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Pollen tube (PT) elongation is important for double fertilization in angiosperms and affects the seed-setting rate and, therefore, crop productivity. Compared to Arabidopsis (Arabidopsis thaliana L.), information on PT elongation in rice (Oryza sativa L.) is limited by the difficulty in obtaining homozygous mutants. In a screen of T-DNA insertional mutants, we identified a mutant in the Tethering protein of actomyosin transport in pollen tube elongation (TAPE) gene with an unusual segregation ratio by genotyping analysis. A CRISPR/Cas9 knockout mutant of TAPE that produced a short PT was sterile, and TAPE was expressed specifically in pollen grains. TAPE is a homolog of a myosin XI adaptor in Arabidopsis with three tetratricopeptide repeat and Phox and Bem1 protein domains. TAPE showed latrunculin B-sensitive, actin-dependent localization to the endoplasmic reticulum. Yeast two-hybrid screening and transcriptome analysis revealed that TAPE interacted with pollen-specific LIM protein 2b and elongation factor 1-alpha. Loss of TAPE affected transcription of 1,259 genes, especially genes related to cell organization, which were downregulated. In summary, TAPE encodes a myosin XI adaptor essential for rice PT elongation.

Cytochrome P450 monooxygenase 704B (CYP704B), a member of the CYP86 clan, was found to be needed in Arabidopsis and rice to biosynthesize precursors of sporopollenin through oxidizing fatty acids. In the present study, we cloned and characterized a CYP704B gene in Panax ginseng, named PgCYP704B1. It shared high sequence identity (98-99%) with CYP704 of Arabidopsis, Theobroma cacao, and Morus notabilis. The phylogenetic comparison of ginseng and higher plants between the members of CYP86 clan revealed that ginseng CYP704 was categorized as a group of CYP704B with dicot plants. The expression of PgCYP704B1 is low in the stem, leaf, and fruit, and high in flower buds, particularly detected in the young gametic cell and tapetum layer of the developing anther. Arabidopsis plants overexpressing PgCYP704B1 improved plant biomass such as plant height, siliques and seed number and size. A cytological observation by transverse and longitudinal semi-thin sections of the siliques cuticles revealed that the cell length increased. Furthermore a chemical analysis showed that PgCYP704B1ox lines increased their cutin monomers contents in the siliques. Our results suggest that PgCYP704B1 has a conserved role during male reproduction for fatty acid biosynthesis and its overexpression increases cutin monomers in siliques that eventually could be used for seed production.

Cytochrome P450 (CYP) catalyzes a wide variety of monooxygenation reactions in plant primary and secondary metabolisms. Land plants contain CYP703, belonging to the CYP71 clan, which catalyzes the biochemical pathway of fatty acid hydroxylation, especially in male reproductive tissues. Korean/Asian ginseng (

Rice is an important food staple that is consumed by half of the human population. Therefore, understanding the regulatory mechanism of male fertility in rice can improve production by enhancing the efficiency of hybrid seed production. However, information on the control mechanism of male fertility by anther dehiscence or wall development in rice is very limited. To further understand the regulatory mechanism for anther dehiscence in rice, we carried out transcriptome analysis for two tissues: the anther wall and pollen at the anthesis stage. With the anatomical meta-expression data, in addition to these tissues, the differentially expressed genes (DEGs) between the two tissues were further refined to identify 1,717 pollen-preferred genes and 534 anther wall-preferred genes. A GUS transgenic line and RT-qPCR analysis for anther wall-preferred genes supported the fidelity of our gene candidates for further analysis. The refined DEGs were functionally classified through Gene Ontology (GO) enrichment and MapMan analyses. Through the analysis of cis-acting elements and alternative splicing variants, we also suggest the feature of regulatory sequences in promoter regions for anther wall-preferred expression and provide information of the unique splicing variants in anther walls. Subsequently, it was found that hormone signaling and the resulting transcriptional regulation pathways may play an important role in anther dehiscence and anther wall development. Our result could provide useful insight for future research to broaden the molecular mechanism of anther dehiscence or anther wall development in rice.

Olá! Me chamo Dayane, sou natural de Caruaru, Pernambuco, um pedacinho de terra no agreste pernambucano conhecido como “A Capital do Forró” e que tem seu ápice cultural durante o mês de junho. Como cidadã caruaruense, o São João sempre foi uma data importante. Fazia questão de participar das quadrilhas juninas do meu colégio. Alugar vestido, sapato, tiara, faixa e preparar todos os detalhes para o momento da minha apresentação fazia parte dos gastos anuais da minha mãe. Desde do ensino médio, não dancei mais em quadrilhas e aquela paixão foi esfriando e se fixou como um marco de uma infância e adolescência. Após esse período, a dança continuou muito ligada a mim. Meu interesse nunca foi me profissionalizar nessa área. Mas, sempre foi deixar meu físico me guiar em uma zona que eu descobria a cada dia. E em 2022, eu buscava por um desafio físico e social. Em abril deste mesmo ano resolvi começar os ensaios e não esperava que uma mera vontade fosse se transformar em um dever. Os meus fins de semana foram tomados, meu dinheiro destinado à comprar maquiagens, figurino e pagar passagem entre uma cidade e outra. Foram incontáveis as vezes que eu tremia de tristeza e raiva, pensando no sufoco que eu estava me metendo. Porém, eu não conseguia sair. Nas apresentações há uma completa entrega, um envolvimento e um propósito que só quem compartilha do amor por uma quadrilha junina pode enxergar. A luta para manter essa tradição era diária, e coletiva. Conto hoje minhas agonias, mas ela não foi a única. Com mais de 50 pessoas envolvidas (entre dançarinos e apoio técnico), os problemas econômicos e pessoais também apareceram para elas. Uma vontade superior de fazer a cultura acontecer e ser expandida para outros locais podia ser enxergada no olhar de cada um. Incluindo no meu. Mais do que nunca, hoje sei o peso que é manter uma tradição cultural, sem apoio do governo e dependendo de um fio forte, porém estreito, chamado amor. E agora faço parte dessa Flor. Ela brotou e se enraizou no meu peito, mostrando que fazer arte é principalmente se deixar levar.

Neste trabalho, analisaremos como o jornal A Verdade constrói seu discurso político em volta dos termos “direita” e “esquerda”. Tomaremos como objeto o editorial O papel do jornalismo operário (2021) que reflete a ideologia e objetivos do jornal. Para este fim, utilizaremos a Análise de Discurso, pois o objetivo é entender a construção de sentido dos termos trabalhados dentro do meio de comunicação.

Pollen in angiosperms plays a critical role in double fertilization by germinating and elongating pollen tubes rapidly in one direction to deliver sperm. In this process, the secretory vesicles deliver cell wall and plasma membrane materials, and excessive materials are sequestered via endocytosis. However, endocytosis in plants is poorly understood. AP180 N-terminal homology (ANTH) domain-containing proteins function as adaptive regulators for clathrin-mediated endocytosis in eukaryotic systems. Here, we identified 17 ANTH domain-containing proteins from rice based on a genome-wide investigation. Motif and phylogenomic analyses revealed seven asparagine-proline-phenylalanine (NPF)-rich and 10 NPF-less subgroups of these proteins, as well as various clathrin-mediated endocytosis-related motifs in their C-terminals. To investigate their roles in pollen germination, we performed meta-expression analysis of all genes encoding ANTH domain-containing proteins in Oryza sativa (OsANTH genes) in anatomical samples, including pollen, and identified five mature pollen-preferred OsANTH genes. The subcellular localization of four OsANTH proteins that were preferentially expressed in mature pollen can be consistent with their role in endocytosis in the plasma membrane. Of them, OsANTH3 represented the highest expression in mature pollen. Functional characterization of OsANTH3 using T-DNA insertional knockout and gene-edited mutants revealed that a mutation in OsANTH3 decreased seed fertility by reducing the pollen germination percentage in rice. Thus, our study suggests OsANTH3-mediated endocytosis is important for rice pollen germination.

Although cell wall dynamics, particularly modification of homogalacturonan (HGA, a major component of pectin) during pollen tube growth, have been extensively studied in dicot plants, little is known about how modification of the pollen tube cell wall regulates growth in monocot plants. In this study, we assessed the role of HGA modification during elongation of the rice pollen tube by adding a pectin methylesterase (PME) enzyme or a PME-inhibiting catechin extract (Polyphenon 60) to in vitro germination medium. Both treatments led to a severe decrease in the pollen germination rate and elongation. Furthermore, using monoclonal antibodies toward methyl-esterified and de-esterified HGA epitopes, it was found that exogenous treatment of PME and Polyphenon 60 resulted in the disruption of the distribution patterns of low- and high-methylesterified pectins upon pollen germination and during pollen tube elongation. Eleven PMEs and 13 PME inhibitors (PMEIs) were identified by publicly available transcriptome datasets and their specific expression was validated by qRT-PCR. Enzyme activity assays and subcellular localization using a heterologous expression system in tobacco leaves demonstrated that some of the pollen-specific PMEs and PMEIs possessed distinct enzymatic activities and targeted either the cell wall or other compartments. Taken together, our findings are the first line of evidence showing the essentiality of HGA methyl-esterification status during the germination and elongation of pollen tubes in rice, which is primarily governed by the fine-tuning of PME and PMEI activities.

Panax ginseng Meyer is one of the important medicinal plants in the world, particularly in Asian countries. Ginseng encounters many stress exposure during its long cultivation period. However, the molecular mechanism of stress resistance is still poorly understood in spite of its importance. In this study, pathogenesis-related protein 6 (PR6), also called proteinase inhibitor (PI), was isolated from ginseng embryogenic callus, named PgPR6. The small size of PR6, containing an open reading frame of 219 bp encoding 72 amino acids, the typical characteristic of PR6 protein, shares the highest sequence similarity to PR6 of Theobroma cacao (69% identity). Sequence and structural analysis indicated that PgPR6 belongs to class Kunitz-type PI family. This is the first report pertaining to the identification of PR6 gene from the P. ginseng genome. The high-level expression of PgPR6 was observed in root as revealed by quantitative real-time PCR. The temporal expression analysis demonstrated that PgPR6 expression was highly up-regulated by signaling molecules, heavy metals, mechanical wounding, chilling, salt, sucrose, and mannitol stress, indicating that PgPR6 may play an important role in the molecular defense response of ginseng to a various range of environmental stresses.

Transcription factors (TFs) are essential for gene regulation in all living organisms, including plants, where among numerous other functions, they control temporal and spatial gene expression in response to environmental stimuli. The v-myb avian myeloblastosis viral oncogene homolog (MYB) family is the largest TF family in plants, with its members involved in developmental processes as well as secondary metabolism. Little is known about MYB genes in Panax ginseng Meyer, despite ginseng’s importance as a widely-used medicinal plant. In this study, we isolated nine MYB genes from P. ginseng (PgMYBs). Phylogenetic comparison of these genes with the MYB genes of other plant species revealed that the PgMYBs clustered into different families based on their putative functions including terpene regulation; five PgMYBs clustered in the R2R3 family, and four PgMYBs clustered in the MYB-related protein group. Further expression analysis of five PgMYBs showed consistently high expression in flower and leaf tissue, suggesting that these PgMYB genes may be involved in development of the above-mentioned tissues. Four PgMYBs were downregulated in response to methyl jasmonate (MJ) and salicylic acid (SA), whereas PgMYB3 was up-regulated, suggesting a role for all these genes in stress response. This is the first comprehensive study of the MYB gene family in P. ginseng, and the information provided here will facilitate further exploration of the functions of these TFs.

Ascorbate peroxidase (APX) plays an essential role in the antioxidant defense mechanism in the plant, serving in the ascorbate–glutathione cycle for the cellular H2O2 metabolism process. As the perennial plant, Panax ginseng Meyer encounters a lot of unfavorable growth conditions, and among them soil salinity significantly decreases the yield. Two APX genes from Panax ginseng were isolated and designated as PgAPX1 and PgAPX2, which are most similar with previously characterized cytosolic APX of Daucus carota and Spuriopimpinella brachycarpa, as revealed by sequence analysis of their deduced amino acid sequences. PgAPXs transcripts are most abundant in leaf tissue, whereas PgAPX1 expression level was higher compared to PgAPX2. Consistent with higher PgAPX1 expression during salt stress in ginseng, PgAPX1-overexpressing Arabidopsis lines (PgAPX1ox) increased the germination rate and root length compared with wild-type (WT) under 200 mM NaCl stress treatment. Furthermore, higher chlorophyll content, relative water content, total APX activity, proline content, and lower H2O2 accumulation were shown in PgAPX1ox plants compared to WT under 200 mM NaCl stress condition. Transcription analysis showed that PgAPX1oxArabidopsis lines were more salt-tolerant by upregulating the ion homeostasis mechanism.

Panax ginseng Meyer, commonly known as ginseng, is considered one of the most important herbs with pharmaceutical values due to the presence of ginsenosides and is cultivated for its highly valued root for medicinal purposes. Recently, it has been recognized that ginseng fruit contains high contents of triterpene such as ginsenoside Re as pharmaceutical compounds. However, it is unclear how carpel, the female reproductive tissue of flowers, is formed during the three-year-old growth before fruit is formed in ginseng plants. Here, we report P. ginseng carpel development at the cytological level, starting from the initial stage of ovule development to seed development. The carpel of P. ginseng is composed of two free stigmas, two free styles, and one epigynous bilocular ovary containing one ovule in each locule. Based on our cytological study, we propose that the female reproductive development in P. ginseng can be classified into seven stages: early phase of ovule development, megasporogenesis, megagametogenesis, pre-fertilization, fertilization, post-fertilization, and seed development. We also describe the correlation of the female and male gametophyte development and compare morphological differences in carpel development between ginseng and other higher plants. One unique feature for ginseng seed development is that it takes 40 days for the embryo to develop to the early torpedo stage and that the embryo is small relative to the seed size, which could be a feature of taxonomic importance. This study will provide an integral tool for the study of the reproductive development and breeding of P. ginseng.

Cytochrome P450 monooxygenase 704B (CYP704B), a member of the CYP86 clan, was found to be needed in Arabidopsis and rice to biosynthesize precursors of sporopollenin through oxidizing fatty acids. In the present study, we cloned and characterized a CYP704B gene in Panax ginseng, named PgCYP704B1. It shared high sequence identity (98-99%) with CYP704 of Arabidopsis, Theobroma cacao, and Morus notabilis. The phylogenetic comparison of ginseng and higher plants between the members of CYP86 clan revealed that ginseng CYP704 was categorized as a group of CYP704B with dicot plants. The expression of PgCYP704B1 is low in the stem, leaf, and fruit, and high in flower buds, particularly detected in the young gametic cell and tapetum layer of the developing anther. Arabidopsis plants overexpressing PgCYP704B1 improved plant biomass such as plant height, siliques and seed number and size. A cytological observation by transverse and longitudinal semi-thin sections of the siliques cuticles revealed that the cell length increased. Furthermore a chemical analysis showed that PgCYP704B1ox lines increased their cutin monomers contents in the siliques. Our results suggest that PgCYP704B1 has a conserved role during male reproduction for fatty acid biosynthesis and its overexpression increases cutin monomers in siliques that eventually could be used for seed production.

Background: In plants, the key roles played by RopGEF-mediated ROP signaling in diverse processes, including polar tip growth, have been identified. Despite their important roles in reproduction, a comprehensive analysis of RopGEF members has not yet been performed in rice (Oryza sativa). To determine whether RopGEF regulators are involved in rice pollen tube growth, we performed genome-wide analysis of this family in rice. Results: Phylogenomic and meta-expression analysis of eleven RopGEFs in rice showed that four genes were preferentially expressed in mature pollen. These four genes contain the plant-specific Rop nucleotide exchanger (PRONE) domain and possible phosphorylated residues, suggesting a conserved role in polar tip growth with Arabidopsis. In subcellular localization analysis of the four RopGEFs through tobacco (Nicotiana benthamiana) infiltration was identified in various cellular organelles, indicating their diverse roles during pollen germination in rice. Moreover, these candidate genes possessing unique cis-acting elements in their promoters compared with the other genes. Conclusions: In this study, four RopGEF genes were identified as pollen-specific gene in eleven members of rice, and the expression pattern, promoter analysis, and evolutionary relationship of the RopGEF family were studied compared with Arabidopsis. Our study indicated that four RopGEF genes might function during pollen germination in distinct subcellular localization. Our study could provide valuable information on the functional study of RopGEF in rice.

Objetivo: Conhecer as práticas de matriciamento em saúde mental desenvolvidas na atenção primária à saúde Métodos: Trata-se de uma revisão integrativa de literatura realizada por meio das bases de dados secundários Scielo, Lilacs e Biblioteca Virtual em Saúde. Resultados: Foram encontrados 42 artigos na Biblioteca Virtual em Saúde, 16 na Scielo e 33 na Lilacs, após aplicação dos critérios de inclusão e exclusão foram selecionados 13 artigos para a amostra final. Algumas barreiras foram indicadas para realização do matriciamento: fragilização da política nacional de atenção básica, heterogeneidade de concepções e modelos de prática, ausência de suporte das gestões e gerência dos serviços, ausência de periodicidade do processo e pouca interferência na organização dos fluxos na rede Rede de Atenção Psicossocial, agenda diversas e problemas entre a Estratégia Saúde da Família e equipes apoiadoras, pouca presença do psiquiatra nas ações de apoio matricial, lógica inserida no modelo ambulatorial com foco na prática de encaminhamento e marcação de consultas, bem como a maneira de implantação da proposta em muitas cidades do país. Considerações finais: Na análise das práticas de apoio matricial desenvolvidas nos cenários da atenção primária à saúde em caráter nacional, reconhece-se um processo embrionário, ainda a ser intensificado e fortalecido.

The chlorophyll a/b-binding protein (CAB) serves in both photosystems (PS), I and II, as a coordinator of antenna pigments in the light-harvesting complex (LHC). The CABs constitute abundant and important proteins in the thylakoid membrane of higher plants. In our study, five CAB genes, which contained full-length cDNA sequences from the 4-year-old ginseng leaves (Panax ginseng Meyer), were isolated and named PgCAB. Phylogenetic comparison of the members of the subfamily between ginseng and higher plants, including Arabidopsis, revealed that the putative functions of these ginseng CAB proteins were clustered into the different family of Arabidopsis CABs; two PgCABs in LHCII family and three PgCABs in LHCI family. The expression analysis of PgCABs consistently showed dark-dependent inhibition in leaves. Expression analysis during abiotic stress identified that PgCAB genes responded to heavy metal, salinity, chilling, and UV stresses differently, suggesting their specific function during photosynthesis. This is the first comprehensive study of the CAB gene family in P. ginseng.

Lipoxygenase (LOX) belongs to a family of non-heme-iron-containing fatty acid dioxygenases that are widely distributed in plants and animals. LOX involved in the synthesis of jasmonic acid and six-carbon (C6) volatiles which is necessary for plant growth and responses to a wide range of biotic and abiotic stresses. We have isolated and characterized LOX cDNA clones from Panax ginseng Meyer. From their deduced amino acid sequences, two diverse classes of 9-LOX (LOX1, LOX2, and LOX3) and 13-LOX (LOX4, LOX5) are defined in P. ginseng. A GenBank Blast X search revealed that the deduced amino acid of PgLOXs share a high degree of homology with LOX from other plants and mammals especially in three distinct motifs; motif1 harboring iron binding regions, motif2 and motif3. Chloroplast localization was predicted for PgLOX5. PgLOXs displayed organ-specific expression, highly expressed in aerial parts of the plant such as 3-year old flower, stem and leaf tissues. PgLOXs mRNAs were elevated strongly by bacterial infection. Expression of PgLOXs was differentially induced in ginseng not only by mechanical damage and methyl jasmonate but also after exposure to withholding water. Ginseng 13-LOXs positively respond to wounding that may involve in production of C6 volatiles and jasmonic acid at the wounded sites. However, the higher expression of PgLOX3 by water deficit and 82 % of the nucleotide sequence identity with the EST from severe drought-stressed leaves of Populus (CU229089.1) at +6371 bp downstream of PgLOX3 genomic DNA structure can suggest drought tolerance role for PgLOX3. Ginseng LOX genes have different expression pattern which may suggest different specific function against various environmental stresses.

Ginseng (Panax ginseng), a valued medicinal herb, is a slow-growing plant that flowers after 3 years of growth with the formation of a solitary terminal umbel inflorescence. However, little is known about cytological events during ginseng reproduction, such as the development of the male organ, the stamen. To better understand the mechanism controlling ginseng male reproductive development, here, we investigated the inflorescence and flower structure of ginseng. Moreover, we performed cytological analysis of anther morphogenesis and showed the common and specialized cytological events including the formation of four concentric cell layers surrounding male reproductive cells followed by subsequent cell differentiation and degeneration of tapetal cells, as well as the formation of mature pollen grains via meiosis and mitosis during ginseng anther development. Particularly, our transverse section and microscopic observations showed that the ginseng tapetal layer exhibits obvious nonsynchronous cell division evidenced by the observation of one or two tapetal layers frequently observed in one anther lobe, suggesting the unique control of cell division. To facilitate the future study on ginseng male reproduction, we grouped the anther development into 10 developmental stages according to the characterized cytological events.

Acaricide resistance is a major obstacle to the control of Rhipicephalus microplus. Historically, the indiscriminate use of chemical compounds has contributed to the selection of populations resistant to different classes of acaricides. Therefore, multiple acaricide resistance is an important threat to the chemical control of the cattle tick. To investigate the occurrence and extent of multiple resistance to acaricides in Southern Brazil we performed larval tests with cypermethrin, chlorpyriphos, amitraz, fipronil and ivermectin on 104 cattle tick field samples from different ranches in Rio Grande do Sul, between the years 2013 and 2015. Adult immersion tests with a commercial formulation mixture of chlorpyriphos and cypermethrin were performed on 75 samples. Four levels of resistance were established according to the mortality of larvae: Level I: mortality between 82% and 95%; Level II: mortality between 57% and 82%; Level III: mortality between 25% and 57%; and Level IV: mortality lower than 25%. Resistance to cypermethrin was detected in 98.08% of the samples evaluated, mostly at resistance level IV. The frequency of samples resistant to amitraz, chlorpyriphos, ivermectin and fipronil was 76.92%, 60.58%, 60.58% and 53.85% respectively. Multiple resistance to three or more compounds was found in 78.85% of the samples. The results obtained in this study are alarming and reveal a new scenario for the challenge of tick control using chemicals. This is an issue of high importance to cattle production systems where this tick is responsible for a high economic impact.

Interspecific hybrids between Panax ginseng and P. quinquefolius results in hybrid vigor and higher ginsenoside contents. Ginseng is one of the most important herbs with valued pharmaceutical effects contributing mainly by the presence of bioactive ginsenosides in the roots. However, ginseng industry is impeded largely by its biological properties, because ginseng plants are slow-growing perennial herbs with lower yield. To increase the ginseng yield and amounts of ginsenosides, we developed an effective ginseng production system using the F(1) progenies obtained from the interspecific reciprocal cross between two Panax species: P. ginseng and P. quinquefolius. Although hybrid plants show reduced male fertility, F(1) hybrids with the maternal origin either from P. ginseng or P. quinquefolius displayed heterosis; they had larger roots and higher contents of ginsenosides as compared with non-hybrid parental lines. Remarkably, the F(1) hybrids with the maternal origin of P. quinquefolius had much higher ginsenoside contents, especially ginsenoside Re and Rb1, than those with the maternal origin of P. ginseng. Additionally, non-targeted metabolomic profiling revealed a clear increase of a large number of primary and secondary metabolites including fatty acids, amino acids and ginsenosides in hybrid plants. To effectively identify the F(1) hybrids for the large-scale cultivation, we successfully developed a molecular marker detection system for discriminating F(1) reciprocal hybrids. In summary, this work provided a practical system for reciprocal hybrid ginseng production, which would facilitate the ginseng production in the future.