Multi-omic analysis of the extension of broccoli quality during storage by folic acid.

INTRODUCTION: Folic acid (FA) is a critical metabolite in all living organisms and an important nutritional component of broccoli. Few studies have been conducted on the impact of an exogenous application of FA on the postharvest physiology of fruits and vegetables during storage. In this regard, the mechanism by which an exogenous application of FA extends the postharvest quality of broccoli is unclear. OBJECTIVE: This study utilized a multicomponent analysis to investigate how an exogenous application of FA effects the postharvest quality of broccoli. METHODS: Broccoli was soaked in 5 mg/L FA for 10 min and the effect of the treatment on the appearance and nutritional quality of broccoli was evaluated. These data were combined with transcriptomic, metabolomic, and DNA methylation data to provide insight into the potential mechanism by which FA delays senescence. RESULTS: The FA treatment inhibited the yellowing of broccoli during storage. CHH methylation was identified as the main type of methylation that occurs in broccoli and the FA treatment was found to inhibit DNA methylation, promote the accumulation of endogenous FA and chlorophyl, and inhibit ethylene biosynthesis in stored broccoli. The FA treatment also prevented the formation of off-odors by inhibiting the degradation of glucosinolate. CONCLUSIONS: FA treatment inhibited the loss of nutrients during the storage of broccoli, delayed its yellowing, and inhibited the generation of off-odors. Our study provides deeper insight into the mechanism by which the postharvest application of FA delays postharvest senescence in broccoli and provides the foundation for further studies of postharvest metabolism in broccoli.

Ozone micro-nano bubble water preserves the quality of postharvest parsley.

The production and use of ozone micro-nano bubble water (O3-MNBW) is an innovative technology that prolongs the reactivity of aqueous-phase ozone and maintains the freshness and quality of fruits and vegetables by removing pesticides, mycotoxins, and other contaminants. The quality of parsley treated with different concentrations of O3-MNBW was investigated during storage at 20 ℃ for 5 d, and found that a ten-minute exposure of parsley to 2.5 mg·L-1 O3-MNBW effectively preserved the sensory quality of parsley, and resulted in lower weight loss, respiration rate, ethylene production, MDA levels, and a higher level of firmness, vitamin C, and chlorophyll content, relative to untreated parsley. The O3-MNBW treatment also increased the level of total phenolics and flavonoids, enhanced peroxidase and ascorbate peroxidase activity, and inhibited polyphenol oxidase activity in stored parsley. Five volatile signatures identified using an electronic nose (W1W, sulfur-compounds; W2S, ethanol; W2W, aromatic- and organic- sulfur compounds; W5S, oxynitride; W1S, methane) exhibited a significant decrease in response to the O3-MNBW treatment. A total of 24 major volatiles were identified. A metabolomic analysis identified 365 differentially abundant metabolites (DMs). Among them, 30 and 19 DMs were associated with characteristic volatile flavor substance metabolism in O3-MNBW and control groups, respectively. The O3-MNBW treatment increased the abundance of most DMs related to flavor metabolism and reduced the level of naringin and apigenin. Our results provide insight into the mechanisms that are regulated in response to the exposure of parsley to O3-MNBW, and confirmed the potential use of O3-MNBW as a preservation technology.

Whole-transcriptome RNA sequencing reveals changes in amino acid metabolism induced in harvested broccoli by red LED irradiation.

Whole-transcriptomic profiling combined with amino acid analysis were conducted in order to gain a better understanding of global changes in amino acid metabolism induced in broccoli by red LED irradiation. The results showed that the contents of almost all 16 amino acids in postharvest broccoli were maintained under red LED illumination. The red LED irradiation enhanced the anabolism of amino acid, including the biosynthesis of aromatic amino acids by upregulating the genes' expression in the shikimate pathway, as well as by upregulating the genes' expression which encoding biosynthetic enzymes in the branched-chain amino acid biosynthesis pathway. Red LED irradiation induced the expression of genes encoding aspartate aminotransferase, which plays a role in Asp synthesis, aspartate kinase, which functions in aspartate metabolism, and a cytoplasmic aspartate aminotransferase that converts 2-Oxoglutarate into Glu. Genes encoding imidazole glycerol-phosphate synthase and histidinol-phosphatase, which function in the His biosynthesis pathway, were also upregulated. According to our results, red LED irradiation delays broccoli's yellowing and senescence by regulating amino acid metabolism. These results enhance our understanding of the role of amino acid metabolism in the senescence of broccoli and the mechanism of red LED irradiation to alter amino acid metabolism in harvested broccoli.

The effect of BVOCs produced by Lysinibacillus fusiformis and LED irradiation on pigment metabolism in stored broccoli.

Volatile organic compounds produced by bacteria (BVOCs) have been proven to effect the postharvest metabolism of fruits and vegetables. The quality, color and antioxidant capacity of membrane lipids of broccoli in storage were effectively maintained by fumigation with BVOCs produced by Lysinibacillus fusiformis combined with white light emitting diode (LED) technology. An analysis of the transcriptome and metabolome of broccoli treated with the combined LED-BVOCs technology resulted in the identification of 49 differentially expressed genes (DEGs) and 13 differentially abundant metabolites (DAMs) involved in photosynthesis (32/0 DEGs upregulated/downregulated; 0/0 DAMs with increased/decreased abundance), chlorophyll (7/0; 1/2), carotenoid (5/0; 1/4) and flavonoid (3/3; 3/2) metabolism. The maintenance of green color in harvested broccoli treated by LED-BVOCs was associated with DEGs and DAMs that inhibited chlorophyll degradation and carotenoid accumulation. Our study provides a theoretical basis for understanding the delayed senescence of broccoli during storage using BVOCs-LED technology.

Palmitic Acid Regulation of Stem Browning in Freshly Harvested Mini-Chinese Cabbage (Brassica pekinensis (Lour.) Rupr.).

The effect of palmitic acid (PA) on stem browning was investigated in freshly harvested mini-Chinese cabbage (Brassica pekinensis). Results indicated that concentrations of PA ranging from 0.03 g L-1 to 0.05 g L-1 inhibited stem browning and decreased the rate of respiration, electrolyte leakage, and weight loss, as well as the level of malondialdehyde (MDA) in freshly harvested mini-Chinese cabbage stored at 25 °C for 5 d. The PA treatment enhanced the activity of antioxidant enzymes (ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), 4-coumarate:CoA ligase (4CL) and phenylalamine ammonia lyase (PAL)), and inhibited the activity of polyphenol oxidase (PPO). The PA treatment also increased the level of several phenolics (chlorogenic acid, gallic acid, catechin, p-coumaric acid, ferulic acid, p-hydroxybenzoic acid, and cinnamic acid) and flavonoids (quercetin, luteolin, kaempferol, and isorhamnetin). In summary, results indicate that treatment of mini-Chinese cabbage with PA represents an effective method for delaying stem browning and maintaining the physiological quality of freshly harvested mini-Chinese cabbage due to the ability of PA to enhance antioxidant enzyme activity and the level of phenolics and flavonoids during 5 d.

Potential virulence factors of Nocardia seriolae AHLQ20-01 based on whole-genome analysis and its pathogenicity to largemouth bass (Micropterus salmoides).

Nocardia seriolae is a major causative agent of fish nocardiosis that results in serious economic losses in the aquaculture industry. However, the virulence factors and pathogenic mechanisms of the bacterium are poorly understood. Here, a new N. seriolae strain AHLQ20-01 was isolated from the diseased Micropterus salmoides and identified by phenotypic examination combined with 16S rRNA sequencing. Subsequently, the potential virulence factors of the strain were analysed at genome level by whole-genome sequencing. The results showed that the whole-genome sequence derived from N. seriolae AHLQ20-01 circular chromosome contains 8,129,380 bp DNA with G + C content of 68.14%, and encompasses 7650 protein-coding genes, 114 pseudo-genes, 3 rRNAs, 66 tRNAs and 36 non-coding RNAs. More importantly, a total of 139 genes, which mainly involved in adhesion, invasion, resistance to oxidative and nitrosative stress, phagosome arresting, iron acquisition system, toxin production and bacterial secretion systems, were identified as core virulence-associated genes. Furthermore, the pathogenicity of N. seriolae AHLQ20-01 to M. salmoides was further investigated through experimental infection. It was found that the LD50 value of the strain to M. salmoides was 9.3 × 106  colony forming unit/fish. Histopathological examination demonstrated typical granuloma with varying sizes in the liver, head kidney, spleen and heart of the experimentally infected fish. Terminal deoxynucleotidyl transferase dUTP nick end labelling assay and 4',6-diamidino-2-phenylindole staining showed that there were distinctly more apoptotic cells in all the tested tissues in the infection group, but not in the control group. Together, these findings provide the foundation to further explore the pathogenic mechanism of N. seriolae, which might contribute to the prevention and treatment of fish nocardiosis.

First identification of Nocardia seriolae GapA adhesion function and its three B-cell epitopes with cell-binding activity.

Fish nocardiosis mainly caused by Nocardia seriolae (N. seriolae) is a serious threat to aquaculture. Bacterial adhesion to host cells mediated by adhesin is an initial step of pathogenesis. But it is not clear whether glyceraldehyde-3-phosphate dehydrogenase (GapA) is an adhesin of N. seriolae. Here, recombinant GapA protein (rGapA) was prokaryotic expressed, and its role in the bacterial adhesion to Ctenopharyngodon idella kidney cells was investigated by indirect immunofluorescence, protein-binding assay and adhesion inhibition assay. The results showed that an obvious green fluorescence was observed on the surface of the cells co-incubated with rGapA protein; the cytomembrane proteins of the cells pretreated with rGapA could react with anti-rGapA antibody; and the antibody significantly inhibited the adhesion ability of the bacteria. Subsequently, B-cell linear epitopes of GapA protein were identified by using a immunoinformatics approach combined with peptide ELISA and Western blot for the first time. It was found that four predicted epitopes (Ep58-69 , Ep139-150 , Ep186-197 , Ep318-329 ) could all react with anti-rGapA antibody and obviously inhibit the immunoreactivity between rGapA and anti-rGapA antibody, and they were confirmed as indeed B-cell linear epitopes of the protein. Furthermore, flow cytometry analysis found the percentage of positive cells co-incubated with FITC-labelled epitope peptides (Ep139-150 , Ep186-197 , Ep318-329 ) was significantly higher than those in the FITC-labelled Ep58-69 , unrelated control peptide and cell control. Collectively, GapA is an adhesin of N. seriolae, and epitope peptides (Ep139-150 , Ep186-197 , Ep318-329 ) possess cell-binding activity, which are potential candidates for developing a multiple epitopes-based adhesin vaccine against fish nocardiosis.

The Adaxial/Abaxial Patterning of Auxin and Auxin Gene in Leaf Veins Functions in Leafy Head Formation of Chinese Cabbage.

Leaf curling is an essential prerequisite for the formation of leafy heads in Chinese cabbage. However, the part or tissue that determines leaf curvature remains largely unclear. In this study, we first introduced the auxin-responsive marker DR5::GUS into the Chinese cabbage genome and visualized its expression during the farming season. We demonstrated that auxin response is adaxially/abaxially distributed in leaf veins. Together with the fact that leaf veins occupy considerable proportions of the Chinese cabbage leaf, we propose that leaf veins play a crucial supporting role as a framework for heading. Then, by combining analyses of QTL mapping and a time-course transcriptome from heading Chinese cabbage and non-heading pak choi during the farming season, we identified the auxin-related gene BrPIN5 as a strong candidate for leafy head formation. PIN5 displays an adaxial/abaxial expression pattern in leaf veins, similar to that of DR5::GUS, revealing an involvement of BrPIN5 in leafy head development. The association of BrPIN5 function with heading was further confirmed by its haplo-specificity to heading individuals in both a natural population and two segregating populations. We thus conclude that the adaxial/abaxial patterning of auxin and auxin genes in leaf veins functions in the formation of the leafy head in Chinese cabbage.

The Microbial Metagenome of Eluates Obtained From the Surface of Broccoli Heads Subjected to Different Light Treatments.

Foodborne illnesses present a major threat to public health and are frequently attributed to foodborne pathogens present on fresh produce. Some opportunistic pathogens of broccoli are also responsible for causing head rot. Three different light treatments, UV-C, red LED (50 µml/m2/s), and UV-C + LED were used to treat broccoli prior to or during storage. Following the light treatments, microorganisms present in eluates obtained from the surface of broccoli heads were characterized using a metagenomic approach. Metagenomic DNA libraries were subjected to high-throughput sequencing on an Illumina Hiseq platform. Results indicated that the combined treatment of LED red light and UV-C provided the best sensory preservation of broccoli, followed by LED red light and then UV-C. The bacterial communities in the eluates obtained from the surface of broccoli heads in all three light treatments were primarily represented at the phylum level by Proteobacteria and Firmicutes, while fungal communities were primarily represented by Ascomycota and Basidiomycota. Further analysis indicated that the all three light treatments reduced the presence of foodborne pathogens and bacterial taxa responsible for broccoli spoilage. While UV-C had a significant inhibitory effect on Botrytis cinerea, the light treatments increased the relative abundance of Pseudomonas fluorescens. Results indicate that a metagenomic approach can be used to detect pathogenic bacteria and fungi on fresh vegetables and assess the impact of management practices, such as light treatments, designed to maintain postharvest quality, on the composition of the microbiome present on the surface of harvested produce.

Effect of a refined nursing model based on nursing quality feedback on the postoperative mental state of patients with laryngeal cancer.

OBJECTIVE: To evaluate the impact and effect of a refined nursing model based on nursing quality feedback on the postoperative mental state of patients with laryngeal cancer. METHODS: According to the principle of a randomized controlled trial, 102 patients undergoing laryngeal cancer surgery were divided into two groups with 51 patients in each group. The control group received a conventional nursing model, and the research group received a refined nursing model based on nursing quality feedback. Patients' mental status, illness perception, finding of benefits, cancer-related fatigue, changes in quality of life before and 3 months after operation were observed, and postoperative complications and nursing satisfaction were collected. RESULTS: The scores of the Self-Rating Anxiety Scale (SAS), the Self-Rating Depression Scale (SDS), the Brief Illness perception Questionnaire (BIPQ), and Revised Piper Fatigue Scale in the research group were lower than those in the control group at 3 months after operation, and the scores of benefit finding, EORTC QLQ-C30, and La Monica-Oberst Patient Satisfaction Scale (LOPSS) were higher than those in the control group (P < 0.05); and the complication rate of the research group (7.84%) was lower than that of the control group (23.53%) (P < 0.05). CONCLUSION: For patients undergoing laryngeal cancer surgery, the use of the refined nursing model based on nursing quality feedback can reduce patients' negative emotions, increase the benefits found, reduce their negative perceptions and cancer-related fatigue, improve their quality of life, reduce complications, and enhance the nurse-patient relationship.

Identification and fine mapping of qSB.A09, a major QTL that controls shoot branching in Brassica rapa ssp. chinensis Makino.

KEY MESSAGE: QTL mapping plus bulked segregant analysis revealed a major QTL for shoot branching in non-heading Chinese cabbage. The candidate gene was then identified using sequence alignment and expression analysis. Shoot branching is a complex quantitative trait that contributes to plant architecture and ultimately yield. Although many studies have examined branching in grain crops, the genetic control of shoot branching in vegetable crops such as Brassica rapa L. ssp. chinensis remains poorly understood. In this study, we used bulked segregant analysis (BSA) of an F2 population to detect a major quantitative trait locus (QTL) for shoot branching, designated shoot branching 9 (qSB.A09) on the long arm of chromosome A09 in Brassica rapa L. ssp. chinensis. In addition, traditional QTL mapping of the F2 population revealed six QTLs in different regions. Of these, the mapping region on chromosome A09 was consistent with the results of BSA-seq analysis, as well as being stable over the 2-year study period, explaining 19.37% and 22.18% of the phenotypic variation across multiple genetic backgrounds. Using extreme recombinants, qSB.A09 was further delimited to a 127-kb genomic region harboring 28 annotated genes. We subsequently identified the GRAS transcription factor gene Bra007056 as a potential candidate gene; Bra007056 is an ortholog of MONOCULM 1 (MOC1), the key gene that controls tillering in rice. Quantitative RT-PCR further revealed that expression of Bra007056 was positively correlated with the shoot branching phenotype. Furthermore, an insertion/deletion marker specific to Bra007056 co-segregated with the shoot branching trait in the F2 populations. Overall, these results provide the basis for elucidating the molecular mechanism of shoot branching in Brassica rapa ssp. chinensis Makino.

AtMOB1 Genes Regulate Jasmonate Accumulation and Plant Development.

The MOB1 proteins are highly conserved in yeasts, animals, and plants. Previously, we showed that the Arabidopsis (Arabidopsis thaliana) MOB1A gene (AtMOB1A/NCP1) plays critical roles in auxin-mediated plant development. Here, we report that AtMOB1A and AtMOB1B redundantly and negatively regulate jasmonate (JA) accumulation and function in Arabidopsis development. The two MOB1 genes exhibited similar expression patterns, and the MOB1 proteins displayed similar subcellular localizations and physically interacted in vivo. Furthermore, the atmob1a atmob1b (mob1a/1b) double mutant displayed severe developmental defects, which were much stronger than those of either single mutant. Interestingly, many JA-related genes were up-regulated in mob1a/1b, suggesting that AtMOB1A and AtMOB1B negatively regulate the JA pathways. mob1a/1b plants accumulated more JA and were hypersensitive to exogenous JA treatments. Disruption of MYC2, a key gene in JA signaling, in the mob1a/1b background partially alleviated the root defects and JA hypersensitivity observed in mob1a/1b. Moreover, the expression levels of the MYC2-repressed genes PLT1 and PLT2 were significantly decreased in the mob1a/1b double mutant. Our results showed that MOB1A/1B genetically interact with SIK1 and antagonistically modulate JA-related gene expression. Taken together, our findings indicate that AtMOB1A and AtMOB1B play important roles in regulating JA accumulation and Arabidopsis development.

Arabidopsis AGC protein kinases IREH1 and IRE3 control root skewing.

AGC protein kinases play important roles in plant growth and development. Several AGC kinases in Arabidopsis have been functionally characterized. However, the "AGC Other" subfamily, including IRE, IREH1, IRE3 and IRE4, has not been well understood. Here, we reported that ireh1 mutants displayed a root skewing phenotype, which can be enhanced by ire3 mutation. IREH1 and IRE3 were expressed in roots, consistent with their function in controlling root skewing. The fluorescence intensities of the microtubule marker KNpro:EGFP-MBD were decreased in ireh1, ire3 and ireh1 ire3 mutants compared to wild type. The microtubule arrangements in ireh1 and ireh1 ire3 mutants were also altered. IREH1 physically interacted with IRE3 in vitro and in planta. Thus, our findings demonstrate that IREH1 and IRE3 protein kinases play important roles in controlling root skewing, and maintaining microtubule network in Arabidopsis.

Ultrasonic vibration seeds showed improved resistance to cadmium and lead in wheat seedling.

Heavy metals have long-term adverse impacts on the health of soil ecosystems and even exhibit hazardous influences on human health. Literatures have shown that heavy metals could result in the reduction of crops growth and development and finally result in crops production decline. To determine whether or not ultrasonic vibration alleviate damage induced by cadmium and lead in crops, the wheat seeds, which is one of the most important agriculture crops in China and other countries in the world, were exposed to 10 min ultrasonic vibration and then the toxicological effects were investigated. Wheat seeds were soaked for 3 h with water and then the seeds were placed in clean beaker with some water, the beaker were placed in ultrasonic apparatus to vibrate (model, KQ-200VDV; frequency, 45 KHz; power, 160 W). Pretreatment seeds of 80 were sown in dishes (Ø 15 cm). After seeds emergence, the seedlings were thinned to 60 per dish. The dishes with seedlings were placed in a growth chamber maintained at 25 °C, 70% relative humidity and 380 µmol mol(-1) CO2 under dark condition. A 400 µmol m(-2) s(-1) photosynthetically active radiation was provided for 8 h (dark for 16 h) after the seed germination. When the seedlings were 2 days old, the seedlings were subjected to cadmium and lead for 4 days and then some selective biochemical and physiological parameters were measured. (1) Although each doses of ultrasonic vibration could improve seed germination, enhance biosynthesis of protein and chlorophyll and seedlings growth, the optimum dosage of ultrasonic vibration was 10 min. (2) Compared with the controls, cadmium and lead stress led to significant increase in the concentrations of malondialdehyde (MDA) and O(-2) and in the conductivity of electrolyte leakage, but the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), the glutathione concentration, and the shoot weight were decreased by Cd and Pb stress. In the case of the seeds exposed to ultrasonic vibration and the seedlings followed by cadmium lead stress, the concentrations of MDA and O(-2), and the conductivity of electrolyte leakage were significantly lower than those in cadmium and lead stress; the activities of CAT, SOD, and GR and the shoot weight were significantly higher (except for glutathione (GSH) concentration) than those in cadmium and lead stress seedlings. The membrane is responsible for the selective inflow and outflow of molecules, ions, and water, and is a dynamic structure that performs a variety of functions. Cellular membrane systems play an important role in the compartmentalization of cells and maintaining intercellular homeostasis. Abiotic and biotic stress can induce functional impairments to the cellular membrane systems through triggering an increased formation of reactive oxygen species (ROS), such as superoxide (O2 (-)), hydrogen peroxide (H2O2), and hydroxyl radicals. There are several pathways that can be utilized to eliminate ROS in plants, e.g., CAT, SOD, and GR and GSH, etc. compared with controls, cadmium, and lead enhanced the concentrations of ROS; decreased the SOD, CAT, and GR activities; the GSH concentration, and the seedling growth. In the case of ultrasonic pretreatment followed by cadmium and lead stresses, the activities of CAT, SOD, and GR were significant higher, and the conductivity of electrolyte leakage and the concentrations of MDA and O2 (-) were significant lower than that of those subjected by cadmium and lead stress. This phenomenon demonstrated ultrasonic pretreatment can help plant eliminate the ROS by enhance the activities of antioxidant enzymes. These results suggested that ultrasonic vibration can alleviate the toxicological effect induced by heavy mental.

Ethylene mediates UV-B-induced stomatal closure via peroxidase-dependent hydrogen peroxide synthesis in Vicia faba L.

Ultraviolet B (UV-B) radiation is an important environmental signal for plant growth and development, but its signal transduction mechanism is unclear. UV-B is known to induce stomatal closure via hydrogen peroxide (H(2)O(2)), and to affect ethylene biosynthesis. As ethylene is also known to induce stomatal closure via H(2)O(2) generation, the possibility of UV-B-induced stomatal closure via ethylene-mediated H(2)O(2) generation was investigated in Vicia faba by epidermal strip bioassay, laser-scanning confocal microscopy, and assays of ethylene production. It was found that H(2)O(2) production in guard cells and subsequent stomatal closure induced by UV-B radiation were inhibited by interfering with ethylene biosynthesis as well as ethylene signalling, suggesting that ethylene is epistatic to UV-B radiation in stomatal movement. Ethylene production preceded H(2)O(2) production upon UV-B radiation, while exogenous ethylene induced H(2)O(2) production in guard cells and subsequent stomatal closure, further supporting the conclusion. Inhibitors for peroxidase but not for NADPH oxidase abolished H(2)O(2) production upon UV-B radiation in guard cells, suggesting that peroxidase is the source of UV-B-induced H(2)O(2) production. Taken together, our results strongly support the idea that ethylene mediates UV-B-induced stomatal closure via peroxidase-dependent H(2)O(2) generation.