Cuticle deposition ceases during strawberry fruit development.

BACKGROUND: Ideally, the barrier properties of a fruit's cuticle persist throughout its development. This presents a challenge for strawberry fruit, with their rapid development and thin cuticles. The objective was to establish the developmental time course of cuticle deposition in strawberry fruit. RESULTS: Fruit mass and surface area increase rapidly, with peak growth rate coinciding with the onset of ripening. On a whole-fruit basis, the masses of cutin and wax increase but on a unit surface-area basis, they decrease. The decrease is associated with marked increases in elastic strain. The expressions of cuticle-associated genes involved in transcriptional regulation (FaSHN1, FaSHN2, FaSHN3), synthesis of cutin (FaLACS2, FaGPAT3) and wax (FaCER1, FaKCS10, FaKCR1), and those involved in transport of cutin monomers and wax constituents (FaABCG11, FaABCG32) decreased until maturity. The only exceptions were FaLACS6 and FaGPAT6 that are presumably involved in cutin synthesis, and FaCER1 involved in wax synthesis. This result was consistent across five strawberry cultivars. Strawberry cutin consists mainly of C16 and C18 monomers, plus minor amounts of C19, C20, C22 and C24 monomers, ω-hydroxy acids, dihydroxy acids, epoxy acids, primary alcohols, carboxylic acids and dicarboxylic acids. The most abundant monomer is 10,16-dihydroxyhexadecanoic acid. Waxes comprise mainly long-chain fatty acids C29 to C46, with smaller amounts of C16 to C28. Wax constituents are carboxylic acids, primary alcohols, alkanes, aldehydes, sterols and esters. CONCLUSION: The downregulation of cuticle deposition during development accounts for the marked cuticular strain, for the associated microcracking, and for their high susceptibility to the disorders of water soaking and cracking.

Involvement of the GH38 Family Exoglycosidase α-Mannosidase in Strawberry Fruit Ripening.

Exoglycosidase enzymes hydrolyze the N-glycosylations of cell wall enzymes, releasing N-glycans that act as signal molecules and promote fruit ripening. Vesicular exoglycosidase α-mannosidase enzymes of the GH38 family (EC 3.2.1.24; α-man) hydrolyze N-glycans in non-reduced termini. Strawberry fruit (Fragaria × ananassa) is characterized by rapid softening as a result of cell wall modifications during the fruit ripening process. Enzymes acting on cell wall polysaccharides explain the changes in fruit firmness, but α-man has not yet been described in F. × ananassa, meaning that the indirect effects of N-glycan removal on its fruit ripening process are unknown. The present study identified 10 GH38 α-man sequences in the F. × ananassa genome with characteristic conserved domains and key residues. A phylogenetic tree built with the neighbor-joining method and three groups of α-man established, of which group I was classified into three subgroups and group III contained only Poaceae spp. sequences. The real-time qPCR results demonstrated that FaMAN genes decreased during fruit ripening, a trend mirrored by the total enzyme activity from the white to ripe stages. The analysis of the promoter regions of these FaMAN genes was enriched with ripening and phytohormone response elements, and contained cis-regulatory elements related to stress responses to low temperature, drought, defense, and salt stress. This study discusses the relevance of α-man in fruit ripening and how it can be a useful target to prolong fruit shelf life.

Unraveling the key step in the aroma puzzle: Insights into alcohol acyltransferases in strawberries.

Alcohol acyltransferases (AATs) play a crucial role in catalyzing the transfer of acyl groups, contributing to the diverse aroma of fruits, including strawberries. In this research we identified nine AAT genes in strawberries through a comprehensive analysis involving phylogenetics, gene structure, conserved motifs, and structural protein model examinations. The study used the 'Camarosa' strawberry genome database, and experiments were conducted with fruits harvested at different developmental and ripening stages. The transcriptional analysis revealed differential expression patterns among the AAT genes during fruit ripening, with only four genes (SAAT, FaAAT2, FaAAT7, and FaAAT9) showing increased transcript accumulation correlated with total AAT enzyme activity. Additionally, the study employed in silico methods, including sequence alignment, phylogenetic analysis, and structural modeling, to gain insights into the AAT protein model structures with increase expression pattern during fruit ripening. The four modeled AAT proteins exhibited structural similarities, including conserved catalytic sites and solvent channels. Furthermore, the research investigated the interaction of AAT proteins with different substrates, highlighting the enzymes' promiscuity in substrate preferences. The study contributes with valuable information to unveil AAT gene family members in strawberries, providing scientific background for further exploration of their biological characteristics and their role in aroma biosynthesis during fruit ripening.

Analysis of Methylesterase Gene Family in Fragaria vesca Unveils Novel Insights into the Role of FvMES2 in Methyl Salicylate-Mediated Resistance against Strawberry Gray Mold.

Methylesterases (MESs) hydrolyze carboxylic ester and are important for plant metabolism and defense. However, the understanding of MES' role in strawberries against pathogens remains limited. This study identified 15 FvMESs with a conserved catalytic triad from the Fragaria vesca genome. Spatiotemporal expression data demonstrated the upregulated expression of FvMESs in roots and developing fruits, suggesting growth involvement. The FvMES promoter regions harbored numerous stress-related cis-acting elements and transcription factors associated with plant defense mechanisms. Moreover, FvMES2 exhibited a significant response to Botrytis cinerea stress and showed a remarkable correlation with the salicylic acid (SA) signaling pathway. Molecular docking showed an efficient binding potential between FvMES2 and methyl salicylate (MeSA). The role of FvMES2 in MeSA demethylation to produce SA was further confirmed through in vitro and in vivo assays. After MeSA was applied, the transient overexpression of FvMES2 in strawberries enhanced their resistance to B. cinerea compared to wild-type plants.

Preparation and characterization of chitosan-based corn protein composites constructed with TG enzyme and their preservation performance on strawberries.

This study describes the synthesis of Chitosan - corn protein (CSZ-TG) composites using TG enzyme (TG) as a cross-linking agent and the preparation of chitosan-based composite membrane material (CSZEO-TG) by blending citrus essential oil (EO) with the synthesized CSZ-TG. The prepared composite membrane material was used for fresh strawberry preservation and characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, X-spectral diffraction, tensile properties, and water vapor and CO2 permeability. Scanning electron microscopy results showed a smooth surface of the composite membrane material after the addition of TG enzyme, while Fourier transforms infrared spectroscopy results showed a structural change of the composite membrane material after the addition of corn protein (Z). The tensile results showed an increase in the tensile strength of the composite membrane material after the addition of TG enzyme, while the flexibility of the composite membrane material was enhanced after the addition of EO. Compared with the pure chitosan membrane (CS), the water vapor and CO2 barrier properties of the composite membrane material after the addition of Z, TG, and EO did not change much, and they all showed better water vapor barrier properties. The results of the antioxidant analysis of the solution of the CSZEO-TG composite membrane material showed that the composite membrane material had efficient antioxidant properties. The effects of the composite film material on the storage period and quality of strawberries were evaluated by the indicators of weight loss, hardness, decay rate, soluble solids, titratable acid content, MDA content, and the content of four enzymes, SOD, POD, PPO and CAT. Comprehensive freshness data analysis showed that CSZEO-TG had the best freshness preservation performance and effectively extended the shelf life of strawberries.

Effect of gum Arabic, xanthan and carrageenan coatings containing antimicrobial agent on postharvest quality of strawberry: Assessing the physicochemical, enzyme activity and bioactive properties.

Effect of edible coatings of gum Arabic, carrageenan and xanthan gum containing lemon grass essential oil 1% w/v on postharvest quality of strawberry was studied under refrigeration for a period of 12 days. Results showed all the three coatings maintained fruit quality parameters during storage compared to control. Among all the coatings, carrageenan coated fruits showed delayed weight loss (10.1 to 8%), decay percentage (78.42 to 14.29%), retained ascorbic acid (0.15 to 0.27 g kg-1), antioxidant activity (18.17 to 25.85%), firmness (9.07 to 12.43 N), L* (32.38 to 40.42), a* (16.08 to 17.22) and b* (27.36 to 33.54). Carrageenan gum also showed lowest cellulase activity (0.03 units h-1 mg protein-1), pectin methylesterase activity (1.13 A620 min-1 mg protein-1) and ß-galactosidase activity (0.51 µmol min-1 mg protein-1), while showed maximum reduction in polygalacturonase activity (0.07 units h-1 mg protein-1) at the end of storage. Carrageenan gum was found effective in retention of anthocyanins and phenolic compounds during storage. Coatings loaded with antimicrobial agent inhibited psychrophilic bacteria, yeast and mold growth. It is concluded that carrageenan gum could better retain strawberry quality up to 12 days under refrigeration.

Exogenous phytosulfokine α (PSKα) applying delays senescence and relief decay in strawberry fruits during cold storage by sufficient intracellular ATP and NADPH availability.

Herein, we employed exogenous phytosulfokine α (PSKα) for delaying senescence and lessening decay in strawberry fruits during storage at 4 °C for 18 days. Our results showed that the strawberry fruits treated with 150 nM PSKα exhibited lower expression of poly-ADP-ribose polymerase 1 (PARP1) gene, leading to a higher intracellular NAD+ availability, beneficial for a sufficient provision of intracellular NADP+ with the activity of NAD kinase (NADK). Moreover, higher activities of glucose 6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), and methylenetetrahydrofolate dehydrogenase (MTHFD) may be the reason for the sufficient intracellular availability of NADPH in strawberry fruits treated with 150 nM PSKα. In addition, strawberry fruits treated with 150 nM PSKα exhibited a sufficient availability of ATP resulted from higher activities of succinate dehydrogenase (SDH) and cytochrome c oxidase (CCO). Therefore, our results indicate that exogenous PSKα could be beneficial for delaying senescence and reducing decay in strawberry fruits during cold storage.

Expression profiling of endo-xylanases during ripening of strawberry cultivars with contrasting softening rates. Influence of postharvest and hormonal treatments.

BACKGROUND: Softening is one of the main features that determine fruit quality during strawberry (Fragaria x ananassa, Duch.) ripening and storage. Being closely related to textural changes, the molecular and biochemical bases underlying strawberry cell-wall metabolism is a matter of interest. Here we investigated the abundance of transcripts encoding putative strawberry endo-xylanases in plant tissues, during fruit ripening and under postharvest and hormonal treatments. Total xylanase activity and expression of related genes in strawberry varieties with contrasting firmness were analyzed. RESULTS: FaXynA and FaXynC mRNA abundance was significantly higher than FaXynB in each plant tissue studied. Higher total xylanase activity was detected at the end of the ripening of the softer cultivar ('Toyonoka') in comparison with the firmer one ('Camarosa'), correlating with the abundance of FaXynA and FaXynC transcripts. Postharvest 1-methylcyclopropene treatment up-regulated FaXynA and FaXynC expressions. FaXynC mRNA abundance decreased with heat treatment but the opposite was observed for FaXynA. Calcium chloride treatment down-regulated FaXynA and FaXynC expression. Both genes responded differently to plant growth regulators' exposure. FaXynC expression was down-regulated by auxins and gibberellins treatment and up-regulated by abscisic acid. FaXynA was up-regulated by auxins, while no changes in mRNA levels were evident by abscisic acid and gibberellins treatment. Ethephon exposure did not change FaXynA and FaXynC expressions. CONCLUSION: New knowledge about the presence of xylanases in ripening strawberry fruit and their response to postharvest and hormonal treatments is provided. Our findings suggest a role for endo-xylanases in hemicelluloses depolymerization and possibly in strawberry fruit softening. © 2020 Society of Chemical Industry.

Contrasting dynamics in abscisic acid metabolism in different Fragaria spp. during fruit ripening and identification of the enzymes involved.

Abscisic acid (ABA) is a key hormone in non-climacteric Fragaria spp, regulating multiple physiological processes throughout fruit ripening. Its concentration increases during ripening, and it promotes fruit (receptacle) development. However, its metabolism in the fruit is largely unknown. We analyzed the concentrations of ABA and its catabolites at different developmental stages of strawberry ripening in diploid and octoploid genotypes and identified two functional ABA-glucosyltransferases (FvUGT71A49 and FvUGT73AC3) and two regiospecific ABA-8'-hydroxylases (FaCYP707A4a and FaCYP707A1/3). ABA-glucose ester content increased during ripening in diploid F. vesca varieties but decreased in octoploid F.×ananassa. Dihydrophaseic acid content increased throughout ripening in all analyzed receptacles, while 7'-hydroxy-ABA and neo-phaseic acid did not show significant changes during ripening. In the studied F. vesca varieties, the receptacle seems to be the main tissue for ABA metabolism, as the concentration of ABA and its metabolites in the receptacle was generally 100 times higher than in achenes. The accumulation patterns of ABA catabolites and transcriptomic data from the literature show that all strawberry fruits produce and metabolize considerable amounts of the plant hormone ABA during ripening, which is therefore a conserved process, but also illustrate the diversity of this metabolic pathway which is species, variety, and tissue dependent.

Identification of the Cytosolic Glucose-6-Phosphate Dehydrogenase Gene from Strawberry Involved in Cold Stress Response.

Glucose-6-phosphate dehydrogenase (G6PDH) plays an important role in plant stress responses. Here, five FaG6PDH sequences were obtained in strawberry, designated as FaG6PDH-CY, FaG6PDH-P1, FaG6PDH-P1.1, FaG6PDH-P2 and FaG6PDH-P0, which were divided into cytosolic (CY) and plastidic (P) isoforms based on the bioinformatic analysis. The respective FaG6PDH genes had distinct expression patterns in all tissues and at different stages of fruit development. Notably, FaG6PDH-CY was the most highly expressed gene among five FaG6PDH members, indicating it encoded the major G6PDH isoform throughout the plant. FaG6PDH positively regulated cold tolerance in strawberry. Inhibition of its activity gave rise to greater cold-induced injury in plant. The FaG6PDH-CY transcript had a significant increase under cold stress, similar to the G6PDH enzyme activity, suggesting a principal participant in response to cold stress. Further study showed that the low-temperature responsiveness (LTR) element in FaG6PDH-CY promoter can promote the gene expression when plant encountered cold stimuli. Besides, FaG6PDH-CY was involved in regulating cold-induced activation of antioxidant enzyme genes (FaSOD, FaCAT, FaAPX and FaGR) and RBOH-dependent ROS generation. The elevated FaG6PDH-CY enhanced ROS-scavenging capability of antioxidant enzymes to suppress ROS excessive accumulation and relieved the oxidative damage, eventually improving the strawberry resistance to cold stress.

Characterization of FcXTH2, a Novel Xyloglucan Endotransglycosylase/Hydrolase Enzyme of Chilean Strawberry with Hydrolase Activity.

Xyloglucan endotransglycosylase/hydrolases (XTHs) are cell wall enzymes with hydrolase (XEH) and/or endotransglycosylase (XET) activities. As they are involved in the modification of the xyloglucans, a type of hemicellulose present in the cell wall, they are believed to be very important in different processes, including growth, development, and fruit ripening. Previous studies suggest that XTHs might play a key role in development and ripening of Fragaria chiloensis fruit, and its characterization is pending. Therefore, in order to provide a biochemical characterization of the FcXTH2 enzyme to explain its possible role in strawberry development, the molecular cloning and the heterologous expression of FcXTH2 were performed. The recombinant FcXTH2 was active and displayed mainly XEH activity. The optimal pH and temperature are 5.5 and 37 °C, respectively. A KM value of 0.029 mg mL-1 was determined. Additionally, its protein structural model was built through comparative modeling methodology. The model showed a typically ß-jelly-roll type folding in which the catalytic motif was oriented towards the FcXTH2 central cavity. Using molecular docking, protein-ligand interactions were explored, finding better interaction with xyloglucan than with cellulose. The data provided groundwork for understanding, at a molecular level, the enzymatic mechanism of FcXTH2, an important enzyme acting during the development of the Chilean strawberry.

Cytosolic/Plastid Glyceraldehyde-3-Phosphate Dehydrogenase Is a Negative Regulator of Strawberry Fruit Ripening.

Cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC) and plastid glyceraldehyde-3-phosphate dehydrogenase (GAPCp) are key enzymes in glycolysis. Besides their catalytic function, GAPC/GAPCp participates in the regulation of plant stress response and growth and development. However, the involvement of GAPC/GAPCp in the regulation of fruit ripening is unclear. In this study, FaGAPC2 and FaGAPCp1 in strawberries were isolated and analyzed. FaGAPC2 and FaGAPCp1 transcripts showed high transcript levels in the fruit. Transient overexpression of FaGAPC2 and FaGAPCp1 delayed fruit ripening, whereas RNA interference promoted fruit ripening and affected fruit anthocyanins and sucrose levels. Change in the expression patterns of FaGAPC2 and FaGAPCp1 also influenced the expression of several glycolysis-related and ripening-related genes such as CEL1, CEL2, SS, ANS, MYB5, NCED1, ABI1, ALDO, PK, and G6PDH, and H2O2 level and reduced glutathione (GSH)/glutathione disulfide (GSSG) redox potential. Meanwhile, metabolomics experiments showed that transient overexpression of FaGAPCp1 resulted in a decrease in anthocyanins, flavonoids, organic acid, amino acids, and their derivatives. In addition, abscisic acid (ABA) and sucrose treatment induced the production of large amounts of H2O2 and inhibited the expression of FaGAPC2/FaGAPCp1 in strawberry fruit. These results revealed that FaGAPC2/FaGAPCp1 is a negative regulator of ABA and sucrose mediated fruit ripening which can be regulated by oxidative stress.

Enzymatic Synthesis of Modified Alternaria Mycotoxins Using a Whole-Cell Biotransformation System.

Reference standards for Alternaria mycotoxins are rarely available, especially the modified mycotoxins alternariol-3-glucoside (AOH-3-G), alternariol-9-glucoside (AOH-9-G), and alternariol monomethylether-3-glucoside (AME-3-G). To obtain these three glucosides as analytical standards for method development and method validation, alternariol and alternariol monomethylether were enzymatically glycosylated in a whole-cell biotransformation system using a glycosyltransferase from strawberry (Fragaria x ananassa), namely UGT71A44, expressed in Escherichia coli (E. coli). The formed glucosides were isolated, purified, and structurally characterized. The exact amount of the isolated compounds was determined using high-performance liquid chromatography with UV-detection (HPLC-UV) and quantitative nuclear resonance spectroscopy (qNMR). This method has proved to be highly effective with biotransformation rates of 58% for AOH-3-G, 5% for AOH-9-G, and 24% for AME-3-G.

Genome-Wide Characterization of Snf1-Related Protein Kinases (SnRKs) and Expression Analysis of SnRK1.1 in Strawberry.

The plant sucrose nonfermenting 1 (SNF1)-related protein kinases (SnRKs) are key regulators in the interconnection of various signaling pathways. However, little is known about the SnRK family in strawberries. In this study, a total of 26 FvSnRKs including one FvSnRK1, nine FvSnRK2s and 16 FvSnRK3s were identified from the strawberry genome database. They were respectively designated as FvSnRK1.1, FvSnRK2.1 to FvSnRK2.9 and FvSnRK3.1 to FvSnRK3.16, according to the conserved domain of each subfamily and multiple sequence alignment with Arabidopsis. FvSnRK family members were unevenly distributed in seven chromosomes. The number of exons or introns varied among FvSnRK1s, FvSnRK2s and FvSnRK3s, but highly conserved in the same subfamily. The FvSnRK1.1 had 10 exons. Most of FvSnRK2s had nine exons or eight introns, except FvSnRK2.4, FvSnRK2.8 and FvSnRK2.9. FvSnRK3 genes were divided into intron-free and intron-harboring members, and the number of introns in intron-harboring group ranged from 11 to 15. Moreover, the phylogenetic analysis showed SnRK1, SnRK2 and SnRK3 subfamilies respectively clustered together in spite of the different species of strawberry and Arabidopsis, indicating the genes were established prior to the divergence of the corresponding taxonomic lineages. Meanwhile, conserved motif analysis showed that FvSnRK sequences that belonged to the same subgroup contained their own specific motifs. Cis-element in promoter and expression pattern analyses of FvSnRK1.1 suggested that FvSnRK1.1 was involved in cold responsiveness, light responsiveness and fruit ripening. Taken together, this comprehensive analysis will facilitate further studies of the FvSnRK family and provide a basis for the understanding of their function in strawberry.

Nitrate reductase rather than nitric oxide synthase activity is involved in 24-epibrassinolide-induced nitric oxide synthesis to improve tolerance to iron deficiency in strawberry (Fragaria × annassa) by up-regulating the ascorbate-glutathione cycle.

Involvement of nitrate reductase (NR) and nitric oxide synthase (NOS)-like enzyme in 24-epibrassinolide (EB)-triggered nitric oxide (NO) synthesis to improve iron deficiency (ID) tolerance in strawberry plants was studied. EB was sprayed to strawberry plants every two days for two weeks. Then, the EB-treated plants were pre-treated with inhibitors of NR, tungstate, or NOS, L-NAME for 3 h. During the first three weeks, Fe was supplied as 100 µM EDTA-Fe or FeSO4 to Fe-sufficient or Fe-deficient plants, respectively. Thereafter, plants were subjected for further three weeks to control (100 µM EDTA-Fe) and Fe deficiency (ID; without Fe). ID reduced biomass, chlorophyll, and chlorophyll fluorescence, while increased oxidative stress parameters, ascorbate (AsA), glutathione (GSH), endogenous NO, and the activities of NR, NOS, and antioxidant enzymes. Pre-treatments with EB and EB + SNP improved ID tolerance of strawberry by improving leaf Fe2+, plant growth, and antioxidant enzyme activities, and causing a further elevation in AsA, GSH, NO, NR and NOS. L-NAME application reversed NOS activity, but it did not eliminate NO, however, tungstate application reversed both NR activity and NO synthesis in plants exposed to ID + EB, suggesting that NR is the main contributor of EB-induced NO synthesis to improve ID tolerance in strawberry plants.

The protein kinase FaSnRK1α regulates sucrose accumulation in strawberry fruits.

In strawberry, sucrose is the major form of carbohydrate translocated from the leaves to the fruits and plays an important role in fruit ripening. As a conserved energy sensor, sucrose nonfermenting-1 (SNF1)-related kinase 1 (SnRK1) plays an important role in plant carbon metabolism. However, evidence that SnRK1 regulates sucrose accumulation in fruits is lacking. In this study, we transiently expressed FaSnRK1α in strawberry fruits and found that overexpression (OE) of the FaSnRK1α gene significantly increased the sucrose content, whereas repression of FaSnRK1α by RNA interference (RNAi) decreased the sucrose content. Further analysis revealed that FaSnRK1α increased the expression of FaSUS1 and FaSUS3 as well as the activity of sucrose synthase (SUS; EC 2.4.1.13) and that FaSPS1 expression and sucrose phosphate synthase (SPS; EC 2.4.1.14) activity were strongly downregulated, which decreased the accumulation of sucrose. However, the expression of FaSPS3, which is reported to contribute to sucrose accumulation, was induced by FaSnRK1α, and FaNI expression and invertase (INV; EC 3.2.1.26) activity were upregulated by FaSnRK1α. In addition, FaSnRK1α positively upregulated the expression of the sucrose transporter (SUT) genes FaSUT1 and FaSUT5 and interacted with FaSUS1, FaSPS1 and FaSPS3 proteins but not with FaSUS3, FaNI, FaSUT1 or FaSUT5 proteins. Overall, FaSnRK1α systematically regulates the expression of the genes and activities of key enzymes involved in the sucrose metabolic pathway and promotes the long-distance transport of sucrose, thereby increasing sucrose accumulation and ultimately promoting fruit ripening. However, the mechanisms by which sucrose transport and degradation are regulated by SnRK1 warrant additional research.

Discovery and characterization of tannase genes in plants: roles in hydrolysis of tannins.

Plant tannins, including condensed tannins (CTs) and hydrolyzable tannins (HTs), are widely distributed in the plant kingdom. To date, tannase (TA) - is a type of tannin acyl-hydrolase hydrolyzing HTs, CT monomer gallates and depsides - has been reported in microbes only. Whether plants express TA remains unknown. Herein, we report plant TA genes. A native Camellia sinensis TA (CsTA) is identified from leaves. Six TAs are cloned from tea, strawberry (Fragaria × ananassa, Fa) and four other crops. Biochemical analysis shows that the native CsTA and six recombinant TAs hydrolyze tannin compounds, depsides and phenolic glycosides. Transcriptional and metabolic analyses reveal that the expression of CsTA is oppositely associated with the accumulation of galloylated catechins. Moreover, the transient overexpression and RNA interference of FaTA are positively associated with the accumulation of ellagitannins in strawberry fruit. Phylogenetic analysis across different kingdoms shows that 29 plant TA homologs are clustered as a plant-specific TA clade in class I carboxylesterases. Further analysis across the angiosperms reveals that these TA genes are dispersed in tannin-rich plants, which share a single phylogenetic origin c. 120 million yr ago. Plant TA is discovered for the first time in the plant kingdom and is shown to be valuable to improve tannin compositions in plants.

Higher expression of the strawberry xyloglucan endotransglucosylase/hydrolase genes FvXTH9 and FvXTH6 accelerates fruit ripening.

Fruit softening in Fragaria (strawberry) is proposed to be associated with the modification of cell wall components such as xyloglucan by the action of cell wall-modifying enzymes. This study focuses on the in vitro and in vivo characterization of two recombinant xyloglucan endotransglucosylase/hydrolases (XTHs) from Fragaria vesca, FvXTH9 and FvXTH6. Mining of the publicly available F. vesca genome sequence yielded 28 putative XTH genes. FvXTH9 showed the highest expression level of all FvXTHs in a fruit transcriptome data set and was selected with the closely related FvXTH6 for further analysis. To investigate their role in fruit ripening in more detail, the coding sequences of FvXTH9 and FvXTH6 were cloned into the vector pYES2 and expressed in Saccharomyces cerevisiae. FvXTH9 and FvXTH6 displayed xyloglucan endotransglucosylase (XET) activity towards various acceptor substrates using xyloglucan as the donor substrate. Interestingly, FvXTH9 showed activity of mixed-linkage glucan:xyloglucan endotransglucosylase (MXE) and cellulose:xyloglucan endotransglucosylase (CXE). The optimum pH of both FvXTH9 and FvXTH6 was 6.5. The prediction of subcellular localization suggested localization to the secretory pathway, which was confirmed by localization studies in Nicotiana tabacum. Overexpression showed that Fragaria × ananassa fruits infiltrated with FvXTH9 and FvXTH6 ripened faster and showed decreased firmness compared with the empty vector control pBI121. Thus FvXTH9 and also FvXTH6 might promote strawberry fruit ripening by the modification of cell wall components.

Dissipation and the effects of thidiazuron on antioxidant enzyme activity and malondialdehyde content in strawberry.

BACKGROUND: Increasing numbers of fruit swelling agents have been used to improve the fruit rate and production yield of strawberries in recent years. The abuse of fruit swelling agents could lead to an increase in the deformation rate and abnormal coloration of strawberry and a decrease in quality at harvest. Therefore, understanding the harmful effects of fruit swelling agents on strawberry will provide guidance for their reasonable use. RESULTS: The residual determination method for measuring thidiazuron (TDZ) in strawberry was developed and validated by liquid chromatography and tandem mass spectrometry (LC-MS/MS). The recoveries of TDZ in strawberry were 97.9-108.5% with relative standard deviations of 0.9% to 5.3%. The dissipation rates of TDZ were different in strawberries cultivated under field and indoor conditions due to the differences in temperature and humidity. The ascorbic acid content increased when TDZ was applied at 2 mg kg-1 . The SOD (superoxide dismutase), POD (peroxidase) and CAT (catalase) activities of strawberry tended to decrease and subsequently increase following the application of TDZ, and the opposite changes occurred on the malondialdehyde (MDA) content of TDZ-treated strawberry. CONCLUSIONS: The analytical method for measuring TDZ in strawberry that was developed was suitable for dissipation studies on this compound. Antioxidant enzyme activities and the MDA content of strawberry were altered, and some reverse effects, such as membrane damage, were inhibited when TDZ was applied. The data obtained in this study might provide suggestions to reduce the adverse effects of TDZ on strawberry and may help to guide the safe and proper use of TDZ in strawberry. © 2019 Society of Chemical Industry.

Genome-wide identification, evolution, and molecular characterization of the PP2C gene family in woodland strawberry.

The protein phosphatase 2C (PP2C) gene family is one of the momentous and conserved plant-specific gene families, known to participate in cellular processes via reversible protein phosphorylation and regulates signal transduction in eukaryotic organisms. Recently, PP2Cs were identified in Arabidopsis and maize, however, the whole-genome analysis of PP2C in strawberry has not yet been reported. In the current research, we found 62 PP2C-encoding genes in total from the strawberry genome. Further, the phylogenetic analysis categorized FvPP2C genes into twelve subgroups with significant structural conservation based on conserved domain and amino acid sequence. Moreover, we observed a strong signature of purifying selection between the comparison of orthologous gene pairs of strawberry and Arabidopsis. The comparison of RNA-sequence (RNA-seq) data published on various vegetative and reproductive tissues of strawberry plant suggested the significant role of FvPP2C genes in organ development. The qRT-PCR validation of thirty FvPP2C genes indicated their critical tolerance-related role under abiotic stress stimuli in strawberry. Finally, the subcellular localization of FvPP2C51 gene proves that it resides and stimulates its function in the nucleus. Our findings provide an overview of the identification of strawberry FvPP2C gene family and demonstrate their critical role in tissue-specific response and abiotic stress-tolerance, thereby, intimating their significance in the strawberry molecular breeding for the resistant cultivars.