Effect of abscisic acid on galactomannan degradation and endo-β-mannanase activity in seeds of Sesbania virgata (Cav.) Pers. (Leguminosae)

Seeds of Sesbania virgata (Cav.) Pers. (Leguminosae) have an endosperm which accumulates galactomannan as a storage polysaccharide in the cell walls. After germination, it is hydrolysed by three enzymes: α-galactosidase (EC 3.2.1.22), endo-β-mannanase (EC 3.2.1.78) and β-mannosidase (EC 3.2.1.25). This work aimed at studying the effect of abscisic acid (ABA) on galactomannan degradation during and after germination. Seeds were imbibed in water or in 10⁻⁴ M ABA, and used to evaluate the effect of exogenous and endogenous ABA. Tissue printing was used to follow biochemical events by detecting and localising endo-β-mannanase in different tissues of the seed. The presence of exogenous ABA provoked a delay in the cellular disassembly of the endosperm and disappearance of endo-β-mannanase in the tissue. This led to a delay in galactomannan degradation. The testa (seed coat) of S. virgata contains endogenous ABA, which decreases ca. fourfold during storage mobilisation after germination, permitting the galactomannan degradation in the endosperm. Furthermore, endo-β-mannanase was immunolocalised in the testa, which has a living cell layer. The ABA appears to modulate storage mobilisation in the legume seed of S. virgata, and a cause–effect relationship between ABA (probably through testa) and activities of hydrolases is proposed.     

Cell wall polysaccharides from fern leaves: evidence for a mannan-rich Type III cell wall in Adiantum raddianum

Primary cell walls from plants are composites of cellulose tethered by cross-linking glycans and embedded in a matrix of pectins. Cell wall composition varies between plant species, reflecting in some instances the evolutionary distance between them. In this work the monosaccharide compositions of isolated primary cell walls of nine fern species and one lycophyte were characterized and compared with those from Equisetum and an angiosperm dicot. The relatively high abundance of mannose in these plants suggests that mannans may constitute the major cross-linking glycan in the primary walls of pteridophytes and lycophytes. Pectin-related polysaccharides contained mostly rhamnose and uronic acids, indicating the presence of rhamnogalacturonan I highly substituted with galactose and arabinose. Structural and fine-structural analyses of the hemicellulose fraction of leaves of Adiantum raddianum confirmed this hypothesis. Linkage analysis showed that the mannan contains mostly 4-Man with very little 4,6-Man, indicating a low percentage of branching with galactose. Treatment of the mannan-rich fractions with endo-β-mannanase produced characteristic mannan oligosaccharides. Minor amounts of xyloglucan and xylans were also detected. These data and those of others suggest that all vascular plants contain xyloglucans, arabinoxylans, and (gluco)mannans, but in different proportions that define cell wall types. Whereas xyloglucan and pectin-rich walls define Type I walls of dicots and many monocots, arabinoxylans and lower proportion of pectin define the Type II walls of commelinoid monocots. The mannan-rich primary walls with low pectins of many ferns and a lycopod indicate a fundamentally different wall type among land plants, the Type III wall.     

Focus on Metabolism: Changes in Whole-Plant Metabolism during the Grain-Filling Stage in Sorghum Grown under Elevated CO2 and Drought

Projections indicate an elevation of the atmospheric CO₂ concentration ([CO₂]) concomitant with an intensification of drought for this century, increasing the challenges to food security. On the one hand, drought is a main environmental factor responsible for decreasing crop productivity and grain quality, especially when occurring during the grain-filling stage. On the other hand, elevated [CO₂] is predicted to mitigate some of the negative effects of drought. Sorghum (Sorghum bicolor) is a C₄ grass that has important economical and nutritional values in many parts of the world. Although the impact of elevated [CO₂] and drought in photosynthesis and growth has been well documented for sorghum, the effects of the combination of these two environmental factors on plant metabolism have yet to be determined. To address this question, sorghum plants (cv BRS 330) were grown and monitored at ambient (400 µmol mol⁻¹) or elevated (800 µmol mol⁻¹) [CO₂] for 120 d and subjected to drought during the grain-filling stage. Leaf photosynthesis, respiration, and stomatal conductance were measured at 90 and 120 d after planting, and plant organs (leaves, culm, roots, prop roots, and grains) were harvested. Finally, biochemical composition and intracellular metabolites were assessed for each organ. As expected, elevated [CO₂] reduced the stomatal conductance, which preserved soil moisture and plant fitness under drought. Interestingly, the whole-plant metabolism was adjusted and protein content in grains was improved by 60% in sorghum grown under elevated [CO₂].Global food demand is projected to increase up to 110% by the middle of this century (Tilman et al., 2011; Alexandratos and Bruinsma, 2012), particularly due to a rise in world population that is likely to plateau at about 9 billion people (Godfray et al., 2010). Additionally, the average concentration of atmospheric CO₂ ([CO₂]) has increased 1.75 µmol mol⁻¹ per year between 1975 and today, reaching 400 µmol mol⁻¹ in April 2015 (NOAA, 2015). According to the A2 emission scenario from the U.S. Environmental Protection Agency, in the absence of explicit climate change policy, atmospheric CO₂ concentrations will reach 800 µmol mol⁻¹ by the end of this century. The increasing atmospheric [CO₂] is resulting in global climate changes, such as reduction in water availability and elevation in temperature. These factors are expected to heavily influence food production in the next years (Godfray and Garnett, 2014; Magrin et al., 2014).Sorghum (Sorghum bicolor) is a C₄ grass, considered a staple food grain for millions of the poorest and most food-insecure people in the semiarid tropics of Africa, Asia, and Central America, serving as an important source of energy, proteins, vitamins, and minerals (Taylor et al., 2006). Moreover, this crop is used for animal feed and as industrial raw material in developed countries such as the United States, which is the main world producer (FAO, 2015). With a fully sequenced genome (Paterson et al., 2009) and over 45,000 accessions representing a large geographic and genetic diversity, sorghum is a good model system in which to study the impact of global climate changes in C₄ grasses.The increase in [CO₂] in the atmosphere, which is the main driver of global climate changes (Meehl et al., 2007), is predicted to boost photosynthesis rates and productivity in a series of C₃ legumes and cereals, mainly due to a decrease in the photorespiration process (Grashoff et al., 1995; Long et al., 2006). On the contrary, due to their capacity to concentrate CO₂ in bundle sheath cells and reduce photorespiration to virtually zero, C₄ plants are unlikely to respond to the elevation of atmospheric [CO₂] (Leakey, 2009). However, even for C₄ plants, elevated [CO₂] can ameliorate the effects caused by drought, maintaining higher photosynthetic rates. This is due to an improvement in the efficiency of water use that is achieved by the reduction in stomatal conductance (Leakey et al., 2004; Markelz et al., 2011).The rate of photosynthesis as well as the redistribution of photoassimilates accumulated in different plant tissues during the day and/or during vegetative growth are crucial to grain development, and later, to its filling (Schnyder, 1993). Due to this relationship, any environmental stress such as drought occurring during the reproductive phase has the potential to result in poor grain filling and losses in yield (Blum et al., 1997). For instance, postanthesis drought can cause up to 30% decrease in yield (Borrell et al., 2000). It is also known that elevated [CO₂], drought, high temperature, and any combinations of these stresses can lead to significant changes in grain composition (Taub et al., 2008; Da Matta et al., 2010; Uprety et al., 2010; Madan et al., 2012), suggesting diverse metabolic alterations and/or adaptations that occur in the plant when it is cultivated in such conditions.Although the impacts of elevated [CO₂] and drought on photosynthesis and the growth of sorghum have been well documented (Conley et al., 2001; Ottman et al., 2001; Wall et al., 2001), no attention has been given to the impact of the combination of these two environmental changes on plant metabolism and composition. Regarding physiology, studies on the growth of sorghum under elevated [CO₂] and drought showed an increase of the net assimilation rate of 23% due to a decrease of 32% in stomatal conductance (Wall et al., 2001). This resulted in sorghum’s ability to use water 17% more efficiently (Conley et al., 2001). An improvement in the final overall biomass under elevated [CO₂] and drought has also been described (Ottman et al., 2001), but without a significant effect in grain yield (Wall et al., 2001).Few studies have been monitoring metabolic pathways in plants under elevated [CO₂] (Li et al., 2008; Aranjuelo et al., 2013) and drought (Silvente et al., 2012; Nam et al., 2015; Wenzel et al., 2015). Furthermore, to our knowledge, there are only two reports in which metabolite profiles or metabolic pathways were investigated under the combination of these two environmental conditions (Sicher and Barnaby, 2012; Zinta et al., 2014). Although it is widely accepted that whole-plant metabolism and composition can impact grain filling and yield, metabolic studies conducted so far have focused on a specific plant organ. For instance, Sicher and Barnaby (2012) analyzed the metabolite profile of leaves from maize (Zea mays) plants that were grown under elevated [CO₂] and drought, but they did not show how those environmental changes could have affected the metabolism of other tissues (e.g. culm and roots) or how they might have influenced the biomass or grain composition.In order to address how the combination of elevated [CO₂] and drought can modify whole-plant metabolism as well as biomass composition in sorghum, this study aimed to (1) evaluate photosynthesis, growth, and yield; (2) underline the differences in biomass composition and primary metabolite profiles among leaves, culm, roots, prop roots, and grains; and (3) determine the effect of elevated [CO₂] and drought on the primary metabolism of each organ.     

The role of exo-(1-->4)-beta-galactanase in the mobilization of polysaccharides from the cotyledon cell walls of Lupinus angustifolius following germination

BACKGROUND AND AIMS: The cotyledons of Lupinus angustifolius contain large amounts of cell wall storage polysaccharide (CWSP) composed mainly of (1-->4)-beta-linked D-galactose residues in the form of branches attached to a rhamnogalacturonan core molecule. An exo-(1-->4)-beta-galactanase with a very high specificity towards (1-->4)-beta-linked D-galactan has been isolated from L. angustifolius cotyledons, and shown to vary (activity and specific protein) in step with CWSP mobilization. This work aimed to confirm the hypothesis that galactan is the main polymer retrieved from the wall during mobilization at the ultrastructural level, using the purified exo-galactanase as a probe. METHODS: Storage mesophyll cell walls ('ghosts') were isolated from the cotyledons of imbibed but ungerminated lupin seeds, and also from cotyledons of seedlings after the mobilization of the CWSP. The pure exo-(1-->4)-beta-galactanase was coupled to colloidal gold particles and shown to be a specific probe for (1-->4)-beta-D-galactan. They were used to localize galactan in ultrathin sections of L. angustifolius cotyledonary mesophyll tissue during CWSP mobilization. KEY RESULTS: On comparing the morphologies of isolated cell walls, the post-mobilization 'ghosts' did not have the massive wall-thickenings of pre-mobilization walls. Compositional analysis showed that the post-mobilization walls were depleted in galactose and, to a lesser extent, in arabinose. When pre-mobilization ghosts were treated with the pure exo-galactanase, they became morphologically similar to the post-mobilization ghosts. They were depleted of approximately 70% of the galactose residues that would have been mobilized in vivo, and retained all the other sugar residues originally present. Sharply defined electron-transparent wall zones or pockets are associated with CWSP mobilization, being totally free of galactan, whereas wall areas immediately adjacent to them were apparently undepleted. CONCLUSIONS: The exo-(1-->4)-beta-galactanase is the principal enzyme involved in CWSP mobilization in lupin cotyledons in vivo. The storage walls dramatically change their texture during mobilization as most of the galactan is hydrolysed during seedling development.