Chemical Composition in Juvenile and Mature Wood of Branch and Main Trunk of Leucaena leucocephala (Lam.) de Wit.

Secondary growth is the most dynamic developmental aspect during the terrestrialization of plants. The development of secondary xylem tissue composed of thick-walled cells with characteristic changes in its structure and chemistry facilitates the growth and development of woody plants. In the present study, the chemical composition of the secondary xylem of juvenile and mature wood from the branch and main trunk of Leucaena leucocephala, has been investigated and the differences established. The biochemical analysis of different cell wall components in the mature wood of the main trunk revealed high holocellulose and α-cellulose and less lignin content in the juvenile wood while its syringyl/guaiacyl (S/G) ratio was less than for the mature wood. As compared to the branch xylem, concentration of cell wall polysaccharides and lignin content was higher in both juvenile and mature wood collected from the main trunk. Thioacidolysis and GC-MS analysis of wood lignin from juvenile and mature wood showed that an increased concentration in lignin content in mature wood is associated with a corresponding increase in S/G ratio. The structural information of the acetylated lignin was investigated by 1H NMR spectroscopy. Our results indicate that the mature wood from the main trunk is superior in pulp yielding and lignin degradability as compared to the juvenile wood of the branch and trunk.

Flexure wood formation via growth reprogramming in hybrid aspen involves jasmonates and polyamines and transcriptional changes resembling tension wood development.

Stem bending in trees induces flexure wood but its properties and development are poorly understood. Here, we investigated the effects of low-intensity multidirectional stem flexing on growth and wood properties of hybrid aspen, and on its transcriptomic and hormonal responses. Glasshouse-grown trees were either kept stationary or subjected to several daily shakes for 5 wk, after which the transcriptomes and hormones were analyzed in the cambial region and developing wood tissues, and the wood properties were analyzed by physical, chemical and microscopy techniques. Shaking increased primary and secondary growth and altered wood differentiation by stimulating gelatinous-fiber formation, reducing secondary wall thickness, changing matrix polysaccharides and increasing cellulose, G- and H-lignin contents, cell wall porosity and saccharification yields. Wood-forming tissues exhibited elevated jasmonate, polyamine, ethylene and brassinosteroids and reduced abscisic acid and gibberellin signaling. Transcriptional responses resembled those during tension wood formation but not opposite wood formation and revealed several thigmomorphogenesis-related genes as well as novel gene networks including FLA and XTH genes encoding plasma membrane-bound proteins. Low-intensity stem flexing stimulates growth and induces wood having improved biorefinery properties through molecular and hormonal pathways similar to thigmomorphogenesis in herbaceous plants and largely overlapping with the tension wood program of hardwoods.

Impact of xylan on field productivity and wood saccharification properties in aspen.

Xylan that comprises roughly 25% of hardwood biomass is undesirable in biorefinery applications involving saccharification and fermentation. Efforts to reduce xylan levels have therefore been made in many species, usually resulting in improved saccharification. However, such modified plants have not yet been tested under field conditions. Here we evaluate the field performance of transgenic hybrid aspen lines with reduced xylan levels and assess their usefulness as short-rotation feedstocks for biorefineries. Three types of transgenic lines were tested in four-year field tests with RNAi constructs targeting either Populus GT43 clades B and C (GT43BC) corresponding to Arabidopsis clades IRX9 and IRX14, respectively, involved in xylan backbone biosynthesis, GATL1.1 corresponding to AtGALT1 involved in xylan reducing end sequence biosynthesis, or ASPR1 encoding an atypical aspartate protease. Their productivity, wood quality traits, and saccharification efficiency were analyzed. The only lines differing significantly from the wild type with respect to growth and biotic stress resistance were the ASPR1 lines, whose stems were roughly 10% shorter and narrower and leaves showed increased arthropod damage. GT43BC lines exhibited no growth advantage in the field despite their superior growth in greenhouse experiments. Wood from the ASPR1 and GT43BC lines had slightly reduced density due to thinner cell walls and, in the case of ASPR1, larger cell diameters. The xylan was less extractable by alkali but more hydrolysable by acid, had increased glucuronosylation, and its content was reduced in all three types of transgenic lines. The hemicellulose size distribution in the GALT1.1 and ASPR1 lines was skewed towards higher molecular mass compared to the wild type. These results provide experimental evidence that GATL1.1 functions in xylan biosynthesis and suggest that ASPR1 may regulate this process. In saccharification without pretreatment, lines of all three constructs provided 8-11% higher average glucose yields than wild-type plants. In saccharification with acid pretreatment, the GT43BC construct provided a 10% yield increase on average. The best transgenic lines of each construct are thus predicted to modestly outperform the wild type in terms of glucose yields per hectare. The field evaluation of transgenic xylan-reduced aspen represents an important step towards more productive feedstocks for biorefineries.

Root and Leaf Anatomy, Ion Accumulation, and Transcriptome Pattern under Salt Stress Conditions in Contrasting Genotypes of Sorghum bicolor.

Roots from salt-susceptible ICSR-56 (SS) sorghum plants display metaxylem elements with thin cell walls and large diameter. On the other hand, roots with thick, lignified cell walls in the hypodermis and endodermis were noticed in salt-tolerant CSV-15 (ST) sorghum plants. The secondary wall thickness and number of lignified cells in the hypodermis have increased with the treatment of sodium chloride stress to the plants (STN). Lignin distribution in the secondary cell wall of sclerenchymatous cells beneath the lower epidermis was higher in ST leaves compared to the SS genotype. Casparian thickenings with homogenous lignin distribution were observed in STN roots, but inhomogeneous distribution was evident in SS seedlings treated with sodium chloride (SSN). Higher accumulation of K+ and lower Na+ levels were noticed in ST compared to the SS genotype. To identify the differentially expressed genes among SS and ST genotypes, transcriptomic analysis was carried out. Both the genotypes were exposed to 200 mM sodium chloride stress for 24 h and used for analysis. We obtained 70 and 162 differentially expressed genes (DEGs) exclusive to SS and SSN and 112 and 26 DEGs exclusive to ST and STN, respectively. Kyoto Encyclopaedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis unlocked the changes in metabolic pathways in response to salt stress. qRT-PCR was performed to validate 20 DEGs in each SSN and STN sample, which confirms the transcriptomic results. These results surmise that anatomical changes and higher K+/Na+ ratios are essential for mitigating salt stress in sorghum apart from the genes that are differentially up- and downregulated in contrasting genotypes.

Pectin from Fruit- and Berry-Juice Production by-Products: Determination of Physicochemical, Antioxidant and Rheological Properties.

Plums (Prunus domestica); red currants (Ribes rubrum); black currants (Ribes nigrum); gooseberries (Ribes uva-crispa); sour cherries (Prunus cerasus); pumpkins (Cuccurbita spp.) are sources for valuable fruit- and berry-juice and cider production. This process leaves a large number of by-products (BP) in the form of pomace, which accounts for up to 80% of the raw material. This by-product represents a rich source of biologically active compounds, especially in the form of different pectic polysaccharides. The pectin extracted from commercial fruits such as citric fruits and apples has high medicinal properties, can be used as edible films and coatings, and is also useful in texture improvement and gel production in the food industry. However, many under-utilized fruits have received little attention regarding the extraction and characterization of their high/value pectin from their by-products. Moreover, the commercial extraction process involving strong acids and high temperature to obtain high-purity pectin leads to the loss of many bioactive components, and these lost components are often compensated for by the addition of synthetic antioxidants and colorants. The aim of the research is to extract pectin from juice production by-products with hot-water extraction using weak organic (0.1 N) citric acid, thus minimizing the impact on the environment. The yield of pectin (PY = 4.47-17.8% DM), galacturonic acid content (47.22-83.57 g 100-1), ash content (1.42-2.88 g 100 g-1), degree of esterification (DE = 45.16-64.06%), methoxyl content (ME = 4.27-8.13%), the total content of phenolic compounds (TPC = 2.076-4.668 µg mg-1, GAE) and the antiradical scavenging activity of the pectin samples (DPPH method (0.56-37.29%)) were determined. Free and total phenolic acids were quantified by saponification using high-pressure liquid chromatography (HPLC). The pectin contained phenolic acids-benzoic (0.25-0.92 µg mg-1), gallic (0.14-0.57 µg mg-1), coumaric (0.04 µg mg-1), and caffeic (0.03 µg mg-1). The pectin extracts from by-products showed glucose and galactose (3.89-21.72 g 100 g-1) as the main neutral sugar monosaccharides. Pectin analysis was performed using FT-IR, and the rheological properties of the pectin gels were determined. The quality of the obtained pectin from the fruit and berry by-products in terms of their high biological activity and high content of glucuronic acids indicated that the products have the potential to be used as natural ingredients in various food products and in pharmaceutical products.

Ultrastructural changes during nectar secretion from extrafloral nectaries of Pithecellobium dulce Benth.

The structural changes in the secretory cells are important to understand the ontogeny and nectar secretion process from the nectaries. In this study, we investigated the ultrastructural changes during different developmental/secretion stages of extrafloral nectaries (EFNs) of Pithecellobium dulce. The dense cytoplasm with active biosynthesis mechanisms such as ribosomes, mitochondria, large nucleus, and plastids with accumulated starch grains characterized the pre-secretion stage of young nectariferous cells. During the secretory phase, the cytoplasm showed distinct changes associated with endomembrane transport such as the predominant occurrence of Golgi, secretory vesicles, and ER resulting in the subsequent appearance of secretions in the intercellular and subcuticular spaces. Cell wall loosening following the dissolution of middle lamellae leading to the formation of subcuticular spaces was evident during advanced stages of nectar secretion. The characteristic cytoplasmic and apoplastic changes associated with cell death were noticed during the post-secretory stages. The structural evidence from the present study suggests the occurrence of two modes of secretion (merocrine and holocrine) during the early and late stages of secretion in the EFNs of P. dulce.

Overexpression of a Plasma Membrane Bound Na+/H+ Antiporter-Like Protein (SbNHXLP) Confers Salt Tolerance and Improves Fruit Yield in Tomato by Maintaining Ion Homeostasis.

A Na+/H+ antiporter-like protein (NHXLP) was isolated from Sorghum bicolor L. (SbNHXLP) and validated by overexpressing in tomato for salt tolerance. Homozygous T2 transgenic lines when evaluated for salt tolerance, accumulated low Na+ and displayed enhanced salt tolerance compared to wild-type plants (WT). This is consistent with the amiloride binding assay of the protein. Transgenics exhibited higher accumulation of proline, K+, Ca2+, improved cambial conductivity, higher PSII, and antioxidative enzyme activities than WT. Fluorescence imaging results revealed lower Na+ and higher Ca2+ levels in transgenic roots. Co-immunoprecipitation experiments demonstrate that SbNHXLP interacts with a Solanum lycopersicum cation proton antiporter protein2 (SlCHX2). qRT-PCR results showed upregulation of SbNHXLP and SlCHX2 upon treatment with 200 mM NaCl and 100 mM potassium nitrate. SlCHX2 is known to be involved in K+ acquisition, and the interaction between these two proteins might help to accumulate more K+ ions, and thus maintain ion homeostasis. These results strongly suggest that plasma membrane bound SbNHXLP involves in Na+ exclusion, maintains ion homeostasis in transgenics in comparison with WT and alleviates NaCl stress.