Measurement of expansin activity and plant cell wall creep by using a commercial texture analyzer

Background: Expansins play an important role in cell wall metabolism and fruit softening. Determination of expansin activity is a challenging problem since it depends on measuring cell wall properties by using ad hoc extensometers, a fact that has strongly restricted its study. Then, the objective of the work was to adapt a methodology to measure cell wall creep and expansin activity using a commercial texture meter, equipped with miniature tensile grips and an ad hoc cuvette of easy construction. Results: It was possible to measure hypocotyls acid growth and expansin activity in a reliable and reproducible way, using a commercial texture meter, common equipment found in laboratories of food science or postharvest technology. Expansin activity was detected in protein extracts from cucumber hypocotyls, tomato and strawberry fruits, and statistical differences in expansin activity were found in both fruit models at different ripening stages. Conclusions: The possibility of measuring expansin activity following this adapted protocol with a commercial texture meter could contribute to ease and increase the analysis of expansin in different systems, leading to a better understanding of the properties of these proteins under different experimental conditions.

Bioensayo para determinar el efecto de cuatro categorías de maca amarilla (Lepidium meyenii) provenientes de la localidad de Huallanca (Ancash) sobre el recuento de espermatozoides en testículo, epidídimo y conducto deferente en animales experimentales / Bioassay to determine the effect of four categories of yellow maca (Lepidium meyenii) from the locality of Huallanca (Ancash) on the sperm count in the testicle, epididymis and vas deferens in experimental animals

INTRODUCCIÓN: El consumo de hipocótilos de Lepidium meyenii (Maca), una planta nativa de los Andes centrales del Perú, tiene importantes efectos en la fertilidad. El motivo del presente trabajo es evaluar el efecto de las diferentes categorías (catalogadas así desde la primera calidad hasta la cuarta calidad de acuerdo al tamaño de los hipocótilos); el efecto de diferentes pH (ácido, natural y alcalino) y el efecto de las vías de administración (orogástrica vs intraperitoneal). Como respuesta biológica se evaluará el recuento espermático en ratones de la cepa Swiss. MÉTODOS: Se utilizaron cuatro tamaños diferentes de maca amarilla provenientes del distrito de Huallanca, Departamento de Ancash, ubicado a 4250 msnsm. Se realizaron tres bioensayos considerando el pH acidificado con ácido clorhídrico (pH=4), el pH normal, y alcalinizado con hidróxido de sodio (pH=8). Para cada bioensayo se utilizaron 35 ratones, los cuales fueron administrados con agua destilada (control), un extracto atomizado estandarizado de maca negra (control positivo) y extractos acuosos de las cuatro categorías con una cantidad de 5mg de maca durante 3 días en un volumen de 0.5mL por animal y también se evaluó el efecto de dos vías de administración (orogástrica e intraperitoneal). Después del tratamiento los ratones fueron sacrificados evaluándose la producción diaria de espermatozoides en testículo, y el conteo espermático en epidídimo y conducto deferente. Se consideró un P<0.05 como significante. RESULTADOS: La primera y segunda categoría, ambas tuvieron los valores más ácidos de pH y la mejor actividad biológica. Los extractos acuosos de maca de primera y segunda categoría con el pH acidificado presentaron un efecto significativo sobre el conteo de espermatozoides en todos los órganos estudiados, seguido del extracto acuoso de pH natural, mientras el extracto acuoso alcalinizado no presentó efecto (P>0.05). La vía de tratamiento más efectiva fue la orogástrica, obteniéndose un elevado recuento espermático (P<0.05) a diferencia de la vía intraperitoneal (P>0.05). CONCLUSIÓN: El extracto acuoso de maca amarilla acidificado de primera categoría mostró el mejor efecto sobre el recuento de espermatozoides en los diferentes órganos reproductivos y la mejor vía de administración fue la orogástrica, lo que sugiere que el pasaje gastro-intestinal promueve la producción de principios activos.

Cambios anatómicos en raíces e hipocótilos de plántulas de Prosopis ruscifolia (Fabaceae) sometidas a estrés salino / Anatomical changes in roots and hypocotyls of Prosopis ruscifolia (Fabaceae) seedlings exposed to saline stress

ResumenProsopis ruscifolia es una especie arbórea pionera en áreas inundadas o salinas. El objetivo de este trabajo fue determinar cambios anatómicos en raíces e hipocótilos de plántulas de P. ruscifolia sometidas a estrés salino, bajo condiciones controladas. Las semillas se recolectaron en bosques nativos de la Región Chaqueña Occidental de Argentina. Las semillas se sembraron sobre toallas de papel humedecidas con soluciones salinas de 100, 200 y 300 mM de NaCl y un control humedecido con agua destilada. Se sembraron cuatro repeticiones de 50 semillas cada una, correspondientes a cada tratamiento, se ubicaron en cajas plásticas herméticas dentro de cámara de siembra a 27 ºC y con fotoperíodo de 12 horas. Doce días después de la siembra, se extrajeron plántulas para estudios anatómicos. Se estudiaron 35 plántulas correspondientes a cada tratamiento. Se midieron en raíces e hipocótilos las siguientes variables anatómicas: diámetro de la raíz principal e hipocótilo (µm), espesor de la corteza (µm), número de estratos celulares en la corteza, diámetro del cilindro central (µm), diámetro de la médula (µm), número de estratos celulares en el periciclo y diámetro tangencial de los vasos (µm). Se realizó ANOVA con diámetro de la raíz o hipocótilo como variable dependiente y espesor de la corteza, número de estratos celulares en la corteza, diámetro del cilindro central, diámetro de la médula, número de estratos celulares en el periciclo, diámetro tangencial de los vasos y concentración salina como variables independientes. El diámetro de la raíz disminuyó significativamente con el aumento de la concentración salina (P < 0.0001). El espesor de la corteza redujo su espesor a 100 mM (P < 0.0001) e incrementó el número de estratos celulares que la componen (P < 0.0002). El diámetro del cilindro central se redujo a la concentración salina de 100 mM (P < 0.0001) y el diámetro de la médula y el número de estratos celulares del periciclo (P < 0.0003) disminuyó progresivamente hasta 300 mM. El diámetro tangencial de los vasos (P < 0.0001) se redujo recién a 300 mM de NaCl. Estos cambios anatómicos podrían estar relacionados con la alteración de la expansión y división celular causada por la salinidad y comprometer la formación de raíces laterales y el almacenamiento de reservas. Los hipocótilos no mostraron cambios anatómicos significativos en respuesta al incremento en la salinidad, con excepción de la variación en la posición de estomas y un incremento en el espesor de la hipodermis. Estos cambios parecen indicar el estrés hídrico impuesto por el bajo potencial osmótico causado por las sales. Las plántulas de P. ruscifolia experimentaron cambios anatómicos en respuesta a las concentraciones salinas analizadas, en rasgos vinculados al almacenamiento de reservas, a la absorción y la conducción de agua y la formación de raíces laterales.

Abstract:Prosopis ruscifolia is a pioneer tree species in flooding or saline areas. The aim of this work was to assess anatomical changes in roots and hypocotyls of P. ruscifolia seedlings induced to saline stress under controlled conditions. Seeds, collected in natural forests of Western Chaco region in Argentina, were sown on paper towels moisturized with saline solutions of 100, 200 and 300 mM of NaCl, and a control group with distilled water. Four repetitions of 50 seeds per treatment were sown, located in hermetic polystyrene boxes, and included in a seeding chamber, at 27 ºC and 12 hours photoperiod. Were studied 35 seedlings from each saline concentration; these seedlings were processed 12 days after sown to obtain microscopic samples. The anatomical variables measured in roots and hypocotyls were the following: main root diameter (µm), bark thickness (µm), number of cell strata in bark, central cylinder diameter (µm), pith diameter (µm), number of cell strata in the pericycle and the tangential diameter of vessels (µm). ANOVA analysis were performed with hypocotyl and root diameters as the dependent variable, and bark thickness (µm), number of cell strata in the bark, the central cylinder diameter (µm), the pith diameter (µm), number of cell strata in the pericycle, the tangential diameter of vessels and the saline concentration as independent variables. Results showed that the root diameter decreased with increasing saline concentrations (P < 0.0001). The bark thickness decreased at 100 mM (P < 0.0001) and the number of cell strata of bark increased to 300 mM (P < 0.0002). The central cylinder diameter decreased at 100 mM saline concentration (P < 0.0001) and the number of cell strata of the pericycle and the pith diameter reduced progressively until 300 mM. The tangential diameter of vessels decreased at 300 mM. These anatomical changes suggested alterations in the expansion and cell division caused by the salinity, and could limit lateral roots formation and reserves storage. Hypocotyls did not show significant anatomical changes in response to increasing salinity, with exception of stomata position and an increase of the hypodermis thickness. These changes indicated that the water stress imposed by low osmotic potential is caused by increasing saline concentration. The seedlings of P. ruscifolia experienced anatomical changes in response to tested saline concentrations in traits related to reserve storage, the absorption and conduction of water, and lateral roots formation. Rev. Biol. Trop. 64 (3): 1007-1017. Epub 2016 September 01.

Morphoanatomy of Serjania communis Cambess. seedling (Sapindaceae) / Morfoanatomia da plântula de Serjania communis Cambess. (Sapindaceae)

Serjania communis Cambess. (Sapindaceae) is a plant with climbing habit and occurs relatively often in Paraná State, Brazil. The fruits were collected at the 'Parque dos Pioneiros' a remnant of subtropical forest in Maringá, Paraná State, Brazil. The seedlings obtained in the greenhouse were described according to traditional techniques in plant morphology. Seedlings were embedded in historesin and sectioned in rotation microtome. The fruit is the samaroid type, the seeds have about 3 mm in length and brown color. Seedlings are epigeal phanerocotylar. The seedlings have a hairy hypocotyl, foliaceous cotyledons, reduced epicotyl, and two opposite eophylls. The root is diarch, the hypocotyl shows root-stem transition structure, stem epicotyl, and dorsiventral and hypostomatous cotyledons and eophylls. 'Tirodendros' with 45 days of age do not develop cambial variant.

Serjania communis Cambess. (Sapindaceae) é uma planta de hábito lianescente e ocorre com relativa frequência no Estado do Paraná, Brasil. A coleta dos frutos para a obtenção de sementes foi realizada no parque florestal dos Pioneiros (bosque II), um remanescente de floresta subtropical de Maringá, Estado do Paraná, Brasil. As plântulas, obtidas em casa de vegetação foram descritas conforme técnicas usuais em morfologia vegetal. Após a germinação, as plântulas foram descritas morfoanatomicamente e também foi verificada se ocorria a instalação de variante cambial nesta fase inicial de desenvolvimento. O fruto é do tipo samaroide. As sementes têm cerca de 3 mm de comprimento, de cor marrom e com germinação epigeia e as plântulas são fanerocotiledonares. As plântulas foram emblocadas em historresina e secionadas em micrótomo de rotação. As plântulas apresentam o hipocótilo piloso, cotilédones foliáceos, o epicótilo é reduzido, os dois primeiros eofilos são opostos e os demais eofilos apresentam filotaxia alterna helicoidal. A raiz é diarca. O hipocótilo é uma região de transição raiz-caule. O epicótilo tem estrutura caulinar eustélica. Os cotilédones e os eofilos são hipostomáticos e dorsiventrais. Tirodendros com 45 dias de idade não desenvolvem variante cambial.

Seedling structure of Euphorbia L. and Chamaesyce Gray species / Estrutura da plântula de espécies de Euphorbia L. e Chamaesyce Gray

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. and Euphorbia graminea Jacq. are weedy species that occur with relative frequency in the region of Maringá, Paraná State, Brazil. Data about the morphology and anatomy of seedlings of these species were obtained, with emphasis on significant structural differences of seedlings between species of Euphorbia and Chamaesyce. Seedlings are phanerocotylar and epigeal. Root is axial and triarch. Cotyledons are leaf-like and dorsiventral. Eophylls and prophylls are simple and dorsiventral. Morphological variations occur in the cotyledon shape of Euphorbia seedlings. It is remarkable the Kranz structure of leaves that occurs only in Chamaesyce.

Chamaesyce prostrata (Aiton) Small, Euphorbia heterophylla L. e E. graminea Jacq. (Euphorbiaceae) são espécies invasoras frequentes na região de Maringá, Paraná. Foram obtidos dados sobre a morfologia e anatomia das plântulas dessas espécies, com ênfase se há caracteres estruturais significativos das plântulas entre as espécies de Euphorbia e Chamaesyce. As plântulas são fanerocotiledonares e epígeas. A raiz é axial e triarca. Os cotilédones são foliáceos e dorsiventrais. Os eofilos e profilos são folhas simples e dorsiventrais. As variações mofológicas ocorrem no formato dos cotilédones das plântulas de Euphorbia. Sob o aspecto anatômico é especialmente notável a estrutura Kranz das folhas que ocorrem somente em Chamaesyce.

Somatic embryogenesis in wild relatives of cotton (Gossypium Spp.) / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypocotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods.

Lepidium meyenii (Maca) reduces spermatogenic damage induced by a single dose of malathion in mice / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To observe the effect of the aqueous extract of hypocotyls of the plant Lepidium meyenii (Maca) on spermatogenic damage induced by the organophosphate insecticide malathion in mice.</p><p><b>METHODS</b>Mice were treated with 80 mg/kg of malathion in the presence or absence of an aqueous extract of Maca, which was orally administered 7, 14 or 21 days after injection of the malathion. Stages of the seminiferous epithelium were assessed by transillumination on days 0, 7, 14 and 21.</p><p><b>RESULTS</b>The administration of Maca increased significantly the length of stage VIII on days 7, 14 and 21 of treatment compared with the controls. An increase in the length of stage IX occurred on day 14 of treatment. Malathion affected spermatogenesis by reducing the lengths of stage IX on day 7, stages VII and IX-XI on day 14 and a recovery of stages IX-XII on day 21. The magnitude of alteration in the length of stage IX produced by malathion was significantly reduced by Maca on days 7 and 14. The length of stage VIII was increased when Maca was administered to mice treated with malathion. Assessment of the relative length of stages of the seminiferous epithelium showed that Maca treatment resulted in rapid recovery of the effect of malathion.</p><p><b>CONCLUSION</b>Maca enhances spermatogenesis following spermatogenic damage caused by the organophosphorous pesticide.</p>

Callus formation and its isoflavonoid accumulation in Maackia amurensis / 中国中药杂志

<p><b>OBJECTIVE</b>To obtain Maackia amurensis callus and investigate some factors influencing total isoflavonoid production in callus.</p><p><b>METHOD</b>Different media were used for callus induction from cotyledonary and hypocotyl explants. Total isoflavonoid content was determined by UV spectrophotometer.</p><p><b>RESULT</b>Medium types and hormone combinations influenced both callus formation and their developmental states. MS medium supplemented with NAA/BA or containing 2,4-D was suitable for callus induction from cotyledonary explants. Among the basal media tested, MS, N6 and B5 were suitable for cotyledonary callus formation and SH for hypocotyl callus formation. The formed callus could synthesize isoflavones. Media containing NAA were suitable for isoflavonoid production in cotyledonary callus and media containing 2,4-D/BA were suitable for hypocotyl callus, but 2,4-D/KT at high concentration inhibited the isoflavonoid accumulation in hypocotyl and coytledonary calli. According to the developmentally morphological characters, the formed callus could be classified into four types. II-type callus with yellow in color and friability in texture showed the highest accumulation of isoflavones.</p><p><b>CONCLUSION</b>The influences of medium type and hormone combinations on Maackia amurensis callus formation are reflected in both developmentally morphological characters and isoflavonoid accumulation of calli. Yellow, friable callus induced from cotyledonary and hypocotyl explants in MS or N6 medium supplemented with 2,4-D and BA exhibit the optimum growth and isoflavonoid production.</p>

Involvement of active oxygen species and peroxidases in phytoalexin production induced in soybean hypocotyls by an elicitor from a saprophytic fungus

The involvement of active oxygen species and apoplastic peroxidases in the mechanism of phytoalexin induction were evaluated in soybean hypocotyls using an eliciting extract from the saprophytic fungus Mucor ramosissimus. Hydrogen peroxide and ferric chloride-ascorbic acid a .OH radical generating system, promoted glyceollin production and increase in peroxidase activity similar to those observed with the fungal elicitor. Addition of catalase or potassium ferrocyanide (an inhibitor of peroxidase) inhibited both processes. It is suggested that there is a cause-effect relationship between peroxidase activity and phytoalexin induction both triggered by oxidative processes.