SND2, a NAC transcription factor gene, regulates genes involved in secondary cell wall development in Arabidopsis fibres and increases fibre cell area in Eucalyptus

<p>Abstract</p> <p>Background</p> <p>NAC domain transcription factors initiate secondary cell wall biosynthesis in <it>Arabidopsis </it>fibres and vessels by activating numerous transcriptional regulators and biosynthetic genes. NAC family member <it>SND2 </it>is an indirect target of a principal regulator of fibre secondary cell wall formation, SND1. A previous study showed that overexpression of <it>SND2 </it>produced a fibre cell-specific increase in secondary cell wall thickness in <it>Arabidopsis </it>stems, and that the protein was able to transactivate the <it>cellulose synthase8 </it>(<it>CesA8</it>) promoter. However, the full repertoire of genes regulated by <it>SND2 </it>is unknown, and the effect of its overexpression on cell wall chemistry remains unexplored.</p> <p>Results</p> <p>We overexpressed <it>SND2 </it>in <it>Arabidopsis </it>and analyzed homozygous lines with regards to stem chemistry, biomass and fibre secondary cell wall thickness. A line showing upregulation of <it>CesA8 </it>was selected for transcriptome-wide gene expression profiling. We found evidence for upregulation of biosynthetic genes associated with cellulose, xylan, mannan and lignin polymerization in this line, in agreement with significant co-expression of these genes with native <it>SND2 </it>transcripts according to public microarray repositories. Only minor alterations in cell wall chemistry were detected. Transcription factor <it>MYB103</it>, in addition to <it>SND1</it>, was upregulated in <it>SND2</it>-overexpressing plants, and we detected upregulation of genes encoding components of a signal transduction machinery recently proposed to initiate secondary cell wall formation. Several homozygous T4 and hemizygous T1 transgenic lines with pronounced <it>SND2 </it>overexpression levels revealed a negative impact on fibre wall deposition, which may be indirectly attributable to excessive overexpression rather than co-suppression. Conversely, overexpression of <it>SND2 </it>in <it>Eucalyptus </it>stems led to increased fibre cross-sectional cell area.</p> <p>Conclusions</p> <p>This study supports a function for <it>SND2 </it>in the regulation of cellulose and hemicellulose biosynthetic genes in addition of those involved in lignin polymerization and signalling. SND2 seems to occupy a subordinate but central tier in the secondary cell wall transcriptional network. Our results reveal phenotypic differences in the effect of <it>SND2 </it>overexpression between woody and herbaceous stems and emphasize the importance of expression thresholds in transcription factor studies.</p

Eucalyptus grandis AUX/INDOLE‑3‑ACETIC ACID 13 (EgrIAA13) is a novel transcriptional regulator of xylogenesis

Auxin is a crucial phytohormone regulating multiple aspects of plant growth and diferentiation, including regulation of vascular cambium activity, xylogenesis and its responsiveness towards gravitropic stress. Although the regulation of these biological processes greatly depends on auxin and regulators of the auxin signalling pathway, many of their specifc functions remain unclear. Therefore, the present study aims to functionally characterise Eucalyptus grandis AUX/INDOLE3-ACETIC ACID 13 (EgrIAA13), a member of the auxin signalling pathway. In Eucalyptus and Populus, EgrIAA13 and its orthologs are preferentially expressed in the xylogenic tissues and downregulated in tension wood. Therefore, to further investigate EgrIAA13 and its function during xylogenesis, we conducted subcellular localisation and Induced Somatic Sector Analysis experiments using overexpression and RNAi knockdown constructs of EgrIAA13 to create transgenic tissue sectors on growing stems of Eucalyptus and Populus. Since Aux/IAAs interact with Auxin Responsive Factors (ARFs), in silico predictions of IAA13-ARF interactions were explored and experimentally validated via yeast-2-hybrid experiments. Our results demonstrate that EgrIAA13 localises to the nucleus and that downregulation of EgrIAA13 impedes Eucalyptus xylem fbre and vessel development. We also observed that EgrIAA13 interacts with Eucalyptus ARF2, ARF5, ARF6 and ARF19A. Based on these results, we conclude that EgrIAA13 is a regulator of Eucalyptus xylogenesis and postulate that the observed phenotypes are likely to result from alterations in the auxin-responsive transcriptome via IAA13-ARF modules such as EgrIAA13-EgrARF5. Our results provide the frst insights into the regulatory role of EgrIAA13 during xylogenesis.Open Access funding enabled and organized by CAUL and its Member Institutions. Funding for this project was secured by GB. During this work, NK was supported by a Melbourne Research Scholarship and the Albert Shimmins Fund provided by the University of Melbourne.A Melbourne Research Scholarship and the Albert Shimmins Fund provided by the University of Melbourne. Open Access funding enabled and organized by CAUL and its Member Institutions.http://link.springer.com/journal/11103dm2022BiochemistryForestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

Induced somatic sector analysis of cellulose synthase (CesA) promoter regions in woody stem tissues

The increasing focus on plantation forestry as a renewable source of cellulosic biomass has emphasized the need for tools to study the unique biology of woody genera such as Eucalyptus, Populus and Pinus. The domestication of these woody crops is hampered by long generation times, and breeders are now looking to molecular approaches such as marker-assisted breeding and genetic modification to accelerate tree improvement. Much of what is known about genes involved in the growth and development of plants has come from studies of herbaceous models such as Arabidopsis and rice. However, transferring this information to woody plants often proves difficult, especially for genes expressed in woody stems. Here we report the use of induced somatic sector analysis (ISSA) for characterization of promoter expression patterns directly in the stems of Populus and Eucalyptus trees. As a case study, we used previously characterized primary and secondary cell wall-related cellulose synthase (CesA) promoters cloned from Eucalyptus grandis. We show that ISSA can be used to elucidate the phloem and xylem expression patterns of the CesA genes in Eucalyptus and Populus stems and also show that the staining patterns differ in Eucalyptus and Populus stems. These findings show that ISSA is an efficient approach to investigate promoter function in the developmental context of woody plant tissues and raise questions about the suitability of heterologous promoters for genetic manipulation in plant species.This work was supported through funding provided by Mondi and Sappi to the Forest Molecular Genetics (FMG) Programme, the Technology and Human Resources for Industry Programme (THRIP) and the National Research Foundation (NRF) of South Africa as well as a Linkage Grant from the Australian Research Council (LP0776563) to GB, AAM and AVS.http://link.springer.com/journal/425hb201

O ventre sacia-se, os olhos não O suporte social em adolescentes que prosseguiram a gravidez e mulheres que recorreram a interrupção voluntária da gravidez na adolescência.

O presente artigo relata um estudo que versou sobre a temática da Gravidez na Adolescência, quer na escolha do Prosseguimento da Gravidez pela jovem, quer pela sua Interrupção Voluntária. Assim, consistiu em averiguar quais as características associadas às adolescentes que mantêm a gravidez e quais as das adolescentes que optam por interrompê-la voluntariamente e, principalmente, em aferir do Suporte Social e suas características em ambas as situaçõesThe present paper presents a study concerning adolescent pregnancy decision. Its purpose was to investigate the characteristics associated with adolescents who maintained their pregnancy and those who chose induced abortion, mainly regarding social support and its characteristics.info:eu-repo/semantics/publishedVersio

The use of induced somatic sectors analysis for the elucidation of gene function and developmental patterns in xylogenic tissue

© 2006 Dr. Antanas Vytas SpokeviciusThe genetic manipulation of perennial woody tree species presents a range of additional challenges compared to that of annual weedy crop species. These include long generation times and reproductive cycle, the heterogeneity of plants under investigation and, when investigating xylogenesis, a number of physical and biochemical limitations to microscopic and molecular experimentation. Efforts have been made to understand molecular aspects of xylogenesis and have involved functional gene testing using transgenic approaches. These methods involve the production of plantlets from a variety of plant tissues using in vitro full plant regeneration techniques. Although these systems are effective, the time taken from transformation event, to plant establishment and growth, then finally to secondary wood production can take up to several years and requires high labor and technical inputs. (For complete abstract open document

Microanalytical techniques for phenotyping secondary xylem

The products of secondary xylem are of significant biological and commercial importance, and as a result, the biology of secondary growth and how intrinsic and extrinsic factors influence this process have been the subject of intense investigation. Studies into secondary xylem range in scale from the cellular to the forest stand level, with phenotypic analyses often involving the assessment of traits relating to cell morphology and cell wall chemical composition. While numerous techniques are currently available for phenotypic analyses of samples containing abundant amounts of secondary tissue, only a few of them (microanalytical techniques) are suitable when working with limiting amounts of secondary tissue or where a fine-scale resolution of morphological features or cell wall chemical composition is required. While polarised light microscopy, scanning electron microscopy, field emission-scanning electron microscopy and X-ray scattering and micro-tomography techniques serve as the most frequently used microanalytical techniques in morphotyping, techniques such as scanning ultraviolet microspectrophotometry, X-ray photoelectron spectroscopy, gas chromatography, Fourier-transform infrared spectroscopy and matrix-assisted laser desorption ionisation mass spectrometry serve as the most commonly used microanalytical techniques in chemotyping. Light microscopy, fluorescence microscopy, confocal laser scanning microscopy, transmission electron microscopy and Raman spectroscopy serve as dual micro morphotyping and chemotyping techniques. In this review, we summarise and discuss these techniques in the light of their applicability as microanalytical techniques to study secondary xylem.http://booksandjournals.brillonline.com/content/journals/22941932hj2020BiochemistryForestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

The Cytoskeleton and Its Role in Determining Cellulose Microfibril Angle in Secondary Cell Walls of Woody Tree Species

<b> </b>Recent advances in our understanding of the molecular control of secondary cell wall (SCW) formation have shed light on<b> </b>molecular mechanisms that underpin domestication traits related to wood formation. One such trait is the cellulose microfibril angle (MFA), an important wood quality determinant that varies along tree developmental phases and in response to gravitational stimulus. The cytoskeleton, mainly composed of microtubules and actin filaments, collectively contribute to plant growth and development by participating in several cellular processes, including cellulose deposition. Studies in Arabidopsis have significantly aided our understanding of the roles of microtubules in xylem cell development during which correct SCW deposition and patterning are essential to provide structural support and allow for water transport. In contrast, studies relating to SCW formation in xylary elements performed in woody trees remain elusive. In combination, the data reviewed here suggest that the cytoskeleton plays important roles in determining the exact sites of cellulose deposition, overall SCW patterning and more specifically, the alignment and orientation of cellulose microfibrils. By relating the reviewed evidence to the process of wood formation, we present a model of microtubule participation in determining MFA in woody trees forming reaction wood (RW)