Neuronal guidance genes in health and diseases

Neurons migrate from their birthplaces to the destinations, and extending axons navigate to their synaptic targets by sensing various extracellular cues in spatiotemporally controlled manners. These evolutionally conserved guidance cues and their receptors regulate multiple aspects of neural development to establish the highly complex nervous system by mediating both short- and long-range cell-cell communications. Neuronal guidance genes (encoding cues, receptors, or downstream signal transducers) are critical not only for development of the nervous system but also for synaptic maintenance, remodeling, and function in the adult brain. One emerging theme is the combinatorial and complementary functions of relatively limited classes of neuronal guidance genes in multiple processes, including neuronal migration, axonal guidance, synaptogenesis, and circuit formation. Importantly, neuronal guidance genes also regulate cell migration and cell-cell communications outside the nervous system. We are just beginning to understand how cells integrate multiple guidance and adhesion signaling inputs to determine overall cellular/subcellular behavior and how aberrant guidance signaling in various cell types contributes to diverse human diseases, ranging from developmental, neuropsychiatric, and neurodegenerative disorders to cancer metastasis. We review classic studies and recent advances in understanding signaling mechanisms of the guidance genes as well as their roles in human diseases. Furthermore, we discuss the remaining challenges and therapeutic potentials of modulating neuronal guidance pathways in neural repair.

Micropropagación de Calibanus hookeri (lem.) trel (1911). Una especie amenazada / Micropropagation of Calibanus hookeri (lem.) trel (1911). A threatened species

RESUMEN La especie Calibanus hookerii perteneciente a la familia Asparagaceae, está registrada en la NOM-059-SEMARNAT-2010 catalogada como planta amenazada. Sus poblaciones naturales se han visto reducidas de manera importante debido a una explotación excesiva y destrucción de su hábitat, por lo que se requiere de métodos de propagación eficaz que aseguren su conservación. La propagación in vitro es una alternativa viable para especies vegetales amenazadas. En la presente investigación se reporta el protocolo para la micropropagación de Calibanus hookerii mediante la germinación de semilla sin testa inoculada en medio MS complementado con 2.5, 5.0 y 7.0 mg L-1 de 6 benciladenina (BA), cinetina (K), 2-isopentil-adenina (2iP) y tidiazuron (TDZ). Las variables a medir fueron porcentaje de germinación, número y longitud de brotes producidos por semilla. El tratamiento más eficiente fue de 5.0 mg L-1 de BA produciéndose un promedio de 26 brotes por semilla; el tratamiento menos eficaz fue con 2.5 mg L-1 K en el cual solamente se obtuvieron dos brotes por semilla. De las tres concentraciones de 2iP solamente en la concentración de 7 mg. L-1 mostró resultados produciendo 6 brotes por semilla. En lo que respecta a la longitud del brote ningún tratamiento superó al testigo (8.07cm). La eficiencia la germinación in vitro fue de 56-97%.

ABSTRACT The species Calibanus hookerii belonging to the family Aspagaceae is registered in the NOM-059-SEMARNAT-2010 cataloged in danger of extinction and therefore is necessary of propagation methods that assure its conservation and its propagation. In vitro propagation is a viable alternative for endangered plant species. The present investigation reports the protocol for the micropropagation of Calibanus hookerii. This was achieved by seed germination without test in MS medium supplemented with 2.5, 5.0 and 7.0 mg L-1 of 6-benzyladenine (BA), kinetin (K), 2-isopentyl-adenine (2iP) and tidiazuron (TDZ). The variables to be measured were percentage of germination, number and length of shoots produced by seed. The most efficient treatment was 5 mg L-1 of BA producing an average of 26 shoots per seed, the worst treatment was with 2.5 K only produced 2 shoots per seed, of the four cytokinins used 2ip treatment in only one study of the Performed showed results (7 mg L-1) producing 6 shoots per seed. Regarding the length of the shoot, no treatment exceeded the control (8.07cm). Finally, in vitro germination was high (56-97%) in all treatments.

Proportional vascularization along the fallopian tubes and ovarian fimbria: assessment by confocal microtomography / Proporção vascular ao longo das tubas uterinas e fímbria ovariana: avaliação pela microtomografia confocal

Abstract Objective: To evaluate and reconstruct three-dimensional images of vascularization along the fallopian tube (FT), as well as to determine its relationship with the ovary and ovarian fimbria, and to quantify the blood vessels along the FT according to its anatomical segments, using confocal microtomography (micro-CT). Materials and Methods: Nine specimens (six FTs and three FTs with ovaries) were fixed in a solution of 10% formalin for > 24 h at room temperature. Iodine staining was performed by soaking the specimens in 10% Lugol's solution for 24 h. All specimens were evaluated using micro-CT. A morphometric analysis was performed on the reconstructed images to quantify the vascular distribution along the FT. Results: In the FTs evaluated, the density of blood vessels was significantly greater in the fimbrial segments than in the isthmic segments (p < 0.05). The ovarian fimbria was clearly identified, demonstrating the important relationship between these vessels and the FT fimbriae. Conclusion: We believe that the vascularization in the fimbriae is greater than and disproportional that in the other segments of FT, and that the ovarian fimbria plays an important role in the development of that difference.

Resumo Objetivo: Avaliar e reconstruir imagens tridimensionais da vascularização ao longo da trompa de Falópio (TF) e sua relação com o ovário e fímbria do ovário, além de quantificar os vasos sanguíneos ao longo da TF de acordo com seus segmentos anatômicos usando microtomografia confocal (micro-TC). Materiais e Métodos: Nove espécimes (seis com TFs e três com TFs e ovários) foram fixados em solução de formol a 10% por mais de 24 h à temperatura ambiente. A coloração com iodo foi realizada usando solução de Lugol a 10% durante 24 h. Todos os espécimes foram avaliados usando micro-TC. Uma análise morfométrica foi realizada nas imagens reconstruídas para quantificar a distribuição vascular ao longo da TF. Resultados: A densidade dos vasos sanguíneos nas fímbrias foi significativamente aumentada em comparação ao segmento ístmico da TF (p < 0,05). A fímbria ovariana foi claramente identificada, demonstrando importante relação entre esses vasos e as fímbrias da TF. Conclusão: Acreditamos que as fímbrias apresentam vascularização aumentada e desproporcional em comparação com os demais segmentos da TF e que a fímbria ovariana desempenha importante papel nessa diferença.

Callus induction, Indirect Organogenesis and Plantlet Regeneration from different explants of Vernonia anthelmintica (L.) Willd

Present study was conducted to standardize callus development and indirect organogenesis from differentexplants of Vernonia anthelmintica (L.) Willd. Among, the various hormonal combinations tested: IndoleAcetic Acid (IAA) and BA and IAA and Kinetin combinations were found to be optimum for inducing callusingwithin a time span of 10 days. Best callus response was observed in 1.5 mg l−1 IAA and 1.5 mg l−1 BA,which produced white friable callus. Best indirect shoot organogenesis was observed in 4 mg l−1 BA and6 mg l−1 kinetin. Elongated shoots when transferred on to half strength Murashige and Skoog (MS) Mediumsupplemented with Indole-3-butyric acid (IBA), rooting was observed. MS medium fortified with 2 mg l−1 IBAshowed better rooting than all other concentrations tested. This concentration produced maximum number ofroots and maximum percentage of rooting. Plantlets developed by indirect organogenesis of V. anthelminticawere successfully acclimatized to field conditions

Direct in vitro regeneration of castor bean plants (Ricinus communis) using epicotyls / Regeneração direta in-vitro de plantas de mamona ( Ricinus communis ) usando epicótilos

A regeneration protocol for castor bean plant (Ricinus communis) was successfully developed using epicotyl sections obtained from in vitro seedlings. The maximum number of induced shoots (4.3 shoots/explant) and highest shoots frequency (75,56%) was obtained in WPM medium supplemented with TDZ (1 mg/L) and zeatin (0.5 mg/L), whereas the minimum number (0.8 shoots/explant) and lowest shoots frequency (37,78%) was obtained in medium containing TDZ (1 mg/L) and BAP (0.5 mg/L). The highest percentage of rooting (93.3%) was obtained in a medium containing IBA (1 mg/L). These plants were transplanted in a mesh house and achieved a high adaptability to acclimatization, reaching 77% survival. On the other hand, the maximum elongation (height) during this stage was 7.9 cm in plants supplemented with WPM nutrients, whereas it was only 4.38 cm in control plants

Foi desenvolvido com sucesso um protocolo de regeneração para a planta de Mamona (Ricinus communis) utilizando seções de epicótilos, obtidas a partir de mudas in vitro. O número máximo de brotações induzidas (4.3 brotos/explante), assim como a maior frequência de brotações (75,56%), foi obtido em meio WPM suplementado com TDZ (1 mg/L) e zeatina (0,5 mg/L). Enquanto que o número mínimo (0,8 brotos/explante), como a menor freqüência de rebentos (37,78%), foi obtido em meio contendo TDZ (1 mg/L) e BAP (0,5 mg/L). Adicionalmente, a maior percentagem de enraizamento (93,3%) foi obtida em um meio contendo IBA (1 mg/L). Depois da regeneração, as plantas foram transplantadas em casa de vegetação e conseguiram uma alta adaptabilidade e aclimatização, atingindo 77% de sobrevivência. Por outro lado, oalongamento máximo (altura) durante este estágio foi de 7,9 cm em plantas suplementadas com nutrientes de WPM, enquanto as plantas de controle presentaram apenas 4,38 cm

In Vitro Organogenesis from Root Explants of Passiflora miniata Mast, an Amazonian Species with Ornamental Potential

ABSTRACT The present study reports a shoot organogenesis-based system for in vitro regeneration of Passiflora miniata, an Amazonia passion fruit species. Root segments were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations (range 2-9 µM) of 6-benzyladenine (BA); thidiazuron (TDZ) or kinetin (KIN). Plant growth regulators were not added to the control treatment. Root explants have showed a high regenerative potential. After 30 days of in vitro culture, the root explants showed several shoots formed direct and indirectly. TDZ provided the best response in the differentiation adventitious shoots, mainly in the presence of 6.8 µM. The cytokinins BA and KIN responded producing a reduced number of shoots. After 120 days, rooted regenerated plants were transferred to a greenhouse for acclimatization. This regeneration system opens new perspectives for micropropagation and conservation of this wild Amazonic passion fruit species.

Relationship between cellular rejection and development of rat transplanted metanephroi from different gestational ages / 中华器官移植杂志

Objective To explore the relationship between cellular rejection and the development of allo-or xenografted primordia from different gestational ages .Methods Whole rat metanephroi from embryonic day E14～ E19 were transplanted into omenta of outbred (SD → SD ,6 groups ,n≥10 each ;E15-E17SDCsA ,3 groups ,n=15 each) ,syngeneic (Lewis→Lewis ,5 groups ,n=8 each) ,allogeneic (Lewis→BN ,E15BN n= 6 each E15BNCsA n= 10 each ,E16BNCsA n= 10 each) rats and xenogeneic (Lewis→C57groups ,E15C57 n=10 each ,E15C57CsA ,n=8 each ;Lewis→Balb/c nude mice ,3 groups ,n=10 each) recipients .Histopathology ,Banff's grading and electron microscopy (EM ) were utilized for assessing the graft development .Similarly ,biochemical indicators and creatinine clearances were measured .Results At 4 weeks post-transplantation , in SD → SD groups ,E14-E17SD metanephroi developed with Banff ' s rejections . E14/E15SD was significantly lighter than E16/E17SD ( P< 0 .01 );E18/E19SD barely developed . After cyclosporine A (CsA , 8 mg·kg -1·d-1 )dosing ,Banff's rejection of E15-E17SDCsA group lessened significantly .In Lewis→BN ,E15BN metanephroi were completely rejected .After dosing CsA (12 mg·kg -1·d-1 ) ,E15BNCsA and E16BNCsA Banff ' s rejections became alleviated . Upon a discontinuation of CsA , both metanephroi were rejected . In Lewis → Lewis , E15 ～ E17Lewis metanephroi developed well . No significant difference existed in Banff's classification (P>0 .05) .E14Lewis and E18Lewis rats had significantly poorly differentiated metanephroi than those in E16 Lewis group .In Lewis→C57BL/6 , E15 metanephroi were rejected at Day 14 post-transplantation (n= 10) and no improvement was evident after CsA dosing (15 mg·kg -1·d-1 ,n=8) .In Lewis→Balb/c nude mice ,all E15～E17Balb/c metanephroi developed well .Both light microscopy and EM examination showed normal nephrons and collecting ducts and wet weight ,creatinine or urea nitrogen of effusion showed no significant difference (P>0 .05) .E15Lewis and E16Lewis had significantly different values of wet weight and creatinine clearances from those of E15SDCsA and E16SDCsA .E15SDCsA had the greatest wet weight and the lowest creatinine clearance rate (P< 0 .01) .Conclusions After controlling rejection during allo-and xenotransplantations ,E15 ,E16 and E17 rat metanephros have similar development characteristics . And cellular immunogenic factors still remain the major barriers to their developments .

Micropropagación y aclimatación de Maguey Pitzometl (Agave marmorata Roezl) en la Mixteca Poblana / Micropropagation and acclimatization of Maguey Pitzometl (Agave marmorata Roezl) in the poblana Mixteca

RESUMEN Agave marmorata R. es una planta que se adapta a terrenos someros y fertilidad baja, también sirve para la retención y conservación de agua de lluvia reduciendo la erosión del suelo. Actualmente se encuentra de forma silvestre y escasa en la Mixteca Poblana (México) donde es intensamente aprovechada en gastronomía y como planta medicinal. Por ello, se propuso la propagación in vitro como estrategia de rescate y conservación a partir de brotes in vitro de Agave marmorata en un medio de cultivo básico de Murashige y Skoog (1962) 100 %, suplementado con azúcar de caña 3 %, myoinositol 100 mg-L-1, agar 0.7 % y 0.40 mg-L-1 de tiamina-HCl, donde se agregaron por separado diferentes concentraciones de 6-Bencil-adenina (BA) y ácido indol-3-acético (AIA) mediante un arreglo factorial 5x5. Se evaluó el número y longitud de brotes y de raíces obtenidos in vitro. Para [as plantas en aclimatación sólo se evaluó el porcentaje de sobrevivencia en diferentes sustratos. Los datos de las variables se sometieron a un análisis de varianza y a una prueba de medias de Tukey (p=0.05). La mejor respuesta para longitud de brotes, número y longitud de raíces fue en el medio MS 100 % adicionando 10 mg-L-1 de AIA y la proliferación de nuevos brotes fue promovida con la adición de BA y AIA en igual concentración. Finalmente, se obtuvo el 100 % de sobrevivencia de las plántulas en los sustratos de peat moss más agrolita y peat moss más arena de río.

ABSTRACT Agave marmorata R. is a plant that adapts to shallow lands and low fertility, it also serves for the retention and conservation of rainwater reducing soil erosion. It is currently found in a wild and scarce form in the Mixteca Poblana (Mexico) where it is intensively used in gastronomy and as a medicinal plant. For this reason, in vitro propagation was proposed as a rescue and conservation strategy from in vitro shoots of Agave marmorata in a basic culture medium of Murashige and Skoog (1 962) 100 %, supplemented with 3 % sugar cane, myo- inositol 100 mg-L-1, agar 0.7 % and 0.40 mg-L-1 thiamine-HCl, where different concentrations of 6-Benzyl-adenine (BA) and indole-3-acetic acid (IAA) were added separately by a 5x5 factorial arrangement. The number and length of shoots and roots obtained in vitro were evaluated. For the plants in acclimation only the percentage of survival in different substrates was evaluated. Los datos de las variables se sometieron a un análisis de varianza y a una prueba de medias de Tukey (p=0.05). The best response for shoot length, number and length of roots was in the MS 100 % medium adding 10 mg-L-1 of IAA and the proliferation of new shoots was promoted with the addition of BA and IAA in the same concentration. Finally, 100 % survival of the seedlings was obtained in the substrates of peat moss plus agrolita and peat moss plus river sand.

Inmunolocalización de Sonic hedgehog en el desarrollo embrio-fetal de ratones ( Mus musculus ) / Immunolocalization of Sonic hedgehog in embryo-fetal mouse ( Mus musculus ) development

Sonic hedgehog (Shh) es un morfógeno esencial para el desarrollo de diversas estructuras, tales como notocorda, placa del piso del tubo neural, miembros, entre otros. Se buscó determinar la inmunolocalización de Shh en embriones y fetos de ratón. Para ello, se eutanasiaron 10 ratones gestantes (Mus musculus) BALB/c, un grupo de 5 animales a los 12,5 días post-coito (dpc), y otro grupo a los 17,5 dpc. Los embriones y fetos obtenidos fueron fijados en formalina al 10 % tamponada en PBS e incluidos en paraplast. Se realizaron cortes transversales seriados. Se utilizó anticuerpo policlonal Shh (Santa Cruz Biotechnology, H-160, conejo), dilución 1:100. Se identificó y describió la inmunolocalización de las muestras marcadas positivamente. La expresión de Shh en los embriones de 12,5 dpc fue inmunopositiva en notocorda, placa del piso del tubo neural, precartílago de radio y ulna, y prácticamente todos los epitelios: bronquial, intestinal, vejiga y uretra. En la etapa fetal, a los 17,5 dpc la inmunopositividad desaparece en el cartílago a excepción de zonas de osificación, disminuye en la epidermis pero aparece en folículos pilosos. La mucosa intestinal se ha diferenciado en segmentos, mostrando una inmunotinción mayor a nivel de las vellosidades intestinales. Shh actúa en distintos estadios del periodo gestacional, siendo clave en la diferenciación de distintas estructuras. En etapas embrionaria, es vital en la formación del sistema nervioso, organogénesis y formación de miembros, por lo que su expresión se encuentra en estas zonas. Sin embargo, en la etapa fetal la expresión cambia a estructuras de mayor especialización como folículo piloso y vellosidades intestinales.

Sonic hedgehog (Shh) is an essential morphogen for the development of various structures, such as notochord, neural tube floor plate, limbs, among others. We sought to determine the immunolocalization of Shh in embryos and mouse fetuses. To do this, 10 pregnant mice (Mus musculus) BALB /c were euthanized, a group of 5 animals at 12.5 days postcoitus (dpc), and another group at 17.5 dpc. Embryos and fetuses obtained were fixed in 10 % formalin buffered in PBS and embedded in paraplast. Serial cross sections were made. Polyclonal antibody Shh (Santa Cruz Biotechnology, H-160, rabbit), dilution 1:100 was used. The immunolocalization of the positively labeled samples was identified and described. Shh expression in 12.5 dpc embryos was immunopositive in notochord, neural tube floor plate, radius precartilage and ulna, and practically all epithelia: bronchial, intestinal, bladder and urethra. In the fetal stage, at 17.5 dpc the immunopositivity disappears in the cartilage except for areas of ossification, decreases in the epidermis but appears in hair follicles. The intestinal mucosa has differentiated into segments, showing greater immunostaining at the level of the intestinal villi. Shh acts in different stages of the gestational period, being key in the differentiation of different structures. In embryonic stages, it is vital in the formation of the nervous system, organogenesis and formation of limbs, so its expression is found in these areas. However, in the fetal stage the expression changes to more specialized structures such as hair follicles and intestinal villi.

Transitando de embrión a feto: la metamorfosis de los cordados / Transiting from embryo to fetus: the metamorphosis of the chordata

Durante el período del desarrollo conocido como prefetal, el embrión cambia sus características ictiomórficas comunes a todos los vertebrados y adquiere gradualmente las formas propias de la especie que representa. Durante este período se forma la cara, involucionan los arcos faríngeos (branquiales) formándose el cuello, y aparecen los miembros. Se constituye, además, la hernia umbilical fisiológica, que consiste en la presencia de asas intestinales dentro del cordón umbilical. El sistema nervioso origina las vesículas telencefálicas, el diencéfalo, mesencéfalo, metencéfalo, y mielencéfalo. Este periodo corresponde a una etapa de máxima susceptibilidad ante los teratógenos que pueden generar malformaciones en todas las especies animales. El objetivo del presente trabajo es presentar los principales eventos acontecidos durante el periodo prefetal, además de una visión y opinión de los autores, proponiendo una nueva denominación a la etapa: periodo metamórfico.

During the period of development known as prefetal, the embryo changes its ictiomorphic characteristics common to all vertebrates and gradually acquires the proper forms of the species it represents. During this period the face is formed, the pharyngeal arches (branchial) involute forming the neck, and the limbs appear. In addition, the physiological umbilical hernia is constituted, which consists of the presence of intestinal loops inside the umbilical cord. The nervous system originates the telencephalic vesicles, the diencephalon, mesencephalon, metencephalon, and myelencephalon. This period corresponds to a stage of maximum susceptibility to teratogens that can generate malformations in all animal species. The objective of this paper is to present the main events that took place during the preferential period, as well as a vision and opinion of the authors, proposing a new name for the stage: metamorphic period.

Morphogenesis of the rhea (Rhea americana) respiratory system in different embryonic and foetal stages / Morfogênese do sistema respiratório da ema (Rhea americana) em diferentes estágios embrionários e fetais

The rhea (Rhea americana) is an important wild species that has been highlighted in national and international livestock. This research aims to analyse embryo-foetal development in different phases of the respiratory system of rheas. Twenty-three embryos and foetuses were euthanized, fixed and dissected. Fragments of the respiratory system, including the nasal cavity, larynx, trachea, syrinx, bronchi and lungs, were collected and processed for studies using light and scanning electron microscopy. The nasal cavity presented cubic epithelium in the early stages of development. The larynx exhibited typical respiratory epithelium between 27 and 31 days. The trachea showed early formation of hyaline cartilage after 15 days. Syrinx in the mucous membrane of 18-day foetuses consisted of ciliated epithelium in the bronchial region. The main bronchi had ciliated epithelium with goblet cells in the syringeal region. In the lung, the parabronchial stage presented numerous parabronchi between 15 and 21 days. This study allowed the identification of normal events that occur during the development of the rhea respiratory system, an important model that has not previously been described. The information generated here will be useful for the diagnosis of pathologies that affect this organic system, aimed at improving captive production systems.(AU)

A ema (Rhea americana) representa importante espécie silvestre que vem se destacando na pecuaria nacional e internacional. Esta pesquisa objetiva analisar o desenvolvimento embrionário-fetal, em diferentes fases, do sistema respiratório de emas. Vinte e três embriões e fetos foram eutanasiados, fixados e dissecados. Fragmentos do sistema respiratório: cavidade nasal, laringe, traqueia, siringe, brônquios e pulmões, foram coletados e processados para estudos por meio de microscopia de luz e microscopia eletrônica de varredura. A cavidade nasal apresentou, nas primeiras fases de desenvolvimento, epitélio estratificado cúbico. A laringe exibiu epitélio respiratório típico entre 27 e 31 dias. A traqueia aos 15 dias apresentou início de formação da cartilagem hialina. Na siringe a túnica mucosa de fetos de 18 dias e formada por epitélio estratificado ciliado na região bronquial. Os brônquios principais apresentavam epitélio estratificado ciliado com células caliciformes na região siringeal. No pulmão, o estágio parabronquial apresentou numerosos parabrônquios entre 15 a 21 dias. Este estudo permitiu a identificação de eventos normais que ocorrem durante o desenvolvimento do sistema respiratório de emas, importante modelo ainda não descrito. As informações geradas serão úteis para o diagnóstico de patologias que acometem este sistema orgânico, visando a melhoria dos sistemas de produção em cativeiro.(AU)

Analysis of histone modification of MtSERK1 during in vitro regeneration in Medicago truncatula / 生物工程学报

Epigenetic modification, especially histone modification, plays an important role in maintaining plant genome stability, regulating gene expression and promoting regeneration in vitro. MtSERK1 is an important marker gene involved in establishing of embryogenic callus during in vitro regeneration of Medicago truncatula. In order to understand the regulation Epigenetic modification, especially histone modification, plays an important role in maintaining plant genome stability, regulating gene expression and promoting regeneration in vitro. MtSERK1 is an important marker gene involved in establishing of embryogenic callus during in vitro regeneration of Medicago truncatula. In order to understand the regulation relationship between dynamic histone modification and MtSERK1s expression during the processes of in vitro organogenesis, the expression of MtSERK1 was analyzed by qRT-PCR, and the modification status of H3K9me2, H3K4me3 and H3K9ac in the promoter region and different regions included in the gene body was analyzed by chromatin immunoprecipitation (ChIP). We found expression activation of MtSERK1 was related to the dynamic changes of histone H3K4me3 and H3K9ac in the 5' and 3' regions. This study will provide important theoretical guidance for understanding of the regulatory mechanism of MtSERK1 and also for establishing efficient genetic transformation system of Medicago truncatula.

Date palm micropropagation: Advances and applications / Micropropagação de tamareira: Avanços e aplicações

ABSTRACT Date palm (Phoenix dactylifera L.) is a fruit tree resilient to adverse climatic conditions predominating in hot arid regions of the Middle East and North Africa. The date fruit contains numerous chemical components that possess high nutritional and medicinal values. Traditional propagation by offshoots is inefficient to satisfy current demands for date palm trees. Alternatively, micropropagation provides an efficient means for large-scale propagation of date palm cultivars. Both somatic embryogenesis and organogenesis, either directly or indirectly though the callus phase, have been demonstrated in date palm in vitro regeneration. Culture initiation commonly utilizes shoot-tip explants isolated from young offshoots. Recently, the immature inflorescences of adult trees were utilized as an alternative nondestructive source of explants. In addition to the nature of the explant used, successful plant regeneration depends on the cultivar, composition of the culture medium and physical status. Challenges of date palm micropropagation include long in vitro cycle, latent contamination, browning, somaclonal variation as well as ex vitro acclimatization and transplanting. A remarkable amount of research investigating these factors has led to optimized protocols for the micropropagation of numerous commercially important cultivars. This has encouraged the development of several international commercial tissue culture laboratories. Molecular characterization provides an assurance of genetic conformity of regenerated plantlets, a key feature for commercial production. This article describes date palm micropropagation protocols and also discusses recent achievements with respect to somaclonal variation, molecular markers, cryopreservation and future prospects.

RESUMO A tamareira (Phoenix dactylifera L.) é uma arvore frutífera adaptada à condições climáticas adversas predominantemente em regiões áridas do Oriente Médio e Norte Africano. As tâmaras possuem vários componentes químicos com alto valor medicinal e nutricional. A propagação tradicional por estacas não é suficiente para satisfazer a demanda por mudas e assim, a micropropagação apresenta-se como uma alternativa eficiente para a produção de mudas em larga escala. Embriogênese somática e organogênese, tanto direta quanto indireta via calos, tem sido usada para obter a regeneração in vitro de tamareira. O inicio do cultivo in vitro normalmente utiliza meristemas excisados de brotações jovens. Recentemente, inflorescências imaturas de árvores adultas são usadas como fonte alternativa de explantes não destrutiva. Além da origem do explante, o sucesso da regenerção depende do cultivar, da composição do meio de cultura e de condições físicas. Desafios na micropropagação de tamareira incluem um longo ciclo in vitro, contaminação, escurecimento do tecido, variação somaclonal além do enraizamento e aclimatização ex vitro. Diversos estudos investigando esses fatores tem conduzido à otimização de protocolos de micropropagação de inúmeros cultivares comerciais proporcionando o estabelecimento de vários laboratórios de cultura de tecidos de plantas. A caracterização molecular permite uma segura conformidade genética do material regenerado, considerado uma característica chave na produção comercial. Essa revisão descreve protocolos de micropropagação de tamareira e aborda as mais recentes conquistas relacionadas à variação somaclonal, marcadores moleculatres, criopreservação e perspectivas futuras.

Regeneración in vitro de plantas de piña ( Ananas comosus ) ecotipo amazónico Gobernadora / In vitro regeneration of Amazonian pineapple ( Ananas comosus ) plants ecotype Gobernadora

Resumen En Venezuela existen cultivares y ecotipos de piña (A. comosus) de importancia local, entre ellos los amazónicos, cultivados principalmente por los aborígenes Piaroa. Ellos siembran los propágulos lo cual restringe la disponibilidad de material para el cultivo a gran escala. Se abordó la limitación recurriendo al cultivo de tejidos vegetales para la propagación in vitro de plantas de piña, ecotipo amazónico Gobernadora, mediante embriogénesis somática (ES) y organogénesis adventicia (OA). El material vegetal empleado correspondió a secciones basales e intermedias de hojas. Sólo las secciones de base foliar (SBF) fueron morfogénicamente inducidas. El mayor número de vitroplantas (1,58 plantas/explante) se obtuvo del callo embriogénico inducido en medio MS con Picloram 10 mg.L-1 + Tidiazuron 2 mg.L-1, transferido a MS sin hormonas. En el proceso organogénico, se obtuvo el mayor número de plantas/explante (5) por vía directa en MS con ácido naftalenoacético 5 mg.L-1 + bencilaminopurina 0,25 mg.L-1, transferido a MS. Siendo este último el mejor sistema de cultivo in vitro por su productividad y por ser una ruta que minimiza la variación somaclonal.

Abstract There are a number of pineapple (Ananas comosus) cultivars and ecotypes of local commercial importance in Venezuela, among them the Amazonian ones, cultivated mainly by the aboriginal Piaroa, are of relevance. They sow the propagules, which restricts the availability of material for large-scale cultivation. This limitation was approached by plant tissue culture for in vitro propagation of Amazonian pineapple plants, Gobernadora ecotype, through somatic embryogenesis (ES) and adventitious organogenesis (OA). Basal and intermediate sections of leaves were tested. Only the leaf base sections (FBS) were morphogenically induced. The highest number of vitroplants (1.58 plants / explant) was obtained from the embryogenic callus induced in MS medium with Picloram 10 mg.L-1 + Thidiazuron 2 mg.L-1, transferred to MS medium without hormones. In the organogenic process, the highest number of plants / explants (5) was obtained directly in MS with naphthaleneacetic acid 5 mg.L-1 + benzylaminopurine 0.25 mg.L-1, transferred to MS. The latter being the best in vitro culture system due to its productivity and for being a method that minimizes somaclonal variation.

Efficient regeneration and genetic transformation platform applicable to five Musa varieties

Background: Banana (Musa spp.) is an important staple food, economic crop, and nutritional fruit worldwide. Conventional breeding has been seriously hampered by their long generation time, polyploidy, and sterility of most cultivated varieties. Establishment of an efficient regeneration and transformation system for banana is critical to its genetic improvement and functional genomics. Results: In this study, a vigorous and repeatable transformation system for banana using direct organogenesis was developed. The greatest number of shoots per explant for all five Musa varieties was obtained using Murashige and Skoog medium supplemented with 8.9 µM benzylaminopurine and 9.1 µM thidiazuron. One immature male flower could regenerate 380­456, 310­372, 200­240, 130­156, and 100­130 well-developed shoots in only 240­270 d for Gongjiao, Red banana, Rose banana, Baxi, and Xinglongnaijiao, respectively. Longitudinal sections of buds were transformed through particle bombardment combined with Agrobacterium-mediated transformation using a promoterless ß-glucuronidase (GUS) reporter gene; the highest transformation efficiency was 9.81% in regenerated Gongjiao plantlets in an optimized selection medium. Transgenic plants were confirmed by a histochemical assay of GUS, polymerase chain reaction, and Southern blot. Conclusions: Our robust transformation platform successfully generated hundreds of transgenic plants. Such a platform will facilitate molecular breeding and functional genomics of banana.

Induction of in vitro shoots of Billbergia euphemiae E. Morren (Bromeliaceae) from leaf explants / Indução in vitro de brotos de Billbergia euphemiae E. Morren (Bromeliaceae) a partir de explantes foliares

Bromeliads are an important group for the maintenance of the Atlantic Forest, with many threatened species due to exacerbated extraction and destruction of their natural habitats. Considering the need of developing protocols for the conservation of these species, the aim of this work was to evaluate the effect of different growth regulators in the in vitro induction of shoots of Billbergia euphemiae. Leaf explants were excised from seedlings derived from in vitro germination and grown on MS medium supplemented with NAA (0, 1 or 2 µM) and BA (0, 2, 4 or 6 µ M) combinations. The evaluation of the number of shoots per explant, shoot length, number of leaves per shoot and longest leaf length average was carried out after 30 and 60 days of culture. The best in vitro responses were observed in the presence of 1 µM NAA after 60 days of culture, which induced the best production of shoots per explant (16.39), as well as the highest rates of shoot length (1.08 cm), number of leaves per shoot (5.00) and the longest leaf length (0.56 cm). This work determined the best conditions for shoot production from leaf explants of B. euphemiae, being the first report on micropropagation of this species.

As bromélias constituem um importante grupo para a manutenção da Floresta Atlântica, com várias espécies ameaçadas de extinção pelo extrativismo exacerbado e a destruição dos habitats naturais. Considerando a necessidade do desenvolvimento de protocolos para a conservação destas espécies, este trabalho teve como objetivo avaliar o efeito de diferentes reguladores de crescimento na indução de brotos de Billbergia euphemiae. Explantes foliares foram excisados de plântulas derivadas da germinação in vitro e inoculados em meio MS suplementado com combinações de ANA (0, 1 ou 2 µM) e BA (0, 2, 4 ou 6 µM). O número de brotos por explantes, comprimento dos brotos, número médio de folhas por broto e comprimento médio da maior folha foram avaliados após 30 e 60 dias de cultura. As melhores respostas foram observadas na presença de ANA a 1 µM, após 60 dias de cultura, que induziu a maior produção de brotos por explante (16,39), assim como as maiores taxas de comprimento dos brotos (1,08 cm), número médio de folhas por broto (5,00) e comprimento da maior folha (0,56 cm). Este trabalho determinou as melhores condições para produção de brotos de B. euphemiae a partir de explantes foliares, representando o primeiro relato de micropropagação desta espécie.

β-defensins and the epididymis: Contrasting influences of prenatal, postnatal, and adult scenarios / 亚洲男科学杂志(英文版)

β-defensins are components of host defense, with antimicrobial and pleiotropic immuno-modulatory properties. Research over the last 15 years has demonstrated abundant expression of a variety of β-defensins in the postnatal epididymis of different species. A gradient of region- and cell-specific expression of these proteins is observed in the epithelium of the postnatal epididymis. Their secretion into the luminal fluid and binding to spermatozoa as they travel along the epididymis has suggested their involvement in reproduction-specific tasks. Therefore, continuous attention has been given to various β-defensins for their role in sperm function and fertility. Although β-defensins are largely dependent on androgens, the underlying mechanisms regulating their expression and function in the epididymis are not well understood. Recent investigation has pointed out to a new and interesting scenario where β-defensins emerge with a different expression pattern in the Wolffian duct, the embryonic precursor of the epididymis, as opposed to the adult epididymis, thereby redefining the concept concerning the multifunctional roles of β-defensins in the developing epididymis. In this review, we summarize some current views of β-defensins in the epididymis highlighting our most recent data and speculations on their role in the developing epididymis during the prenatal-to-postnatal transition, bringing attention to the many unanswered questions in this research area that may contribute to a better understanding of epididymal biology and male fertility.

POTENCIAL DE REGENERACIÓN DE Psychotria ipecacuanha (Rubiaceae) A PARTIR DE CAPAS DELGADAS DE CÉLULAS / Regeneration Potential of Psychotria ipecacuanha (Rubiaceae)from Thin Cell Layers

Conociendo las propiedades medicinales de la especie vegetal Psychotria ipecacuanha (Brot.) Stokes, su crítico estado de conservación, así como las dificultades que presenta para la propagación efectiva, el presente estudio tuvo como objetivo evaluar su potencial de propagación por los sistemas de regeneración in vitro, organogénesis y embriogénesis somática. Para este propósito, capas delgadas de células (CDCs) de tallos y de hojas, así como segmentos foliares fueron sometidos a diferentes tratamientos con reguladores de crecimiento y condiciones de luz. Además se estableció el efecto de diferentes longitudes de onda vía diodos emisores de luz (LEDs), sobre la regeneración en estos explantes y nudos provenientes de plantas in vitro. Los resultados obtenidos mostraron que los segmentos de hoja y las CDCs de tallo sembrados en el medio de cultivo MS suplementado con las combinaciones de los reguladores de crecimiento IBA + BAP e IBA + TDZ formaron embriones somáticos y brotes. Los cortes histológicos realizados corroboraron estos dos tipos de origen. Se encontró que bajo la condición lumínica 16/8, se alcanzaron los mejores resultados de inducción de brotes y embriones. En cuanto al efecto de las diferentes longitudes de onda de luz, se encontró que las correspondientes al rojo, verde y blanca, favorecieron el crecimiento y desarrollo de brotes y la inducción de embriones somáticos. El desarrollo de los brotes a partir de los nudos no presentó diferencias estadísticas entre los tratamientos con LEDs, por lo que se recomienda el uso de la luz blanca continua y con fotoperiodo durante el proceso de multiplicación y desarrollo de éstos.

Knowing the medicinal properties of the plant specie Psychotria ipecacuanha (Brot.) Stokes, its critical condition and the difficulties for its effective propagation, the present study aimed to assess the potential of propagation of P. ipecacuanha by in vitro systems of regeneration, organogenesis and somatic embryogenesis. For this purpose, thin cell layers (TCL) of stems and leaves, as well as leaf segments were subjected to different treatments of plant growth regulators and light conditions. Furthermore, the effect of different wavelengths via light emitting diodes (LED's), was established for the regeneration in these explants and nodal explants from in vitro plants. The results showed that leaf segments and stem TCL, cultured in MS medium supplemented with the combinations of growth regulators IBA + BAP and IBA + TDZ, formed somatic embryos and shoots. The histological sections supported the two types of source. It was found that the best results in shoot induction and embryos were achieved under the light condition 16/8-h light/ dark. Regarding the effect of different wavelengths, it was found that those corresponding to red, green, and white supported the growth and shoot development as well as somatic embryos induction. The shoots development from the nodal explants did not show statistical differences between LEDs treatments, so the use of a continuous white light and photoperiod is recommended during their multiplication and development.

Somaclones em batata identificados pelos descritores mínimos de broto / Potato somaclones identified by minimum descriptors sprout

Este estudo objetivou verificar a estabilidade fenotípica das cultivares de batata 'Asterix' e 'Macaca', avaliar o efeito do tipo de explante (organogênese direta e indireta) e do tempo de subcultivo (12 e 70 meses) em meio nutritivo MS sobre a ocorrência de somaclones nas duas cultivares na produção de batata semente, mediante o emprego de sete descritores mínimos de broto. Em 'Asterix' e 'Macaca' ocorreram somaclones em quatro dos sete descritores, contudo, apenas no formato e pubescência da base do broto houve variação, simultaneamente, em ambas. Os dois genótipos são suscetíveis à ocorrência de variação somaclonal. Registrou-se somaclonesnos dois tempos de subcultivo nas duas cultivares. Diferente do amplamente registrado, identificaram-se somaclones em segmentos apicais caulinares e nodais originados de organogênese direta em 'Asterix' e 'Macaca'.

It was examined the phenotypic stability of potato cultivars 'Asterix' and 'Macaca', evaluated the effect of explant type (direct and indirect organogenesis) and subculture time (12 and 70 months) in MS nutrient medium on the occurrence of somaclonal variation in both cultivars in seed potato production through the use of seven minimum descriptors sprout. In 'Asterix' and 'Macaca' somaclones have occurred in four of the seven descriptors, however, only in the shape and in the base of the bud pubescence that somaclonal variation occurred simultaneously in both cultivars. Both genotypes are susceptible to the occurrence of somaclonal variation. It was identify the occurrence of somaclones both at 12 months and 70 months of subculture in both genotypes. Unlike the widely recorded, somaclones were identified in shoot apical segments and nodal segments derived from direct organogenesis in 'Asterix' and 'Macaca'.

Towards Efficient In vitro Regeneration of Cowpea (Vigna unguiculata L. Walp): A Review.

Cowpea is a crop of tremendous economic and ecological values particularly in sub-Saharan Africa, where over 80% of the crop is produced and consumed. Due to heavy attack by pests and diseases, actual yield does not exceed 20% of the crop’s potential in most of the production regions. Shortages of genes to combat biotic stresses in the germplasm and sexual incompatibility with wild relatives are major impediments in cowpea improvement. Genetic modification of cowpea with relevant genes can address these problems. Establishment of reproducible regeneration system is a prerequisite for genetic transformation of cowpea using transgenic technology. Cowpea is among the most recalcitrant crops for manipulation under In vitro condition especially via de novo process. However, strategies to regenerate cowpea under In vitro conditions have evolved steadily in the last three decades. In this review, we give a summary of cowpea regeneration work carried out so far and discussed approaches employed as well as challenges of developing efficient regeneration systems in cowpea.