Cell Wall Disassembly Research Articles

Storage temperature affects metabolism of sweet corn

Temperature is one of the critical factors affecting the postharvest storage quality of sweet corn, but effects of storage temperatures have not been elucidated at the molecular level. In this study, the effects of storage at 0 °C, 4 °C and 20 °C, on quality and associated effects on the transcriptome and metabolome were investigated. Compared with storage at 20 °C, cobs stored at 0 °C and 4 °C had up-regulated expression of genes associated with cell wall disassembly (PG, PE, BMY, β-Gal) and down-regulated expression of genes involved in lignin synthesis, the up-regulation of SPP, SPS and SUS involved in sucrose synthesis, and the down-regulation of IN, FK, HXK and PFK involved in sucrose catabolism. Most of the genes involved in ethylene biosynthesis and signaling were inhibited at 0 °C, except for ERFs, while abscisic acid synthesis and signal transduction were most active at this temperature. Near-freezing temperature also maintains the integrity of the cell membrane by inhibiting the expression of PLD, PLC and LOX, which encode membrane lipid-degrading enzymes. Metabolomics and combined analysis showed that the contents of lysophospholipid, glutathione and L-ascorbic acid in sweet corn increased at low temperatures. These results provide insight into the effects of low temperatures on sweet corn quality.

Characterization of FchAGL9 and FchSHP, two MADS-boxes related to softening of Fragaria chiloensis fruit

Fragaria chiloensis is a Chilean native species that softens intensively during its ripening. Its softening is related to cell wall disassembly due to the participation of cell wall degrading enzymes. Softening of F. chiloensis fruit can be accelerated by ABA treatment which is accompanied by the increment in the expression of key cell wall degrading genes, however the molecular machinery involved in the transcriptional regulation has not been studied until now. Therefore, the participation of two MADS-box transcription factors belonging to different subfamilies, FchAGL9 and FchSHP, was addressed. Both TFs are members of type-II MADS-box family (MIKC-type) and localized in the nucleus. FchAGL9 and FchSHP are expressed only in flower and fruit tissues, rising as the fruit softens with the highest expression level at C3–C4 stages. EMSA assays demonstrated that FchAGL9 binds to CArG sequences of RIN and SQM, meanwhile FchSHP interacts only with RIN. Bimolecular fluorescence complementation and yeast two-hybrid assays confirmed FchAGL9-FchAGL9 and FchAGL9-FchSHP interactions. Hetero-dimer structure was built through homology modeling concluding that FchSHP monomer binds to DNA. Functional validation by Luciferase-dual assays indicated that FchAGL9 transactivates FchRGL and FchPG's promoters, meanwhile FchSHP transactivates those of FchEXP2, FchRGL and FchPG. Over-expression of FchAGL9 in C2 F. chiloensis fruit rises FchEXP2 and FchEXP5 transcripts, meanwhile the over-expression of FchSHP also increments FchXTH1 and FchPL; in both cases there is a down-regulation of FchRGL and FchPG. In summary, we provided evidence of FchAGL9 and FchSHP participating in the transcription regulation associated to F. chiloensis's softening.

Cell wall disassembly events accompanying softening in the core and external zones of the strawberry fruit receptacle

Softening is one of the most important changes determining the quality and postharvest life of strawberry. Though some studies have characterized the cell wall modifications accompanying firmness loss during storage, the reports on polysaccharide disassembly are widely variable and in some cases contradictory. To further understand strawberry postharvest softening, conducted a comprehensive cell wall analysis including polymer solubilization, depolymerization and composition of the Core and External zones of fruit´s receptacle during storage. Core tissues were firmer and stiffer and softened more than the External receptacle zone. Wall dismantling was remarkably different depending on the region evaluated. Core tissues showed greater losses and size downshift of hemicelluloses as well as more extensive depolymerization of pectin. Instead, softening in the External region occurred with limited depolymerization and with massive loss of arabinose and xylose. Xyloglucans showed active turnover in the Core, whereas xylans were the most degraded hemicelluloses in the External region. Results show that each discrete region of the strawberry fruit receptacle has a unique sequence of biochemical wall dismantling events leading to postharvest softening.

Dicyclohexylcarbodiimide and disodium succinate regulate the pulp softening and breakdown in fresh longan by modulating the metabolism of cell wall polysaccharides

Cell wall polysaccharides (CWP) are an important component of cell structure, and its content variation is a primary factor affecting the texture of fresh longan. The influences of dicyclohexylcarbodiimide (DCC) and disodium succinate (DS) treatments on the pulp softening and pulp breakdown, and their relation to CWP metabolism in pulp of longan fruit were explored. Compared with the control longan, DCC-treated fruit showed lower value of pulp firmness, higher index of pulp breakdown, higher expression levels of cell wall disassembly enzymes (CWDEs)-related genes (DlPME1, DlPME3, DlPG1, Dlβ-Gal1, Dlβ-Gal16, DlCx7, DlXET3, DlXET27), and higher activities of CWDEs including pectin methyl esterase (PME), polygalacturonase (PG), β-galactosidase (β-Gal), cellulase (Cx), xyloglucan endotransglycosylase (XET), lower values of cell wall materials, and lower levels of CWP including ionic-soluble pectin, covalent-soluble pectin, cellulose, and hemicellulose. Whereas, the opposite effects were exhibited in DS-treated fruit. The above data indicate that DCC-hastened longan pulp softening and pulp breakdown was due to the up-regulating expressions of CWDEs-related genes, the activating activities of CWDEs, and the accelerating CWP disassembly by DCC treatment. Whereas, DS repressed longan pulp softening and pulp breakdown through down-regulating the expressions of CWDEs-related genes, inhibiting CWDEs activities, and reducing CWP degradation.

Transcriptome analysis integrated with changes in cell wall polysaccharides of different fresh-cut chili pepper cultivars during storage reveals the softening mechanism

Cell wall disassembly and transcriptomic changes during storage of two fresh-cut chili pepper cultivars displaying contrasting softening rates were investigated. Results showed that Hangjiao No. 2 (HJ-2) softened more rapidly than Lafeng No. 3 (LF-3). Compared with LF-3, HJ-2 had a higher content of WSP, more side chains of RG-I in three pectin fractions, and higher activities of PME, PL, and β-Gal at day-0. During storage, HJ-2 showed more markable pectin solubilization, more severe degradation in CSP and NSP, and greater loss of side chains from RG-I in three pectin fractions, which were correlated with increased activities of PG and α-L-Af. Furthermore, the higher up-regulation of PG (LOC107870605, LOC107851416) and α-L-Af (LOC107848776, LOC107856612) were screened in HJ-2. In conclusion, the different softening rate between cultivars was not only due to the fundamental differences in pectin structure but also pectin degradation regulated by related enzymes and gene expression levels.

Parasitic dodder expresses an arsenal of secreted cellulases with multi-substrate specificity during host invasion

Cuscuta campestris is a common and problematic parasitic plant which relies on haustoria to connect to and siphon nutrients from host plants. Glycoside hydrolase family 9 (GH9) cellulases (EC 3.2.1.4) play critical roles in plant cell wall biosynthesis and disassembly, but their roles during Cuscuta host invasion remains underexplored. In this study, we identified 22 full-length GH9 cellulase genes in C. campestris genome, which encoded fifteen secreted and seven membrane-anchored cellulases that showed distinct phylogenetic relationships. Expression profiles suggested that some of the genes are involved in biosynthesis and remodeling of the parasite's cell wall during haustoriogenesis, while other genes encoding secreted B- and C-type cellulases are tentatively associated with degrading host cell walls during invasion. Transcriptomic data in a host-free system and in the presence of susceptible or partially resistant tomato hosts, showed for especially GH9B7, GH9B11 and GH9B12 a shift in expression profiles in the presence of hosts, being more highly expressed during host attachment, indicating that Cuscuta can tune cellulase expression in response to a host. Functional analyses of recombinant B- and C-type cellulases showed endoglucanase activities over wide pH and temperature conditions, and activities towards multiple cellulose and hemicellulose substrates. These findings improve our understanding of host cell wall disassembly by Cuscuta, and cellulase activity towards broad substrate range potentially explain its wide host range. This is the first study to provide a broad biochemical insight into Cuscuta GH9 cellulases, which based on our study may have potential applications in industrial bioprocessing.

γ-Aminobutyric acid delays fruit softening in postharvest kiwifruit by inhibiting starch and cell wall degradation

The effects of 10 mM γ-aminobutyric acid (GABA) treatment on fruit softening, starch degradation, cell wall disassembly in cold-stored ‘Hongyang’ kiwifruit after harvest were investigated. The results depicted that GABA treatment maintained firmness and delayed fruit softening during storage by retaining high starch content due to the inhibition of the transcription levels of related starch-degrading genes, such as AcGWD, AcPWD, and AcBAMs. In addition, compared with the control, higher level of chelated-soluble pectin, Na2CO3-soluble pectin, hemicellulose and cellulose content but lower water-soluble pectin content were observed in the treated kiwifruit as GABA suppressed cell wall breakdown by down-regulating the expression of AcPE, AcPG, AcBGALs, and AcXTHs. In summary, our results indicated that exogenous GABA contributes to maintaining fruit firmness, delaying softening in postharvest kiwifruit by inhibiting starch degradation and preserving cell wall metabolism.

Expansin SlExp1 and endoglucanase SlCel2 synergistically promote fruit softening and cell wall disassembly in tomato.

Fruit softening, an irreversible process that occurs during fruit ripening, can lead to losses and waste during postharvest transportation and storage. Cell wall disassembly is the main factor leading to loss of fruit firmness, and several ripening-associated cell wall genes have been targeted for genetic modification, particularly pectin modifiers. However, individual knockdown of most cell wall-related genes has had minimal influence on cell wall integrity and fruit firmness, with the notable exception of pectate lyase. Compared to pectin disassembly, studies of the cell wall matrix, the xyloglucan-cellulose framework, and underlying mechanisms during fruit softening are limited. Here, a tomato (Solanum lycopersicum) fruit ripening-associated α-expansin (SlExpansin1/SlExp1) and an endoglucanase (SlCellulase2/SlCel2), which function in the cell wall matrix, were knocked out individually and together using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9-mediated genome editing. Simultaneous knockout of SlExp1 and SlCel2 enhanced fruit firmness, reduced depolymerization of homogalacturonan-type pectin and xyloglucan, and increased cell adhesion. In contrast, single knockouts of either SlExp1 or SlCel2 did not substantially change fruit firmness, while simultaneous overexpression of SlExp1 and SlCel2 promoted early fruit softening. Collectively, our results demonstrate that SlExp1 and SlCel2 synergistically regulate cell wall disassembly and fruit softening in tomato.

Aminoethoxyvinylglycine Reduces Preharvest Fruit Drop and Fruit Ethylene Evolution in ‘Red Delicious’ Apple but Affects Fruit Size and Quality Inconsistently

Aminoethoxyvinylglycine (AVG) is widely used in commercial apple (Malus ×domestica Borkh.) production to reduce preharvest fruit drop (PFD) and delay ripening for harvest management. Recently, the maximum allowable concentration of AVG was doubled (up to 264 mg⋅L−1). Reports of the relationship between the AVG concentration and fruit growth, size, and quality have been contradictory. We evaluated the relationship between the AVG concentration and PFD, fruit size, fruit quality, and expression of ethylene signaling-related and cell wall modification-related genes. Experiments were conducted in 2019 and 2020 using mature ‘Red Delicious’ in western North Carolina. The AVG treatments [0 and 132 (AVG-1x) and 264 mg⋅L−1 (AVG-2x)] were applied 3 weeks before the expected harvest. The AVG treatments reduced fruit drop and internal ethylene concentration relative to the control in both years. There was no difference in drop between AVG-1x and AVG-2x applications. Only in 2020 did AVG treatments delay fruit softening and starch hydrolysis and reduce soluble solids concentration. There were no effects on red fruit color development. Fruit size was unaffected by AVG in 2019, but it was reduced in 2020 with the AVG-2x application. AVG reduced ethylene synthesis and altered signaling, evidenced by decreased relative expression of genes related to ethylene signaling (ARGOS1, ARGOS2). AVG applications also reduced the expression of EXPA8;1, suggesting that reduced cell wall disassembly was associated with a reduction in fruit softening. These results indicate that preharvest applications of 132 mg⋅L−1 AVG effectively reduced PFD via altering ethylene evolution and signaling. Use of a higher AVG concentration was of limited benefit.

Insights into the Genes Involved in ABA Biosynthesis and Perception during Development and Ripening of the Chilean Strawberry Fruit.

Hormones act as master ripening regulators. In non-climacteric fruit, ABA plays a key role in ripening. Recently, we confirmed in Fragaria chiloensis fruit that in response to ABA treatment the fruit induces ripening-associated changes such as softening and color development. In consequence of these phenotypic changes, transcriptional variations associated with cell wall disassembly and anthocyanins biosynthesis were reported. As ABA stimulates the ripening of F. chiloensis fruit, the molecular network involved in ABA metabolism was analyzed. Therefore, the expression level of genes involved in ABA biosynthesis and ABA perception was quantified during the development of the fruit. Four NCED/CCDs and six PYR/PYLs family members were identified in F. chiloensis. Bioinformatics analyses confirmed the existence of key domains related to functional properties. Through RT-qPCR analyses, the level of transcripts was quantified. FcNCED1 codifies a protein that displays crucial functional domains, and the level of transcripts increases as the fruit develops and ripens, in parallel with the increment in ABA. In addition, FcPYL4 codifies for a functional ABA receptor, and its expression follows an incremental pattern during ripening. The study concludes that FcNCED1 is involved in ABA biosynthesis; meanwhile, FcPYL4 participates in ABA perception during the ripening of F. chiloensis fruit.

CRISPR/Cas9 editing of the polygalacturonase FaPG1 gene improves strawberry fruit firmness.

Firmness is one of the most important fruit quality traits in strawberries. The postharvest shelf life of this soft fruit is highly limited by the loss of firmness, where cell wall disassembly plays an important role. Previous studies demonstrated that the polygalacturonase FaPG1 has a key role in remodelling pectins during strawberry softening. In this study, FaPG1 knockout strawberry plants have been generated using the CRISPR/Cas9 system delivered via Agrobacterium tumefaciens. Ten independent lines, cv. "Chandler", were obtained, and all of them were successfully edited as determined by PCR amplification and T7 endonuclease assay. The targeted mutagenesis insertion and deletion rates were analyzed using targeted deep sequencing. The percentage of edited sequences varied from 47% up to almost 100%, being higher than 95% for seven of the selected lines. Phenotypic analyses showed that 7 out of the eight lines analyzed produced fruits significantly firmer than the control, ranging from 33 to 70% increase in firmness. There was a positive relationship between the degree of FaPG1 editing and the rise in fruit firmness. Minor changes were observed in other fruit quality traits, such as colour, soluble solids, titratable acidity or anthocyanin content. Edited fruits showed a reduced softening rate during postharvest, displayed a reduced transpirational water loss, and were less damaged by Botrytis cinerea inoculation. The analysis of four potential off-target sites revealed no mutation events. In conclusion, editing the FaPG1 gene using the CRISPR/Cas9 system is an efficient method for improving strawberry fruit firmness and shelf life.

Cell wall disassembly, metabolome and transcriptome analysis in sweet cherry fruit with induced surface pitting

Surface pitting is the main quality problem that develops during prolonged storage of sweet cherries. It appears as one or more depressions on the surface of the fruit and is associated with the collapse of cells under the skin of the fruit. However, this physiological disorder is not shown in all cultivars with the same intensity. This research aims to analyze the cell wall disassembly of two cultivars displaying contrasting susceptibility to damage after pitting induction. In addition, we evaluated the metabolomic and transcriptomic changes in sweet cherries during cold storage. Our results show that damage to ‘Sweetheart’ fruit was more severe than damage to ‘Bing’ fruit. No differences were observed in the cell wall composition between nonpitted and pitted cherries; however, the varietal differences during cold storage were the most significant. The resistant cultivar ‘Bing’ showed a longer sidechain of RG-I; instead, ‘Sweetheart’ PME and PG activity was more marked at the beginning of cold storage. The metabolomics analysis revealed several compounds related to the abiotic stress response, such as 3-O-coumaroyl-D-quinic acid, chlorogenic acid, GABA and β-sitosterol. Furthermore, transcriptomics showed a higher expression of stress-related hormones in the susceptible cultivar and cell wall remodeling-related genes in the resistant cultivar. In conclusion, the contrasting susceptivity to surface pitting in sweet cherries can be attributed to the varietal response to cold storage rather than the mechanical stress of pitting induction.

Pectate Lyase Gene VvPL1 Plays a Role in Fruit Cracking of Table Grapes

Fruit cracking seriously affects the commercial value of table grapes. To explore whether cell wall disassembly influences grape berry cracking, first, the differences in the cell wall metabolism were compared between cracking-resistant "Shennongjinhuanghou" (SN) and cracking-susceptible "Xiangfei" (XF) varieties. Our results showed that cell wall disassembly events were extremely different between "SN" and "XF." The cracking-resistant "SN" had a higher pectinmethylesterase activity in the early stage and lower polygalacturonase, β-galactosidase, pectate lyase, and cellulase activities from veraison, cooperatively yielding higher ionically bound pectin, covalently bound pectin, hemicellulose, and lower water-soluble pectin, leading to a stronger skin break force and elasticity and conferring "SN" with higher cracking resistance. Furthermore, the function of the VvPL1 gene in fruit cracking was verified by heterologously transforming tomatoes. The transgenic experiment showed that overexpressed fruits had a higher activity of pectate lyase from the breaking stage and a lower level of covalently bound pectin, ionically bound pectin, cellulose, and hemicellulose and a higher level of water-soluble pectin at the red ripe stage, which resulted in a significantly reduced skin break force and flesh firmness and increased fruit cracking incidences. In conclusion, our results demonstrated that the cracking susceptibility of the grape berry is closely related to cell wall disassembly events and VvPL1 plays an important role in fruit cracking.

The difference of the cell wall metabolism between ‘Fuyan’ and ‘Dongbi’ longans and its relationship with the pulp breakdown

The aims of present works were to explore the difference in pulp breakdown of ‘Fuyan’ and ‘Dongbi’ longans and its relationship with cell wall metabolism. Comparison with ‘Fuyan’ longan fruit, postharvest ‘Dongbi’ longan fruit showed lower pulp breakdown index, lower activities of PE, PG, cellulase, β-Gal, XET, and lower expression levels of their corresponding genes. In addition, higher levels of cell wall polysaccharides including ISP, CSP, cellulose and hemicellulose were exhibited in ‘Dongbi’ longan pulp. These findings implied that, the reduced activities of enzymes and the down-regulated expressions of genes-involved in cell wall disassembly were shown in ‘Dongbi’ longan pulp, which might reduce the dissolution of polysaccharides and maintain a higher structural integrity in ‘Dongbi’ longan pulp cell wall, and consequently the mitigated pulp breakdown was displayed in ‘Dongbi’ longan during storage.

Elucidating the role of SlXTH5 in tomato fruit softening

Fruit softening in tomato (Solanum lycopersicum) is closely associated with cell wall disassembly, which is brought about through the action of a range of cell wall structure-related enzymes and other proteins such as expansins. Xyloglucan endotransglucosylase/hydrolase (XTH) (EC 2.4.1.207 and/or EC 3.2.1.151) has been proposed to be key player involved in xyloglucan metabolism. SlXTH5 showed the highest expression level among all SlXTHs during tomato ripening. In this study, the role of SlXTH5 involved in tomato softening was investigated in CRISPR-based knockout mutants of SlXTH5. Loss-of-function of SlXTH5 in transgenic tomato lines resulted in slightly firmer fruit pericarp, but significantly decreased their color index compared with azygous wild type (WT) control fruits. Increased paste viscosity was detected in CRISPR mutants, indicating that the activity of SlXTH5 is responsible for maintaining cell wall structural integrity. Immunocytochemistry studies were performed using the monoclonal antibody probe LM25 to examine the localization and distribution of xyloglucan in the pericarp cells of the CRISPR mutant fruits. The data indicated more xyloglucan was retained in the pericarp of CRISPR mutant fruit than in WT control fruit. This study revealed the link between SlXTH5 and xyloglucan metabolism and indicated the potential of manipulating SlXTH5 to regulate fruit softening. .

Botrytis cinerea infection accelerates ripening and cell wall disassembly to promote disease in tomato fruit.

Postharvest fungal pathogens benefit from the increased host susceptibility that occurs during fruit ripening. In unripe fruit, pathogens often remain quiescent and unable to cause disease until ripening begins, emerging at this point into destructive necrotrophic lifestyles that quickly result in fruit decay. Here, we demonstrate that one such pathogen, Botrytis cinerea, actively induces ripening processes to facilitate infections and promote disease in tomato (Solanum lycopersicum). Assessments of ripening progression revealed that B. cinerea accelerated external coloration, ethylene production, and softening in unripe fruit, while mRNA sequencing of inoculated unripe fruit confirmed the corresponding upregulation of host genes involved in ripening processes, such as ethylene biosynthesis and cell wall degradation. Furthermore, an enzyme-linked immunosorbent assay (ELISA)-based glycomics technique used to assess fruit cell wall polysaccharides revealed remarkable similarities in the cell wall polysaccharide changes caused by both infections of unripe fruit and ripening of healthy fruit, particularly in the increased accessibility of pectic polysaccharides. Virulence and additional ripening assessment experiments with B. cinerea knockout mutants showed that induction of ripening depends on the ability to infect the host and break down pectin. The B. cinerea double knockout Δbc polygalacturonase1 Δbc polygalacturonase2 lacking two critical pectin degrading enzymes was incapable of emerging from quiescence even long after the fruit had ripened at its own pace, suggesting that the failure to accelerate ripening severely inhibits fungal survival on unripe fruit. These findings demonstrate that active induction of ripening in unripe tomato fruit is an important infection strategy for B. cinerea.

Hydrogen-rich water delays fruit softening and prolongs shelf life of postharvest okras

The effect of hydrogen-rich water (HRW) treatment on softening, cell wall components and cell wall metabolic genes in okras after harvest was studied. The results showed that HRW treatment could maintain fruit firmness and delay softening, thereby prolonging shelf life in okras during storage. The treated okras displayed significantly lower levels water- and chelate-soluble pectins while higher contents of Na2CO3-soluble pectin, hemicellulose and cellulose. The cell wall biosynthesis was maintained by HRW treatment via up-regulating genes involved in biosynthesis of pectin, hemicellulose and cellulose at the beginning of storage. On the contrary, the treatment could inhibit the cell wall disassembly due to the down-regulation of numerous cell wall degradative genes including AePME, AeGAL and AeCX at the end of storage. Taken together, our results suggested that HRW treatment delayed softening and extended shelf life in postharvest okras through modifying cell wall biosynthesis and disassembly at different times of storage.

Transcriptomic analysis of CO2-treated strawberries (Fragaria vesca) with enhanced resistance to softening and oxidative stress at consumption.

One of the greatest threats to wild strawberries (Fragaria vesca Mara des Bois) after harvest is the highly perishability at ambient temperature. Breeders have successfully met the quality demands of consumers, but the prevention of waste after harvest in fleshy fruits is still pending. Most of the waste is due to the accelerated progress of senescence-like process after harvest linked to a rapid loss of water and firmness at ambient temperature. The storage life of strawberries increases at low temperature, but their quality is limited by the loss of cell structure. The application of high CO2 concentrations increased firmness during cold storage. However, the key genes related to resistance to softening and cell wall disassembly following transference from cold storage at 20°C remain unclear. Therefore, we performed RNA-seq analysis, constructing a weighted gene co-expression network analysis (WGCNA) to identify which molecular determinants play a role in cell wall integrity, using strawberries with contrasting storage conditions, CO2-cold stored (CCS), air-cold stored (ACS), non-cold stored (NCS) kept at ambient temperature, and intact fruit at harvest (AH). The hub genes associated with the cell wall structural architecture of firmer CO2-treated strawberries revealed xyloglucans stabilization attributed mainly to a down-regulation of Csl E1, XTH 15, Exp-like B1 and the maintenance of expression levels of nucleotide sugars transferases such as GMP and FUT as well as improved lamella integrity linked to a down-regulation of RG-lyase, PL-like and PME. The preservation of cell wall elasticity together with the up-regulation of LEA, EXPA4, and MATE, required to maintain cell turgor, is the mechanisms controlled by high CO2. In stressed air-cold stored strawberries, in addition to an acute softening, there is a preferential transcript accumulation of genes involved in lignin and raffinose pathways. Non-cold stored strawberries kept at 20°C after harvest are characterized by an enrichment in genes mainly involved in oxidative stress and up-expression of genes involved in jasmonate biosynthesis. The present results on transcriptomic analysis of CO2-treated strawberries with enhanced resistance to softening and oxidative stress at consumption will help to improve breeding strategies of both wild and cultivated strawberries.

Characteristics of histological alterations and hormone-variations in floral tissues of edible daylily (Hemerocallis citrina) buds during postharvest senescence

Daylily (Hemerocallis citrina Baroni.) buds suffer rapid senescence after harvest. In order to understand morphological and physiological changes of daylily buds during postharvest senescence, histological alterations and hormonal variations of different floral organs (receptacles, sepals, petals, stamens and pistils) were characterized during ambient storage (21 ± 1 °C). The external tissues (receptacles, sepals) turned yellowing and crimpling, while the internal tissues (petals, stamens) get autolyzed and collapsed in advance. Ultrastructural observations suggested that the rapid disassembly of cell walls, dismission of cell organelles (in particular, chloroplasts, mitochondrial, vacuoles), and disruption of cytoplasm occurred in floral segments. Results from biochemical measurements revealed variations of phytohormones including abscisic acid (ABA), salicylic acid, gibberellin, indole-3-acetic acid, 1-aminocyclopropane-1-carboxylic acid, jasmonic acid, and brassinosteroids in different floral organs. Hierarchical clustering analysis revealed associations of variations of these phytohormones with the senescence of different floral organs. ABA was found as the most abundant phytohormone and mostly positively corelated with the senescence of daylily buds. Pearson correlation analysis further unraveled that hormone-crosstalking co-works on the regulation of the senescence of daylily buds. The fundamental understanding on the postharvest senescence would help to establish novel postharvest strategies to maintain quality of daylily flower buds and extend shelf life.

Transcriptome profiling of cashew apples (Anacardium occidentale) genotypes reveals specific genes linked to firmness and color during pseudofruit development

We found 34 and 71 key genes potentially involved in flavonoid biosynthesis and cell wall disassembly, respectively, which could be associated with specific peel coloration and softening of each genotype. Cashew apple (Anacardium occidentale) has a great economic importance worldwide due to its high nutritional value, peculiar flavor and aroma. During ripening, the peduncle develops different peel color and becomes quickly fragile due to its oversoftening, impacting its consumers' acceptance. In view of this, the understanding about its transcriptional dynamics throughout ripening is imperative. In this study, we performed a transcriptome sequencing of two cashew apple genotypes (CCP 76 and BRS 265), presenting different firmness and color peel, in the immature and ripe stages. Comparative transcriptome analysis between immature and ripe cashew apple revealed 4374 and 3266 differentially expressed genes (DEGs) to CCP 76 and BRS 265 genotypes, respectively. These genes included 71 and 34 GDEs involved in the cell wall disassembly and flavonoid biosynthesis, respectively, which could be associated with firmness loss and anthocyanin accumulation during cashew apple development. Then, softer peduncle of CCP 76 could be justified by down-regulated EXP and up-regulation of genes involved in pectin degradation (PG, PL and PAE) and in cell wall biosynthesis. Moreover, genes related to flavonoid biosynthesis (PAL, C4H and CHS) could be associated with early high accumulation of anthocyanin in red-peel peduncle of BRS 265. Finally, expression patterns of the selected genes were tested by real-time quantitative PCR (qRT-PCR), and the qRT-PCR results were consistent with transcriptome data. The information generated in this work will provide insights into transcriptome responses to cashew apple ripening and hence, it will be helpful for cashew breeding programs aimed at developing genotypes with improved quality traits.