scholarly journals The in vivo impact of MsLAC1, a Miscanthus laccase isoform, on lignification and lignin composition contrasts with its in vitro substrate preference

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Feng He ◽  
Katja Machemer-Noonan ◽  
Philippe Golfier ◽  
Faride Unda ◽  
Johanna Dechert ◽  
...  
Keyword(s):  
Cell Wall ◽  
Transcription Factors ◽  
Lignin Content ◽  
Lignin Biosynthesis ◽  
Lignin Composition ◽  
Xylem Fibers ◽  
Breeding Target

Abstract Background Understanding lignin biosynthesis and composition is of central importance for sustainable bioenergy and biomaterials production. Species of the genus Miscanthus have emerged as promising bioenergy crop due to their rapid growth and modest nutrient requirements. However, lignin polymerization in Miscanthus is poorly understood. It was previously shown that plant laccases are phenol oxidases that have multiple functions in plant, one of which is the polymerization of monolignols. Herein, we link a newly discovered Miscanthus laccase, MsLAC1, to cell wall lignification. Characterization of recombinant MsLAC1 and Arabidopsis transgenic plants expressing MsLAC1 were carried out to understand the function of MsLAC1 both in vitro and in vivo. Results Using a comprehensive suite of molecular, biochemical and histochemical analyses, we show that MsLAC1 localizes to cell walls and identify Miscanthus transcription factors capable of regulating MsLAC1 expression. In addition, MsLAC1 complements the Arabidopsis lac4–2 lac17 mutant and recombinant MsLAC1 is able to oxidize monolignol in vitro. Transgenic Arabidopsis plants over-expressing MsLAC1 show higher G-lignin content, although recombinant MsLAC1 seemed to prefer sinapyl alcohol as substrate. Conclusions In summary, our results suggest that MsLAC1 is regulated by secondary cell wall MYB transcription factors and is involved in lignification of xylem fibers. This report identifies MsLAC1 as a promising breeding target in Miscanthus for biofuel and biomaterial applications.

scholarly journals Molecular Characterization of HOXA2 and HOXA3 Binding Properties

2021 ◽  
Vol 9 (4) ◽  
pp. 55
Author(s):  
Joshua Mallen ◽  
Manisha Kalsan ◽  
Peyman Zarrineh ◽  
Laure Bridoux ◽  
Shandar Ahmad ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Molecular Characterization ◽  
Sequence Motif ◽  
Body Parts ◽  
Binding Affinities ◽  
Binding Properties ◽  
Functional Consequences

The highly conserved HOX homeodomain (HD) transcription factors (TFs) establish the identity of different body parts along the antero–posterior axis of bilaterian animals. Segment diversification and the morphogenesis of different structures is achieved by generating precise patterns of HOX expression along the antero–posterior axis and by the ability of different HOX TFs to instruct unique and specific transcriptional programs. However, HOX binding properties in vitro, characterised by the recognition of similar AT-rich binding sequences, do not account for the ability of different HOX to instruct segment-specific transcriptional programs. To address this problem, we previously compared HOXA2 and HOXA3 binding in vivo. Here, we explore if sequence motif enrichments observed in vivo are explained by binding affinities in vitro. Unexpectedly, we found that the highest enriched motif in HOXA2 peaks was not recognised by HOXA2 in vitro, highlighting the importance of investigating HOX binding in its physiological context. We also report the ability of HOXA2 and HOXA3 to heterodimerise, which may have functional consequences for the HOX patterning function in vivo.

Prediction of forage digestibility from some laboratory procedures. 2. Comparison of in vivo digestibility at two institutes.

1971 ◽  
Vol 19 (2) ◽  
pp. 106-113
Author(s):  
B. Deinum
Keyword(s):  
Cell Wall ◽  
Organic Matter ◽  
Sulfuric Acid ◽  
Lignin Content ◽  
Forage Digestibility ◽  
Significant Difference ◽  
Laboratory Procedures ◽  
Sulfuric Acid Lignin

For part 1 see Abst. 1564, Vol. 40. 2. The relation between digestibilities of organic matter in vivo and in vitro did not differ significantly between the institutes at Hoorn and Wageningen. No difference was found in digestibility of the cell wall constituents in relation to the lignin content, nor was there a significant difference in the relation between the percentage of digestible cell contents and the percentage of cell contents in forage from well managed pastures. Twelve forages of highly digestible perennial ryegrass deviated from these farm forages at Wageningen, showing smaller excretion of bacterial and endogenous residue. Comparison of the 2 lignin procedures showed that 72% sulfuric acid lignin gave consistent residual standard deviations of digestibility of cell wall constituents; permanganate lignin gave smaller errors in forages at Wageningen but greater errors in forages at Hoorn. (Abstract retrieved from CAB Abstracts by CABI’s permission)

Characterization of Cell Wall Degrading Enzymes of Rhizoctonia solani AG-3 from Tobacco Target Spot

2014 ◽  
Vol 1010-1012 ◽  
pp. 1161-1164
Author(s):  
Yan Qin Zhao ◽  
Yuan Hua Wu ◽  
Xiu Xiang Zhao ◽  
Meng Nan An ◽  
Jian Guang Chen ◽  
...  
Keyword(s):  
Cell Wall ◽  
Rhizoctonia Solani ◽  
Cell Wall Degrading Enzymes ◽  
Degrading Enzymes ◽  
Yield And Quality ◽  
Target Spot ◽  
Causal Pathogen

Rhizoctonia solaniKühn is a causal pathogen responsible for many types of plant disease worldwide and a major soilborne fungal pathogen that severely impairs yield and quality of tobacco worldwide. Activities, pathogenicity of the cell wall-degrading enzymes produced by theRhizoctoniasolanifrom tobacco target spot disease both in liquid medium and in tobacco tissue were studied. The result showed thatR.solanifrom tobacco can produce pectinase and cellulase both in vitro and vivo, and the activity of PG and PMG was the highest in vitro. The activity of Cx and β-glucosidase was the highest in vivo, and enzyme production ability of strong pathogenicity strains is stronger than the weak pathogenicity strains in vitro.

scholarly journals LACCASE14 is required for the deposition of guaiacyl lignin and affects cell wall digestibility in poplar

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Shifei Qin ◽  
Chunfen Fan ◽  
Xiaohong Li ◽  
Yi Li ◽  
Jian Hu ◽  
...  
Keyword(s):  
Cell Wall ◽  
Genetic Manipulation ◽  
Lignin Content ◽  
Biomass Conversion ◽  
Enzymatic Saccharification ◽  
Lignin Biosynthesis ◽  
Populus Tomentosa ◽  
Wall Structure ◽  
Biomass Saccharification

Abstract Background The recalcitrance of lignocellulosic biomass provided technical and economic challenges in the current biomass conversion processes. Lignin is considered as a crucial recalcitrance component in biomass utilization. An in-depth understanding of lignin biosynthesis can provide clues to overcoming the recalcitrance. Laccases are believed to play a role in the oxidation of lignin monomers, leading to the formation of higher-order lignin. In plants, functions of only a few laccases have been evaluated, so little is known about the effect of laccases on cell wall structure and biomass saccharification. Results In this study, we screened a gain-of-function mutant with a significant increase in lignin content from Arabidopsis mutant lines overexpressing a full-length poplar cDNA library. Further analysis confirmed that a Chinese white poplar (Populus tomentosa) laccase gene PtoLAC14 was inserted into the mutant, and PtoLAC14 could functionally complement the Arabidopsis lac4 mutant. Overexpression of PtoLAC14 promoted the lignification of poplar and reduced the proportion of syringyl/guaiacyl. In contrast, the CRISPR/Cas9-generated mutation of PtLAC14 results in increased the syringyl/guaiacyl ratios, which led to integrated enhancement on biomass enzymatic saccharification. Notably, the recombinant PtoLAC14 protein showed higher oxidized efficiency to coniferyl alcohol (precursor of guaiacyl unit) in vitro. Conclusions This study shows that PtoLAC14 plays an important role in the oxidation of guaiacyl deposition on cell wall. The reduced recalcitrance of the PtoLAC14-KO lines suggests that PtoLAC14 is an elite target for cell wall engineering, and genetic manipulation of this gene will facilitate the utilization of lignocellulose.

scholarly journals Distinct and Overlapping Functions of Miscanthus sinensis MYB Transcription Factors SCM1 and MYB103 in Lignin Biosynthesis

2021 ◽  
Vol 22 (22) ◽  
pp. 12395
Author(s):  
Philippe Golfier ◽  
Olga Ermakova ◽  
Faride Unda ◽  
Emily K. Murphy ◽  
Jianbo Xie ◽  
...  
Keyword(s):  
Cell Wall ◽  
Transcription Factors ◽  
Secondary Cell Wall ◽  
Target Genes ◽  
Lignin Content ◽  
Ectopic Expression ◽  
Lignin Biosynthesis ◽  
Miscanthus Sinensis ◽  
Growth Stages ◽  
Transcriptional Responses

Cell wall recalcitrance is a major constraint for the exploitation of lignocellulosic biomass as a renewable resource for energy and bio-based products. Transcriptional regulators of the lignin biosynthetic pathway represent promising targets for tailoring lignin content and composition in plant secondary cell walls. However, knowledge about the transcriptional regulation of lignin biosynthesis in lignocellulosic feedstocks, such as Miscanthus, is limited. In Miscanthus leaves, MsSCM1 and MsMYB103 are expressed at growth stages associated with lignification. The ectopic expression of MsSCM1 and MsMYB103 in N. benthamiana leaves was sufficient to trigger secondary cell wall deposition with distinct sugar and lignin compositions. Moreover, RNA-seq analysis revealed that the transcriptional responses to MsSCM1 and MsMYB103 overexpression showed an extensive overlap with the response to the NAC master transcription factor MsSND1, but were distinct from each other, underscoring the inherent complexity of secondary cell wall formation. Furthermore, conserved and previously described promoter elements as well as novel and specific motifs could be identified from the target genes of the three transcription factors. Together, MsSCM1 and MsMYB103 represent interesting targets for manipulations of lignin content and composition in Miscanthus towards a tailored biomass.

scholarly journals Characterization of a sweet basil acyltransferase involved in eugenol biosynthesis

2020 ◽  
Vol 71 (12) ◽  
pp. 3638-3652 ◽  
Author(s):  
Niha Dhar ◽  
Sreelatha Sarangapani ◽  
Vaishnavi Amarr Reddy ◽  
Nadimuthu Kumar ◽  
Deepa Panicker ◽  
...  
Keyword(s):  
Lignin Content ◽  
Glandular Trichomes ◽  
Lignin Biosynthesis ◽  
Coniferyl Alcohol ◽  
Sweet Basil ◽  
Transgenic Lines ◽  
Common Substrate ◽  
Total Lignin

Abstract Sweet basil (Ocimum basilicum) plants produce its characteristic phenylpropene-rich essential oil in specialized structures known as peltate glandular trichomes (PGTs). Eugenol and chavicol are the major phenylpropenes produced by sweet basil varieties whose synthetic pathways are not fully elucidated. Eugenol is derived from coniferyl acetate by a reaction catalysed by eugenol synthase. An acyltransferase is proposed to convert coniferyl alcohol to coniferyl acetate which is the first committed step towards eugenol synthesis. Here, we perform a comparative next-generation transcriptome sequencing of different tissues of sweet basil, namely PGT, leaf, leaf stripped of PGTs (leaf–PGT), and roots, to identify differentially expressed transcripts specific to PGT. From these data, we identified a PGT-enriched BAHD acyltransferase gene ObCAAT1 and functionally characterized it. In vitro coupled reaction of ObCAAT1 with eugenol synthase in the presence of coniferyl alcohol resulted in eugenol production. Analysis of ObCAAT1-RNAi transgenic lines showed decreased levels of eugenol and accumulation of coniferyl alcohol and its derivatives. Coniferyl alcohol acts as a common substrate for phenylpropene and lignin biosynthesis. No differences were found in total lignin content of PGTs and leaves of transgenic lines, indicating that phenylpropene biosynthesis is not coupled to lignification in sweet basil.

scholarly journals The elaborate route for UDP-arabinose delivery into the Golgi of plants

2017 ◽  
Vol 114 (16) ◽  
pp. 4261-4266 ◽  
Author(s):  
Carsten Rautengarten ◽  
Devon Birdseye ◽  
Sivakumar Pattathil ◽  
Heather E. McFarlane ◽  
Susana Saez-Aguayo ◽  
...  
Keyword(s):  
Cell Wall ◽  
Ring Configuration ◽  
Signaling Peptides ◽  
Transport Assay ◽  
Mutant Lines ◽  
Identification And Characterization ◽  
Nucleotide Sugar Transporter

In plants, L-arabinose (Ara) is a key component of cell wall polymers, glycoproteins, as well as flavonoids, and signaling peptides. Whereas the majority of Ara found in plant glycans occurs as a furanose ring (Araf), the activated precursor has a pyranose ring configuration (UDP-Arap). The biosynthesis of UDP-Arap mainly occurs via the epimerization of UDP-xylose (UDP-Xyl) in the Golgi lumen. Given that the predominant Ara form found in plants is Araf, UDP-Arap must exit the Golgi to be interconverted into UDP-Araf by UDP-Ara mutases that are located outside on the cytosolic surface of the Golgi. Subsequently, UDP-Araf must be transported back into the lumen. This step is vital because glycosyltransferases, the enzymes mediating the glycosylation reactions, are located within the Golgi lumen, and UDP-Arap, synthesized within the Golgi, is not their preferred substrate. Thus, the transport of UDP-Araf into the Golgi is a prerequisite. Although this step is critical for cell wall biosynthesis and the glycosylation of proteins and signaling peptides, the identification of these transporters has remained elusive. In this study, we present data demonstrating the identification and characterization of a family of Golgi-localized UDP-Araf transporters in Arabidopsis. The application of a proteoliposome-based transport assay revealed that four members of the nucleotide sugar transporter (NST) family can efficiently transport UDP-Araf in vitro. Subsequent analysis of mutant lines affected in the function of these NSTs confirmed their role as UDP-Araf transporters in vivo.

scholarly journals Prediction of forage digestibility from some laboratory procedures.

1969 ◽  
Vol 17 (2) ◽  
pp. 119-127
Author(s):  
B. Duinum ◽  
P.J. Van Soest
Keyword(s):  
Cell Wall ◽  
Chemical Analysis ◽  
Lignin Content ◽  
Rumen Fluid ◽  
Grass Silage ◽  
Forage Digestibility ◽  
The Poor ◽  
Laboratory Procedures

Digestibility of 106 samples of hay, grass, grass silage and legume forage was estimated by regression from cell contents and cell wall constituents estimated chemically. Cell wall constituents were cellulose, hemicelluloses, lignin, cutin, plant silica and soil contamination. Values were not as closely related to digestibility in vivo by sheep as were values obtained by digestion with rumen fluid in vitro. Correlation between digestibility of cell wall constituents and lignin content was poor, and this and other reasons for the poor results from chemical analysis are discussed.-T. D. B. (Abstract retrieved from CAB Abstracts by CABI’s permission)

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

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