Myo-inositol oxygenase CgMIOX3 alleviates S-RNase-induced inhibition of incompatible pollen tubes in pummelo.

Pummelo (Citrus grandis L. Osbeck) exhibits S-RNase-based self-incompatibility (SI), during which S-RNase cytotoxicity inhibits pollen tubes in an S-haplotype specific manner. The entry of S-RNase into self-pollen tubes triggers a series of reactions. However, these reactions are still poorly understood in pummelo. In the present study, we used S-RNases as baits to screen a pummelo pollen cDNA library and characterized a myo-inositol oxygenase (CgMIOX3) that physically interacts with S-RNases. CgMIOX3 is highly expressed in pummelo pollen tubes and its down-regulation leads to a reduction in pollen tube growth. Upon entering pollen tubes, S-RNases increase the expression of CgMIOX3 and enhance its activity by directly binding to it in an S-haplotype-independent manner. CgMIOX3 improves pollen tube growth under oxidative stress through ascorbic acid accumulation and increases the length of self-pollen tubes. Furthermore, over-expression of CgMIOX3 increases the relative length of self-pollen tubes growing in the style of petunia (Petunia hybrida). This study provides intriguing insights into the pumelo SI system, revealing a regulatory mechanism mediated by CgMIOX3 that plays an important role in the resistance of pollen tubes to S-RNase cytotoxicity.

Fruit Quality Assessment of Novel Hybrid Pummelo × Sweet Orange and Its Molecular Characterization Using Acidity Specific Markers.

Research background: There is considerable diversity in newly developed pummelo × sweet orange citrus hybrids. Most hybrids showed lower peel thickness and high juice yield but there is a lack of information on fruit quality parameters and molecular characterization. Therefore, the aim of the current study is to determine the content of antioxidants and properties of the fresh juice of 24 new pummelo × sweet orange citrus hybrids (Citrus maxima [Burm. f.] Osbeck × Citrus sinensis [L.] Osbeck) and the parental genotypes along with molecular characteristics determined using acidity specific markers. Experimental approach: The correlation and estimate of inheritance of the fruit juice properties: ascorbic acid, total phenol, total flavonoid, total antioxidant, total soluble solid and sugar contents, pH, titratable acidity, along with sensory evaluation was performed. Molecular characterization of these hybrids was carried out using de novo generated acidity specific simple sequence repeat (SSR) markers. Results and conclusions: The main constituents of the fruit juice of pummelo × sweet orange hybrids were observed in the range of w(ascorbic acid)=40.00-58.13 mg/100 g, total phenols expressed as gallic acid equivalents w(GAE)=40.67-107.33 mg/100 g, total antioxidants expressed as Trolox equivalents b(Trolox)=2.03-5.49 µmol/g, total flavonoids expressed as quercetin equivalents w(QE)=23.67-59.33 mg/100 g, along with other properties: total soluble solids=7.33-11.33 %, w(total sugar)=2.10-5.76 %, w(reducing sugar)=1.69-2.78 %, w(non-reducing sugar)=0.39-3.17 % and titratable acidity 1.00-2.11 %. The above parameters differed significantly in the fruit juice of the evaluated pummelo × sweet orange hybrids. Considering these parameters, the hybrids SCSH 17-9, SCSH 13-13, SCSH 11-15 and SCSH 3-15 had superior antioxidant properties in terms of these parameters. A higher heritability (≥80 %) was also observed for all juice properties. Molecular characterization of pummelo × sweet orange hybrids showed that >50 % of the hybrids were grouped with medium acidity parents. Both molecular and biochemical parameter-based clustering showed that interspecific hybrids exhibit transgressive segregation with increased antioxidants that help alleviate the health problems. Novelty and scientific contribution: These newly developed pummelo × sweet orange citrus hybrids are a valuable source of high-quality antioxidants for a healthy diet. The identification of trait markers that enable selection at the seedling stage is of great benefit to citrus breeders, as the characteristic features of a mature tree are not yet visible at the juvenile stage.

Evolution-guided multiomics provide insights into the strengthening of bioactive flavone biosynthesis in medicinal pummelo.

Pummelo (Citrus maxima or Citrus grandis) is a basic species and an important type for breeding in Citrus. Pummelo is used not only for fresh consumption but also for medicinal purposes. However, the molecular basis of medicinal traits is unclear. Here, compared with wild citrus species/Citrus-related genera, the content of 43 bioactive metabolites and their derivatives increased in the pummelo. Furthermore, we assembled the genome sequence of a variety for medicinal purposes with a long history, Citrus maxima 'Huazhouyou-tomentosa' (HZY-T), at the chromosome level with a genome size of 349.07 Mb. Comparative genomics showed that the expanded gene family in the pummelo genome was enriched in flavonoids-, terpenoid-, and phenylpropanoid biosynthesis. Using the metabolome and transcriptome of six developmental stages of HZY-T and Citrus maxima 'Huazhouyou-smooth' (HZY-S) fruit peel, we generated the regulatory networks of bioactive metabolites and their derivatives. We identified a novel MYB transcription factor, CmtMYB108, as an important regulator of flavone pathways. Both mutations and expression of CmtMYB108, which targets the genes PAL (phenylalanine ammonia-lyase) and FNS (flavone synthase), displayed differential expression between Citrus-related genera, wild citrus species and pummelo species. This study provides insights into the evolution-associated changes in bioactive metabolism during the origin process of pummelo.

Cell wall modification and lignin biosynthesis involved in disease resistance against Diaporthe citri in harvested pummelo fruit elicited by carvacrol.

BACKGROUND: Phomopsis stem-end rot caused by Diaporthe citri, causes significant commercial postharvest losses of pummelo fruit during storage. Carvacrol (CVR) is a known generally recognized as safe and has the ability to prolong the preservation of harvested fruits. In the present study, the inhibitory effects of CVR treatment at the appropriate concentration on Phomopsis stem-end rot development of harvested pummelo fruit inoculated with D. citri were evaluated by the amounts of cell wall components, the activities and gene expressions of related enzymes involved in cell wall modification and lignin biosynthesis. RESULTS: Results indicated that CVR completely inhibited D. citri growth in vitro at 200 mg L-1 and significantly controlled Phomopsis stem-end rot development in harvested pummelo. The CVR treatment delayed peel softening and browning, and retarded electrolyte leakage, superoxide radical (O2 •- ) production, and malondialdehyde content. The CVR-treated fruit maintained higher amounts of cell wall material, protopectin, hemicelluloses, and cellulose, but exhibited lower water-soluble pectin amount. Moreover, in D. citri-inoculated fruit, CVR treatment suppressed the activities and gene expressions of cell wall disassembling-enzymes, including pectin methylesterase, polygalacturonase, cellulase, and ß-galactosidase, while the development of cell wall degradation was reduced. Meanwhile, the CVR treatment enhanced the lignin biosynthesis by increasing the activities and up-regulating the gene expressions of phenylalanine ammonialyase, cinnamic alcohol dehydrogenase, and peroxidase accompanied with elevated level of lignin in pummelo fruit. CONCLUSION: The disease resistance to D. citri in pummelo fruit elicited by CVR treatment is related to delaying cell wall degradation and enhancing lignin biosynthesis. © 2021 Society of Chemical Industry.

Grapefruit and pomelo peel extracts as natural antioxidants for improved storage stability of Turkey patties during refrigerated storage.

The study investigated the effect of albedo and flavedo extracts of grapefruit and pomelo on storage stability of turkey patties during refrigerated storage. Five different types of products were developed depending on the addition of extracts viz. control (C), albedo and flavedo extract of grapefruit (GA and GF), and albedo and flavedo extract of pomelo (PA and PF). The products were stored for 10 days and evaluated for lipid and protein oxidation, pH, colour and sensory properties. The extracts improved the lipid oxidative stability by decreasing (P < 0.05) the TBARS values of the samples compared to control group. No effect of the extract treatments (P > 0.05) was observed on pH and colour values of the products (a* and b*). The lowest TBARS values were found for the products containing GF (0.78 mg MA/kg) on day 4. The treatments showed a significant impact on flavour of the products and the highest scores were obtained for PA containing products.

Disinfectant of pummelo (Citrus Grandis L. Osbeck) fruit juice using gaseous ozone.

This work studied the effectiveness of gaseous ozone disinfection on pummelo (Citrus Grandis L. Osbeck) fruit juice components. Unfiltered and filtered pummelo fruit juices were treated with gaseous ozone for up to 50 min with ozone concentration fixed at 600 mg/h. A microbiological and physicochemical properties analysis were conducted on the ozone-treated fruit juices samples. It was found that the survival rate of aerobic bacteria, yeast and mold in unfiltered pummelo fruit juice were higher compared to filtered juice, as the juice components acted as protective barriers to the microorganisms. The microorganisms' inactivation in pummelo fruit juices was also observed to have increased as the ozone treatment time increased. Significant effects on total colour difference, ascorbic acid content, and total phenolic content were also observed over increased ozone-treatment time. However, ozone was shown to be ineffective in activating PME activity in both types of juice. The experimental results of this study indicated that pummelo fruit juice components had significant effects on the effectiveness of gaseous ozone, however, the degree of the effects depends on the different fruit components (total soluble solids, total phenolic content). As a conclusion, filtered juice showed better quality characteristics in comparison to unfiltered juice post-ozone treatment.

Volatile Composition in Two Pummelo Cultivars (Citrus grandis L. Osbeck) from Different Cultivation Regions in China.

This study investigated the composition of volatile compounds in two pummelo cultivars, including 'Shatian' and 'Guanxi', cultivated in different regions of China with the aim of studying the effect of cultivar and cultivation condition on biosynthesis of volatile compounds in pummelo. Volatile compounds were extracted from pummelo juice using head-space microextraction and then analyzed using gas chromatography coupled with mass spectrometry. Results showed that a total of 49 volatile compounds was detected in the study, including 11 aldehydes, 7 alcohols, 3 ketones, 7 esters, 19 terpenes and 2 other volatiles. The 'Guanxi' pummelo cultivar possessed a more complex composition of volatile compounds compared with the 'Shatian' cultivar. Meanwhile, the volatile compounds appeared to exhibit a higher concentration in the 'Guanxi' cultivar samples than the 'Shatian' cultivar. Cluster analysis revealed that the 'Guanxi' cultivar samples from the different regions were grouped together, whereas the 'Shatian' cultivar samples were assembled. Principal component analysis showed that an obvious separation was observed between the 'Guanxi' and 'Shatian' cultivar. However, the 'Shatian-SC15' was significantly separated from the other 'Shatian' cultivar samples. These indicated that cultivar genotype was the primary factor that determined the volatile profile of the pummelo cultivar. Cultivation region might affect the biosynthesis of volatile compounds, resulting in the differentiation of the volatile composition in each pummelo cultivar.

Phenolic compositions and antioxidant activities of grapefruit (Citrus paradisi Macfadyen) varieties cultivated in China.

The phenolic compounds in different fruit parts including the flavedos, albedos, segment membranes, juice vesicles and seeds of nine grapefruit varieties cultivated in China were determined and their antioxidant capacities were evaluated using three methods. Naringin and neohesperedin were the dominant flavonoids in all grapefruit tested. Fenghongtangmuxun and Jiwei flavedo had the highest contents of naringin (5666.82 µg/g DW) and neohesperedin (1022 µg/g DW), respectively. Gallic acid was the major phenolic acid in all grapefruit tested, and Jiwei juice vesicles had the highest content of gallic acid (343.7 µg/g DW). Fenghongtangmuxun juice vesicles were rich in chlorogenic acid (110.23 µg/g DW), caffeic acid (53.86 µg/g DW) and ferulic acid (23.12 µg/g DW). Overall, the flavedo was rich in flavonoid, while juice vesicle had high amounts of phenolic acid. The Jiwei, Fenghongtangmuxun, Maxu, Huoyan and Hongmaxu grapefruit cultivars contained more phenolics and exhibited higher antioxidant capacities than Shatianyou and Liangpingyou pummelos, and were good sources of natural phytochemical antioxidants.

Effects of pectinase clarification treatment on phenolic compounds of pummelo (Citrus grandis l. Osbeck) fruit juice.

The purpose of this study is to investigate the changes occured on phenolic compounds between two Malaysian varieties of pummelo fruit juice: Ledang (PO55) and Tambun (PO52) post-enzymatic clarification. The changes in polyphenols composition were monitored using High Performance Liquid Chromatography Diode Array Detection and Folin Ciocalteu's method. Clarification treatment of pummelo fruit juice with a commercial pectinase was optimized based on incubation temperature, time and enzyme concentration. Both varieties of pummelo fruit juice were treated with different optimized variables which produced the highest clarities with the least effect to the juice physical quality. Tambun variety was found to have significantly more total phenolic compounds (p <0.05) in comparison to Ledang variety, possibly due to the amount of naringin. Three types of hydroxycinnamic acids (chlorogenic, caffeic and coumaric acid) and three compounds of flavanones (naringin, hesperidin and narirutin) were found in both fruit juices, where naringin and chlorogenic acid were the major contributor to the total phenolic content. Naringin, which gave out bitter aftertaste to the juice, was found to decrease, 1.6 and 0.59 % reduction in Ledang and Tambun respectively, post-enzymatic treatment. The decrease in naringin, albeit nominal, could be a potential benefit to the juice production in reducing the bitterness of the juice. Post-enzymatic analysis furthermore resulted in no significance differences (p <0.05) on the total phenolic compounds of both varieties. This study in summary provides a compositional database for Malaysian pummelo fruit juice of various phenolic compounds, which can provide useful information for evaluating the authenticity and the health benefits from the juice.

Efficacy of various techniques on biochemical characteristics and bitterness of pummelo juice.

The consumer acceptability of pummelo juice is affected badly due to the presence of bitter principles in it. Therefore in order to avoid such bitterness development, the extracted juice from pummelo was subjected to five different treatments like juice diffused into syrup (70°Brix), lye peeling of segments in boiling NaOH for 2-3 min, increasing the pH of juice, hot water treatment (50 °C) prior to peeling for 20 min and without any treatment (control) for suppressing the development of bitterness in the juice. Based on bio-chemical analysis, diffusion of juice into syrup (70°Brix) showed better result as compared to other treatments. The maximum amount of TSS was found in juice diffused into syrup (i. e. 45, 30 and 15°Brix) along with highest TSS/acid ratio (92.21, 49.87 and 17.53). Higher amount of acidity was observed in pH adjusted samples with 4.25, 4.50 and 4.75 respectively. However, control samples showed higher amount of ascorbic acid (73.97 mg/100 ml juice) content followed by pH adjusted samples. The highest organoleptic score for taste (8.00), colour (8.83), aroma (8.66), overall acceptability (7.88) and extent of debittering (7.50) were recorded in juice diffused into syrup 70°Brix and achieved final TSS of juice at 45, 30 and 15°Brix respectively. Moreover, the above treatment (juice diffused into syrup 70°Brix) showed promising low cost and easy to adopt technique of debittering in respect of extent of debittering and maintaining sensory quality during storage of pummel juice.

Nutritional and volatile profiles of pulp and flavedo from four local pummelo cultivars grown in Fujian province of China.

The nutritional and volatile profiles of pulp and flavedo samples from four distinct local pummelo landraces ("Siji," "Pingshan," "Wendan," and "Guanxi") cultivated in Fujian province of China were investigated. "Guanxi" pummelo exhibited relatively high contents of vitamin C (42.01 mg/100 mL) and phenols (360.61 mg/L) and displayed a robust antioxidant capacity (41.15 mg/100 mL). Conversely, the red pulp from "Pingshan" demonstrated relatively high values of carotenoids (55.96 µg/g) and flavonoids (79.79 mg/L). Considerable differences were observed in volatile compositions between the two fruit tissues and among the four genotypes. A total of 166 and 255 volatile compounds were detected in the pulp and flavedo samples, respectively. Notably, limonene and ß-myrcene were identified as the principal volatile compounds in flavedo, whereas hexanal was highly abundant in the pulp of "Siji," "Pingshan," and "Guanxi." "Wendan" displayed distinct separation from the other three pummelo cultivars in principal component analysis based on the pulp volatile compositions. This distinction was attributed to the higher number and content of volatile compounds in "Wendan" pulp, particularly the remarkable enrichment of ß-myrcene. The newly characterized pummelo landraces and genotype/tissue-dependent variations in volatiles provide essential information for the genetic improvement of pummelo aroma, as well as for fruit processing and utilization.

Properties of an active film based on glutenin/tamarind gum and loaded with binary microemulsion of melatonin/pummelo essential oil and its preservation for Agaricus bisporus.

In order to improve the effectiveness of the active packaging, we aimed to develop an active packaging film with unidirectional sustained release, high barrier protection, and seamless attachment between the layers. An active film based on glutenin/tamarind gum loaded with the binary microemulsion of melatonin/pummelo essential oil (G/T-M-E) with sustained release and combination effects of internal and external layers was prepared. The outer barrier layer exerted an excellent protective barrier effect after adding (3-chloropropyl) triethoxysilane, which effectively reduced external interference and the ineffective diffusion of active substances in the inner layer. The effective attachment of melatonin and essential oil layer in the G/T-M-E film enhanced antioxidation, microorganism inhibition, and free-radical-scavenging properties, which effectively delayed the senescence of post-harvest white mushrooms. Furthermore, the G/T-M-E exhibited excellent tensile strength, barrier capacity, and load-bearing strength, which had a potential, positive effect on food preservation. Therefore, this film is highly recommended for packaging purposes.

Aqueous extract of pummelo pulp (Citrus maxima) improves the biochemical profile and reduces the inflammation process in Wistar rats with non-alcoholic fatty liver disease.

This study investigated the effect of pummelo extract (Citrus maxima) on biochemical, inflammatory, antioxidant and histological changes in NAFLD rats. Forty male Wistar rats divided into four groups were used: (1) control group; (2) fructose associated with high-fat diet - DHF; (3) normal diet + pummelo extract (50 mg/kg); and (4) FHD + pummelo extract. This was administered at dose of 50 mg/kg of the animal's weight, by gavage, for 45 days. Significant improvement in lipid profile, liver and kidney function, inflammation, oxidative stress markers was identified in group 4 compared to group 2. Regarding TNF-α and IL-1ß, group 2 showed higher values (respectively 142, 5 ± 0.7 and 560.5 ± 2.7 pg/mg protein) compared to group 4 (respectively 91.4 ± 0.9 and 402.1.4 ± 0.9 pg/mg protein), p < 0.05. Significant increases were found in SOD and CAT activities, respectively 0.10 ± 0.06 and 8.62 ± 1.67 U/mg protein for group 2 and respectively 0.28 ± 0.08 and 21.52 ± 2.28 U/mg of protein for group 4. Decreases in triglycerides, hepatic cholesterol and fat droplets in hepatic tissue were observed in group 4 compared to group 2. Results highlight that pummelo extract may be useful for prevent the development of NAFLD.

Anti-diabetic effect of modified 'Guanximiyou' pummelo peel pectin on type 2 diabetic mice via gut microbiota.

This study aimed to investigate the mechanisms of nature and modified 'Guanximiyou' pummelo peel pectin (GGP and MGGP) in alleviating T2DM through in vitro and in vivo. After modification, pectin was transformed from high methoxy pectin (HMP) to low methoxy pectin (LMP), and the content of galacturonic acid was increased. These made MGGP have stronger antioxidant capacity and better inhibition effect on corn starch digestion in vitro. In vivo experiments have shown that both GGP and MGGP inhibited the development of diabetes after 4 weeks of ingestion. However, MGGP can more effectively reduce blood glucose and regulate lipid metabolism, and has significant antioxidant capacity and the ability to promote SCFAs secretion. In addition, 16S rRNA analysis showed that MGGP changed the composition of intestinal microbiota in diabetic mice, decreased the abundance of Proteobacteria, and increased the relative abundance of Akkermansia, Lactobacillus, Oscillospirales and Ruminococcaceae. The phenotypes of the gut microbiome also changed accordingly, indicating that MGGP can inhibit the growth of pathogenic bacteria, alleviate intestinal functional metabolic disorders and reverse the potential risk of related complications. Altogether, our findings demonstrate that MGGP, as a dietary polysaccharide, may inhibit the development of diabetes by reversing the imbalance of gut microbiota.

Thermal conditions and active substance stability affect the isomerization and degradation of lycopene.

This study evaluated the effects of thermal treatment conditions (100-120 °C, 0-5 h) and active substance stability on the isomerization and degradation of lycopene in the "Sanhong" pummelo (C. grandis cv. Sanhongmiyou) matrix. The results revealed that the degradation of (all-E)-lycopene and total lycopene in pummelo could be well fit by the first-order kinetic model under thermal treatment. The total Z-lycopene ratio was maintained at nearly 40 % (120 °C, 4 h). The behavior of lycopene was related to the degradation of active substances, such as the degradation of ascorbic acid and polysaccharides. The stability of polysaccharides was temperature-dependent, and with the increasing temperature (100-120 °C), a significant negative correlation was observed between the content of Z-isomers and polysaccharides. The enhancement of lycopene isomerization was attributed to the degradation of polysaccharides under thermal treatment. These findings facilitate the functional development and efficient utilization of lycopene in fruit processing.

Structural elucidation of flavonoids from Shatianyu (Citrus grandis L. Osbeck) pulp and screening of key antioxidant components.

The Citrus genus is a good source of dietary flavonoids, which have many health benefits. As a representative citrus fruit, the flavonoids composition in Shatianyu (Citrus grandis L. Osbeck) pulp remains to be investigated. In the present study, 11 flavonoids were isolated and identified from Shatianyu pulp flavonoid extracts (SPFEs). Among them, 4 flavonoids were previously undescribed and 2 flavonoids were firstly isolated from pummelo. The cellular antioxidant activity (CAA) and oxygen radical absorbance capacity (ORAC) of isolated compounds were evaluated. Naringin and rhoifolin showed the highest ORAC activity, and the presence of a 3-hydroxy-3-methylglutaryl or a 4'-glucose decreased the ORAC activity of flavonoids. The contribution of isolated flavonoids to the holistic antioxidant activity of SPFEs was determined by an online knockout method. Melitidin, bergamjuicin and naringin contributed most to ORAC activity, while bergamjuicin, melitidin and apigenin-4'-O-ß-d-glucopyranosyl-7-O-α-l-rhamnopyranosyl-(1 â†’ 2)-[6″-O-(3- hydroxy-3-methylgltaryl)]-ß-d-glucopyranoside contributed most to CAA activity.

Carvacrol delays Phomopsis stem-end rot development in pummelo fruit in relation to maintaining energy status and antioxidant system.

Pummelo fruit rapidly depreciate in commodity value due to postharvest fungal decay and fruit quality deterioration. Here, we used carvacrol (CVR) to control Phomopsis stem-end rot (SER) caused by Diaporthe citri in pummelo fruit stored at 25 °C. Antifungal activity of CVR inhibited D. citri growth and Phomopsis SER development. Harvested pummelo fruit treated with CVR delayed firmness loss and lowered electrolyte leakage, and retarded hydrogen peroxide (H2O2) and malondialdehyde (MDA) accumulation. Unlike the control fruit, the CVR-treated fruit maintained higher levels of adenosine triphosphate and energy charge, and increased ATPase, succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and cytochrome C oxidase (CCO) activities, along with up-regulated expression levels of the respective genes. CVR improved the antioxidant capacity, as evidenced by higher non-enzymatic antioxidants amounts, higher activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR), and up-regulated expression levels of ROS-scavenging-related genes. Collectively, CVR treatment maintained the energy status and antioxidant capacity in D. citri-infected pummelo fruit, which revealed antifungal mechanisms critical for controlling postharvest fungal diseases.

Apple polyphenols delay postharvest senescence and quality deterioration of 'Jinshayou' pummelo fruit during storage.

Introduction: Apple polyphenols (AP), derived from the peel of mature-green apples, are widely used as natural plant-derived preservatives in the postharvest preservation of numerous horticultural products. Methods: The goal of this research was to investigate how AP (at 0.5% and 1.0%) influences senescence-related physiological parameters and antioxidant capacity of 'Jinshayou' pummelo fruits stored at 20°C for 90 d. Results: The treating pummelo fruit with AP could effectively retard the loss of green color and internal nutritional quality, resulting in higher levels of total soluble solid (TSS) content, titratable acidity (TA) content and pericarp firmness, thus maintaining the overall quality. Concurrently, AP treatment promoted the increases in ascorbic acid, reduced glutathione, total phenols (TP) and total flavonoids (TF) contents, increased the scavenging rates of 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and hydroxyl radical (•OH), and enhanced the activities of superoxide dismutase (SOD), catalase, peroxidase, ascorbate peroxidase (APX), and glutathione reductase (GR) as well as their encoding genes expression (CmSOD, CmCAT, CmPOD, CmAPX, and CmGR), reducing the increases in electrolyte leakage, malondialdehyde content and hydrogen peroxide level, resulting in lower fruit decay rate and weight loss rate. The storage quality of 'Jinshayou' pummelo fruit was found to be maintained best with a 1.0% AP concentration. Conclusion: AP treatment can be regarded as a promising and effective preservative of delaying quality deterioration and improving antioxidant capacity of 'Jinshayou' pummelo fruit during storage at room temperature.

Identification of a delayed leaf greening gene from a mutation of pummelo.

Delayed greening of young leaves is an unusual phenomenon of plants in nature. Citrus are mostly evergreen tree species. Here, a natural mutant of "Guanxi" pummelo (Citrus maxima), which shows yellow leaves at the young stage, was characterized to identify the genes underlying the trait of delayed leaf greening in plants. A segregating population with this mutant as the seed parent and a normal genotype as the pollen parent was generated. Two DNA pools respectively from the leaves of segregating seedlings with extreme phenotypes of normal leaf greening and delayed leaf greening were collected for sequencing. Bulked segregant analysis (BSA) and InDel marker analysis demonstrated that the delayed leaf greening trait is governed by a 0.3 Mb candidate region on chromosome 6. Gene expression analysis further identified a key candidate gene (Citrus Delayed Greening gene 1, CDG1) in the 0.3 Mb region, which showed significantly differential expression between the genotypes with delayed and normal leaf greening phenotypes. There was a 67 bp InDel region difference in the CDG1 promoter and the InDel region contains a TATA-box element. Confocal laser-scanning microscopy revealed that the CDG1-GFP fusion protein signals were co-localized with the chloroplast signals in the protoplasts. Overexpression of CDG1 in tobacco and Arabidopsis led to the phenotype of delayed leaf greening. These results suggest that the CDG1 gene is involved in controlling the delayed leaf greening phenotype with important functions in chloroplast development.

A sensitive and practical ELISA for analyzing naringenin in pummelo and herb samples.

Naringenin, a flavonoid compound found in pummelo, is a key biological active compound in some traditional Chinese medicines, including Citri reticulatae pericarpium, Citri reticulatae pericarpium viride, Aurantii fructus immaturus, and Aurantii fructus. These Chinese medicinal preparations are the peels or immature fruits of certain citrus species. Aiming at detecting naringenin in complex matrices such as pummelo and traditional Chinese medicines, we put forward a sensitive and practical indirect competitive enzyme-linked immunosorbent assay (icELISA) based on anti-naringenin monoclonal antibodies (anti-Nar-mAbs). The median inhibitory concentration (IC50) was 4.43 ng/mL, and the working range was 1.15-15.81 ng/mL. The findings of the icELISA for the analysis of naringenin in pummelo and herb samples had a good correlation with the ultra performance liquid chromatography (UPLC) methodology and showed good accuracy and reproducibility. These data demonstrated that the developed icELISA is reliable, accurate, and suitable for detecting naringenin in pummelo and traditional Chinese medicines.