Chemical and genetic basis of orange flavor.

Sweet orange (Citrus sinensis) exhibits limited genetic diversity and high susceptibility to Huanglongbing (HLB). Breeding HLB-tolerant orange-like hybrids is in dire need. However, our understanding of the key compounds responsible for orange flavor and their genetic regulation remains elusive. Evaluating 179 juice samples, including oranges, mandarins, Poncirus trifoliata, and hybrids, distinct volatile compositions were found. A random forest model predicted untrained samples with 78% accuracy and identified 26 compounds crucial for orange flavor. Notably, seven esters differentiated orange from mandarin flavor. Cluster analysis showed six esters with shared genetic control. Differential gene expression analysis identified C. sinensis alcohol acyltransferase 1 (CsAAT1) responsible for ester production in orange. Its activity was validated through overexpression assays. Phylogeny revealed the functional allele was inherited from pummelo. A SNP-based DNA marker in the coding region accurately predicted phenotypes. This study enhances our understanding of orange flavor compounds and their biosynthetic pathways and expands breeding options for orange-like cultivars.

Application of Thymol Vapors to Control Postharvest Decay Caused by Penicillium digitatum and Lasiodiplodia theobromae in Grapefruit.

Two of the major postharvest diseases impacting grapefruit shelf life and marketability in the state of Florida (USA) are stem-end rot (SER) caused by Lasiodiplodia theobromae and green mold (GM) caused by Penicillium digitatum. Here, we investigated the in vitro and in vivo efficacy of vapors of thymol, a natural compound found in the essential oil of various plants and the primary constituent of thyme (Thymus vulgaris) oil, as a potential solution for the management of GM and SER. Thymol vapors at concentrations lower than 10 mg L-1 significantly inhibited the mycelial growth of both pathogens, causing severe ultrastructural damage to P. digitatum conidia. In in vivo trials, the incidence and lesion area of GM and SER on inoculated grapefruit were significantly reduced after a 5 d exposure to 50 mg L-1 thymol vapors. In addition, the in vitro and in vivo sporulation of P. digitatum was suppressed by thymol. When applied in its vapor phase, thymol had no negative effect on the fruit, neither introducing perceivable off-flavor nor causing additional weight loss. Our findings support the pursuit of further studies on the use of thymol, recognized as safe for human health and the environment, as a promising strategy for grapefruit postharvest disease management.

Physical, Chemical, and Sensory Properties of a Turmeric-Fortified Pineapple Juice Beverage.

Beverage mixtures based on pineapple (Ananas comosus) and turmeric (Curcuma longa) juice as a ready-to-drink product were developed, and their physicochemical, nutritional, and sensory properties were evaluated. Four different concentrations of turmeric juice (5%, 10%, 15%, and 20% (v/v)) were added to pineapple juice to make turmeric-fortified pineapple (TFP) juice samples. Pineapple juice without turmeric was the control. The L*, a*, b*, titratable acidity (TA), total antioxidant capacity, and %DPPH scavenging values, as well as the concentrations of the phenolic compounds curcumin and demethoxycurcumin, were significantly increased with increasing turmeric concentration. Thirty volatile compounds were detected in the mixed juice samples with turmeric. Most of the turmeric-specific compounds, including monoterpenes, sesquiterpenes and turmerones, were detected in the TFP juice samples. While the antioxidant activity of the juice samples increased with increasing turmeric concentration, the pineapple juice fortified with 10% turmeric (10%T) had the best overall quality as determined by panelists. Greater concentrations of turmeric were associated with decreased palatability due to reduced mouthfeel and sweetness and increased aftertaste and sourness. These results suggest that the 10%T juice could be developed into a commercial functional beverage with increased overall flavor and nutritional quality.

Sensory quality of Citrus scion hybrids with Poncirus trifoliata in their pedigrees.

Hybrids of Poncirus trifoliata L. Raf. with Citrus have shown degrees of tolerance to the deadly citrus greening disease, hence prompting interest as potential commercial varieties. Although P. trifoliata is known to produce fruit that is inedible, fruit from many advanced hybrid trees have not been evaluated for their quality potential. The sensory quality of selected Citrus hybrids with varying degrees of P. trifoliata in their pedigrees is reported herein. Four Citrus × P. trifoliata hybrids developed through the USDA Citrus scion breeding program-1-76-100, 1-77-105, 5-18-24, and 5-18-31-had acceptable eating quality and sweet and sour taste, with mandarin, orange, fruity-noncitrus, and floral flavors. On the other hand, hybrids with higher proportion of P. trifoliata in their pedigrees, US 119 and 6-23-20, produced a juice characterized by green, cooked, bitter, and Poncirus-like flavor and aftertaste. Partial least square regressions revealed that the Poncirus-like off-flavor is likely due to a combination of higher than typical amounts of sesquiterpene hydrocarbons (woody/green odor), monoterpenes (citrus/pine), and terpene esters (floral) and a lack of aldehydes with typical citrus odor (octanal, nonanal, and decanal). Sweetness and sourness were mostly explained by high sugars and acids, respectively. Further, carvones and linalool contributed to sweetness in the samples from early and late seasons, respectively. In addition to highlighting chemical contributors to sensory descriptors in Citrus × P. trifoliata hybrids, this study provides useful information on sensory quality for future citrus breeding efforts. PRACTICAL APPLICATION: The relationships between the sensory quality and secondary metabolites of Citrus × P. trifoliata hybrids described in this study help identify disease-resistant Citrus scion hybrids with acceptable flavor and help mobilize this resistance in future breeding efforts. It also shows potential of such hybrids to be commercialized.

Effect of Huanglongbing on the Volatile Organic Compound Profile of Fruit Juice and Peel Oil in 'Ray Ruby' Grapefruit.

Along with orange and mandarin, grapefruit production in Florida has declined sharply due to Huanglongbing (HLB), or citrus greening disease, caused by Candidatus Liberibacter asiaticus (CLas). HLB affects the volatile profiles of juice and peel oil in oranges, but there is limited information on grapefruit. In this research, 'Ray Ruby' grapefruit were harvested in 2020 and 2021 from healthy (HLB-) and HLB-affected (HLB+) trees. Peel oil was extracted by hydrodistillation, and the volatiles were analyzed by direct injection of the oil samples into gas chromatography-mass spectrometry (GC-MS). Volatiles in the juice were analyzed by headspace (HS)-solid-phase microextraction (SPME) coupled with GC-MS. HLB significantly altered the volatile profiles of peel oil and juice in 'Ray Ruby' grapefruit. Juice samples of HLB+ fruits had lower decanal, nonanal, and octanal, important citrus juice flavor compounds. HLB+ samples also showed reduced content of nonterpene compounds, other aliphatic and terpene aldehydes, and terpene ketones. Ethanol, acetaldehyde, ethyl acetate, and ethyl butanoate were increased in HLB+ juice samples, indicating an HLB-induced stress response. The most abundant compounds D-limonene and ß-caryophyllene, as well as other sesquiterpenes, were increased in HLB+ juice and peel oil samples. On the other hand, the oxidative/dehydrogenated terpenes were increased by HLB in peel oil but decreased in the juice sample. Nootkatone, the key grapefruit volatile was consistently reduced by HLB in both peel oil and juice samples. The impact of HLB on nootkatone deteriorated the quality of both juice and peel oil in grapefruits.

Soil Amendment and Storage Effect the Quality of Winter Melons (Benincasa hispida (Thunb) Cogn.) and Their Juice.

Winter melon fruits were grown in the field using anaerobic soil disinfestation (ASD) and conventional fertilizer alone as the control treatment. Fruits were harvested and stored at 20 °C for 120 d, the juice was processed on day one and day 120, and the effects of soil amendment and 120 d storage on the juice's physical and chemical (sugars, acids, volatile and nutritional compounds) properties were evaluated. Fruit juice extracted from ASD-grown fruit had greater magnitude of zeta potential than the control juice, indicating it was physically more stable than the juice obtained from the control conditions. ASD fruit juice had lower soluble solids content (SSC), and lower volatile compounds that contribute green, grass, and sulfur notes, and negatively influence flavor quality. ASD fruit juice had higher vitamin B5 and cytidine. Juice processed from 120 d stored fruit had less yield due to 12.4-15.6% weight loss. The non-soluble solids content was higher and particle size was larger, and the SSC and individual sugars decreased. However, titratable acidity (TA) increased primarily due to increased citric acid. Out of 16 free amino acids, 6 increased and only 1 decreased. However, three out of five nucleosides decreased; vitamins B1 and B6 increased; vitamins B2, B3 and C decreased. Overall, juice derived from fruit produced using ASD was physically more stable and had less SSC and off-odor volatiles than the control, while the fruit juice of those stored for 120 d had lower SSC and higher TA and nutritional profiles, comparable to freshly harvested fruit.

Nano- and micro-sized carnauba wax emulsions-based coatings incorporated with ginger essential oil and hydroxypropyl methylcellulose on papaya: Preservation of quality and delay of post-harvest fruit decay.

Carnauba wax nano and micro-sized emulsions and hydroxypropyl methylcellulose coatings, alone or combined with ginger essential oils (GEO) were applied on papayas and evaluated under several storage conditions. In a first experiment, storage parameters were: 6 days at 22 °C, and 9 days at 13 °C followed by 5 days at 22 °C. In a second experiment, storage was: 5 days at 22 °C, and 10 days at 16 °C followed by 3 days at 22 °C. Coating effects were dependent on storage conditions. While fruits were in cold storage, there were few changes; however, at 22 °C, the differences between coatings became more evident. Nanoemulsions maintained papaya quality during storage by retarding firmness loss, color changes, and reducing respiration rates, resulting in delayed ripening. GEO exhibited some positive effect on fungal disease control. Nanoemulsion-based coatings improved shelf life by reducing weight loss, color development, and slowing ripening of papaya fruit.

Nanoemulsions as Edible Coatings: A Potential Strategy for Fresh Fruits and Vegetables Preservation.

Fresh fruits and vegetables are perishable commodities requiring technologies to extend their postharvest shelf life. Edible coatings have been used as a strategy to preserve fresh fruits and vegetables in addition to cold storage and/or controlled atmosphere. In recent years, nanotechnology has emerged as a new strategy for improving coating properties. Coatings based on plant-source nanoemulsions in general have a better water barrier, and better mechanical, optical, and microstructural properties in comparison with coatings based on conventional emulsions. When antimicrobial and antioxidant compounds are incorporated into the coatings, nanocoatings enable the gradual and controlled release of those compounds over the food storage period better than conventional emulsions, hence increasing their bioactivity, extending shelf life, and improving nutritional produce quality. The main goal of this review is to update the available information on the use of nanoemulsions as coatings for preserving fresh fruits and vegetables, pointing to a prospective view and future applications.

Effects of Harvest Maturity, Refrigeration and Blanching Treatments on the Volatile Profiles of Ripe "Tasti-Lee" Tomatoes.

The interactive effects of six maturity stages and refrigerated storage (chilling)/blanching (heating) treatments on the volatile profiles of ripe tomatoes were studied. A total of 42 volatiles were identified, of which 19 compounds had odor activity values equal to or greater than 1. Of those, "green" and "leafy" aroma volatiles were most abundant. Chilling and heating treatments both suppressed overall volatile production, with chilling having the greater impact, regardless of harvest maturity. However, fruit harvested at the turning stage had the least volatile suppression by chilling and heating treatments in comparison with fruit harvested earlier or later, mostly in the fatty acid- and phenylalanine-derived volatiles. Volatiles derived from amino acids were promoted by heat treatment for fruit harvested at all maturities, and those derived from carotenoid and phenylalanine pathways and harvested at advanced harvest maturities were stimulated by chilling treatment. Volatile production is generally believed to be improved by delayed harvest, with vine-ripe being optimum. However, opposite results were observed possibly because the later-harvested fruit had longer exposure to open-field weather stress. The best harvest maturity recommendation is the turning stage where fruit developed abundant volatiles and were least impacted by chilling and heating treatments.

The Effect of Controlled-Release Carvacrol on Safety and Quality of Blueberries Stored in Perforated Packaging.

The objective of this research was to evaluate the use of a controlled-release carvacrol powder to delay storage decay and maintain the safety of blueberries. The controlled-release carvacrol powder was a microcapsule of carvacrol (11% (w/w) active carvacrol) surrounded by a pectin/sodium alginate matrix. The microcapsules were packed in an air-permeable pouch, and then attached to the top of a clamshell filled with blueberries. The blueberries, inoculated with Escherichia coli or Colletotrichum acutatum, or non-inoculated control, were monitored for microbial growth and quality for 10 days at 10 °C and 5 days at 20 °C. Three treatments were compared: controlled-release microencapsulated carvacrol, non-encapsulated carvacrol, and control. The results showed that both the microencapsulated carvacrol and the non-encapsulated carvacrol treatments significantly reduced the populations of yeast and mold, and of E. coli and mesophilic aerobic bacteria. The microencapsulated carvacrol treated berries retained better quality due to significantly lower weight loss than control after 10 days at 10 °C. Sensory panelists found that the microencapsulated carvacrol berries had significantly higher overall blueberry flavor and lower discernible off-flavor in comparison with the non-encapsulated treatment after 3 days at 20 °C. The fruit internal quality, including total soluble solids content (SSC), and titratable acidity (TA), was not significantly affected by any treatment. These results indicate that pectin/sodium alginate controlled-release microencapsulated carvacrol can be used for the preservation of blueberries or other small fruit.

Color biogenesis data of tomatoes treated with hot-water and high temperature ethylene treatments.

Controlled postharvest stresses were used to induce the synthesis of carotenoids in tomato fruit. The accumulation of carotenoids was observed by the change of color of the tomato fruit from green to red. This change of color was monitored by the a* value and hue of the CIELAB* color coordinates in which the a* value increased following a sigmoidal curve and hue decreased in a similar trend. This sigmoidal curve marked the transition from chloroplasts to chromoplasts; in other words, the change of color tracked the disorganization or degreening, which was simultaneously accompanied by chromoplast biogenesis or red color development when tomatoes were at the Turning stage of development. The color data and photographic images provides information on how heat stress affected the synchronicity of chloroplast disorganization and chromoplast biogenesis in the early developmental stages of tomato ripening.

Genomic Characterization of the Fruity Aroma Gene, FaFAD1, Reveals a Gene Dosage Effect on γ-Decalactone Production in Strawberry (Fragaria × ananassa).

Strawberries produce numerous volatile compounds that contribute to the unique flavors of fruits. Among the many volatiles, γ-decalactone (γ-D) has the greatest contribution to the characteristic fruity aroma in strawberry fruit. The presence or absence of γ-D is controlled by a single locus, FaFAD1. However, this locus has not yet been systematically characterized in the octoploid strawberry genome. It has also been reported that the volatile content greatly varies among the strawberry varieties possessing FaFAD1, suggesting that another genetic factor could be responsible for the different levels of γ-D in fruit. In this study, we explored the genomic structure of FaFAD1 and determined the allele dosage of FaFAD1 that regulates variations of γ-D production in cultivated octoploid strawberry. The genome-wide association studies confirmed the major locus FaFAD1 that regulates the γ-D production in cultivated strawberry. With the hybrid capture-based next-generation sequencing analysis, a major presence-absence variation of FaFAD1 was discovered among γ-D producers and non-producers. To explore the genomic structure of FaFAD1 in the octoploid strawberry, three bacterial artificial chromosome (BAC) libraries were developed. A deletion of 8,262 bp was consistently found in the FaFAD1 region of γ-D non-producing varieties. With the newly developed InDel-based codominant marker genotyping, along with γ-D metabolite profiling data, we revealed the impact of gene dosage effect for the production of γ-D in the octoploid strawberry varieties. Altogether, this study provides systematic information of the prominent role of FaFAD1 presence and absence polymorphism in producing γ-D and proposes that both alleles of FaFAD1 are required to produce the highest content of fruity aroma in strawberry fruit.

Huanglongbing and Foliar Spray Programs Affect the Chemical Profile of "Valencia" Orange Peel Oil.

Florida orange trees have been affected by huanglongbing (HLB) for more than a decade. To alleviate disease-caused tree decline, maintain fruit productivity, and reduce disease transmission, enhanced foliar spray programs combining vector control and nutritional supplementation have been applied to healthy and diseased trees. The aim of this research was to discover if the various foliar sprays affect fruit peel oil chemical components. In this study, "Valencia" orange trees, with or without HLB (HLB±), were treated with the grower standard program (control, C) or one of four proprietary enhanced foliar spray programs (N1, N2, N3, and N4) over 16 months. Compared with HLB-, HLB+ samples had lower concentrations of typical peel oil components, including valencene, octanal, and decanal, and were abundant in oxidative/dehydrogenated terpenes, such as carvone and limonene oxide. However, limonene, the dominant component, was not affected by any treatment. Control and three out of four enhanced foliar spray programs, N2, N3, and N4, had very little influence on the chemical profiles of both HLB- and HLB+ samples, while N1 treatment greatly altered the chemical profile of HLB+ samples, resulting in peel oil similar to that of HLB- samples.

Volatiles Influencing Sensory Attributes and Bayesian Modeling of the Soluble Solids-Sweetness Relationship in Strawberry.

Descriptive analysis via trained sensory panels has great power to facilitate flavor improvement in fresh fruits and vegetables. When paired with an understanding of fruit volatile organic compounds, descriptive analysis can help uncover the chemical drivers of sensory attributes. In the present study, 213 strawberry samples representing 56 cultivars and advanced selections were sampled over seven seasons and subjected to both sensory descriptive and chemical analyses. Principal component analysis and K-cluster analyses of sensory data highlighted three groups of strawberry samples, with one classified as superior with high sweetness and strawberry flavor and low sourness and green flavor. Partial least square models revealed 20 sweetness-enhancing volatile organic compounds and two sweetness-reducing volatiles, many of which overlap with previous consumer sensory studies. Volatiles modulating green, sour, astringent, overripe, woody, and strawberry flavors were also identified. The relationship between soluble solids content (SSC) and sweetness was modeled with Bayesian regression, generating probabilities for sweetness levels from varying levels of soluble solids. A hierarchical Bayesian model with month effects indicated that SSC is most correlated to sweetness toward the end of the fruiting season, making this the best period to make phenotypic selections for soluble solids. Comparing effects from genotypes, harvest months, and their interactions on sensory attributes revealed that sweetness, sourness, and firmness were largely controlled by genetics. These findings help formulate a paradigm for improvement of eating quality in which sensory analyses drive the targeting of chemicals important to consumer-desired attributes, which further drive the development of genetic tools for improvement of flavor.

Extraction Method Affects Contents of Flavonoids and Carotenoids in Huanglongbing-Affected "Valencia" Orange Juice.

A previous study using healthy "Valencia" orange fruit found that juicing extraction methods dramatically changed the orange juice (OJ) flavor and phytochemical profiles. The present study was conducted to confirm whether the same changes occur when Huanglongbing (HLB)-affected oranges were used. HLB has extensively spread to most OJ processing regions around the world, substantially deteriorating fruit and juice flavor quality and altering the phytochemical profiles. In this study, the effect of two major juice extractor types, a shear force extractor (SFE) and a reamer extractor (RE), on the juice quality and physiochemical profile was assessed using HLB-affected orange fruit. Juice extracted via SFE resulted in a lower yield with lower peel oil and higher pellet (peel tissue particles) content compared to juice obtained via RE. The SFE juice also had higher levels of hesperidin and other flavonoid glycosides, mainly due to plentiful peel tissue particles. The SFE juice was also abundant in carotenoids due to a large amount of flavedo particles in the juice. On the other hand, polymethoxylated flavones occurred at higher concentrations in the RE juice, and this may be due to the higher peel oil content in this juice. The SFE juice was rich in flavonoid glycosides and carotenoids, which are associated with potential antioxidant properties; however, the extra portion of the phytonutrients mostly existed in the pellets and possibly had low bioavailability. The results obtained from the HLB-affected oranges are in agreement with the previous observations of healthy oranges.

A brief hot-water treatment alleviates chilling injury symptoms in fresh tomatoes.

BACKGROUND: Reducing the negative effects of chilling injury (CI) in tomatoes after harvest is essential to ensure good quality and to minimize losses. CI is a postharvest disorder associated with the generation of reactive oxygen species (ROS) in the fruit. Therefore, antioxidant accumulation can counteract ROS, alleviating CI symptoms. In this sense, it has been confirmed that a brief hot-water (HW) immersion promotes the synthesis of antioxidants. RESULTS: HW treatment at 52 °C for 5 min significantly reduced chilling-associated decay, from 66.7% to 17.2% in breaker turning (BT) and from 55.8% to 9.8% in mature green (MG) 'BHN-602' tomatoes stored at 5 °C for 2 weeks and from 26.7% to 6.7% in BT tomatoes stored at 5 °C for 1 week. Also, HW treatment significantly increased lycopene content by 17% in BT tomatoes stored at 5 °C for 2 weeks, as well as ascorbic acid by 11%, lipophilic phenolics by 18% and total phenolics by 6.5% in BT tomatoes stored at 12.5 °C for 1 week. Despite the increase of antioxidants, HW treatment did not enhance the sensory aromatic profile, color and antioxidant capacity. Interestingly, HW treatment reduced ripening time by 3 days in MG tomatoes stored at 5 °C for 2 weeks or at 12.5 °C for 1 week. CONCLUSION: HW treatment applied to MG or BT 'BHN-602' tomatoes can alleviate the development of some CI symptoms, particularly decay, possibly by increasing antioxidants that scavenge ROS. © 2020 Society of Chemical Industry.

Effect of fruit maturity on volatiles and sensory descriptors of four mandarin hybrids.

Mandarins (or tangerines) are mainly consumed as fresh fruits due to the ease of peeling and desirable flavor. Sweetness, acidity, and flavor of mandarin are the most important criteria for consumer preference. The objective of this study was to evaluate the effects of harvest date on sensory and chemical components of four mandarin cultivars (Murcott, 411, Temple, and 'LB8-9' Sugar Belle®). Volatiles were extracted from the headspace of juice samples with solid phase microextraction (SPME) and analyzed using gas chromatography-mass spectrometry (GC-MS). The optimum harvest window for eating quality of 411 was late January to mid-February (soluble solids content [SSC]/titratable acidity [TA]: 11.3 to 14.0), Sugar Belle® fruits were best tasted when harvested from mid- to end of January (SSC/TA: 14.1 to 16.1), and February was the best month for harvesting Murcott (SSC/TA: 13.10 to 18.0) and Temple (SSC/TA:10.3 to 12.50). Sensory perception of sweetness, ripeness, and juiciness increased as SSC/TA increased while sourness and bitterness decreased. Pumpkin flavor, an indicator of overripe fruit, was mainly noticed late in the season. Tangerine flavor tended to decrease, whereas fruity-noncitrus flavor tended to increase with fruit maturity. Monoterpenes were the most abundant volatiles and tended to decrease with fruit maturity, whereas alcohols, esters, and aldehydes increase. Aldehydes, esters, and alcohols were positively correlated with sweetness, ripeness, juiciness, and fruity characteristics, and negatively with sourness and bitterness. On the other hand, monoterpenes were positively correlated with bitterness and tangerine flavor, and negatively correlated with sweetness and fruity-noncitrus flavor. The highest number of esters was found in Temple, whereas Murcott and 411 were high in aldehydes.

Mitigation of Off-Flavor in Huanglongbing-Affected Orange Juice Using Natural Citrus Non-Volatile Compounds.

Huanglongbing (HLB) disease has reduced orange yield and flavor quality in Florida. Orange juice (OJ) and peel molasses (PM) compounds are "from the named fruit" (FTNF) and were tested for their effects on HLB-affected OJ taste. These and other "target" compounds, reportedly found in citrus, were spiked into off-flavored, HLB-affected reference juices. The spiked juice was evaluated by a sensory panel in comparison with unspiked juice for "sweetness", "sourness", "bitterness", "astringency", and "aftertaste". Of the target compounds, feruloyl putrescine, taxifolin, and neodiosmin most effectively reduced "bitterness", feruloyl putrescine reduced "astringency" and "aftertaste", while feruloyl putrescine and neodiosmin enhanced perceived "sweetness", perhaps due to reduced bitterness. OJ and PM fractions containing reported bitter limonoids, hydroxycinnamates, hesperidin, and/or polymethoxylated flavones enhanced "sourness', "bitterness", "astringency", and "aftertaste", and/or suppressed perceived "sweetness" (perhaps due to enhanced bitterness). Other fractions, containing feruloyl putrescine and/or vicenin-2 enhanced "sweetness" and reduced "bitterness", "sourness", "astringency", and/or "aftertaste". These potentially FTNF compounds may be useful for management of HLB-affected OJ juice flavor.

Comparative analysis of the transcriptomes of the calyx abscission zone of sweet orange insights into the huanglongbing-associated fruit abscission.

Citrus greening disease or huanglongbing (HLB) is associated with excessive pre-harvest fruit drop. To understand the mechanisms of the HLB-associated fruit abscission, transcriptomes were analyzed by RNA sequencing of calyx abscission zones (AZ-C) of dropped "Hamlin" oranges from HLB-diseased trees upon shaking the trees (Dd), retained oranges on diseased trees (Rd), dropped oranges from healthy shaken trees (Dh), and retained oranges on healthy trees (Rh). Cluster analysis of transcripts indicated that Dd had the largest distances from all other groups. Comparisons of transcriptomes revealed 1047, 1599, 813, and 764 differentially expressed genes (DEGs) between Dd/Rd, Dd/Dh, Dh/Rh, and Rd/Rh. The gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses indicated hormone signaling, defense response, and secondary metabolism were involved in HLB-associated fruit abscission. Ethylene (ET) and jasmonic acid (JA) synthesis/signaling-related genes were upregulated in Dd, while other phytohormone-related genes were generally downregulated. In addition, genes related to JA/ET-activated defense response were upregulated in Dd as well. Consistent with the phytohormone gene expression data, increased levels (p < 0.05) of ET and JA, and a decreased level (p < 0.05) of abscisic acid were found in Dd compared with Rd, Dh or Rh. Lasiodiploidia theobromae level in Dd AZ-C was higher than the other fruit types, confirmed by qPCR, indicating AZ-C secondary fungal infection of HLB fruit may exacerbate their abscission. This information will help formulate effective strategies to control HLB-related abscission.

Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene.

BACKGROUND: Valencene is a major sesquiterpene in citrus oil and biosynthesized by valencene synthase (Cstps1; EC: 4.2.3.73) from the 15-carbon substrate farnesyl diphosphate. It is abundant in juice of some mandarins (e.g. Citrus reticulata Blanco cv. Fortune), however, it is undetectable in others (e.g. C. reticulata Blanco cv. Murcott), We have discovered that the Murcott mandarin Cstps1 gene expression is severely reduced. A previous genetic mapping study using an F1 population of Fortune × Murcott found that the segregation of valencene production in fruit exhibited a Mendelian inheritance ratio of 1:1. There was only one dominant locus associated with valencene content detected on the mandarin genetic map. The goal of this study was to understand the molecular mechanism underlying the valencene deficiency observed in some citrus hybrids. RESULTS: There was a clear relationship between presence or absence of the valencene synthase gene (Cstps1) expression, and presence or absence of valencene among randomly selected mandarin hybrids. Cloning the coding regions of Cstps1 from Fortune and Murcott mandarin, and aligning with previous reported Valencia orange Cstps1 sequence, showed that they both exhibited extremely high similarity with the known Cstps1. By further cloning and analyzing the promoter region of Cstps1 from Valencia, Fortune and Murcott, a 12-nucleotide deletion at approximately - 270 bp from the Cstps1 coding region was only found in Murcott. Three binary vectors, designated as p1380-FortP-GUSin, p1380-MurcP-GUSin and p1380-MurcP(+ 12)-GUSin, were developed for promoter activity analysis. Transient over-expression of Fortune Cstps1 promoter in sweet orange showed notable GUS activity, but the Murcott Cstps1 promoter did not. In addition, by re-inserting the 12-nucleotide fragment, the activity of the Murcott Cstps1 promoter was mostly recovered. CONCLUSION: The deficiency of valencene production in some mandarins is probably due to a 12-nucleotide deletion in the promoter region of the Cstps1, which could be a crucial switch of Cstps1 transcription. Our results further enhanced the understanding of valencene biosynthesis in citrus.