Genome-wide identification of GH9 gene family and the assessment of its role during fruit abscission zone formation in Vaccinium ashei.

KEY MESSAGE: The genomic location and stage-specific expression pattern of GH9 genes reveal their critical roles during fruit abscission zone formation in Vaccinium ashei. Glycosyl hydrolase family 9 (GH9) cellulases play a crucial role in both cellulose synthesis and hydrolysis during plant growth and development. Despite this importance, there is currently no study on the involvement of GH9-encoding genes, specifically VaGH9s, in abscission zone formation of rabbiteye blueberries (Vaccinium ashei). In this study, we identified a total of 61 VaGH9s in the genome, which can be classified into 3 subclasses based on conserved motifs and domains, gene structures, and phylogenetic analyses. Our synteny analysis revealed that VaGH9s are more closely related to the GH9s of Populus L. than to those of Arabidopsis, Vitis vinifera, and Citrus sinensis. In silico structural analysis predicted that most of VaGH9s are hydrophilic, and localized in cell membrane and/or cell wall, and the variable sets of cis-acting regulatory elements and functional diversity with four categories of stress response, hormone regulation, growth and development, and transcription factor-related elements are present in the promoter sequence of VaGH9s genes. Transcriptomic analysis showed that there were 22 differentially expressed VaGH9s in fruit abscission zone tissue at the veraison stage, and the expression of VaGH9B2 and VaGH9C10 was continuously increased during fruit maturation, which were in parallel with the increasing levels of cellulase activity and oxidative stress indicators, suggesting that they are involved in the separation stage of fruit abscission in Vaccinium ashei. Our work identified 22 VaGH9s potentially involved in different stages of fruit abscission and would aid further investigation into the molecular regulation of abscission in rabbiteye blueberries fruit.

Genome-wide characterization, evolution and expression profiling of UDP-glycosyltransferase family in pomelo (Citrus grandis) fruit.

BACKGROUND: Pomelo is one of the three major species of citrus. The fruit accumulates a variety of abundant secondary metabolites that affect the flavor. UDP-glycosyltransferases (UGTs) are involved in the glycosylation of secondary metabolites. RESULTS: In the present study, we performed a genome-wide analysis of pomelo UGT family, a total of 145 UGTs was identified based on the conserved plant secondary product glycosyltransferase (PSPG) motif. These UGT genes were clustered into 16 major groups through phylogenetic analysis of these genes with other plant UGTs (A-P). Pomelo UGTs were distributed unevenly among the chromosomes. At least 10 intron insertion events were observed in these UGT genome sequences, and I-5 was identified to be the highest conserved one. The expression profile analysis of pomelo UGT genes in different fruit tissues during development and ripening was carried out by RNA-seq. CONCLUSIONS: We identified 145 UGTs in pomelo fruit through transcriptome data and citrus genome database. Our research provides available information on UGTs studies in pomelo, and provides an important research foundation for screening and identification of functional UGT genes.

UDP-glucosyltransferase PpUGT85A2 controls volatile glycosylation in peach.

The monoterpene linalool is a major contributor to aroma and flavor in peach (Prunus persica) fruit. It accumulates during fruit ripening, where up to ~40% of the compound is present in a non-volatile glycosylated form, which affects flavor quality and consumer perception by retronasal perception during tasting. Despite the importance of this sequestration to flavor, the UDP-glycosyltransferase (UGT) responsible for linalool glycosylation has not been identified in peach. UGT gene expression during peach fruit ripening and among different peach cultivars was analyzed using RNA sequencing, and transcripts correlated with linalyl-ß-d-glucoside were selected as candidates for functional analysis. Kinetic resolution of a racemic mixture of R,S-linalool was shown for PpUGT85A2, with a slight preference for S-(+)-linalool. PpUGT85A2 was shown to catalyze synthesis of linalyl-ß-d-glucoside in vitro, although it did not exhibit the highest enzyme activity between tested substrates. Subcellular localization of PpUGT85A2 in the cytoplasm and nucleus was detected. Application of linalool to peach leaf disks promoted PpUGT85A2 expression and linalyl-ß-d-glucoside generation. Transient expression in peach fruit and stable overexpression in tobacco and Arabidopsis resulted in significant accumulation of linalyl-ß-d-glucoside in vivo. Taken together, the results indicate that PpUGT85A2 expression is a major control point predicting linalyl-ß-d-glucoside content.

UV-B irradiation differentially regulates terpene synthases and terpene content of peach.

Plants generate protective molecules in response to ultraviolet (UV) light. In laboratory experiments, 48 h UV-B irradiation of peach fruits and leaves reduced the flavour-related monoterpene linalool by 60%. No isoprene was detected, but other terpenoids increased significantly, including a threefold accumulation of the sesquiterpene (E,E)-α-farnesene, which was also increased by jasmonic acid treatment. RNA sequencing revealed altered transcript levels for two terpene synthases (TPSs): PpTPS1, a TPS-g subfamily member, decreased by 86% and PpTPS2, a TPS-b subfamily member, increased 80-fold. Heterologous expression in Escherichia coli and transient overexpression in tobacco and peach fruits showed PpTPS1 was localized in plastids and associated with production of linalool, while PpTPS2 was responsible for (E,E)-α-farnesene biosynthesis in the cytoplasm. Candidate regulatory genes for these responses were identified. Commercial peach production in Asia involves fruit bagging to maintain marketable yield and quality. TPS gene expression and volatile terpenoid production in field experiments, using bags transmitting high UV-B radiation, showed similar effects on peach volatiles to those from laboratory experiments. Bags transmitting less UV-B light ameliorated the reduction in the flavour volatile linalool, indicating that flavour components of peach fruits can be modulated by selecting an appropriate source of environmental screening material.

Melatonin affects cuticular wax profile in rabbiteye blueberry (Vaccinium ashei) during fruit development.

Cuticular wax has been implicated in the first line of plant defense mechanism against external stresses. In this study, cuticular wax on the fruits of two rabbiteye blueberry cultivars cv. Baldwin and Brightwell were examined in terms of the morphology and chemical composition during fruit development in the presence and absence of pre-harvest melatonin (MT) treatment. It revealed that the crystal wax appeared as numerous tubules, and its morphology was not affected by MT treatment. A dominant constituent of triterpenoid at veraison stage was ß-amyrin (45.62% of total wax) in Baldwin and oleanolic acid (22.79% of total wax) in Brightwell. Exogenous MT application significantly promoted fruit quality and increased total content of cuticular wax, however, the effect of MT on individual wax components greatly varied depending on cultivars and fruit developmental stage. Consequently, MT application may enhance sustainability of post-harvest fruit production systems by facilitating accumulation of cuticular wax.

Involvement of MdUGT75B1 and MdUGT71B1 in flavonol galactoside/glucoside biosynthesis in apple fruit.

Apple is rich in flavonol glycosides, which are believed to contribute to putative health benefits associated with apple consumption. Glycosylation, catalyzed by uridine diphospho-glycosyltransferases (UGTs), is the last step in flavonol biosynthesis, which confers molecular stability and solubility to the flavonol. In the present study, the involvement of two UGTs, MdUGT75B1 and MdUGT71B1, in flavonol biosynthesis in apple was investigated. The major flavonols are quercetin 3-O-glycosides, and UV-B and blue light treatment significantly enhanced the accumulation of quercetin 3-O-galactoside, quercetin 3-O-glucoside, and kaempferol 3-O-galactoside. Transcript levels of MdUGT75B1 and MdUGT71B1 in fruit subjected to different treatments were correlated well with flavonol accumulation. MdUGT75B1 showed flavonol-specific activity with a preference for UDP-galactose as the sugar donor, while MdUGT71B1 using UDP-glucose exhibited a wider substrate acceptance. Thus, MdUGT75B1 and MdUGT71B1 are key UGTs involved in flavonol biosynthesis and may have important roles in regulating accumulation of these health-promoting bioactive compounds in apple.

Electromagnetic fields and modal excitations on a thin silver film.

In this paper we extend the fast-all-modes method and the numerical modified steepest-descent-path method to the optical frequency range by finding all modes and solving the total electric field in three dimensions that is due to a point source above a lossy thin metal film with a negative permittivity situated between two dissimilar dielectric materials. We show that up to four proper surface wave modes may propagate on the film surface, including both backward and forward waves. We also solve for the electric field below the lossy thin metal film and verify the existence of superlensing of the electric field, comparing that case to the case of a dielectric film where no superlensing occurs. The CPU time using the fast-all-modes method and the numerical modified steepest-descent-path method is considerably less than that using the conventional method of integration along the Sommerfeld integration path.

Glycosidically bound volatiles as affected by ripening stages of Satsuma mandarin fruit.

Composition and changes in free volatiles have been extensively studied in citrus fruit such as mandarin. However, components of glycosidically bound volatiles and changes during fruit ripening have been rarely investigated. A total of 56 glycosidically-bound volatiles were identified in fruit peel at four ripening stages. The highest concentrations in glycosidically-bound volatiles were observed for methyl salicylate in ripening fruit. Concentration of total bound volatiles increased from color conversion stage at 150days after bloom (DAB), peaked at yellow stage (190DAB), followed by a decrease at orange stage (210DAB). Satsuma mandarin fruit at different ripening stages could be separated in a partial least squares-discriminant analysis (PLS-DA) plot using glycosidically bound volatiles as variables. In total 35 glycosidically bound volatiles were identified with variable importance in projection (VIP) score exceeding 1, which may be potential markers for separating fruit at different ripening stages.

Genome-Wide Identification, Expression Patterns, and Functional Analysis of UDP Glycosyltransferase Family in Peach (Prunus persica L. Batsch).

Peach (Prunus persica L. Batsch) is a commercial grown fruit trees, important because of its essential nutrients and flavor promoting secondary metabolites. The glycosylation processes mediated by UDP-glycosyltransferases (UGTs) play an important role in regulating secondary metabolites availability. Identification and characterization of peach UGTs is therefore a research priority. A total of 168 peach UGT genes that distributed unevenly across chromosomes were identified based on their conserved PSPG motifs. Phylogenetic analysis of these genes with plant UGTs clustered them into 16 groups (A-P). Comparison of the patterns of intron-extron and their positions within genes revealed one highly conserved intron insertion event in peach UGTs. Tissue specificity, temporal expression patterns in peach fruit during development and ripening, and in response to abiotic stress UV-B irradiation was investigated using RNA-seq strategy. The relationship between UGTs transcript levels and concentrations of glycosylated volatiles was examined to select candidates for functional analysis. Heterologous expressing these candidate genes in Escherichia coli identified UGTs that were involved in the in vitro volatile glycosylation. Our results provide an important source for the identification of functional UGT genes to potential manipulate secondary biosynthesis in peach.