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Vale Professor Xiaoxian Liu

Xu F, Binns C, Low WY

Asia Pac J Public Health, 2020 5 16;32(2-3):134-135.
PMID: 32410510 DOI: 10.1177/1010539520915009
Intention to have a third child among millennial parents with two children in eastern China: A cross-sectional survey

Xu F, Jiang SJ, Ge PP, Yang J, Lu S

Nurs Open, 2024 May;11(5):e2178.
PMID: 38757435 DOI: 10.1002/nop2.2178

AIM: This Study Aimed to Assess the Intention to Have a Third Child among Millennial Parents (25-40 years old) with Two Children in a City in Eastern China and to Explore the Influencing Factors Related to Fertility Intention.
DESIGN: A cross-sectional design study.
METHODS: A convenience sampling method was used to enrol participants of childbearing age who visited two tertiary hospitals in Hang zhou, a city in eastern China, from June 2021 to March 2022. We conducted a face-to-face questionnaire survey with 520 participants and calculated the prevalence of intention-related factors. Multivariate logistic regression was used to analyse the independent influencing factors of fertility intention.
RESULTS: In total, 105 (20.2%) participants had the intention to have a third child. The results showed that 'employment status', 'age', 'reasons for wanting a third child', the considered 'biggest barrier to having a third child', 'views on the three-child policy', 'desired free services', 'supporting work policies' and 'assistance policies' were significant independent influencing factors of intention to have a third child (p-value
Sensitive biomolecule detection in lateral flow assay with a portable temperature-humidity control device

Choi JR, Hu J, Feng S, Wan Abas WA, Pingguan-Murphy B, Xu F

Biosens Bioelectron, 2016 May 15;79:98-107.
PMID: 26700582 DOI: 10.1016/j.bios.2015.12.005

Lateral flow assays (LFAs) have currently attracted broad interest for point-of-care (POC) diagnostics, but their application has been restricted by poor quantification and limited sensitivity. While the former has been currently solved to some extent by the development of handheld or smartphone-based readers, the latter has not been addressed fully, particularly the potential influences of environmental conditions (e.g., temperature and relative humidity (RH)), which have not yet received serious attention. The present study reports the use of a portable temperature-humidity control device to provide an optimum environmental requirement for sensitivity improvement in LFAs, followed by quantification by using a smartphone. We found that a RH beyond 60% with temperatures of 55-60°C and 37-40°C produced optimum nucleic acid hybridization and antigen-antibody interaction in LFAs, respectively representing a 10-fold and 3-fold signal enhancement over ambient conditions (25°C, 60% RH). We envision that in the future the portable device could be coupled with a fully integrated paper-based sample-to-answer biosensor for sensitive detection of various target analytes in POC settings.
RpoS differentially affects the general stress response and biofilm formation in the endophytic Serratia plymuthica G3

Liu X, Wu Y, Chen Y, Xu F, Halliday N, Gao K, et al.

Res. Microbiol., 2016 Apr;167(3):168-77.
PMID: 26671319 DOI: 10.1016/j.resmic.2015.11.003

The σ(S) subunit RpoS of RNA polymerase functions as a master regulator of the general stress response in Escherichia coli and related bacteria. RpoS has been reported to modulate biocontrol properties in the rhizobacterium Serratia plymuthica IC1270. However, the role of RpoS in the stress response and biofilm formation in S. plymuthica remains largely unknown. Here we studied the role of RpoS from an endophytic S. plymuthica G3 in regulating these phenotypes. Mutational analysis demonstrated that RpoS positively regulates the global stress response to acid or alkaline stresses, oxidative stress, hyperosmolarity, heat shock and carbon starvation, in addition to proteolytic and chitinolytic activities. Interestingly, rpoS mutations resulted in significantly enhanced swimming motility, biofilm formation and production of the plant auxin indole-3-acetic acid (IAA), which may contribute to competitive colonization and environmental fitness for survival. These findings provide further insight into the strain-specific role of RpoS in the endophytic strain G3 of S. plymuthica, where it confers resistance to general stresses encountered within the plant environment. The heterogeneous functionality of RpoS in rhizosphere and endophytic S. plymuthica populations may provide a selective advantage for better adaptation to various physiological and environmental stresses.
Engineering of microscale three-dimensional pancreatic islet models in vitro and their biomedical applications

Gao B, Wang L, Han S, Pingguan-Murphy B, Zhang X, Xu F

Crit Rev Biotechnol, 2016 Aug;36(4):619-29.
PMID: 25669871 DOI: 10.3109/07388551.2014.1002381

Diabetes now is the most common chronic disease in the world inducing heavy burden for the people's health. Based on this, diabetes research such as islet function has become a hot topic in medical institutes of the world. Today, in medical institutes, the conventional experiment platform in vitro is monolayer cell culture. However, with the development of micro- and nano-technologies, several microengineering methods have been developed to fabricate three-dimensional (3D) islet models in vitro which can better mimic the islet of pancreases in vivo. These in vitro islet models have shown better cell function than monolayer cells, indicating their great potential as better experimental platforms to elucidate islet behaviors under both physiological and pathological conditions, such as the molecular mechanisms of diabetes and clinical islet transplantation. In this review, we present the state-of-the-art advances in the microengineering methods for fabricating microscale islet models in vitro. We hope this will help researchers to better understand the progress in the engineering 3D islet models and their biomedical applications such as drug screening and islet transplantation.
Hydrogel-based methods for engineering cellular microenvironment with spatiotemporal gradients

Wang L, Li Y, Huang G, Zhang X, Pingguan-Murphy B, Gao B, et al.

Crit Rev Biotechnol, 2016 Jun;36(3):553-65.
PMID: 25641330 DOI: 10.3109/07388551.2014.993588

Natural cellular microenvironment consists of spatiotemporal gradients of multiple physical (e.g. extracellular matrix stiffness, porosity and stress/strain) and chemical cues (e.g. morphogens), which play important roles in regulating cell behaviors including spreading, proliferation, migration, differentiation and apoptosis, especially for pathological processes such as tumor formation and progression. Therefore, it is essential to engineer cellular gradient microenvironment incorporating various gradients for the fabrication of normal and pathological tissue models in vitro. In this article, we firstly review the development of engineering cellular physical and chemical gradients with cytocompatible hydrogels in both two-dimension and three-dimension formats. We then present current advances in the application of engineered gradient microenvironments for the fabrication of disease models in vitro. Finally, concluding remarks and future perspectives for engineering cellular gradients are given.
Advances in paper-based point-of-care diagnostics

Hu J, Wang S, Wang L, Li F, Pingguan-Murphy B, Lu TJ, et al.

Biosens Bioelectron, 2014 Apr 15;54:585-97.
PMID: 24333570 DOI: 10.1016/j.bios.2013.10.075

Advanced diagnostic technologies, such as polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), have been widely used in well-equipped laboratories. However, they are not affordable or accessible in resource-limited settings due to the lack of basic infrastructure and/or trained operators. Paper-based diagnostic technologies are affordable, user-friendly, rapid, robust, and scalable for manufacturing, thus holding great potential to deliver point-of-care (POC) diagnostics to resource-limited settings. In this review, we present the working principles and reaction mechanism of paper-based diagnostics, including dipstick assays, lateral flow assays (LFAs), and microfluidic paper-based analytical devices (μPADs), as well as the selection of substrates and fabrication methods. Further, we report the advances in improving detection sensitivity, quantification readout, procedure simplification and multi-functionalization of paper-based diagnostics, and discuss the disadvantages of paper-based diagnostics. We envision that miniaturized and integrated paper-based diagnostic devices with the sample-in-answer-out capability will meet the diverse requirements for diagnosis and treatment monitoring at the POC.
An integrated lateral flow assay for effective DNA amplification and detection at the point of care

Choi JR, Hu J, Gong Y, Feng S, Wan Abas WA, Pingguan-Murphy B, et al.

Analyst, 2016 05 10;141(10):2930-9.
PMID: 27010033 DOI: 10.1039/c5an02532j

Lateral flow assays (LFAs) have been extensively explored in nucleic acid testing (NAT) for medical diagnostics, food safety analysis and environmental monitoring. However, the amount of target nucleic acid in a raw sample is usually too low to be directly detected by LFAs, necessitating the process of amplification. Even though cost-effective paper-based amplification techniques have been introduced, they have always been separately performed from LFAs, hence increasing the risk of reagent loss and cross-contaminations. To date, integrating paper-based nucleic acid amplification into colorimetric LFA in a simple, portable and cost-effective manner has not been introduced. Herein, we developed an integrated LFA with the aid of a specially designed handheld battery-powered system for effective amplification and detection of targets in resource-poor settings. Interestingly, using the integrated paper-based loop-mediated isothermal amplification (LAMP)-LFA, we successfully performed highly sensitive and specific target detection, achieving a detection limit of as low as 3 × 10(3) copies of target DNA, which is comparable to the conventional tube-based LAMP-LFA in an unintegrated format. The device may serve in conjunction with a simple paper-based sample preparation to create a fully integrated paper-based sample-to-answer diagnostic device for point-of-care testing (POCT) in the near future.
Paper-based point-of-care testing for diagnosis of dengue infections

Choi JR, Hu J, Wang S, Yang H, Wan Abas WA, Pingguan-Murphy B, et al.

Crit Rev Biotechnol, 2017 Feb;37(1):100-111.
PMID: 26912259

Dengue endemic is a serious healthcare concern in tropical and subtropical countries. Although well-established laboratory tests can provide early diagnosis of acute dengue infections, access to these tests is limited in developing countries, presenting an urgent need to develop simple, rapid, and robust diagnostic tools. Point-of-care (POC) devices, particularly paper-based POC devices, are typically rapid, cost-effective and user-friendly, and they can be used as diagnostic tools for the prompt diagnosis of dengue at POC settings. Here, we review the importance of rapid dengue diagnosis, current dengue diagnostic methods, and the development of paper-based POC devices for diagnosis of dengue infections at the POC.
Paper-based sample-to-answer molecular diagnostic platform for point-of-care diagnostics

Choi JR, Tang R, Wang S, Wan Abas WA, Pingguan-Murphy B, Xu F

Biosens Bioelectron, 2015 Dec 15;74:427-39.
PMID: 26164488 DOI: 10.1016/j.bios.2015.06.065

Nucleic acid testing (NAT), as a molecular diagnostic technique, including nucleic acid extraction, amplification and detection, plays a fundamental role in medical diagnosis for timely medical treatment. However, current NAT technologies require relatively high-end instrumentation, skilled personnel, and are time-consuming. These drawbacks mean conventional NAT becomes impractical in many resource-limited disease-endemic settings, leading to an urgent need to develop a fast and portable NAT diagnostic tool. Paper-based devices are typically robust, cost-effective and user-friendly, holding a great potential for NAT at the point of care. In view of the escalating demand for the low cost diagnostic devices, we highlight the beneficial use of paper as a platform for NAT, the current state of its development, and the existing challenges preventing its widespread use. We suggest a strategy involving integrating all three steps of NAT into one single paper-based sample-to-answer diagnostic device for rapid medical diagnostics in the near future.
Cryopreservation of Human Mesenchymal Stem Cells for Clinical Applications: Current Methods and Challenges

Yong KW, Wan Safwani WK, Xu F, Wan Abas WA, Choi JR, Pingguan-Murphy B

Biopreserv Biobank, 2015 Aug;13(4):231-9.
PMID: 26280501 DOI: 10.1089/bio.2014.0104

Mesenchymal stem cells (MSCs) hold many advantages over embryonic stem cells (ESCs) and other somatic cells in clinical applications. MSCs are multipotent cells with strong immunosuppressive properties. They can be harvested from various locations in the human body (e.g., bone marrow and adipose tissues). Cryopreservation represents an efficient method for the preservation and pooling of MSCs, to obtain the cell counts required for clinical applications, such as cell-based therapies and regenerative medicine. Upon cryopreservation, it is important to preserve MSCs functional properties including immunomodulatory properties and multilineage differentiation ability. Further, a biosafety evaluation of cryopreserved MSCs is essential prior to their clinical applications. However, the existing cryopreservation methods for MSCs are associated with notable limitations, leading to a need for new or improved methods to be established for a more efficient application of cryopreserved MSCs in stem cell-based therapies. We review the important parameters for cryopreservation of MSCs and the existing cryopreservation methods for MSCs. Further, we also discuss the challenges to be addressed in order to preserve MSCs effectively for clinical applications.
Fulltext Mechanoregulation of cardiac myofibroblast differentiation: implications for cardiac fibrosis and therapy

Yong KW, Li Y, Huang G, Lu TJ, Safwani WK, Pingguan-Murphy B, et al.

Am J Physiol Heart Circ Physiol, 2015 Aug 15;309(4):H532-42.
PMID: 26092987 DOI: 10.1152/ajpheart.00299.2015

Cardiac myofibroblast differentiation, as one of the most important cellular responses to heart injury, plays a critical role in cardiac remodeling and failure. While biochemical cues for this have been extensively investigated, the role of mechanical cues, e.g., extracellular matrix stiffness and mechanical strain, has also been found to mediate cardiac myofibroblast differentiation. Cardiac fibroblasts in vivo are typically subjected to a specific spatiotemporally changed mechanical microenvironment. When exposed to abnormal mechanical conditions (e.g., increased extracellular matrix stiffness or strain), cardiac fibroblasts can undergo myofibroblast differentiation. To date, the impact of mechanical cues on cardiac myofibroblast differentiation has been studied both in vitro and in vivo. Most of the related in vitro research into this has been mainly undertaken in two-dimensional cell culture systems, although a few three-dimensional studies that exist revealed an important role of dimensionality. However, despite remarkable advances, the comprehensive mechanisms for mechanoregulation of cardiac myofibroblast differentiation remain elusive. In this review, we introduce important parameters for evaluating cardiac myofibroblast differentiation and then discuss the development of both in vitro (two and three dimensional) and in vivo studies on mechanoregulation of cardiac myofibroblast differentiation. An understanding of the development of cardiac myofibroblast differentiation in response to changing mechanical microenvironment will underlie potential targets for future therapy of cardiac fibrosis and failure.
Hot air treatment reduces postharvest decay in Chinese bayberries during storage by affecting fungal community composition

Dai K, Han P, Zou X, Jiang S, Xu F, Wang H, et al.

Food Res Int, 2021 02;140:110021.
PMID: 33648251 DOI: 10.1016/j.foodres.2020.110021

Chinese bayberry fruit were treated with hot air (HA) at 48 ℃ for 3 h and then stored at 4 ℃ for 15 d. Changes in fungal communities were analyzed by high-throughput sequencing (HTS), and decay and fruit quality were monitored during storage. The results showed that HA treatment effectively maintains fruit quality and the richness and diversity of fungal communities. Heat treatment inhibited decay development and reduced the growth of fungi in the genera Botryotinia spp., Davidiella spp., Hanseniaspora spp., and Candida spp. Canonical correspondence analysis further revealed that Botryotinia spp. and Davidiella spp. were positively correlated with fruit decay and weight loss. FUNGuild analysis demonstrated that HA-treated bayberries had a lower relative abundance within the plant pathogen guild, but higher relative abundance within the endophyte guild. The results suggest that HA treatment reduces pathogens by favoring the increase of endophytes, providing new insight into the decay development and quality changes during the storage of postharvest Chinese bayberries.
Global genetic diversity of Spirometra tapeworms

Hong X, Liu SN, Xu FF, Han LL, Jiang P, Wang ZQ, et al.

Trop Biomed, 2020 Mar 01;37(1):237-250.
PMID: 33612735

Spirometra larvae are etiological agents of human sparganosis. However, the systematics of spirometrid cestodes has long been controversial. In order to determine the current knowledge on the evolution and genetic structure of Spirometra, an exhaustive population diversity analysis of spirometrid cestodes using the mitochondrial gene: cytochrome c oxidase subunit 1 (cox1) was performed. All publicly available cox1 sequences available in the GenBank and 127 new sequencing genes from China were used as the dataset. The haplotype identify, network, genetic differentiation and phylogenetic analysis were conducted successively. A total of 488 sequences from 20 host species, representing four spirometrid tapeworms (S. decipiens, S. ranarum, S. erinaceieuropaei and Sparganum proliferum) and several unclassified American and African isolates from 113 geographical locations in 17 countries, identified 45 haplotypes. The genetic analysis revealed that there are four clades of spirometrid cestodes: Clade 1 (Brazil + USA) and Clade 2 (Argentina + Venezuela) included isolates from America, Clade 3 contained African isolates and one Korean sample, and the remainders from Asia and Australia belonged to Clade 4; unclassified Spirometra from America and Africa should be considered the separate species within the genus; and the taxonomy of two Korea isolates (S. erinaceieuropaei KJ599680 and S. decipiens KJ599679) was still ambiguous and needs to be further identified. In addition, the demographical analyses supported population expansion for the total spirometrid population. In summary, four lineages were found in the spirometrid tapeworm, and further investigation with deeper sampling is needed to elucidate the population structure.
Hepatocellular BChE as a therapeutic target to ameliorate hypercholesterolemia through PRMT5 selective degradation to restore LDL receptor transcription

Wang D, Tan KS, Zeng W, Li S, Wang Y, Xu F, et al.

Life Sci, 2022 Mar 15;293:120336.
PMID: 35065166 DOI: 10.1016/j.lfs.2022.120336

AIMS: Individuals with nonalcoholic hepatosteatosis (NAFLD) have a worse atherogenic lipoprotein profile and are susceptible to cardiovascular diseases. The MEK-ERK signaling cascades are central regulators of the levels of LDL receptor (LDLR), a major determinant of circulating cholesterol. It is elusive how hepatic steatosis contributes to dyslipidemia, especially hypercholesterolemia.
MAIN METHODS: The effects of BChE on signaling pathways were determined by immunoblotting in a BChE knockout hepatocyte cell line. DiI-LDL probe was used to explore the effect of BChE expression on LDL internalization. Co-immunoprecipitation and LC-MS were used to explore the interacting proteins with BChE. Finally, a hepatocyte-restricted BChE silencing mouse model was established by AAV8-Tbg-shRNA, and the hypercholesterolemia was induced by 65% kcal% high-fat, high-sucrose diet feeding.
MAIN FINDINGS: Here we demonstrate that butyrylcholinesterase (BChE) governs the LDL receptor levels and LDL uptake capacity through the MEK-ERK signaling cascades to promote Ldlr transcription. BChE interacts and co-localizes with PRMT5, a protein methylation modifier controlling the ERK signaling. PRMT5 regulates LDLR-dependent LDL uptake and is a substrate of chaperone-mediated autophagy (CMA). BChE deficiency induces the PRTM5 degradation dependent on CMA activity, possibly through facilitating the HSC70 (Heat shock cognate 71 kDa) recognition of PRMT5. Remarkably, in vivo hepatocyte-restricted BChE silencing reduces plasma cholesterol levels substantially. In contrast, the BChE knockout mice are predisposed to hypercholesterolemia.
SIGNIFICANCE: Taken together, these findings outline a regulatory role for the BChE-PRMT5-ERK-LDLR axis in hepatocyte cholesterol metabolism, and suggest that targeting liver BChE is an effective therapeutic strategy to treat hypercholesterolemia.
Epinecidin-1, a marine antifungal peptide, inhibits Botrytis cinerea and delays gray mold in postharvest peaches

Fan L, Wei Y, Chen Y, Jiang S, Xu F, Zhang C, et al.

Food Chem, 2023 Mar 01;403:134419.
PMID: 36191421 DOI: 10.1016/j.foodchem.2022.134419

This study investigatedthe mechanism of epinecidin-1 against Botrytis cinerea, in vitro, and its effectiveness at inhibiting gray mold on postharvest peach fruit. We found that in vitro, epinecidin-1 had significantly greater antifungal activity against B. cinerea than either clavanin-A or mytimycin, two other marine derived antimicrobial peptides that we tested. Its antifungal activity was heat-resistant (15 min at 40-100 °C) and tolerant to lower concentrations of cations (<100 mM Na+, K+; <10 mM Ca2+). Epinecidin-1 interacted directly with B. cinerea genomic DNA, and that in mycelia, epinecidin-1 exposure induced accumulation of intracellular ROS and increased the permeability of cell membranes resulting in leakage of nucleic acids and aberrant cell morphology. Meanwhile, 200 μM of epinecidin-1 had a significant inhibitory effect on gray mold injected into peach fruit. These results suggested that epinecidin-1 showed promise as a potential method for controlling postharvest gray mold in peaches.
ROS Stress and Cell Membrane Disruption are the Main Antifungal Mechanisms of 2-Phenylethanol against Botrytis cinerea

Zou X, Wei Y, Jiang S, Xu F, Wang H, Zhan P, et al.

J Agric Food Chem, 2022 Nov 16;70(45):14468-14479.
PMID: 36322824 DOI: 10.1021/acs.jafc.2c06187

2-Phenylethanol (2-PE), a common compound found in plants and microorganisms, exhibits broad-spectrum antifungal activity. Using Botrytis cinerea, we demonstrated that 2-PE suppressed mycelium growth in vitro and in strawberry fruit and reduced natural disease without adverse effects to fruit quality. 2-PE caused structural damage to mycelia, as shown by scanning and transmission electron microscopy. From RNA sequencing analysis we found significantly upregulated genes for enzymatic and nonenzymatic reactive oxygen species (ROS) scavenging systems including sulfur metabolism and glutathione metabolism, indicating that ROS stress was induced by 2-PE. This was consistent with results from assays demonstrating an increase ROS and hydrogen peroxide levels, antioxidant enzyme activities, and malondialdehyde content in treated cells. The upregulation of ATP-binding cassette transporter genes, the downregulation of major facilitator superfamily transporters genes, and the downregulation of ergosterol biosynthesis genes indicated a severe disruption of cell membrane structure and function. This was consistent with results from assays demonstrating compromised membrane integrity and lipid peroxidation. To summarize, 2-PE exposure suppressed B. cinerea growth through ROS stress and cell membrane disruption.
Fulltext Lab Scale Extracted Conditions of Polyphenols from Thinned Peach Fruit Have Antioxidant, Hypoglycemic, and Hypolipidemic Properties

Dai K, Wei Y, Jiang S, Xu F, Wang H, Zhang X, et al.

Foods, 2021 Dec 31;11(1).
PMID: 35010225 DOI: 10.3390/foods11010099

Thinned peach polyphenols (TPPs) were extracted by ultrasonic disruption and purified using macroporous resin. Optimized extraction conditions resulted in a TPPs yield of 1.59 ± 0.02 mg GAE/g FW, and optimized purification conditions resulted in a purity of 43.86% with NKA-9 resin. TPPs composition was analyzed by UPLC-ESI-QTOF-MS/MS; chlorogenic acid, catechin, and neochlorogenic acid were the most abundant compounds in thinned peaches. Purified TPPs exhibited scavenging activity on DPPH, ABTS, hydroxyl radical, and FRAP. TPPs inhibited α-amylase and α-glucosidase by competitive and noncompetitive reversible inhibition, respectively. TPPs also exhibited a higher binding capacity for bile acids than cholestyramine. In summary, TPPs from thinned peaches are potentially valuable because of their high antioxidant, hypoglycemic, and hypolipidemic capacities, and present a new incentive for the comprehensive utilization of thinned peach fruit.
Engineering ellipsoidal cap-like hydrogel particles as building blocks or sacrificial templates for three-dimensional cell culture

Zhang W, Huang G, Ng K, Ji Y, Gao B, Huang L, et al.

Biomater Sci, 2018 Mar 07.
PMID: 29511758 DOI: 10.1039/c7bm01186e

Hydrogel particles that can be engineered to compartmentally culture cells in a three-dimensional (3D) and high-throughput manner have attracted increasing interest in the biomedical area. However, the ability to generate hydrogel particles with specially designed structures and their potential biomedical applications need to be further explored. This work introduces a method for fabricating hydrogel particles in an ellipsoidal cap-like shape (i.e., ellipsoidal cap-like hydrogel particles) by employing an open-pore anodic aluminum oxide membrane. Hydrogel particles of different sizes are fabricated. The ability to produce ellipsoidal cap-like magnetic hydrogel particles with controlled distribution of magnetic nanoparticles is demonstrated. Encapsulated cells show high viability, indicating the potential for using these hydrogel particles as structure- and remote-controllable building blocks for tissue engineering application. Moreover, the hydrogel particles are also used as sacrificial templates for fabricating ellipsoidal cap-like concave wells, which are further applied for producing size controllable cell aggregates. The results are beneficial for the development of hydrogel particles and their applications in 3D cell culture.
Melatonin treatment delays postharvest senescence of broccoli with regulation of carotenoid metabolism

Lou J, Wu C, Wang H, Cao S, Wei Y, Chen Y, et al.

Food Chem, 2023 May 15;408:135185.
PMID: 36525725 DOI: 10.1016/j.foodchem.2022.135185

The effect of melatonin treatment on the carotenoid metabolism in broccoli florets during storage was explored. The results indicated that 100 µmol/L of melatonin maintained the sensory quality of broccoli florets, which retarded the increase of the L* value and the decrease of the H value. Melatonin treatment increased the activities of tryptophan decarboxylase (TDC), tryptamine 5-hydroxylase (T5H), serotonin N-acetyltransferase (SNAT) and N-acetylserotonin methyltransferase (ASMT), leading to the enrichment of endogenous melatonin content in broccoli florets. Meanwhile, the treatment inhibited the concentrations of β-carotene, β-cryptoxanthin, zeaxanthin and lutein, which was beneficial in delaying the yellowing of broccoli. In addition, a series of carotenoid biosynthetic genes such as BoPSY, BoPDS, BoZDS, BoLCYβ and BoZEP was also suppressed by melatonin. Further analysis revealed that the lower carotenoid content and the down-regulated BoNCED expression in treated broccoli resulted in less accumulation of abscisic acid precursors, inhibiting abscisic acid production during the yellowing process.

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