Desta forma, a integração de técnicas auxiliares contribui para a compreensão da sistemática e filogenia do grupo. Assim, este estudo teve como objetivo investigar Hypostomus cochliodon e H. boulengeri do riacho Onça (bacia do rio Paraguai) por meio de técnicas aloenzimáticas e citogenéticas. As análises citogenéticas em H. boulengeri mostraram número cromossômico igual a 2n = 68 (14m+14sm+18st+22a), sistema de NOR múltiplo revelado por Ag-NOR e 18S-FISH, um polimorfismo de heterocromatina em acrocêntricos e a presença de microcromossomos Bs. Hypostomus cochliodon mostrou um número diploide de 64 cromossomos (16m+26sm+14st+8a); apesar do sistema de NOR simples, alguns indivíduos apresentaram NOR em ambos os telômeros detectados por Ag-NOR e 18S-FISH. Uma isozima identificou dois loci diagnósticos (Idh-A and Gdh-A) entre as duas espécies e múltiplos loci com alelos únicos em H. boulengeri. A variabilidade genética indicada pela heterozigosidade média (He) foi de 0,2461 e 0,0309 em H. boulengeri e H. cochliodon, respectivamente. Assim, este estudo relata os primeiros dados citogenéticos para H. boulengeri e os primeiros dados isoenzimáticos para H. boulengeri e H. cochliodon. As duas espécies apresentaram evidentes diferenças citogenéticas e isoenzimáticas com a obtenção de marcadores genéticos exclusivos fornecendo suporte para futuros estudos evolutivos no grupo. Sul taxonômicos forma grupo Assim H bacia Paraguai n 6 14m+14sm+18st+22a, 14m14sm18st22a msmsta 14m+14sm+18st+22a , 14m 14sm 18st 22a m sm st (14m+14sm+18st+22a) AgNOR Ag 18SFISH, 18SFISH SFISH 18S FISH, FISH S 18S-FISH Bs 16m+26sm+14st+8a 16m26sm14st8a 16m 26sm 14st 8a (16m+26sm+14st+8a) simples 18SFISH. FISH. IdhA Idh GdhA Gdh Gdh-A He (He 02461 0 2461 0,246 00309 0309 0,030 respectivamente (14m+14sm+18st+22a (16m+26sm+14st+8a 0246 246 0,24 0030 030 0,03 024 24 0,2 003 03 0,0 02 2 0, 00

Abstract Hypostomus is distributed by Central and South America basins, with diverse species with taxonomic conflicts. This way, the integration of auxiliary techniques contributes to understanding the systematics and phylogeny of the group. Thus, this study aimed to investigate the Hypostomus cochliodon and H. boulengeri from the Onça stream (Paraguai River basin) by allozyme and cytogenetic techniques. Hypostomus boulengeri showed a diploid number of 68 chromosomes (14m+14sm+18st+22a), multiple NOR revealed by Ag-NOR and 18S rDNA FISH, a polymorphism of heterochromatin in acrocentrics and the presence of B microchromosome. Hypostomus cochliodon showed a diploid number of 64 chromosomes (16m+26sm+14st+8a); despite the single NOR, some individuals showed NOR in both telomeres detected by Ag-NOR and 18S rDNA FISH. Isozyme identified two diagnostic loci (Idh-A and Gdh-A) between the two species and multiple loci with unique alleles in H. boulengeri. The genetic variability indicated by the mean heterozygosity (He) was 0.2461 and 0.0309 in H. boulengeri and H. cochliodon,respectively.Thus, this study reports the first cytogenetic data for H. boulengeri and the first isozymatic data for H. boulengeri and H. cochliodon. The two species presented evident cytogenetic and isoenzymatic differences with the obtaining of exclusive genetic markers providing support for future evolutionary studies in the group. basins conflicts way group Thus H Paraguai basin 6 14m+14sm+18st+22a, 14m14sm18st22a msmsta 14m+14sm+18st+22a , 14m 14sm 18st 22a m sm st (14m+14sm+18st+22a) AgNOR Ag S FISH microchromosome 16m+26sm+14st+8a 16m26sm14st8a 16m 26sm 14st 8a (16m+26sm+14st+8a) IdhA Idh A GdhA Gdh Gdh-A He (He 02461 0 2461 0.246 00309 0309 0.030 cochliodonrespectivelyThus respectively cochliodon,respectively.Thus (14m+14sm+18st+22a (16m+26sm+14st+8a 0246 246 0.24 0030 030 0.03 024 24 0.2 003 03 0.0 02 2 0. 00

Abstract The current study proposes to investigate the diversity and phylogeny of trypanosomes parasitizing wild birds from the Brazilian Atlantic Forest. Cytological examination was carried out by light microscopy of blood smears and positive birds were selected for amplification of the 18S rDNA sequence through PCR. The resulting amplicons were subjected to purification, cloning, and sequencing analysis. Phylogenetic reconstruction was conducted, including all avian trypanosomes representative’s lineages. A total of ten bird samples from species of Turdus flavipes (N=1/12), T. albicollis (N=1/8), Tachyphonus coronatus (N=6/121), Thamnophilus caerulescens (N=1/22) and Synallaxis spixi (N=1/8) were positive for Trypanosoma spp. In the six specimens of T. coronatus, five distinct lineages of Trypanosoma spp. 18S-rRNA were observed in ninety sequences obtained, and using the strategy of cloning independent PCR, it was possible to observe that two of them were related to T. avium (JB01/JB02), and three were closed related to T. bennetti (JB03/ JB04/JB05). Addionaly, all fifteen sequences obtained from T. caerulescens/ S. spixi/T. flavipes/T. albicollis were identical. The present research is the first study to access molecular diversity and polyparasitism by avian trypanosomes in Brazil. The current research exhibits the wide genetic variability in avian trypanosomes and its non-specific relationship with its avian hosts. Forest S PCR purification analysis conducted representatives representative s N=1/12, N112 N N=1/12 , 1 12 (N=1/12) T N=1/8, N18 N=1/8 8 N=6/121, N6121 N=6/121 6 121 (N=6/121) N=1/22 N122 22 (N=1/22 (N=1/8 spp 18SrRNA SrRNA rRNA JB01/JB02, JB01JB02 JBJB JB01/JB02 JB01 JB02 JB (JB01/JB02) JB03/ JB03 (JB03 JB04/JB05. JB04JB05 JB04/JB05 . JB04 JB05 JB04/JB05) Addionaly spixiT spixi/T flavipesT flavipes/T identical Brazil nonspecific non specific hosts N11 N=1/1 (N=1/12 N1 N=1/ N612 N=6/12 (N=6/121 N=1/2 N12 2 (N=1/2 (N=1/ JB01JB0 JB01/JB0 JB0 (JB01/JB02 (JB0 JB04JB0 JB04/JB0 (N=1/1 N=1 N61 N=6/1 (N=6/12 (N=1 JB01JB JB01/JB (JB01/JB0 (JB JB04JB JB04/JB N= N6 N=6/ (N=6/1 (N= (JB01/JB N=6 (N=6/ (N (N=6