Involvement of Pinus taeda MYB1 and MYB8 in phenylpropanoid metabolism and secondary cell wall biogenesis: a comparative in planta analysis

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Abstract

The involvement of two R2R3- MYB genes from Pinus taeda L., PtMYB1 and PtMYB8, in phenylpropanoid metabolism and secondary cell wall biogenesis was investigated in planta. These pine MYBs were constitutively overexpressed (OE) in Picea glauca (Moench) Voss, used as a heterologous conifer expression system. Morphological, histological, chemical (lignin and soluble phenols), and transcriptional analyses, i.e. microarray and reverse transcription quantitative PCR (RT-qPCR) were used for extensive phenotyping of MYB-overexpressing spruce plantlets. Upon germination of somatic embryos, root growth was reduced in both transgenics. Enhanced lignin deposition was also a common feature but ectopic secondary cell wall deposition was more strongly associated with PtMYB8-OE. Microarray and RT-qPCR data showed that overexpression of each MYB led to an overlapping up-regulation of many genes encoding phenylpropanoid enzymes involved in lignin monomer synthesis, while misregulation of several cell wall-related genes and other MYB transcription factors was specifically associated with PtMYB8-OE. Together, the results suggest that MYB1 and MYB8 may be part of a conserved transcriptional network involved in secondary cell wall deposition in conifers.

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Most cited references 65

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Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

Yoav Benjamini, Yosef Hochberg (1995)

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NAC transcription factors, NST1 and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis.

Nobutaka Mitsuda, Akira Iwase, Hiroyuki Yamamoto … (2007)

Wood is formed by the successive addition of secondary xylem, which consists of cells with a conspicuously thickened secondary wall composed mainly of lignin and cellulose. Several genes involved in lignin and cellulose biosynthesis have been characterized, but the factors that regulate the formation of secondary walls in woody tissues remain to be identified. In this study, we show that plant-specific transcription factors, designated NAC SECONDARY WALL THICKENING PROMOTING FACTOR1 (NST1) and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis thaliana. In nst1-1 nst3-1 double knockout plants, the secondary wall thickenings in interfascicular fibers and secondary xylem, except for vascular vessels, were completely suppressed without affecting formation of cells destined to be woody tissues. Conversely, as shown previously for NST1, overexpression of NST3 induced ectopic secondary wall thickenings in various aboveground tissues. Furthermore, the expression of chimeric repressors derived from NST1 and NST3 suppressed secondary wall thickenings in the presumptive interfascicular fibers. Because putative orthologs of NST1 and NST3 are present in the genome of poplar, our results suggest that they are also key regulators of the formation of secondary walls in woody plants and could be used as a tool for the genetic engineering of wood and its derivatives.

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Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation.

Alan Christensen, Robert A Sharrock, Peter H. Quail (1992)

Two genomic clones (lambda Ubi-1 and lambda Ubi-2) encoding the highly conserved 76 amino acid protein ubiquitin have been isolated from maize. Sequence analysis shows that both genes contain seven contiguous direct repeats of the protein coding region in a polyprotein conformation. The deduced amino acid sequence of all 14 repeats is identical and is the same as for other plant ubiquitins. The use of transcript-specific oligonucleotide probes shows that Ubi-1 and Ubi-2 are expressed constitutively at 25 degrees C but are inducible to higher levels at elevated temperatures in maize seedlings. Both genes contain an intron in the 5' untranslated region which is inefficiently processed following a brief, severe heat shock. The transcription start site of Ubi-1 has been determined and a transcriptional fusion of 0.9 kb of the 5' flanking region and the entire 5' untranslated sequence of Ubi-1 with the coding sequence of the gene encoding the reporter molecule chloramphenicol acetyl transferase (CAT) has been constructed (pUBI-CAT). CAT assays of extracts of protoplasts electroporated with this construct show that the ubiquitin gene fragment confers a high level of CAT expression in maize and other monocot protoplasts but not in protoplasts of the dicot tobacco. Expression from the Ubi-1 promoter of pUBI-CAT yields more than a 10-fold higher level of CAT activity in maize protoplasts than expression from the widely used cauliflower mosaic virus 35S promoter of a 35S-CAT construct. Conversely, in tobacco protoplasts CAT activity from transcription of pUBI-CAT is less than one tenth of the level from p35S-CAT.

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Author and article information

Journal

Journal ID (nlm-ta): J Exp Bot

Journal ID (hwp): jexbot

Journal ID (publisher-id): exbotj

Title: Journal of Experimental Botany

Publisher: Oxford University Press

ISSN (Print): 0022-0957

ISSN (Electronic): 1460-2431

Publication date (Print): October 2008

Publication date (Electronic): 19 September 2008

Publication date PMC-release: 19 September 2008

Volume: 59

Issue: 14

Pages: 3925-3939

Affiliations

[1 ]Centre d’Étude de la Forêt, Université Laval, Québec (QC), Canada G1V 0A6

[2 ]UMR UPS, CNRS 5546, Pôle de Biotechnologies Végétales, 24 chemin de Borde Rouge, BP42617, Auzeville Tolosane, 31326 Castanet Tolosan, France

[3 ]Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, Québec (QC), Canada G1V 4C7

[4 ]Canada Research Chair in Wood and Fibre Quality, Department of Wood Science, University of British Columbia, 4030-2424 Main Mall, Vancouver (BC), Canada V6T 1Z4

[5 ]DPSP, Ministère des Ressources naturelles et de la Faune, 2700, rue Einstein, Québec (QC), Canada G1P 3W8

[6 ]Department of Biological Sciences, CW405 Biological Sciences Building, University of Alberta, Edmonton (AB), Canada T6G 2E9

Author notes

[* ]To whom correspondence should be addressed. E-mail: bomalc@ 123456rsvs.ulaval.ca

Article

DOI: 10.1093/jxb/ern234

PMC ID: 2576632

PubMed ID: 18805909

SO-VID: 0774f299-517d-4165-b440-a9a997674408

License:

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

History

Date received : 2 June 2008

Date revision received : 15 August 2008

Date accepted : 19 August 2008

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Involvement of Pinus taeda MYB1 and MYB8 in phenylpropanoid metabolism and secondary cell wall biogenesis: a comparative in planta analysis

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Plant MYBs

Most cited references 65

Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing

NAC transcription factors, NST1 and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis.

Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation.

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