Secondary Cell Wall Lignification Research Articles

QTL analysis of the developmental changes in cell wall components and forage digestibility in maize (Zea mays L.)

Cell wall architecture plays a key role in stalk strength and forage digestibility. Lignin, cellulose, and hemicellulose are the three main components of plant cell walls, and they can impact stalk quality by affecting the structure and strength of the cell wall. To explore cell wall development during secondary cell wall lignification in maize stalks, conventional and conditional genetic mapping were used to identify the dynamic quantitative trait loci (QTLs) of the cell wall components and digestibility traits during five growth stages after silking. Acid detergent lignin (ADL), cellulose (CEL), acid detergent fiber (ADF), neutral detergent fiber (NDF), and in vitro dry matter digestibility (IVDMD) were evaluated in a maize recombinant inbred line (RIL) population. ADL, CEL, ADF, and NDF gradually increased from 10 to 40 days after silking (DAS), and then they decreased. IVDMD initially decreased until 40 DAS, and then it increased slightly. Seventy-two QTLs were identified for the five traits, and each accounted for 3.48–24.04% of the phenotypic variation. Six QTL hotspots were found, and they were localized in the 1.08, 2.04, 2.07, 7.03, 8.05, and 9.03 bins of the maize genome. Within the interval of the pleiotropic QTL identified in bin 1.08 of the maize genome, six genes associated with cell wall component biosynthesis were identified as potential candidate genes for stalk strength as well as cell wall-related traits. In addition, 26 conditional QTLs were detected in the five stages for all of the investigated traits. Twenty-two of the 26 conditional QTLs were found at 30 DAS conditioned using the values of 20 DAS, and at 50 DAS conditioned using the values of 40 DAS. These results indicated that cell wall-related traits are regulated by many genes, which are specifically expressed at different stages after silking. Simultaneous improvements in both forage digestibility and lodging resistance could be achieved by pyramiding multiple beneficial QTL alleles identified in this study.

UDP-GLYCOSYLTRANSFERASE 72E3 Plays a Role in Lignification of Secondary Cell Walls in Arabidopsis.

Lignin is present in plant secondary cell walls and is among the most abundant biological polymers on Earth. In this work we investigated the potential role of the UGT72E gene family in regulating lignification in Arabidopsis. Chemical determination of floral stem lignin contents in ugt72e1, ugt72e2, and ugt72e3 mutants revealed no significant differences compared to WT plants. In contrast, the use of a novel safranin O ratiometric imaging technique indicated a significant increase in the cell wall lignin content of both interfascicular fibers and xylem from young regions of ugt72e3 mutant floral stems. These results were globally confirmed in interfascicular fibers by Raman microspectroscopy. Subsequent investigation using a bioorthogonal triple labelling strategy suggested that the augmentation in lignification was associated with an increased capacity of mutant cell walls to incorporate H-, G-, and S-monolignol reporters. Expression analysis showed that this increase was associated with an up-regulation of LAC17 and PRX71, which play a key role in lignin polymerization. Altogether, these results suggest that UGT72E3 can influence the kinetics of lignin deposition by regulating monolignol flow to the cell wall as well as the potential of this compartment to incorporate monomers into the growing lignin polymer.

Role of laccase gene in wheat NILs differing at QTL-Fhb1 for resistance against Fusarium head blight

Fusarium head blight (FHB), caused mainly by Fusarium graminearum (Fg), is one of the most severe diseases of wheat. It affects grain yield and quality due to mycotoxin contamination, which is harmful for both human and livestock consumption. Cell wall lignification, following pathogen invasion, is one of the innate defense responses. Plant laccases are known to lignify the secondary cell walls. A metabolo-genomics study identified laccase as one of the candidate genes in QTL-Fhb1 of wheat NILs derived from Sumai 3*5/Thatcher cross. Based on phylogenetics, it was named as TaLAC4. Real-time qPCR revealed a strongly induced expression of TaLAC4 in NIL-R. The VIGS based transient silencing of TaLAC4 in NIL-R resulted in an increased susceptibility leading to Fg spread within the entire spike in 15dpi, contrasting to non-silenced where the infection was limited to inoculated spikelets. Histopathology revealed thickened cell walls, mainly due to G-lignin, in non-silenced NIL-R, relative to silenced, in conjunction with higher total lignin content. Metabolic profiling of TaLAC4 silenced NILs identified the accumulation of several precursor metabolites higher in abundances upstream TaLAC4. These results confirm that the resistance function of TaLAC4 in NIL-R is due to pathogen-induced lignification of secondary cell walls in the rachis.

Transgenic Potato Plants Overexpressing SOD and APX Exhibit Enhanced Lignification and Starch Biosynthesis with Improved Salt Stress Tolerance

Potato (Solanum tuberosum L.) is a major crop worldwide and its productivity is severely reduced by high soil salinity. The aim of the current study was to gain insights into the response and regulation of the antioxidant system in the transgenic potato, overexpressing two genes viz. thermostable CuZn-superoxide dismutase (PaSOD) gene from polyextremophile high-altitude plant Potentilla atrosanguinea and ascorbate peroxide (RaAPX) gene from Rheum australe under salinity stress. Transgenic lines and wild type (WT) were assessed under 50, 100, and 150 mM concentrations of NaCl treatment at the anatomical, transcriptional and metabolic levels. Although superoxide dismutase (SOD) and ascorbate peroxide (APX) activities in transgenic lines were significantly higher than WT under salt stress, the secondary cell wall lignification was profoundly affected by the modulated antioxidant enzymes activities and the corresponding H2O2 levels in transgenic. Expression profiles showed that the several genes and transcription factors directly involved in lignin biosynthesis were profoundly induced in transgenic lines as compared to WT. In addition, transgenic plants harbored increased starch accumulation, improved growth attributes, and reduced accumulation of reactive oxygen species than WT plants. Together, these results indicated that the expression of SOD and APX genes in transgenic potato may function as a positive set of physiological, anatomical, and molecular adjustments in the H2O2 regulated lignin biosynthesis signaling pathway, enabling the plants to withstand severe saline conditions.

The Eucalyptus linker histone variant EgH1.3 cooperates with the transcription factor EgMYB1 to control lignin biosynthesis during wood formation.

Wood, also called secondary xylem, is a specialized vascular tissue constituted by different cell types that undergo a differentiation process involving deposition of thick, lignified secondary cell walls. The mechanisms needed to control the extent of lignin deposition depending on the cell type and the differentiation stage are far from being fully understood. We found that the Eucalyptus transcription factor EgMYB1, which is known to repress lignin biosynthesis, interacts specifically with a linker histone variant, EgH1.3. This interaction enhances the repression of EgMYB1's target genes, strongly limiting the amount of lignin deposited in xylem cell walls. The expression profiles of EgMYB1 and EgH1.3 overlap in xylem cells at early stages of their differentiation as well as in mature parenchymatous xylem cells, which have no or only thin lignified secondary cell walls. This suggests that a complex between EgMYB1 and EgH1.3 integrates developmental signals to prevent premature or inappropriate lignification of secondary cell walls, providing a mechanism to fine-tune the differentiation of xylem cells in time and space. We also demonstrate a role for a linker histone variant in the regulation of a specific developmental process through interaction with a transcription factor, illustrating that plant linker histones have other functions beyond chromatin organization.

Contrasting nitrogen fertilization treatments impact xylem gene expression and secondary cell wall lignification in Eucalyptus.

BackgroundNitrogen (N) is a main nutrient required for tree growth and biomass accumulation. In this study, we analyzed the effects of contrasting nitrogen fertilization treatments on the phenotypes of fast growing Eucalyptus hybrids (E. urophylla x E. grandis) with a special focus on xylem secondary cell walls and global gene expression patterns.ResultsHistological observations of the xylem secondary cell walls further confirmed by chemical analyses showed that lignin was reduced by luxuriant fertilization, whereas a consistent lignin deposition was observed in trees grown in N-limiting conditions. Also, the syringyl/guaiacyl (S/G) ratio was significantly lower in luxuriant nitrogen samples. Deep sequencing RNAseq analyses allowed us to identify a high number of differentially expressed genes (1,469) between contrasting N treatments. This number is dramatically higher than those obtained in similar studies performed in poplar but using microarrays. Remarkably, all the genes involved the general phenylpropanoid metabolism and lignin pathway were found to be down-regulated in response to high N availability. These findings further confirmed by RT-qPCR are in agreement with the reduced amount of lignin in xylem secondary cell walls of these plants.ConclusionsThis work enabled us to identify, at the whole genome level, xylem genes differentially regulated by N availability, some of which are involved in the environmental control of xylogenesis. It further illustrates that N fertilization can be used to alter the quantity and quality of lignocellulosic biomass in Eucalyptus, offering exciting prospects for the pulp and paper industry and for the use of short coppices plantations to produce second generation biofuels.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-014-0256-9) contains supplementary material, which is available to authorized users.

Contrasting nitrogen fertilization treatments impact xylem gene expression and secondary cell wall lignification in Eucalyptus

Contrasting nitrogen fertilization treatments impact xylem gene expression and secondary cell wall lignification in Eucalyptus

Phenols and lignin are involved in the defence response of banana (Musa) plants to Radopholus similis infection

The role of lignin and phenols in plant defence ranges from preformed characteristic to inducible physical and chemical response against nematode infection. Our study shows the involvement of lignin and phenols in the defence of two newly identified resistant banana (Musa) genotypes to burrowing nematode Radopholus similis infection. Results were compared with reference resistant and susceptible banana cultivars. Histochemical analysis of root cross sections showed a more extensive secondary cell wall lignification of vascular bundles in R. similis-infected plants than in the nematode non-infected plants. Increased extensive lignification was not associated with the cortex cells that are directly attacked by the nematode. This showed that the increased lignification is a general defence response to protect the vascular bundle from damage rather than resisting the nematode development and reproduction at the root cortex. Histochemical localisation showed no preformed phenolic cells in the cortex of the non-infected, R. similis-resistant and -susceptible Musa genotypes. By contrast, phenolic substances were the major constituents of the nematode-infected necrotic cells. Phenols and lignin contents were also quantitatively assayed. The Folin-Ciocalteu assay confirmed the increase in phenol content of nematode-infected root cells. Phenol content in nematode-infected plants was twice the amount of phenol content in nematode non-infected plants at 3 weeks after infection. This is possibly due to the biosynthesis or accumulation of secondary metabolites such as phenolic phytoalexins in the nematode infection sites of all the banana genotypes. This study clearly demonstrates that phenols and lignin play an important role in the defence mechanisms of Musa to R. similis infection.

Bringing trees into the fuel line

Bringing trees into the fuel line

Isolation and Characterization of a Novel Peroxidase Gene ZPO-C Whose Expression and Function are Closely Associated with Lignification during Tracheary Element Differentiation

In an attempt to elucidate the regulatory mechanism of vessel lignification, we isolated ZPO-C, a novel peroxidase gene of Zinnia elegans that is expressed specifically in differentiating tracheary elements (TEs). The ZPO-C transcript was shown to accumulate transiently at the time of secondary wall thickening of TEs in xylogenic culture of Zinnia cells. In situ hybridization indicated specific accumulation of the ZPO-C transcript in immature vessels in Zinnia seedlings. Immunohistochemical analysis using anti-ZPO-C antibody showed that the ZPO-C protein is abundant in TEs, especially at their secondary walls. For enzymatic characterization of ZPO-C, 6 x His-tagged ZPO-C was produced in tobacco cultured cells and purified. The ZPO-C:6 x His protein had a peroxidase activity preferring sinapyl alcohol as well as coniferyl alcohol as a substrate, with a narrow pH optimum around 5.25. The peroxidase activity required calcium ion and was elevated by increasing Ca2+ concentration in the range of 0-10 mM. An Arabidopsis homolog of ZPO-C, At5g51890, was examined for expression patterns with transgenic plants carrying a yellow fluorescent protein (YFP) gene under the control of the At5g51890 promoter. The YFP fluorescence localization demonstrated vessel-specific expression of At5g51890 in the Arabidopsis roots. Taken collectively, our results strongly suggest that ZPO-C and its homologs play an important role in lignification of secondary cell walls in differentiating TEs.

Conifers in cold environments synchronize maximum growth rate of tree‐ring formation with day length

Intra-annual radial growth rates and durations in trees are reported to differ greatly in relation to species, site and environmental conditions. However, very similar dynamics of cambial activity and wood formation are observed in temperate and boreal zones. Here, we compared weekly xylem cell production and variation in stem circumference in the main northern hemisphere conifer species (genera Picea, Pinus, Abies and Larix) from 1996 to 2003. Dynamics of radial growth were modeled with a Gompertz function, defining the upper asymptote (A), x-axis placement (beta) and rate of change (kappa). A strong linear relationship was found between the constants beta and kappa for both types of analysis. The slope of the linear regression, which corresponds to the time at which maximum growth rate occurred, appeared to converge towards the summer solstice. The maximum growth rate occurred around the time of maximum day length, and not during the warmest period of the year as previously suggested. The achievements of photoperiod could act as a growth constraint or a limit after which the rate of tree-ring formation tends to decrease, thus allowing plants to safely complete secondary cell wall lignification before winter.

A laccase‐like phenoloxidase is correlated with lignin biosynthesis in Zinnia elegans stem tissues

SummaryStem tissues from different internodes of 4–6 week‐old Zinnia elegans cv. Envy plants were sectioned and stained with chromogenic substrates previously used in studies of laccases (p‐diphenol:O2 oxidoreductases) isolated from tree tissues. The pattern of color development found when stem sections were stained in the presence and absence of H2O2 suggested that p‐diphenol:O2 oxidoreductase activity was tightly correlated spatially and temporally with the lignification of secondary cell walls in developing primary xylem. The correlation between this laccase‐like phenoloxidase activity and lignification appeared tighter than that between lignification and peroxidases stained using the same substrates. Zymogram analysis of the phenoloxidase activities catalyzed by enzymes that were not boiled prior to separation by SDS—PAGE suggested that a single enzyme was predominantly responsible for the laccase‐like phenoloxidase activity in Zinnia stems. Some of this enzyme was released from cell wall residue by washing with high ionic strength buffer; however, substantial amounts of the enzyme could only be recovered after treatment of the residue with cell wall‐degrading enzymes. This phenoloxidase appears to share significant characteristics with the coniferyl alcohol oxidase isolated from developing secondary xylem in pines, which suggests that such enzymes may be widespread in vascular plants.