Production of prebiotic enriched maple syrup through enzymatic conversion of sucrose into fructo-oligosaccharides.

Maple syrup, a popular natural sweetener has a high content of sucrose, whose consumption is linked to different health issues such as obesity and diabetes. Hence, within this paper, the conversion of sucrose to prebiotics (fructo-oligosaccharides, FOS) was proposed as a promising approach to obtaining a healthier, value-added product. Enzymatic conversion was optimized with respect to key experimental factors, and thereafter derived immobilized preparation of fructosyltransferase (FTase) from Pectinex® Ultra SP-L (FTase-epoxy Purolite, 255 IU/g support) was successfully utilized to produce novel functional product in ten consecutive reaction cycles. The product, obtained under optimal conditions (60 °C, 7.65 IU/mL, 12 h), resulted in 56.0% FOS, 16.7% sucrose, and 27.3% monosaccharides of total carbohydrates, leading to a 1.6-fold reduction in caloric content. The obtained products` prebiotic potential toward the probiotic strain Lactobacillus plantarum 299v was demonstrated. The changes in physico-chemical and sensorial characteristics were esteemed as negligible.

Integrated analysis of metabolome, transcriptome, and bioclimatic factors of Acer truncatum seeds reveals key candidate genes related to unsaturated fatty acid biosynthesis, and potentially optimal production area.

BACKGROUND: Lipids found in plant seeds are essential for controlling seed dormancy, dispersal, and defenses against biotic and abiotic stress. Additionally, these lipids provide nutrition and energy and are therefore important to the human diet as edible oils. Acer truncatum, which belongs to the Aceaceae family, is widely cultivated around the world for its ornamental value. Further because its seed oil is rich in unsaturated fatty acids (UFAs)- i.e. α-linolenic acid (ALA) and nervonic acid (NA)- and because it has been validated as a new food resource in China, the importance of A. truncatum has greatly risen. However, it remains unknown how UFAs are biosynthesized during the growth season, to what extent environmental factors impact their content, and what areas are potentially optimal for their production. RESULTS: In this study, transcriptome and metabolome of A. truncatum seeds at three representative developmental stages was used to find the accumulation patterns of all major FAs. Cumulatively, 966 metabolites and 87,343 unigenes were detected; the differential expressed unigenes and metabolites were compared between stages as follows: stage 1 vs. 2, stage 1 vs. 3, and stage 2 vs. 3 seeds, respectively. Moreover, 13 fatty acid desaturases (FADs) and 20 ß-ketoacyl-CoA synthases (KCSs) were identified, among which the expression level of FAD3 (Cluster-7222.41455) and KCS20 (Cluster-7222.40643) were consistent with the metabolic results of ALA and NA, respectively. Upon analysis of the geographical origin-affected diversity from 17 various locations, we found significant variation in phenotypes and UFA content. Notably, in this study we found that 7 bioclimatic variables showed considerable influence on FAs contents in A. truncatum seeds oil, suggesting their significance as critical environmental parameters. Ultimately, we developed a model for potentially ecological suitable regions in China. CONCLUSION: This study provides a comprehensive understanding of the relationship between metabolome and transcriptome in A. truncatum at various developmental stages of seeds and a new strategy to enhance seed FA content, especially ALA and NA. This is particularly significant in meeting the increasing demands for high-quality edible oil for human consumption. The study offers a scientific basis for A. truncatum's novel utilization as a woody vegetable oil rather than an ornamental plant, potentially expanding its cultivation worldwide.

High-quality maple genome reveals duplication-facilitated leaf color diversity.

Acer truncatum is a horticultural tree species with individuals that display either yellow or red leaves in autumn, giving it high ornamental and economic value. 'Lihong' of A. truncatum is an excellent cultivar due to its characteristic of having autumn leaves that turn a bright and beautiful shade of red, while its closely related cultivar 'Bunge' does not. However, the molecular mechanism underlying the color change in the cultivar 'Lihong' is still unclear. Here, we assembled a high-quality genome sequence of Acer truncatum 'Lihong' (genome size = 688 Mb, scaffold N50 = 9.14 Mb) with 28,438 protein-coding genes predicted. Through comparative genomic analysis, we found that 'Lihong' had experienced more tandem duplication events although it's a high degree of collinearity with 'Bunge'. Especially, the expansion of key enzymes in the anthocyanin synthesis pathway was significantly uneven between the two varieties, with 'Lihong' genome containing a significantly higher number of tandem/dispersed duplication key genes. Further transcriptomic, metabolomic, and molecular functional analyses demonstrated that several UFGT genes, mainly resulting from tandem/dispersed duplication, followed by the promoter sequence variation, may contribute greatly to the leaf color phenotype, which provides new insights into the mechanism of divergent anthocyanin accumulation process in the 'Lihong' and 'Bunge' with yellow leaves in autumn. Further, constitutive expression of two UFGT genes, which showed higher expression in 'Lihong', elevated the anthocyanin content. We proposed that the small-scale duplication events could contribute to phenotype innovation.

Quantitative examination of the anatomy of the juvenile sugar maple xylem.

New methodologies have enabled viable sap yields from juvenile sugar maple trees. To further improve yields, a better understanding of sap exudation is required. To achieve this, the anatomy of the xylem must first be fully characterised. We examine juvenile maple saplings using light optical microscopy (LOM) and scanning electron microscopy (SEM), looking at sections cut along differing orientations as well as macerations. From this we measure various cell parameters. We find diameter and length of vessel elements to be 28 ± 8 µm and 200 ± 50 µm, for fibre cells 8 ± 3 µm and 400 ± 100 µm, and for ray parenchyma cells 8 ± 2 µm and 50 ± 20 µm. We also examine pitting present on different cell types. On vessel elements we observe elliptical bordered pits connecting to other vessel elements (with major axis of 2.1 ± 0.7 µm and minor 1.3 ± 0.3 µm) and pits connecting to ray parenchyma (with major axis of 4 ± 2 µm and minor 2.0 ± 0.7 µm). We observe two distinct pit sizes on fibres with circular pits 0.7 ± 0.2 µm in diameter and ellipsoidal pits 1.6 ± 0.4 µm by 1.0 ± 0.3 µm. We do not observe distinct pitting patterns on different fibre types. The various cell and pit measurements obtained generally agree with the limited data available for mature trees, with the exception of vessel element and fibre length, both of which were significantly smaller than reported values.

Acer truncatum leaves extract modulates gut microbiota, improves antioxidant capacity, and alleviates lipopolysaccharide-induced inflammation in broilers.

This study investigated the appropriate way of dietary Acer truncatum leaves (ATL) addition, the effect of disease prevention and its mechanism of action. In experiment 1, 192 Arbor Acres broilers were assigned to 4 treatment groups, fed with basal diets containing 2% bran, replacing it with primary and fermented ATL, and additional 0.3% ATL extract to the basal diet for 42 d, respectively. In experiment 2, 144 broilers were assigned to 3 treatment groups for 21-d trial: (1) C-N group, basal diets, and injected with 0.9% (w/v) sterile saline; (2) C-L group, basal diets, and injected with lipopolysaccharide (LPS); (3) T-L group, ATL diets and injected with LPS. In experiment 1, ATL significantly decreased the index of abdominal fat at 42 d (P < 0.05). ATL extract had a better ability to improve antioxidant capacity and reduce inflammatory levels among all treatment groups, which significantly decreased the content of MDA in the liver and ileum mucosa at 21 d, and increased the expression of IL-10 and Occludin in jejunal mucosa at 42 d (P < 0.05). In experiment 2, ATL significantly increased the level of T-AOC in the liver, decreased the expression of NF-κB in the jejunal mucosa and ileum mucosa (P < 0.05), and restored LPS-induced the changed level of CAT in jejunal mucosa, the expression of IL-6, Claudin-1, and ZO-1 in jejunal mucosa and IL-1ß in ileum mucosa (P < 0.05). Analysis of gut microbiota indicated that ATL enhanced the abundances of Bacteroidota and reduced the proportion of Firmicutes (P < 0.05), and the changed levels of T-AOC in body, IL-1ß, IL-6, IL-10, and NF-κB in jejunum mucosa and propionic acid in cecal were associated with gut microbiota. Collectively, our data showed that the extract of ATL had a better antioxidant and anti-inflammatory effects than primality and fermented. Extraction of ATL modulated intestinal microbiota, and had a protective effect on oxidative stress, inflammation, and intestinal barrier function in broilers challenged with LPS.

1,5-Anhydro-d-glucitol derivative and galloylated flavonoids isolated from the leaves of Acer ginnala Maxim. as dual inhibitors of PTP1B and α-glucosidase enzymes: In vitro and in silico studies.

Four undescribed compounds (two 1,5-anhydro-d-glucitol derivatives and two galloyl derivatives) and fourteen known compounds were isolated and structurally identified from leaves of Acer ginnala Maxim. (Amur maple). Structures and absolute configurations of the four undescribed compounds were determined using extensive analysis of NMR spectroscopic, HRESI-MS, modified Mosher ester method, and comparison with spectroscopic data of known compounds. Bioactivity evaluation revealed that the isolated 1,5-anhydro-d-glucitol derivative, galloylated flavonol rhamnosides, and galloylated flavanols had inhibitory effects on both protein tyrosine phosphatase-1B (PTP1B, IC50 values ranging of 3.46-12.65 µM) and α-glucosidase (IC50 values ranging of 0.88-6.06 µM) in comparison with a positive control for PTP1B (ursolic acid, IC50 = 5.10 µM) or α-glucosidase (acarbose, IC50 = 141.62 µM). A combination of enzyme kinetic analysis and molecular docking provided additional evidence in favor of their inhibitory activities and mechanism. These data demonstrate that A. ginnala Maxim. together with its constituents are promising sources of potent candidates for developing novel anti-diabetic medications.

Patterns of physical, chemical, and metabolic characteristics of sugar maple leaves with depth in the crown and in response to nitrogen and phosphorus addition.

Few previous studies have described the patterns of leaf characteristics in response to nutrient availability and depth in the crown. Sugar maple has been studied for both sensitivity to light, as a shade-tolerant species, and sensitivity to soil nutrient availability, as a species in decline due to acid rain. To explore leaf characteristics from the top to bottom of the canopy, we collected leaves along a vertical gradient within mature sugar maple crowns in a full-factorial nitrogen (N) by phosphorus (P) addition experiment in three forest stands in central New Hampshire, USA. Thirty-two of the 44 leaf characteristics had significant relationships with depth in the crown, with the effect of depth in the crown strongest for leaf area, photosynthetic pigments and polyamines. Nitrogen addition had a strong impact on the concentration of foliar N, chlorophyll, carotenoids, alanine and glutamate. For several other elements and amino acids, N addition changed patterns with depth in the crown. Phosphorus addition increased foliar P and boron (B); it also caused a steeper increase of P and B with depth in the crown. Since most of these leaf characteristics play a direct or indirect role in photosynthesis, metabolic regulation or cell division, studies that ignore the vertical gradient may not accurately represent whole-canopy performance.

Ribosome footprint profiling enables elucidating the systemic regulation of fatty acid accumulation in Acer truncatum.

BACKGROUND: The accumulation of fatty acids in plants covers a wide range of functions in plant physiology and thereby affects adaptations and characteristics of species. As the famous woody oilseed crop, Acer truncatum accumulates unsaturated fatty acids and could serve as the model to understand the regulation and trait formation in oil-accumulation crops. Here, we performed Ribosome footprint profiling combing with a multi-omics strategy towards vital time points during seed development, and finally constructed systematic profiling from transcription to proteomes. Additionally, we characterized the small open reading frames (ORFs) and revealed that the translational efficiencies of focused genes were highly influenced by their sequence features. RESULTS: The comprehensive multi-omics analysis of lipid metabolism was conducted in A. truncatum. We applied the Ribo-seq and RNA-seq techniques, and the analyses of transcriptional and translational profiles of seeds collected at 85 and 115 DAF were compared. Key members of biosynthesis-related structural genes (LACS, FAD2, FAD3, and KCS) were characterized fully. More meaningfully, the regulators (MYB, ABI, bZIP, and Dof) were identified and revealed to affect lipid biosynthesis via post-translational regulations. The translational features results showed that translation efficiency tended to be lower for the genes with a translated uORF than for the genes with a non-translated uORF. They provide new insights into the global mechanisms underlying the developmental regulation of lipid metabolism. CONCLUSIONS: We performed Ribosome footprint profiling combing with a multi-omics strategy in A. truncatum seed development, which provides an example of the use of Ribosome footprint profiling in deciphering the complex regulation network and will be useful for elucidating the metabolism of A. truncatum seed oil and the regulatory mechanisms.

Red pigments in autumn leaves of Norway maple do not offer significant photoprotection but coincide with stress symptoms.

The reasons behind autumn colors, a striking manifestation of anthocyanin synthesis in plants, are poorly understood. Usually, not all leaves of an anthocyanic plant turn red or only a part of the leaf blade turns red. In the present study, we compared green, red and yellow sections of senescing Norway maple leaves, asking if red pigments offer photoprotection, and if so, whether the protection benefits the senescing tree. Green and senescing maple leaves were illuminated with strong white, green or red light in the absence or presence of lincomycin which blocks photosystem II (PSII) repair. Irrespective of the presence of anthocyanins, senescing leaves showed weaker capacity to repair PSII than green leaves. Furthermore, the rate of photoinhibition of PSII did not significantly differ between red and yellow sections of senescing maple leaves. We also followed pigment contents and photosynthetic reactions in individual leaves, from the end of summer until abscission of the leaf. In maple, red pigments accumulated only during late senescence, but light reactions stayed active until most of the chlorophyll had been degraded. PSII activity was found to be lower and non-photochemical quenching higher in red leaf sections, compared with yellow sections of senescing leaves. Red leaf sections were also thicker. We suggest that the primary function of anthocyanin synthesis is not to protect senescing leaves from excess light but to dispose of carbohydrates. This would relieve photosynthetic control, allowing the light reactions to produce energy for nutrient translocation at the last phase of autumn senescence when carbon skeletons are no longer needed.

Transcriptomic and metabolomic analyses reveal how girdling promotes leaf color expression in Acer rubrum L.

BACKGROUND: Acer rubrum L. (red maple) is a popular tree with attractive colored leaves, strong physiological adaptability, and a high ornamental value. Changes in leaf color can be an adaptive response to changes in environmental factors, and also a stress response to external disturbances. In this study, we evaluated the effect of girdling on the color expression of A. rubrum leaves. We studied the phenotypic characteristics, physiological and biochemical characteristics, and the transcriptomic and metabolomic profiles of leaves on girdled and non-girdled branches of A. rubrum. RESULTS: Phenotypic studies showed that girdling resulted in earlier formation of red leaves, and a more intense red color in the leaves. Compared with the control branches, the girdled branches produced leaves with significantly different color parameters a*. Physiological and biochemical studies showed that girdling of branches resulted in uneven accumulation of chlorophyll, carotenoids, anthocyanins, and other pigments in leaves above the band. In the transcriptomic and metabolomic analyses, 28,432 unigenes including 1095 up-regulated genes and 708 down-regulated genes were identified, and the differentially expressed genes were mapped to various KEGG (kyoto encyclopedia of genes and genomes) pathways. Six genes encoding key transcription factors related to anthocyanin metabolism were among differentially expressed genes between leaves on girdled and non-girdled branches. CONCLUSIONS: Girdling significantly affected the growth and photosynthesis of red maple, and affected the metabolic pathways, biosynthesis of secondary metabolites, and carbon metabolisms in the leaves. This resulted in pigment accumulation in the leaves above the girdling site, leading to marked red color expression in those leaves. A transcriptome analysis revealed six genes encoding anthocyanin-related transcription factors that were up-regulated in the leaves above the girdling site. These transcription factors are known to be involved in the regulation of phenylpropanoid biosynthesis, anthocyanin biosynthesis, and flavonoid biosynthesis. These results suggest that leaf reddening is a complex environmental adaptation strategy to maintain normal metabolism in response to environmental changes. Overall, the results of these comprehensive phenotype, physiological, biochemical, transcriptomic, and metabolomic analyses provide a deeper and more reliable understanding of the coevolution of red maple leaves in response to environmental changes.

The relevance of urolithins-based metabotyping for assessing the effects of a polyphenol-rich dietary intervention on intestinal permeability: A post-hoc analysis of the MaPLE trial.

A polyphenol-rich diet reduced intestinal permeability (IP) in older adults. Our aim was to evaluate if participants categorized according to urolithin metabotypes (UMs) exhibited different responses in the MaPLE trial. Fifty-one older adults (mean age: 78 years) completed an 8-week randomized-controlled-crossover trial comparing the effects of a polyphenol-rich vs. a control diet on IP, assessed through zonulin levels. Plasma and urinary metabolomics were evaluated with a semi-targeted UHPLC-MS/MS method. Gut microbiota was characterized by 16S rRNA gene profiling. UMs were determined according to urolithin excretion in 24 h urine samples. Multivariate statistics were used to characterize the differences in metabolomic and metataxonomic responses across UMs. Thirty-three participants were classified as urolithin metabotype A (UMA), 13 as urolithin metabotype B (UMB), and 5 as urolithin metabotype 0 (UM0) according to their urinary excretion of urolithins. Clinical, dietary, and biochemical characteristics at baseline were similar between UMs (all p > 0.05). After the polyphenol-rich diet, UMB vs. UMA participants showed a 2-fold higher improvement of zonulin levels (p for interaction = 0.033). Moreover, UMB vs. UMA participants were characterized for alterations in fatty acid metabolism, kynurenine pathway of tryptophan catabolism, and microbial metabolization of phenolic acids. These changes were correlated with the reduction of zonulin levels and modifications of gut microbes (increased Clostridiales, including, R. lactaris, and G. formicilis). In conclusion, urolithin-based metabotyping identified older adults with a higher improvement of IP after a polyphenol-rich diet. Our results reinforce the concept that UMs may contribute to tailor personalized nutrition interventions.

De novo transcriptome revealed genes involved in anthocyanin biosynthesis, transport, and regulation in a mutant of Acer pseudosieboldianum.

BACKGROUND: Acer pseudosieboldianum is a kind of excellent color-leafed plants, and well known for its red leaves in autumn. At the same time, A. pseudosieboldianum is one of the native tree species in the northeast of China, and it plays an important role in improving the lack of color-leafed plants in the north. In previous study, we found a mutant of the A. pseudosieboldianum that leaves intersect red and green in spring and summer. However, it is unclear which genes cause the color change of mutant leaves. RESULTS: In order to study the molecular mechanism of leaf color formation, we analyzed the leaves of the mutant group and the control group from A. pseudosieboldianum by RNA deep sequencing in this study. Using an Illumina sequencing platform, we obtained approximately 276,071,634 clean reads. After the sequences were filtered and assembled, the transcriptome data generated a total of 70,014 transcripts and 54,776 unigenes, of which 34,486 (62.96%) were successfully annotated in seven public databases. There were 8,609 significant DEGs identified between the control and mutant groups, including 4,897 upregulated and 3,712 downregulated genes. We identified 13 genes of DEGs for leaf color synthesis that was involved in the flavonoid pathway, 26 genes that encoded transcription factors, and eight genes associated with flavonoid transport. CONCLUSION: Our results provided comprehensive gene expression information about A. pseudosieboldianum transcriptome, and directed the further study of accumulation of anthocyanin in A. pseudosieboldianum, aiming to provide insights into leaf coloring of it through transcriptome sequencing and analysis.

Key regulatory pathways, microRNAs, and target genes participate in adventitious root formation of Acer rubrum L.

Red maple (Acer rubrum L.) is a type of colorful ornamental tree with great economic value. Because this tree is difficult to root under natural conditions and the seedling survival rate is low, vegetative propagation methods are often used. Because the formation of adventitious roots (ARs) is essential for the asexual propagation of A. rubrum, it is necessary to investigate the molecular regulatory mechanisms of AR formation in A. rubrum. To address this knowledge gap, we sequenced the transcriptome and small RNAs (sRNAs) of the A. rubrum variety 'Autumn Fantasy' using high-throughput sequencing and explored changes in gene and microRNA (miRNA) expression in response to exogenous auxin treatment. We identified 82,468 differentially expressed genes (DEGs) between the treated and untreated ARs, as well as 48 known and 95 novel miRNAs. We also identified 172 target genes of the known miRNAs using degradome sequencing. Two key regulatory pathways (ubiquitin mediated proteolysis and plant hormone signal transduction), Ar-miR160a and the target gene auxin response factor 10 (ArARF10) were selected based on KEGG pathway and cluster analyses. We further investigated the expression patterns and regulatory roles of ArARF10 through subcellular localization, transcriptional activation, plant transformation, qRT-PCR analysis, and GUS staining. Experiments overexpressing ArARF10 and Ar-miR160a, indicated that ArARF10 promoted AR formation, while Ar-miR160a inhibited AR formation. Transcription factors (TFs) and miRNAs related to auxin regulation that promote AR formation in A. rubrum were identified. Differential expression patterns indicated the Ar-miR160a-ArARF10 interaction might play a significant role in the regulation of AR formation in A. rubrum. Our study provided new insights into mechanisms underlying the regulation of AR formation in A. rubrum.

Transcriptome Sequencing and Screening of Anthocyanin-Related Genes in the Leaves of Acer truncatum Bunge.

Acer truncatum Bunge is generally used as an ornamental tree because of its autumn leaves, although the viewing period is short-approximately 7-15 days. Color improvement of ornamental trees has consistently been an important research topic because color partially determines the value of the commodity; however, a lack of genomic data have limited the progress of molecular breeding research in this area. The purposes of this study were to obtain a transcriptome database for A. truncatum, screen anthocyanin biosynthesis-related genes, and reveal the mechanisms underlying leaf color transformation to provide a basis for increasing the viewing period or breeding cultivars that display red leaves throughout the growing season via gene regulation. In this study, although the use of an Illumina HiSeq 2000 platform and systematic bioinformatics analysis using both young and mature leaves as experimental materials, 233,912,882 clean reads were generated and 121,287 unique transcripts were retrieved. We selected 16 color-related genes (from the transcriptome results) for qRT-PCR to validate the results, and the expression trends of the selected genes were largely consistent with the transcriptome analysis results, with a consistency of 0.875. According to the results of the transcriptome analysis, the validation, and previous studies, we obtained sequences of genes related to anthocyanins, including CHS, CHI, ANS, UFGT, UGT75c1, DFR, BZ1, F3H, F3'H, LAR, ANR, FLS, and those of several transcription factors, including MYB1, BHLH, and WD40. Verifying specific regulation by one or several of these genes in the control of leaf color requires further research. The acquisition of transcriptomic information, especially information concerning anthocyanin biosynthesis-related genes and their base sequences, can provide a theoretical basis for the study of the molecular mechanisms determining changes in leaf color in Acer and is of great importance to the breeding of new cultivars.

Biotransformation of sucrose rich Maple syrups into fructooligosaccharides, oligolevans and levans using levansucrase biocatalyst: Bioprocess optimization and prebiotic activity assessment.

Maple syrup was investigated as a source to produce FOSs and ß-(2-6)-linked-oligolevans/levans. The modulation of this biotransformation was achieved through the control of Maple syrup °Bx and reaction conditions. Reaction time was identified as the most influential factor for the oligolevans/FOSs production in Maple syrup 30°Bx reaction system as well as for the oligolevans/levans synthesis in the 66°Bx one. In the predictive model of oligolevans/levans production in Maple syrup 60°Bx, the interactive effect between levansucrase unit and reaction time was significant (p-value of 0.0008). The optimal conditions for oligolevans/FOSs production (109.20 g/L) in Maple syrup 30°Bx were 3.73 U/mL, pH 6.60 and 23.12 h; while 5 U/mL, pH 6.04 and 29.92 h were identified as the optimal conditions for oligolevans/levans production (147.09 g/L) in Maple syrup 66°Bx. As compared to inulin-type commercial FOSs, the fermentation of oligolevans/FOSs from Maple syrup led to a higher count of Lactobacillus acidophilus and Bifidobacterium lactis and resulted in a higher production of lactic acid. This study lays the foundation for the biotransformation of Maple syrups into functional prebiotic ingredients.

Assembly and comparative analysis of the first complete mitochondrial genome of Acer truncatum Bunge: a woody oil-tree species producing nervonic acid.

BACKGROUND: Acer truncatum (purpleblow maple) is a woody tree species that produces seeds with high levels of valuable fatty acids (especially nervonic acid). The species is admired as a landscape plant with high developmental prospects and scientific research value. The A. truncatum chloroplast genome has recently been reported; however, the mitochondrial genome (mitogenome) is still unexplored. RESULTS: We characterized the A. truncatum mitogenome, which was assembled using reads from PacBio and Illumina sequencing platforms, performed a comparative analysis against different species of Acer. The circular mitogenome of A. truncatum has a length of 791,052 bp, with a base composition of 27.11% A, 27.21% T, 22.79% G, and 22.89% C. The A. truncatum mitogenome contains 62 genes, including 35 protein-coding genes, 23 tRNA genes and 4 rRNA genes. We also examined codon usage, sequence repeats, RNA editing and selective pressure in the A. truncatum mitogenome. To determine the evolutionary and taxonomic status of A. truncatum, we conducted a phylogenetic analysis based on the mitogenomes of A. truncatum and 25 other taxa. In addition, the gene migration from chloroplast and nuclear genomes to the mitogenome were analyzed. Finally, we developed a novel NAD1 intron indel marker for distinguishing several Acer species. CONCLUSIONS: In this study, we assembled and annotated the mitogenome of A. truncatum, a woody oil-tree species producing nervonic acid. The results of our analyses provide comprehensive information on the A. truncatum mitogenome, which would facilitate evolutionary research and molecular barcoding in Acer.

Anatomical adjustment of mature leaves of sycamore maple (Acer pseudoplatanus L.) to increased irradiance.

Trees regenerating in the understory respond to increased availability of light caused by gap formation by undergoing a range of morphological and physiological adjustments. These adjustments include the production of thick, sun-type leaves containing thicker mesophyll and longer palisade cells than in shade-type leaves. We asked whether in the shade-regenerating tree Acer pseudoplatanus, the increase in leaf thickness and expansion of leaf tissues are possible also in leaves that had been fully formed prior to the increase in irradiance, a response reported so far only for a handful of species. We acclimated potted seedlings to eight levels (from 1 to 100%) of solar irradiance and, in late summer, transferred a subset of them to full sunlight. Within 30 days, the shaded leaves increased leaf mass per area and became thicker mostly due to elongation of palisade cells, except for the most shaded individuals which suffered irreversible photo-oxidative damage. This anatomical acclimation was accompanied by partial degradation of chlorophyll and a transient decline in photosynthetic efficiency of PSII (Fv/FM). These effects were related to the degree of pre-shading. The Fv/FM recovered substantially within the re-acclimation period. However, leaves of transferred plants were shed significantly earlier in the fall, indicating that the acclimation was not fully effective. These results show that A. pseudoplatanus is one of the few known species in which mature leaves may re-acclimate anatomically to increased irradiance. This may be a potentially important mechanism enhancing utilization of gaps created during the growing season.

Combined transcriptome and metabolome integrated analysis of Acer mandshuricum to reveal candidate genes involved in anthocyanin accumulation.

The red color formation of Acer mandshuricum leaves is caused by the accumulation of anthocyanins primarily, but the molecular mechanism researches which underlie anthocyanin biosynthesis in A. mandshuricum were still lacking. Therefore, we combined the transcriptome and metabolome and analyzed the regulatory mechanism and accumulation pattern of anthocyanins in three different leaf color states. In our results, 26 anthocyanins were identified. Notably, the metabolite cyanidin 3-O-glucoside was found that significantly correlated with the color formation, was the predominant metabolite in anthocyanin biosynthesis of A. mandshuricum. By the way, two key structural genes ANS (Cluster-20561.86285) and BZ1 (Cluster-20561.99238) in anthocyanidin biosynthesis pathway were significantly up-regulated in RL, suggesting that they might enhance accumulation of cyanidin 3-O-glucoside which is their downstream metabolite, and contributed the red formation of A. mandshuricum leaves. Additionally, most TFs (e.g., MYBs, bZIPs and bHLHs) were detected differentially expressed in three leaf color stages that could participate in anthocyanin accumulation. This study sheds light on the anthocyanin molecular regulation of anthocyanidin biosynthesis and accumulation underlying the different leaf color change periods in A. mandshuricum, and it could provide basic theory and new insight for the leaf color related genetic improvement of A. mandshuricum.

De novo transcriptome sequencing and anthocyanin metabolite analysis reveals leaf color of Acer pseudosieboldianum in autumn.

BACKGROUND: Leaf color is an important ornamental trait of colored-leaf plants. The change of leaf color is closely related to the synthesis and accumulation of anthocyanins in leaves. Acer pseudosieboldianum is a colored-leaf tree native to Northeastern China, however, there was less knowledge in Acer about anthocyanins biosynthesis and many steps of the pathway remain unknown to date. RESULTS: Anthocyanins metabolite and transcript profiling were conducted using HPLC and ESI-MS/MS system and high-throughput RNA sequencing respectively. The results demonstrated that five anthocyanins were detected in this experiment. It is worth mentioning that Peonidin O-hexoside and Cyanidin 3, 5-O-diglucoside were abundant, especially Cyanidin 3, 5-O-diglucoside displayed significant differences in content change at two periods, meaning it may be play an important role for the final color. Transcriptome identification showed that a total of 67.47 Gb of clean data were obtained from our sequencing results. Functional annotation of unigenes, including comparison with COG and GO databases, yielded 35,316 unigene annotations. 16,521 differentially expressed genes were identified from a statistical analysis of differentially gene expression. The genes related to leaf color formation including PAL, ANS, DFR, F3H were selected. Also, we screened out the regulatory genes such as MYB, bHLH and WD40. Combined with the detection of metabolites, the gene pathways related to anthocyanin synthesis were analyzed. CONCLUSIONS: Cyanidin 3, 5-O-diglucoside played an important role for the final color. The genes related to leaf color formation including PAL, ANS, DFR, F3H and regulatory genes such as MYB, bHLH and WD40 were selected. This study enriched the available transcriptome information for A. pseudosieboldianum and identified a series of differentially expressed genes related to leaf color, which provides valuable information for further study on the genetic mechanism of leaf color expression in A. pseudosieboldianum.

Efflux and assimilation of xylem-transported CO2 in stems and leaves of tree species with different wood anatomy.

Determining the fate of CO2 respired in woody tissues is necessary to understand plant respiratory physiology and to evaluate CO2 recycling mechanisms. An aqueous 13 C-enriched CO2 solution was infused into the stem of 3-4 m tall trees to estimate efflux and assimilation of xylem-transported CO2 via cavity ring-down laser spectroscopy and isotope ratio mass spectrometry, respectively. Different tree locations (lower stem, upper stem and leafy shoots) and tissues (xylem, bark and leaves) were monitored in species with tracheid, diffuse- and ring-porous wood anatomy (cedar, maple and oak, respectively). Radial xylem CO2 diffusivity and xylem [CO2 ] were lower in cedar relative to maple and oak trees, thereby limiting label diffusion. Part of the labeled 13 CO2 was assimilated in cedar (8.7%) and oak (20.6%) trees, mostly in xylem and bark tissues of the stem, while limited solution uptake in maple trees hindered the detection of label assimilation. Little label reached foliar tissues, suggesting substantial label loss along the stem-branch transition following reductions in the radial diffusive pathway. Differences in respiration rates and radial xylem CO2 diffusivity (lower in conifer relative to angiosperm species) might reconcile discrepancies in efflux and assimilation of xylem-transported CO2 so far observed between taxonomic clades.