Genome-Wide Identification of the Xyloglucan endotransglucosylase/Hydrolase (XTH) and Polygalacturonase (PG) Genes and Characterization of Their Role in Fruit Softening of Sweet Cherry

Fruit firmness is an important economical trait in sweet cherry (Prunus avium L.) where the change of this trait is related to cell wall degradation. Xyloglucan endotransglycosylase/hydrolase (XTH) and polygalacturonases (PGs) are critical cell-wall-modifying enzymes that occupy a crucial position in fruit ripening and softening. Herein, we identified 18 XTHs and 45 PGs designated PavXTH1-18 and PavPG1-45 based on their locations in the genome of sweet cherry. We provided a systematical overview of PavXTHs and PavPGs, including phylogenetic relationships, conserved motifs, and expression profiling of these genes. The results showed that PavXTH14, PavXTH15 and PavPG38 were most likely to participated in fruit softening owing to the substantial increment in expression during fruit development and ripening. Furthermore, the phytohormone ABA, MeJA, and ethephon significantly elevated the expression of PavPG38 and PavXTH15, and thus promoted fruit softening. Importantly, transient expression PavXTH14, PavXTH15 and PavPG38 in cherry fruits significantly reduced the fruit firmness, and the content of various cell wall components including hemicellulose and pectin significantly changed correspondingly in the transgenic fruit. Taken together, these results present an extensive analysis of XTHs and PGs in sweet cherry and provide potential targets for breeding softening-resistant sweet cherry cultivars via manipulating cell wall-associated genes.     

Monitoring Apricot (Prunus armeniaca L.) Ripening Progression through Candidate Gene Expression Analysis

This study aimed at the monitoring of the apricot (Prunus armeniaca L.) ripening progression through the expression analysis of 25 genes related to fruit quality traits in nine cultivars with great differences in fruit color and ripening date. The level of pigment compounds, such as anthocyanins and carotenoids, is a key factor in food taste, and is responsible for the reddish blush color or orange skin and flesh color in apricot fruit, which are desirable quality traits in apricot breeding programs. The construction of multiple linear regression models to predict anthocyanins and carotenoids content from gene expression allows us to evaluate which genes have the strongest influence over fruit color, as these candidate genes are key during biosynthetic pathways or gene expression regulation, and are responsible for the final fruit phenotype. We propose the gene CHS as the main predictor for anthocyanins content, CCD4 and ZDS for carotenoids content, and LOX2 and MADS-box for the beginning and end of the ripening process in apricot fruit. All these genes could be applied as RNA markers to monitoring the ripening stage and estimate the anthocyanins and carotenoids content in apricot fruit during the ripening process.     

Triacylglycerol and Glycerophospholipid Identification and Accumulation During Ripening of Pistacia lentiscus L. (Lentisc) Fruit

Triacylglycerol (TAG) and phospholipid (PL) compositions of vegetable oils are considered a marker of quality and are often used in industry to control the purity of the oils and to detect adulteration. In this study, the TAG and PL composition of developing fruit of Pistacia lentiscus were investigated for the first time. The total TAG content was found to increase rapidly during fruit ripening from 105 to 966 mg/100 g of oil respectively between the 35th and the 175th day after fructification (DAF). During this period, 16 different molecular species of TAG were identified and quantified. POO was the major TAG from the second stage of maturation. Only four classes of PL were identified in the P. lentiscus oil: the phosphatidic acid (PA), the phosphatidylethanolamine (PE), the phosphatidylglycerol (PG) and the phosphatidylinositol (PI). The mass spectra obtained showed the presence of nine molecular species of PA, five species of PE and seven molecular species for each PG and PI classes. The total phospholipid content decreased rapidly during fruit ripening, from 45.5 % at the 15th DAF to 6.88 % at the 175th DAF.     

Isolation,Expression, and Characterization of a Hydroperoxide Lyase Gene from Cucumber

A full-length cDNA coding for hydroperoxide lyase (CsHPL) was isolated from cucumber fruits of No. 26 (Southern China type) and No.14-1 (Northern China type), which differed significantly in fruit flavor. The deduced amino acid sequences of CsHPL from both lines show the same and significant similarity to known plant HPLs and contain typical conserved domains of HPLs. The recombinant CsHPL was confirmed to have 9/13-HPL enzymatic activity. Gene expression levels of CsHPL were measured in different organs, especially in fruits of different development stages of both lines. The HPL activities of fruit were identified basing on the catalytic action of crude enzyme extracts incubating with 13-HPOD (13-hydroperoxy-(9Z,12E)-octadecadienoic acid) and 13-HPOD + 9-HPOD (9-hydroperoxy-(10E,12Z)-octadecadienoic acid), and volatile reaction products were analyzed by GC-MS (gas chromatography-mass spectrometry). CsHPL gene expression in No. 26 fruit occurred earlier than that of total HPL enzyme activity and 13-HPL enzyme activity, and that in No. 14-1 fruit was consistent with total HPL enzyme activity and 9-HPL enzyme activity. 13-HPL enzyme activities decreased significantly and the 9-HPL enzyme activities increased significantly with fruit ripening in both lines, which accounted for the higher content of C6 aldehydes at 0–6 day post-anthesis (dpa) and higher content of C9 aldehydes at 9–12 dpa.     

Effects of Insect Herbivory on Bilberry Production and Removal of Berries by Frugivores

The evolutionary purpose of a fleshy fruit is to attract seed dispersers and get the seeds dispersed by frugivorous animals. For this reason, fruits should be highly rewarding to these mutualists. However, insect herbivory can alter plant reproductive success e.g. by decreasing fruit yield or affecting the attractiveness of the fruits to mutualistic seed dispersers. Under natural conditions, we tested the effects of experimental larval-defoliation on berry ripening and consumption of a non-cultivated dwarf shrub, the bilberry (Vaccinium myrtillus L.), which produces animal-dispersed berries with high sugar and anthocyanin concentration. Bilberry ramets with high fruit yield were most likely to have their berries foraged, indicating that frugivores made foraging choices based on the abundance of berries. Moreover, the probability for berries being foraged was the lowest for non-defoliated ramets that grew adjacent to larval-defoliated ramets, even though larval-defoliation did not affect the biochemical composition (total concentrations of anthocyanins, sugars and organic acids) or the probability of ripening of berries. We hypothesise that the lower probability for berries being foraged in these ramets may be a consequence of rhizome- or volatile-mediated communication between ramets, resulting in a priming effect of the herbivore defence and lower attractiveness of the non-defoliated ramets.     

A Method for Assaying of Protein Kinase Activity In Vivo and Its Use in Studies of Signal Transduction in Strawberry Fruit Ripening

Strawberry (Fragaria × ananassa) fruit ripening is regulated by a complex of cellular signal transduction networks, in which protein kinases are key components. Here, we report a relatively simple method for assaying protein kinase activity in vivo and specifically its application to study the kinase, FaMPK6, signaling in strawberry fruit. Green fluorescent protein (GFP)-tagged FaMPK6 was transiently expressed in strawberry fruit and after stimuli were applied to the fruit it was precipitated using an anti-GFP antibody. The precipitated kinase activity was measured in vitro using ³²P-ATP and myelin basic protein (MBP) as substrates. We also report that FaMPK6 is not involved in the abscisic acid (ABA) signaling cascade, which is closely associated with FaMPK6 signaling in other plant species. However, methyl jasmonate (MeJA), low temperature, and high salt treatments were all found to activate FaMPK6. Transient manipulation of FaMPK6 expression was observed to cause significant changes in the expression patterns of 2749 genes, of which 264 were associated with MeJA signaling. The data also suggest a role for FaMPK6 in modulating cell wall metabolism during fruit ripening. Taken together, the presented method is powerful and its use will contribute to a profound exploration to the signaling mechanism of strawberry fruit ripening.     

Season-Long Volatile Emissions from Peach and Pear Trees In Situ, Overlapping Profiles, and Olfactory Attraction of an Oligophagous Fruit Moth in the Laboratory

Insect herbivores that have more than one generation per year and reproduce on different host plants are confronted with substantial seasonal variation in the volatile blends emitted by their hosts. One way to deal with such variation is to respond to a specific set of compounds common to all host plants. The oriental fruit moth Cydia (=Grapholita) molesta is a highly damaging invasive pest. The stone fruit peach (Prunus persica) is its primary host, whereas pome fruits such as pear (Pyrus communis) are considered secondary hosts. In some parts of their geographic range, moth populations switch from stone to pome fruit orchards during the growing season. Here, we tested whether this temporal switch is facilitated by female responses to plant volatiles. We collected volatiles from peach and pear trees in situ and characterized their seasonal dynamics by gas chromatography–mass spectrometry. We also assessed the effects of the natural volatile blends released by the two plant species on female attraction by using Y-tube olfactometry. Finally, we related variations in volatile emissions to female olfactory responses. Our results indicate that the seasonal host switch from peach to pear is facilitated by the changing olfactory effect of the natural volatile blends being emitted. Peach volatiles were only attractive early and mid season, whereas pear volatiles were attractive from mid to late season. Blends from the various attractive stages shared a common set of five aldehydes, which are suggested to play an essential role in female attraction to host plants. Particular attention should be given to these aldehydes when designing candidate attractants for oriental fruit moth females.     

Potential use of poly(lactic acid) nanofibers mats as Nano-sachets in postharvest of climacteric fruits and vegetables

This study is reported as the first demonstration of the use of solution blow spun mats in a Nano-sachet system containing a humidity absorber hydrogel and a paraffin wax tablet impregnated with potassium permanganate (KMnO₄) for exogenous ethylene (ex-C₂H₄) oxidation, in order to delay fruit ripening. The tomatoes quality during storage time was characterized by gas chromatography (GC) (ethylene concentration and CO₂ emission analysis—respiration rate), and colorimetry (green/red color parameter). Ex-C₂H₄ production over storage time was reduced for the Nano-sachet system when compared to the control system. Color changes, from 72 to 120 h, displayed fruit still predominantly green in the Nano-Sachet system (a* from −9.1 to −6.9), color transition green-red in the TNT-sachet (−7.5 to −0.3) and increased red color fruit in the control system (−6.1 to 3.7). This work suggests that nanofibrous mats can be used as a packaging system with the ability to protect, potentialize ethylene oxidation and extend climacteric fruits and vegetables quality in shelf life.     

Changes in the Content of Organic Acids and Expression Analysis of Citric Acid Accumulation-Related Genes during Fruit Development of Yellow (Passiflora edulis f. flavicarpa) and Purple (Passiflora edulis f. edulis) Passion Fruits

Organic acids are key components that determine the taste and flavor of fruits and play a vital role in maintaining fruit quality and nutritive value. In this study, the fruits of two cultivars of passion fruit Yellow (Passiflora edulis f. flavicarpa) and purple (Passiflora edulis f. edulis) were harvested at five different developmental stages (i.e., fruitlet, green, veraison, near-mature and mature stage) from an orchard located in subtropical region of Fujian Province, China. The contents of six organic acids were quantified using ultra-performance liquid chromatography (UPLC), activities of citric acid related enzymes were determined, and expression levels of genes involved in citric acid metabolism were measured by quantitative real-time PCR (qRT-PCR). The results revealed that citric acid was the predominant organic acid in both cultivars during fruit development. The highest citric acid contents were observed in both cultivars at green stage, which were reduced with fruit maturity. Correlation analysis showed that citrate synthase (CS), cytosolic aconitase (Cyt-ACO) and cytosolic isocitrate dehydrogenase (Cyt-IDH) may be involved in regulating citric acid biosynthesis. Meanwhile, the PeCS2, PeACO4, PeACO5 and PeIDH1 genes may play an important role in regulating the accumulation of citric acid. This study provides new insights for future elucidation of key mechanisms regulating organic acid biosynthesis in passion fruit.     

Profile of enzyme in drupe of oueslati's cv. olives during ripening phases: A support method implementation in the production of extra virgin olive oil

Quality of virgin olive oil (VOO) depends on phenolic molecules content, which depends on the biochemical characteristics of olive fruits, namely endogenous enzymes. In order to ascertain the influence of olive fruit ripening degree on the phenol content, enzyme activities in olive fruits, and the quality of the corresponding oils were studied during Oueslati olive ripening. In fact, three enzymes were studied: peroxidase (POX) in olive seeds, polyphenoloxidase (PPO), and β-glucosidase (β-GL) in olive fruits mesocarp. Each enzyme showed specific trend: POX activity increased gradually until reaching a maximum (17.061 ± 0.101 U g⁻¹ FW) at ripening index (RI) 3.6 and then decreased slowly at advanced ripening stage. However, the maximum of PPO activity (240.421 ± 0.949 U g⁻¹ FW) was observed earlier at RI of 0.7. Concerning β-glucosidase activity, its maximal was 60.857 ± 1.105 U g⁻¹ FW at RI 2.8, then, it decreased sharply to reach 17.096 ± 0.865 U g⁻¹ FW at RI 3.9. A significant increase of total phenol content as well as the antioxidant activity were observed during Oueslati olive ripening. Moreover, phenolic profile indicated that appropriate harvesting date of Oueslati olives coincided with RI 3.9 given that highest content of most important individuals phenolic compounds responsible for the main VOO biological properties achieved on this date. Furthermore, phenols amount of Oueslati VOO was principally due to PPO enzyme activity as the increase in total phenols coincides with the decrease in PPO activity.     

Do Fruit Ripening Volatiles Enable Resource Specialism in Polyphagous Fruit Flies?

Frugivorous tephritid fruit flies have lineages with high levels of host generalism. These insects use olfaction to locate fruits, but how they are able to recognize the odors of so many different host species is poorly understood. We used a series of behavioral experiments to investigate the role of fruit ripening volatiles as host cues in the Queensland fruit fly, Bactrocera tryoni (Froggatt), a polyphagous pest in Australia. Odors of mature guava (Psidium guajava) attracted female and male flies more strongly than three other ripening stages and guava pulp. We analyzed volatiles from guava odor and selected eleven compounds, all of which elicited an electrophysiological response in the antenna of female flies. Three of these, ethyl acetate, ethyl butyrate, and ethyl propionate, were released at the highest rates from the most attractive ripening stage. In behavioral trials, these three esters were not attractive individually, whereas a combination was necessary and sufficient in attracting female flies. The three-component blend was as attractive as the entire 11-component blend, which without these key volatiles was not attractive. Moreover, injecting low ranking hosts (squash and cucumber) with the three volatiles increased attraction in ovipositing female flies. These fruit flies are classed as generalists, but like many polyphagous insects they could be regarded as resource specialists, preferring specific plant reproductive stages with predictable odor cues. Exploring olfaction from this perspective could improve our understanding of host choice in polyphagous insects, and the selection of volatiles to be used as attractants in insect pest management.     

Identification of Host Blends that Attract the African Invasive Fruit Fly,Bactrocera invadens

Bactrocera invadens, an invasive fruit fly species in the Afro-tropical region belonging to the Bactrocera dorsalis complex, causes considerable damage to fruit production and productivity. We sought to find attractants from hosts of B. invadens that could serve as baits in traps for monitoring and management of this pest. The attractiveness of volatiles from four different fruit species (mango, guava, banana and orange) at two stages of ripeness (ripe or unripe) was tested in an olfactometer assay. All fruits were attractive against a clean air control. Using hexane extracts of volatile collections of fruits, we demonstrated that male flies preferred the volatiles of ripe guava and orange over unripe fruit extracts. There was a slight difference in preference between females and males; females preferred orange to guava and mango, whereas males preferred mango and guava to orange. Gas chromatography/electroantennographic detection (GC/EAD) and GC/mass spectrometry (GC/MS) were used to identify compounds to which B. invadens antennae were sensitive. GC/EAD recordings from distal and medio-central parts of the fly antenna showed responses to a number of compounds from each fruit species, with esters dominating the responses. Synthetic blends were made for each fruit species using the shared antennally active compounds in ratios found in the extracts. In the olfactometer, B. invadens was most attracted to the banana and orange blends, followed by the mango and guava blends. The synthetic banana blend was as attractive as the volatile collection of banana, although both were less attractive than the fruit. The results demonstrate that composing attractive blends from GC/EAD-active constituents shared by host fruits can be effective for formulating attractive synthetic host mimics for generalist fruit fly species, such as B. invadens.