Simple, Fast and Efficient Methods for Analysing the Structural, Ultrastructural and Cellular Components of the Cell Wall.

Plant cell walls are a fundamental component of plant biology and play an essential role in plant growth and development. The metabolic components of the cell wall can be investigated in a fast, simple, and highly efficient manner using various and distinct microscopy techniques. Here, we report implementing a flowchart to analyse tobacco plants' structural, ultrastructural, and metabolic components supplemented with far-red light. In addition, biochemical components, such as lignin, cellulose, phenolic compounds, and reducing sugars, present in the plant cell walls were quantified using light, fluorescence, and electron microscopy. Our data were generated from samples prepared via tissue fixation, incorporation in resins, and slicing using microtomes. Moreover, we have used routine staining and contrast techniques to characterise plant cell walls. Here, we describe several protocols that use classic and modern techniques as well as qualitative and quantitative analytical methods to study cell walls, enabling the plant research community to understand and select the most suitable methods for the microscopic analysis of metabolic components. Finally, we discuss specific ideas aimed at new students of plant anatomy and microscopy. This research not only described the structural, ultrastructural, and metabolic components of the plant cell wall, but also explained the strategies for understanding cellular development.

Complementary chromatic and far-red photoacclimations in Synechococcus ATCC 29403 (PCC 7335). I: The phycobilisomes, a proteomic approach.

Synechococcus ATCC 29403 (PCC 7335) is a unicellular cyanobacterium isolated from Puerto Peñasco, Sonora Mexico. This cyanobacterium performs complementary chromatic acclimation (CCA), far-red light photoacclimation (FaRLiP), and nitrogen fixation. The Synechococcus PCC 7335 genome contains at least 31 genes for proteins of the phycobilisome (PBS). Nine constitutive genes were expressed when cells were grown under white or red lights and the resulting proteins were identified by mass spectrometry in isolated PBS. Five inducible genes were expressed under white light, and phycoerythrin subunits and associated linker proteins were detected. The proteins of five inducible genes expressed under red light were identified, the induced phycocyanin subunits, two rod linkers and the rod-capping linker. The five genes for FaRLiP phycobilisomes were expressed under far-red light together with the apcF gene, and the proteins were identified by mass spectrometry after isoelectric focusing and SDS-PAGE. Based on in silico analysis, Phylogenetic trees, and the observation of a highly conserved amino acid sequence in far-red light absorbing alpha allophycoproteins encoded by FaRLiP gene cluster, we propose a new nomenclature for the genes. Based on a ratio of ApcG2/ApcG3 of six, a model with the arrangement of the allophycocyanin trimers of the core is proposed.

Concerted transcription of auxin and carbohydrate homeostasis-related genes underlies improved adventitious rooting of microcuttings derived from far-red treated Eucalyptus globulus Labill mother plants.

Economically important plant species, such as Eucalyptus globulus, are often rooting recalcitrant. We have previously shown that far-red light enrichment applied to E. globulus donor-plants improved microcutting rooting competence and increased rooting zone/shoot carbohydrate ratio. To better understand this developmental response, the relative expression profiles of genes involved in auxin signaling (ARF6, ARF8, AGO1), biosynthesis (YUC3) and transport (AUX1, PIN1, PIN2); sucrose cleavage (SUS1, CWINV1), transport (SUC5), hexose phosphorylation (HXK1, FLN1) and starch biosynthesis (SS3) were quantified during adventitious rooting of E. globulus microcuttings derived from donor plants exposed to far-red or white light. Expression of auxin transport-related genes increased in the first days of root induction. Far-red enrichment of donor plants induced ARF6, ARF8 and AGO1 in microcuttings. The first two gene products could activate GH3 and other rooting related genes, whereas AGO1 deregulation of the repressor ARF17 may relief rooting inhibition. Increased sink strength at the basal stem with sucrose unloading in root tissue mediated by SUC and subsequent hydrolysis by SUS1 were also supported by gene expression profile. Fructose phosphorylation and starch biosynthesis could also contribute to proper carbon allocation at the site of rooting, as evidenced by increased expression of related genes. These data are in good agreement with increased contents of hexoses and starch at the cutting base severed from far-red exposed donor plants. To sum up, pathways integrating auxin and carbohydrate metabolism were activated in microcuttings derived from donor plants exposed to far red light enrichment, thereby improving rooting response in E. globulus.

Morphology, ecophysiology and germination of seeds of the Neotropical tree Alibertia patinoi (Rubiaceae) / Morfología, ecofisiología y germinación de semillas del árbol neotropical

Alibertia patinoi (Rubiaceae). Alibertia patinoi (Rubiaceae) is of economic and cultural importance for communities in the Colombian Pacific and Amazon regions, where it is cultivated and mature fruits are highly appreciated and consumed. Since there is a lack of knowledge of the seed physiology of this species, we describe here the germination behavior and morphometry of seeds of Alibertia patinoi, and relate them to its habitat. Fruits were collected from a mixed food crop and a commercial plantation in Guaimía village, Buenaventura, Colombia, a tropical rain forest area. We measured length, width, thickness, mass (n=1 400), and moisture content of seeds (n=252). Primary dormancy tests were conducted (n=200), followed by imbibition (n=252) and germination dynamics, under different conditions of light and temperature specific to understory and forest clearings (n=300 seeds). Finally, seed storage behavior was established (n=100 seeds). We observed that size and mass of seeds had a narrow range of values that did not differ within or among fruits and that the species did not exhibit primary dormancy. The seeds are recalcitrant, and recently harvested seeds exhibited higher seed moisture content (ca. 44%) and continuous metabolism. The seed germination percentage was observed to be higher under the specific dense canopy forest light and temperature conditions; furthermore, neither enriched far-red light nor darkness conditions inhibited germination. We concluded that rapid germination could be the establishment strategy of this species. Also, the physiological traits (i.e., rapid germination rate, low germination requirements, absence of primary dormancy, and recalcitrant behavior) and seed size and mass, suggest that A. patinoi is adapted to conditions of mature tropical rain forests.

La germinación y morfometría de las semillas de Alibertia patinoi se describen y se relacionan con el hábitat de la especie, el Bosque Pluvial Tropical Americano. Se midió la longitud, ancho, grosor, peso y contenido de humedad de las semillas. Se realizaron pruebas de latencia, imbibición y dinámica de la germinación con diferentes condiciones de luz y temperatura específicas de claros y de sotobosque cubierto por un dosel denso. Además, se estableció el comportamiento de almacenamiento de las semillas. El tamaño y peso de las semillas no difieren dentro ni entre frutos. Las semillas recién cosechadas no tienen latencia primaria, presentan alto contenido de humedad (ca. 44%), metabolismo continuo y son recalcitrantes. La germinación tuvo porcentajes altos y en condiciones de luz y temperatura específicas de bosque con follaje denso, y ni la luz enriquecida con rojo lejano ni la oscuridad inhibieron la germinación. Se concluye que la germinación rápida podría ser la estrategia de establecimiento de esta especie. Además, los rasgos fisiológicos, el tamaño y peso de las semillas, sugieren que A. patinoi está adaptada a las condiciones del bosque lluvioso tropical maduro y se comporta como una especie que no es pionera.

O afilhamento da aveia afetado pela qualidade da luz em plantas sob competição

Oat plants grown in communities can change their morphology at early stages of development because of differential light absorption on the red and far-red wavelengths. This was studied in open air experiments with oat cultivar UFRGS 15, sown from 300 to 350 plants/m², under natural radiation conditions, in wintertime, in Southern Brazil. In the first experiment, blue filters were placed between plant rows in order to decrease light quality, between Haun stages 1.1 and 3.1. For second experiment, low fluence of red (660nm) e far-red (730nm) light were supplemented during the day to increase (red) and decrease (far-red) respectively the light quality. This was done between Haun stage 1.1 to 3.1, at two distances from the plants. In the third experiment, red and far-red light were also supplemented but for two different periods: emergence to Haun stage 2.1 and Haun stage 2.1 to 4.1. The blue filters and far-red supplementation (expts 1 and 2) induced plants to emit less tillers and tillers with less dry mass but was not seen in experiment 3, when soil chemical conditions were better than the other experiments. With red supplementation, plants emitted more tillers with more dry mass. Under these conditions, the mass ratio mainstem/tiller was less proeminent than under blue filters and far-red supplementation.

As plantas de aveia, quando cultivadas em comunidades, podem mudar sua morfologia nos primeiros estádios de desenvolvimento, devido à absorção diferencial da luz vermelha (V) e vermelha extrema (Ve). Objetivando identificar mudanças no desenvolvimento da aveia pela modificação na razão entre a luz V e Ve, foram conduzidos três experimentos com aveia (cv.UFRGS 15). A densidade variou de 300 a 350p IMG SRC="http:/img/revistas/cr/v31n3/a05img01.gif">/m², sob condições naturais de radiação, durante o inverno, no sul do Brasil. No primeiro experimento, foram colocados filtros azuis entre as linhas de plantas para diminuir a qualidade da luz, entre os estádios 1.1 e 3.1 da escala Haun. No segundo experimento, baixas densidades de fluxo de luz V (660nm) e Ve (730nm) foram suplementadas durante o dia, a diferentes distâncias. No terceiro experimento, baixos fluxos de V e Ve também foram suplementadas, mas em dois períodos distintos: da emergência ao estádio 2.1 e do estádio 2.1 a 4.1. O uso de filtro azul e a suplementação de luz Ve determinaram menor emissão de afilhos e afilhos de menor massa seca (MS). Isso não foi verificado no terceiro experimento, no qual as melhores condições químicas possivelmente diminuíram a magnitude do efeito da luz. Já a suplementação com luz vermelha determinou maior emissão e afilhos de maior MS, devido a menor priorização da acumulação de MS no colmo principal.

A qualidade da luz afeta o afilhamento em plantas de trigo, quando cultivadas sob competição

The selective absortion of red light by foliage changes the light quality in plant communities. This work was conducted to determinate if plants can detect those early modifications by changing tiller emission and dry mass accumulation. Embrapa 16 wheat cultivar was grown at approximatelly 400p- /m² under natural radiation conditions during wintertime, in Southern Brazil. On the first experiment (poor light quality), green and red filters were placed between rows, from Haun stages 1.0 to 3.1. For the second experiment (good light quality), low fluence red light (660nm) was supplemented during the day from Haun stage 1.0 to 3.1. In the third experiment (poor light quality) low fluence far-red light (730nm) was supplemented during the day from Haun stage 1.0 to 3.1. Light quality affected tiller emission, tillers dry mass and mass allocation between mainstem and tillers. Poor light quality (filters that reduced red light and light supplementation) induced plants to emit less tillers with less dry mass per tiller. Poor light quality priorized mainstem against dry mass under community conditions, even if there was not a real competition for the total amount of light received by plants. Good light quality (red light supplementation) had the opposite effect. Tillers were emitted in higher amounts and mass allocation for mainstem was less than for low light quality treatments.

O presente trabalho foi conduzido com o objetivo de verificar os efeitos da mudança na qualidade da luz sobre a emissão de afilhos e na sua acumulação de massa seca. A cultivar de trigo EMBRAPA 16 foi cultivada em comunidades de aproximadamente 400p- /m², em condições naturais de radiação durante o inverno, no Sul do Brasil. No primeiro experimento (baixa qualidade de luz), utilizaram-se filtros verde e vermelho entre as linhas, entre os estádios Haun de 1.0 a 3.1. No segundo experimento (melhor qualidade de luz), as plantas foram suplementadas durante o dia com luz vermelha (V) (660nm), entre os estádios Haun 1.0 e 3.1. Já no terceiro experimento (baixa qualidade de luz), as plantas foram suplementadas durante o dia com luz vermelha extrema (Ve) (730nm), entre os estádios Haun 1.0 e 3.1. A qualidade da luz afetou a emissão de afilhos, sua massa seca e sua distribuição entre o colmo principal e os afilhos. A baixa qualidade da luz (filtros verde e vermelho e luz Ve) induziu as plantas a emitir menos afilhos com menos massa seca/afilho. A baixa qualidade da luz priorizou o colmo principal sob condições de comunidades, antes que tivesse sido estabelecida a competição pela quantidade total de luz recebida pelas plantas. A melhor qualidade de luz (luz V) teve efeito oposto. Os afilhos foram emitidos em maior número e a alocação de massa seca foi menos priorizada para o colmo principal do que nos tratamentos com baixa qualidade.

A qualidade da luz afeta o afilhamento em plantas de trigo, quando cultivadas sob competição

The selective absortion of red light by foliage changes the light quality in plant communities. This work was conducted to determinate if plants can detect those early modifications by changing tiller emission and dry mass accumulation. Embrapa 16 wheat cultivar was grown at approximatelly 400p- /m² under natural radiation conditions during wintertime, in Southern Brazil. On the first experiment (poor light quality), green and red filters were placed between rows, from Haun stages 1.0 to 3.1. For the second experiment (good light quality), low fluence red light (660nm) was supplemented during the day from Haun stage 1.0 to 3.1. In the third experiment (poor light quality) low fluence far-red light (730nm) was supplemented during the day from Haun stage 1.0 to 3.1. Light quality affected tiller emission, tillers dry mass and mass allocation between mainstem and tillers. Poor light quality (filters that reduced red light and light supplementation) induced plants to emit less tillers with less dry mass per tiller. Poor light quality priorized mainstem against dry mass under community conditions, even if there was not a real competition for the total amount of light received by plants. Good light quality (red light supplementation) had the opposite effect. Tillers were emitted in higher amounts and mass allocation for mainstem was less than for low light quality treatments.

O presente trabalho foi conduzido com o objetivo de verificar os efeitos da mudança na qualidade da luz sobre a emissão de afilhos e na sua acumulação de massa seca. A cultivar de trigo EMBRAPA 16 foi cultivada em comunidades de aproximadamente 400p- /m², em condições naturais de radiação durante o inverno, no Sul do Brasil. No primeiro experimento (baixa qualidade de luz), utilizaram-se filtros verde e vermelho entre as linhas, entre os estádios Haun de 1.0 a 3.1. No segundo experimento (melhor qualidade de luz), as plantas foram suplementadas durante o dia com luz vermelha (V) (660nm), entre os estádios Haun 1.0 e 3.1. Já no terceiro experimento (baixa qualidade de luz), as plantas foram suplementadas durante o dia com luz vermelha extrema (Ve) (730nm), entre os estádios Haun 1.0 e 3.1. A qualidade da luz afetou a emissão de afilhos, sua massa seca e sua distribuição entre o colmo principal e os afilhos. A baixa qualidade da luz (filtros verde e vermelho e luz Ve) induziu as plantas a emitir menos afilhos com menos massa seca/afilho. A baixa qualidade da luz priorizou o colmo principal sob condições de comunidades, antes que tivesse sido estabelecida a competição pela quantidade total de luz recebida pelas plantas. A melhor qualidade de luz (luz V) teve efeito oposto. Os afilhos foram emitidos em maior número e a alocação de massa seca foi menos priorizada para o colmo principal do que nos tratamentos com baixa qualidade.

O afilhamento da aveia afetado pela qualidade da luz em plantas sob competição

Oat plants grown in communities can change their morphology at early stages of development because of differential light absorption on the red and far-red wavelengths. This was studied in open air experiments with oat cultivar UFRGS 15, sown from 300 to 350 plants/m², under natural radiation conditions, in wintertime, in Southern Brazil. In the first experiment, blue filters were placed between plant rows in order to decrease light quality, between Haun stages 1.1 and 3.1. For second experiment, low fluence of red (660nm) e far-red (730nm) light were supplemented during the day to increase (red) and decrease (far-red) respectively the light quality. This was done between Haun stage 1.1 to 3.1, at two distances from the plants. In the third experiment, red and far-red light were also supplemented but for two different periods: emergence to Haun stage 2.1 and Haun stage 2.1 to 4.1. The blue filters and far-red supplementation (expts 1 and 2) induced plants to emit less tillers and tillers with less dry mass but was not seen in experiment 3, when soil chemical conditions were better than the other experiments. With red supplementation, plants emitted more tillers with more dry mass. Under these conditions, the mass ratio mainstem/tiller was less proeminent than under blue filters and far-red supplementation.

As plantas de aveia, quando cultivadas em comunidades, podem mudar sua morfologia nos primeiros estádios de desenvolvimento, devido à absorção diferencial da luz vermelha (V) e vermelha extrema (Ve). Objetivando identificar mudanças no desenvolvimento da aveia pela modificação na razão entre a luz V e Ve, foram conduzidos três experimentos com aveia (cv.UFRGS 15). A densidade variou de 300 a 350p IMG SRC="http:/img/revistas/cr/v31n3/a05img01.gif">/m², sob condições naturais de radiação, durante o inverno, no sul do Brasil. No primeiro experimento, foram colocados filtros azuis entre as linhas de plantas para diminuir a qualidade da luz, entre os estádios 1.1 e 3.1 da escala Haun. No segundo experimento, baixas densidades de fluxo de luz V (660nm) e Ve (730nm) foram suplementadas durante o dia, a diferentes distâncias. No terceiro experimento, baixos fluxos de V e Ve também foram suplementadas, mas em dois períodos distintos: da emergência ao estádio 2.1 e do estádio 2.1 a 4.1. O uso de filtro azul e a suplementação de luz Ve determinaram menor emissão de afilhos e afilhos de menor massa seca (MS). Isso não foi verificado no terceiro experimento, no qual as melhores condições químicas possivelmente diminuíram a magnitude do efeito da luz. Já a suplementação com luz vermelha determinou maior emissão e afilhos de maior MS, devido a menor priorização da acumulação de MS no colmo principal.