Expression of cell wall related genes in basal and ear internodes of silking brown-midrib-3, caffeic acid O-methyltransferase (COMT) down-regulated, and normal maize plants

<p>Abstract</p> <p>Background</p> <p>Silage maize is a major forage and energy resource for cattle feeding, and several studies have shown that lignin content and structure are the determining factors in forage maize feeding value. In maize, four natural <it>brown-midrib </it>mutants have modified lignin content, lignin structure and cell wall digestibility. The greatest lignin reduction and the highest cell wall digestibility were observed in the <it>brown-midrib-3 </it>(<it>bm3</it>) mutant, which is disrupted in the caffeic acid <it>O</it>-methyltransferase (COMT) gene.</p> <p>Results</p> <p>Expression of cell wall related genes was investigated in basal and ear internodes of normal, COMT antisens (AS225), and <it>bm3 </it>maize plants of the INRA F2 line. A cell wall macro-array was developed with 651 gene specific tags of genes specifically involved in cell wall biogenesis. When comparing basal (older lignifying) and ear (younger lignifying) internodes of the normal line, all genes known to be involved in constitutive monolignol biosynthesis had a higher expression in younger ear internodes. The expression of the COMT gene was heavily reduced, especially in the younger lignifying tissues of the ear internode. Despite the fact that AS225 transgene expression was driven only in sclerenchyma tissues, COMT expression was also heavily reduced in AS225 ear and basal internodes. COMT disruption or down-regulation led to differential expressions of a few lignin pathway genes, which were all over-expressed, except for a phenylalanine ammonia-lyase gene. More unexpectedly, several transcription factor genes, cell signaling genes, transport and detoxification genes, genes involved in cell wall carbohydrate metabolism and genes encoding cell wall proteins, were differentially expressed, and mostly over-expressed, in COMT-deficient plants.</p> <p>Conclusion</p> <p>Differential gene expressions in COMT-deficient plants highlighted a probable disturbance in cell wall assembly. In addition, the gene expressions suggested modified chronology of the different events leading to cell expansion and lignification with consequences far beyond the phenylpropanoid metabolism. The reduced availability of monolignols and S units in <it>bm3 </it>or AS225 plants led to plants also differing in cell wall carbohydrate, and probably protein, composition. Thus, the deficiency in a key-enzyme of the lignin pathway had correlative effects on the whole cell wall metabolism. Furthermore, the observed differential expression between <it>bm3 </it>and normal plants indicated the possible involvement in the maize lignin pathway of genes which up until now have not been considered to play this role.</p

Biochemical composition and nutritional value of Balanites aegyptiaca (L.) Del fruit pulps from Northern Ferlo in Senegal

Balanites aegyptiaca, a forest species of socio-economic interest for rural people, was chosen to be planted in the frame-work of the Great Green Wall project in Senegal. Although in vastly demand during food scarcity period before crops harvest, the species’ fruit is poorly valued despite its important production. In this study, the fruit pulps were harvested at maturity and the biochemical characteristics were accessed in order to better understand the fruit’s nutritional value. The moisture, sugars, proteins, amino acids, ash, fat, minerals and some vitamins were determined by standard methods of analysis. The results indicated that the fruits are a good alimentary source of sugar, minerals (mainly potassium) and ascorbic acid. The pulp proteins were qualitatively balanced, but were present only in weak quantities. Its caloric value is high due to the high concentration of sugars. Low humidity should allow a fairly good postharvest fruit conservation. The dietary intake of this fruit for local people is very valuable especially in terms of nutrition.Keywords: Balanites aegyptiaca, pulp, biochemical, nutritional valueAfrican Journal of Biotechnology, Vol. 13(2), pp. 336-342, 8 January, 201

Investigating the unusually high cell wall digestibility of the old INRA early flint F4 maize inbred line

The old INRA flint early line F4, which belongs to the northern flint group, is typified by its high cell wall digest- ibility which reaches values as high as those observed in several brown-midrib bm3 mutant lines. The F4 line thus appeared as a model that could contribute to the understanding of genetic mechanisms involved in variation of secondary wall traits. Different strategies and results were thus gathered including especially cell wall biochemical and digestibility investigations, expression approaches, QTL investigations, and colocalizations between QTLs and candidate genes. Lignin content was lower in F4 than in other lines, with a tendency to lower p-coumarate content. The Syringyl/Guaiacyl lignin unit ratio was similar in F4 as in other lines, but this ratio was nearly not reduced in F4bm3, conversely to what is observed in bm3 mutants. In comparison with the INRA F2 control line, expressions of three PAL genes including the ZmPAL, of the ZmF5H1 and the ZmCOMT genes were significantly reduced in F4 lignifying ear internodes at early silking stage. In the F7025 x F4 RIL progeny, seven QTLs were shown with favorable alleles (increasing cell wall digestibility) originating from F4. Two strong QTLs were located in bins 1.03 and 2.03 colocalizing with the ZmMYB019 and ZmSWN6 transcription factors, respectively. Orthologs of ZmMYB019 have been shown to be involved in lignin biosynthesis, and the PpMYB8 ortholog was shown to regu- late PAL gene expression in maritime pine. The ZmSWN6 NAC transcription factor is an upstream master regulator of the secondary wall biosynthetic programs. At the other QTL positions, colocalizations were also shown with other secondary wall related ZmMYB, but also with BAHD genes involved in arabinoxylan feruloylation, and with the position of the bm6 mutation. Three QTL positions were shown with favorable alleles originating from F7025, which colocalized with ZmMYB and ZmNAC transcriptions factors. As a tentative conclusion, the F4 unusually high cell wall digestibility is likely greatly related to the altered working of at least two major transcription factors regulating cell wall biosynthesis and assembly

Disease resistance or growth: the role of plant hormones in balancing immune responses and fitness costs

Plant growth and response to environmental cues are largely governed by phytohormones. The plant hormones ethylene, jasmonic acid, and salicylic acid (SA) play a central role in the regulation of plant immune responses. In addition, other plant hormones, such as auxins, abscisic acid (ABA), cytokinins, gibberellins, and brassinosteroids, that have been thoroughly described to regulate plant development and growth, have recently emerged as key regulators of plant immunity. Plant hormones interact in complex networks to balance the response to developmental and environmental cues and thus limiting defense-associated fitness costs. The molecular mechanisms that govern these hormonal networks are largely unknown. Moreover, hormone signaling pathways are targeted by pathogens to disturb and evade plant defense responses. In this review, we address novel insights on the regulatory roles of the ABA, SA, and auxin in plant resistance to pathogens and we describe the complex interactions among their signal transduction pathways. The strategies developed by pathogens to evade hormone-mediated defensive responses are also described. Based on these data we discuss how hormone signaling could be manipulated to improve the resistance of crops to pathogens

WAT1 (WALLS ARE THIN1) defines a novel auxin transporter in plants and integrates auxin signaling in secondary wall formation in Arabidopsis fibers

International audienceWAT1 (WALLS ARE THIN1) defines a novel auxin transporter in plants and integrates auxin signaling in secondary wall formation in Arabidopsis fibers. IUFRO Tree Biotechnology Conference 2011: From Genomes to Integration and Deliver

Functional characterization of SlscADH1, a fruit-ripening associated short-chain alcohol dehydrogenase of tomato

A tomato short-chain dehydrogenase-reductase (SlscADH1) is preferentially expressed in fruit with a maximum expression at the breaker stage while expression in roots, stems, leaves and flowers is very weak. It represents a potential candidate for the formation of aroma volatiles by interconverting alcohols and aldehydes. The SlscADH1 recombinant protein produced in Escherichia coli exhibited dehydrogenase-reductase activity towards several volatile compounds present in tomato flavour with a strong preference for the NAD/NADH co-factors. The strongest activity was observed for the reduction of hexanal (Km = 0.175 mM) and phenylacetaldehyde (Km = 0.375 mM) in the presence of NADH. The oxidation process of hexanol and 1-phenylethanol was much less efficient (Kms of 2.9 and 23.0 mM, respectively), indicating that the enzyme preferentially acts as a reductase. However activity was observed only for hexanal, phenylacetaldehyde, (E)-2-hexenal and acetaldehyde and the corresponding alcohols. No activity could be detected for other aroma volatiles important for tomato flavour, such as methyl-butanol/methyl-butanal, 5-methyl-6-hepten-2-one/5-methyl-6-hepten-2-ol, citronellal/citronellol, neral/nerol, geraniol. In order to assess the function of the SlscADH1 gene, transgenic plants have been generated using the technique of RNA interference (RNAi). Constitutive down-regulation using the 35S promoter resulted in the generation of dwarf plants, indicating that the SlscADH1 gene, although weakly expressed in vegetative tissues, had a function in regulating plant development. Fruitspecific down-regulation using the 2A11 promoter had no morphogenetic effect and did not alter the aldehyde/alcohol balance of the volatiles compounds produced by the fruit. Nevertheless, SlscADH1-inhibited fruit unexpectedly accumulated higher concentrations of C5 and C6 volatile compounds of the lipoxygenase pathway, possibly as an indirect effect of the suppression of SlscADH1 on the catabolism of phospholipids and/or integrity of membranes

Combined approaches provide an anatomical and transcriptomic fingerprint of maize cell wall digestibility

Understanding cell wall biosynthesis and degradation in grasses has become a major aim in plant biology. Although independent previous reports have focused on specific features that dictate cell wall digestibility, cytological, biochemical, and gene regulation parameters have never been integrated within the same study. Herein, we applied a combination of state-of-the-art technologies and different scales of observation on two maize lines that are characterized by highly contrasted forage digestibility. Comparative image analysis of internode sections allow to get an anatomical fingerprint associated with high digestibility: a thin peripheral rind of lignified parenchyma, small numerous vascular bundles, and low proportion of PeriVascular Sclerenchyma (PVS). This cell type patterning led to enhanced digestibility when internode sections were treated with Celluclast, a commercially cell wall degrading enzyme. At a lower scale of observation, Laser Capture Microdissection (LCM) followed by thioacidolysis of PVS revealed a higher proportion of Syringyl (S) unit lignins in the low digestible line while the high digestible line was p-Hydroxyphenyl (H)-rich. Moreover, cytological observation of internodes of the two lines point out that this difference in composition is associated with a delayed lignification of PVS. At the same time, comparative transcriptomics on internodes indicated differential expression of several genes encoding enzymes along the phenylpropanoid pathway and known cell wall-associated Transcription Factors (TFs). Together, these results give an integrative view of different factors which could aim in designing a maize silage ideotype and provide a novel set of potential regulatory genes controlling lignification in maize

Arabidopsiscell wall composition determines disease resistance specificity and fitness

[EN] Plant cell walls are complex structures subject to dynamic remodeling in response to developmental and environmental cues and play essential functions in disease resistance responses. We tested the specific contribution of plant cell walls to immunity by determining the susceptibility of a set of Arabidopsis cell wall mutants (cwm) to pathogens with different parasitic styles: a vascular bacterium, a necrotrophic fungus, and a biotrophic oomycete. Remarkably, most cwm mutants tested (29/34; 85.3%) showed alterations in their resistance responses to at least one of these pathogens in comparison to wild-type plants, illustrating the relevance of wall composition in determining disease-resistance phenotypes. We found that the enhanced resistance of cwm plants to the necrotrophic and vascular pathogens negatively impacted cwm fitness traits, such as biomass and seed yield. Enhanced resistance of cwm plants is not only mediated by canonical immune pathways, like those modulated by phytohormones or microbeassociated molecular patterns, which are not deregulated in the cwm tested. Pectin-enriched wall fractions isolated from cwm plants triggered immune responses in wild-type plants, suggesting that wall-mediated defensive pathways might contribute to cwm resistance. Cell walls of cwm plants show a high diversity of composition alterations as revealed by glycome profiling that detect specific wall carbohydrate moieties. Mathematical analysis of glycome profiling data identified correlations between the amounts of specific wall carbohydrate moieties and disease resistance phenotypes of cwm plants. These data support the relevant and specific function of plant wall composition in plant immune response modulation and in balancing disease resistance/development trade-offs.SIThis work has been also financially supported by the Severo Ochoa Program for Centers of Excellence in R&D from the Agencia Estatal de Investigación of Spain (Grant SEV-2016-0672 (2017-2021) to the Centro de Biotecnología y Genómica de Plantas). In the frame of this program, H.M. was a postdoctoral fellow. H.M. was also supported by an Individual Fellowship grant (SignWALLINg-624721) from the European Union. E.M. was a Juan de la Cierva Postdoctoral Fellow from MINECO, and L.B. was a Formacion Personal Investigador fellow of MICIU. The generation of the CCRC-series of plant cell glycan-directed monoclonal antibodies used in this work was supported by the US NSF (DBI-0421683 and IOS 0923992) to M.G.H

Characterization of a cinnamoyl-CoA reductase 1 (CCR1) mutant in maize: effects on lignification, fibre development, and global gene expression

Cinnamoyl-CoA reductase (CCR), which catalyses the first committed step of the lignin-specific branch of monolignol biosynthesis, has been extensively characterized in dicot species, but few data are available in monocots. By screening a Mu insertional mutant collection in maize, a mutant in the CCR1 gene was isolated named Zmccr1–. In this mutant, CCR1 gene expression is reduced to 31% of the residual wild-type level. Zmccr1– exhibited enhanced digestibility without compromising plant growth and development. Lignin analysis revealed a slight decrease in lignin content and significant changes in lignin structure. p-Hydroxyphenyl units were strongly decreased and the syringyl/guaiacyl ratio was slightly increased. At the cellular level, alterations in lignin deposition were mainly observed in the walls of the sclerenchymatic fibre cells surrounding the vascular bundles. These cell walls showed little to no staining with phloroglucinol. These histochemical changes were accompanied by an increase in sclerenchyma surface area and an alteration in cell shape. In keeping with this cell type-specific phenotype, transcriptomics performed at an early stage of plant development revealed the down-regulation of genes specifically associated with fibre wall formation. To the present authors’ knowledge, this is the first functional characterization of CCR1 in a grass species