Molecular cloning and characterization of an actindepolymerizing factor gene in Hevea brasiliensis

Actin-depolymerizing factor (ADF) plays important roles in regulating actin dynamics by maintaining the optimum equilibrium between unpolymerized actin molecules and assembled actin filaments in different cellular processes. In this study, the first ADF gene in Hevea brasiliensis designated as HbADF, was isolated. The HbADF contained an open reading frame (ORF) encoding 139 amino acids. The deduced HbADF showed high identities to plants ADF proteins. Besides a conserved ADF domain, HbADF also contained putative actin and specific F-actin binding sites, phosphorylation site and possible CAM (calmodulin) combining region. The phylogenetic analysis indicated that HbADF was clustered in the subclass I. Being consistent with  phylogenetic result, the expression of HbADF was constitutive. The HbADF transcripts were upregulated by ethephon and wounding treatments; whereas, HbADF was firstly induced, and then gradually downregulated by jasmonic acid. The expression profiles and characterizations of HbADF suggested that HbADF might be  associated with latex regeneration and flow in H. brasiliensis.Key words: Actin cytoskeleton, actin-depolymerizing factor, expression analysis, Hevea brasiliensis, semiquantitative reverse-transcription polymerase chain reaction

Poly[[diaqua-μ2-4,4′-bipyridyl-μ2-o-phthalato-nickel(II)] dihydrate]

In the title layer complex, {[Ni(C8H4O4)(C10H8N2)(H2O)2]·2H2O}n, the Ni atom has a distorted octa­hedral environment, defined by the phthalate and 4,4′-bipyridyl ligands which link the Ni atoms, forming a square lattice in the bc plane. This extends into a three-dimensional supra­molecular network through O—H⋯O hydrogen-bonding inter­actions. The Ni atom lies on, and both ligands are bis­ected by, a crystallographic twofold axis

Poly[bis­(2,2′-bipyridine-κ2 N,N′)deca-μ-oxido-dioxidodicopper(II)tetra­vanadium(V)]

The title compound, [Cu2V4O12(C10H8N2)2]n, shows a two-dimensional copper–vanadate layer composed of eight-membered rings, each containing four corner-sharing VO4 tetra­hedra; these are linked through six penta­coordinated CuII atoms with the 2,2′-bipyridine ligands attached and pointing above and below the plane of the layer. The Cu atom is coordinated by two N donors from the 2,2′-bipyridine ligand and three O atoms from three adjacent VO4 units to form a distorted tetragonal pyramid. These layers are further connected by π–π inter­actions between inter­leaving bipyridine ligands of adjacent layers [centroid–centroid distances = 3.63 (1) and 3.68 (1) Å] into a three-dimensional supra­molecular structure

Identification and characterization of genes associated with tapping panel dryness from Hevea brasiliensis latex using suppression subtractive hybridization

<p>Abstract</p> <p>Background</p> <p>Tapping panel dryness (TPD) is one of the most serious threats to natural rubber production. Although a great deal of effort has been made to study TPD in rubber tree, the molecular mechanisms underlying TPD remain poorly understood. Identification and systematical analyses of the genes associated with TPD are the prerequisites for elucidating the molecular mechanisms involved in TPD. The present study is undertaken to generate information about the genes related to TPD in rubber tree.</p> <p>Results</p> <p>To identify the genes related to TPD in rubber tree, forward and reverse cDNA libraries from the latex of healthy and TPD trees were constructed using suppression subtractive hybridization (SSH) method. Among the 1106 clones obtained from the two cDNA libraries, 822 clones showed differential expression in two libraries by reverse Northern blot analyses. Sequence analyses indicated that the 822 clones represented 237 unique genes; and most of them have not been reported to be associated with TPD in rubber tree. The expression patterns of 20 differentially expressed genes were further investigated to validate the SSH data by reverse transcription PCR (RT-PCR) and real-time PCR analysis. According to the Gene Ontology convention, 237 unique genes were classified into 10 functional groups, such as stress/defense response, protein metabolism, transcription and post-transcription, rubber biosynthesis, etc. Among the genes with known function, the genes preferentially expressed were associated with stress/defense response in the reverse library, whereas metabolism and energy in the forward one.</p> <p>Conclusions</p> <p>The genes associated with TPD were identified by SSH method in this research. Systematic analyses of the genes related to TPD suggest that the production and scavenging of reactive oxygen species (ROS), ubiquitin proteasome pathway, programmed cell death and rubber biosynthesis might play important roles in TPD. Therefore, our results not only enrich information about the genes related to TPD, but also provide new insights into understanding the TPD process in rubber tree.</p

Interação adulto-criança: a influência da disposição na sala de aula

Mestrado em Educação Pré-Escolar e Ensino no 1º Ciclo do Ensino BásicoO presente relatório, realizado no âmbito da unidade curricular de Prática Pedagógica Supervisionada do Mestrado em Educação Pré-Escolar e Ensino do 1º Ciclo do Ensino Básico, visa compreender a influência da disposição das mesas na sala de aula na interação professor-aluno. Assim, foi realizado um estudo de caso, tendo como participantes quatro alunos de uma turma do 2º ano, do 1º Ciclo do Ensino Básico. Teve como principal técnica de recolha de dados a observação direta não participante. Relativamente aos dados foi efetuada uma análise de conteúdo, tendo por base a Escala de Empenhamento do Adulto do Manual do Projeto Desenvolvendo a Qualidade em Parcerias (DQP). Nesta escala constam três categorias referentes ao estilo de interação adulto-criança: Sensibilidade, Estimulação e Autonomia. Os resultados demonstraram que a disposição das mesas na sala de aula influenciam a interação do professor com os alunos. Em todas as disposições a categoria Sensibilidade obteve os valores mais elevados, seguindo-se a categoria Estimulação e, com valores mais baixos, a categoria Autonomia. No que respeita às disposições, foi a disposição em “U” que registou os valores mais elevados em todas as categorias e os mais baixos encontraram-se na disposição em “Filas e Colunas”. Em termos de interações, mais uma vez, foi na disposição em “U” onde estas ocorreram em maior número, seguindo-se a disposição “ Filas e Colunas”.This report, carried out within the framework of the Supervised Pedagogical Practice course of the Master's Degree in Pre-primary and Primary Education, aims to understand the influence of the arrangement of the tables in the classroom in the teacher-student interaction. Thus, a case study was carried out, having as participants four students of a class of the 2nd year of the 1st Cycle of Basic Education. The main technique of data collection was direct non-participant observation. Regarding the data, a content analysis was performed, based on the Adult Engagement Scale of the Developing Quality in Partnerships (DQP) Project Manual. In this scale there are three categories referring to the style of adult-child interaction: Sensitivity, Stimulation and Autonomy. The results showed that the arrangement of the tables in the classroom influenced the interaction of the teacher with the students. In all provisions, the Sensitivity category obtained the highest values, followed by the Stimulation category and, with lower values, the Autonomy category. Concerning the provisions, it was the "U" layout that registered the highest values in all categories and the lowest ones were found in the layout in "Rows and Columns". In terms of interactions, once again, it was in the "U" layout where these occurred in greater numbers, following the "Rows and Columns" layout

Efficient isolation of high quality RNA from tropical palms for RNA-seq analysis

Currently, RNA-seq as a high throughput technology is being widely applied to various species to elucidate the complexity of the transcriptome and to discover large number of novel genes. However, the technology has had poor success in elucidating the transcriptome of tropical palms, as it is difficult to isolate high quality RNA from tropical palm tissues due to their high polysaccharide and polyphenol content. Here, we developed an RNA-isolation protocol for tropical palms, the MRIP method (Methods for RNA Isolation from Palms). The integrity of the RNA molecules extracted using this protocol was determined to be of high quality by means of gel electrophoresis and Agilent 2100 Bioanalyzer microfluidic electrophoresis chip examination with a RIN (RNA Integrity Number) value of more than 9, indicating that the mRNAs were of good integrity. Subsequently the isolated RNA was used for transcription analysis without further purification. With Illumina sequencing, we obtained 54.9 million short reads and then conducted de novo assembly to gain 57,304 unigenes with an average length of 752 base pairs. Moreover, the RNA isolated with this protocol was also successfully used for real-time RT-PCR. These results suggested that the RNA isolated was suitable for Illumina RNA sequencing and quantitative real-time RT-PCR. Furthermore, this method was also successful in isolating total RNA from the leaves of various Palmaceae species