Agronomy-Basel
·
2023
·Vol. 13
·期 6
·1489
·
IF 3.7
Yidan Kou,Jinpeng Ren,Yujie Ma,Rongrong Guo,Juane Shang,Dongliang Qiu,Cuilan Ma
In this study, the two peach cultivars ‘Baifeng’ and ‘Weiduanmihong’ were used as experimental materials, and their fruits were sprayed with different concentrations of L-glutamic acid, brassinolide, and sucrose to study the effects of these three exogenous substances on fruit quality and anthocyanin metabolism of peaches. The results showed that the appearance quality (average single fruit weight, fruit firmness, and peel color difference), nutritional quality (soluble solids, soluble sugar, titratable acid, anthocyanins, total phenols, flavonoids, etc.), peel anthocyanin-related enzyme activity, and related gene expression of ‘Baifeng’ and ‘Weiduanmihong’ peaches treated with three different exogenous substances were different from those of the control. Higher-concentration treatments could significantly improve the appearance of peach fruit, the nutritional quality of peach fruit and the activity of anthocyanin-related enzymes in peel, as well as promote the expression of related genes. Treatment with 400 mg/L L-glutamic acid significantly promoted the average fruit weight of ‘Baifeng’ peaches. Treatment with 800 mg/L L-glutamic acid significantly promoted the increase in PAL enzyme activity and the expression ofPpPALandPpF3Hin the two peach varieties and significantly promoted the expression of anthocyanin metabolism genesPpF3′HandPpGST1in ‘Baifeng’ peach peel and anthocyanin metabolism genePpUFGTin ‘Weiduanmihong’ peach peel. Treatment with 34 mg/L sucrose significantly increased the fruit firmness of ‘Baifeng’ peaches and the soluble sugar content of ‘Weiduanmihong’ peaches. Treatment with 51 mg/L sucrose significantly promoted the increase in flavonoid content andPpUFGTexpression in ‘Baifeng’ peach fruit and significantly promoted the expression of anthocyanin metabolism genesPpDFRandPpANSin ‘Weiduanmihong’ peach peel. Treatment with 0.6 mg/L brassinolide significantly promoted the increase in soluble solids (TTS), soluble sugar, anthocyanin, total phenol content, PAL enzyme activity, UFGT enzyme activity, and the expression of anthocyanin metabolism genesPpDFRandPpMYB10.1in ‘Baifeng’ peach fruit, and it significantly increased the average single fruit weight, fruit hardness, anthocyanin content, and UFGT enzyme activity of ‘Weiduanmihong’ peach fruit and promoted the expression of anthocyanin metabolism genesPpF3HandPpGST1in ‘Weiduanmihong’ peach peel. The comprehensive effect of 0.6 mg/L brassinolide treatment on improving peach fruit quality and increasing anthocyanin content produces the best results and could be popularized in production practices.Keywords:peach (Prunus persicaL.);exogenous substance treatment;fruit quality;anthocyanin;qRT-PCR technology
研究领域:
农学
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植物生理学
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分子生物学
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高影响因子
TALANTA
·
2023
·
IF 6.1
Haodong Cui,Wenxing Song,Xiaoling Ru,Wen Fu,Ling Ji,Wenhua Zhou,Zhen Zhao,Guangbo Qu,Xue-Feng Yu,Guibin Jiang
The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based methods are being widely applied for clinical diagnosis, timely and accurate diagnosis towards COVID-19 causing virus, the SARS-CoV-2, is still limited due to labor-intensive and time-consuming operations. Herein, we report a new viral RNA extraction method based on poly-(amino ester) with carboxyl group (PC)-coated magnetic nanoparticles (pcMNPs) for the sensitive detection of SARS-CoV-2. This method combines the lysis and binding steps into one step, and refines multiple washing steps into one step, giving a turnaround time of less than 9 min. Furthermore, the extracted pcMNP-RNA complexes can be directly introduced into subsequent RT-PCR reactions without elution. This simplified viral RNA method could be well adapted in fast manual and automated high-throughput nucleic acids extraction protocols suitable for different scenarios. A high sensitivity down to 100 copies/mL and a linear correlation between 100 and 10 6 copies/mL of SARS-CoV-2 pseudovirus particles are achieved in both protocols. Benefitting from the simplicity and excellent performances, this new method can dramatically improve the efficiency and reduce operational requirements for the early clinical diagnosis and large-scale SARS-CoV-2 nucleic acid screening.
研究领域:
分子生物学
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医学诊断
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纳米技术
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传染病研究
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PubMed
高影响因子
PLANT DISEASE
·
2023
·
IF 4.5
Miss Bairu Ren,Prof. Yi Wang,Dr. Huijie Chen,Mr. Lei Diao,Miss Jiaxin Wang,Miss Shuoyuan Zhang,Mr. Yongzhe Zhang,Prof. Meiping Zhang,Dr. Rui Yin,Dr. Yanfang Wang
Botrytis cinerea is a ubiquitous pathogen that can infect at least 200 dicotyledonous plant species including many agriculturally and economically important crops. In Ginseng, the fungus may cause ginseng gray mold disease, causing great economic losses in ginseng industry. Therefore, the early detection of Botrytis cinerea in the process of ginseng production is necessary for the disease prevention and control of the pathogen’s spread. In this study, a polymerase chain reaction-nucleic acid sensor (PCR-NAS) rapid detection technique was established, and it can be used for field detection of B. cinerea through anti-pollution design and portable integration. The present study showed that the sensitivity of PCR-NAS technology is 10 times higher than that of traditional PCR-electrophoresis, and there is no need for expensive detection equipment or professional technicians. The detection results of nucleic acid sensors can be read by the naked eye under 3 minutes. Meanwhile, the technique has high specificity for the detection of B. cinerea. The testing of 50 field samples showed that the detection results of PCR-NAS were consistent with those of the real-time quantitative PCR (qPCR) method. The PCR-NAS technique established in this study can be used as a novel nucleic acid field detection technique, and it has a potential application in the field detection of B. cinerea to achieve early warning of the pathogen infection.
研究领域:
植物病理学
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分子生物学
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农业科学
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PubMed
高影响因子
JOURNAL OF FOOD PROCESS ENGINEERING
·
2026
·Vol. 49
·期 1
·e70351
·
IF 2.9
Kai Chen,JianZhi Luo,Bin Xiao,XueWei Yuan,Juan Zhang,Nan Wang,LiQiang Liu,AiKe Li,Ailiang Chen
This study established an optimized CTAB-based DNA extraction method for edible oils, which enables stable and reliable DNA recovery from commercially available edible oils for authenticity identification and adulteration detection. Real-time PCR was used to assess DNA precipitation time and organic solvent extraction, revealing that longer precipitation increased inhibitors, hindering PCR amplification, while organic solvents enhanced efficiency. The method was applied to detect adulteration in commercially available soybean and olive oils. A standard curve based on real-time PCR Ct values demonstrated the ability to detect as low as 10% soybean oil in olive oil, with recovery rates of 92% and 80% for samples spiked with 70% and 30% soybean oil, respectively. The coefficients of variation (CV) were 11% and 14%, within acceptable ranges. This reliable method provides a valuable tool for edible oil authentication and adulteration detection.
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高影响因子
FOOD RESEARCH INTERNATIONAL
·
2026
·
IF 8.8
Lu Han,Min Chen,Qian Huang,Weijie Lan,Chong Wang,Leiqing Pan,Kang Tu
Foodborne illnesses caused by Salmonella rank first among bacterial foodborne diseases, representing a significant risk to public health and food safety, which necessitates the development of rapid and sensitive detection technologies. This study developed an upconversion‑tungsten disulfide PCR fluorescence sensor based on a “dual quenching-dual recovery” mechanism for the detection of Salmonella typhimurium ( S. typhimurium ). By utilizing the dual properties of tungsten disulfide, which include peroxidase-like activity and high fluorescence quenching efficiency, the sensor establishes a quantitative correlation between the upconversion fluorescence signal and S. typhimurium concentration, following target-specific signal amplification via PCR. The constructed method demonstrated good specificity and repeatability, with a detection limit as low as 22.5 CFU/mL. Furthermore, by reducing the number of PCR cycles by approximately 12 (from 34 to 22), the detection time was shortened by about 24 min while achieving a detection limit comparable to that of qPCR. The sensor performed excellently in real food matrices (chicken, eggs, fish), showing spike recovery rates of 92.0%–106.3% and relative standard deviations below 10%. In summary, the upconversion fluorescence biosensing strategy integrated with PCR amplification presented in this study provides a highly sensitive, rapid, and reliable solution for the pathogen detection, significantly enhancing the capacity for prevention and control of foodborne diseases.
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PubMed
高影响因子
Jove-Journal of Visualized Experiments
·
2023
·
IF 1.2
Wenwen Dong,Chunxian Tao,Bo Yang,Erika Miyake,Zhenqing Li,Dawei Zhang,Yoshinori Yamaguchi
Polymerase chain reaction (PCR) is a traditional method employed for the amplification of a target gene that has played an important role in biomolecular diagnostics. However, traditional PCR is very time-consuming because of the low-temperature variation efficiency. This work proposes a continuous-flow-PCR (CF-PCR) system based on a microfluidic chip. The amplification time can be greatly reduced by running the PCR solution into a microchannel placed on heaters set at different temperatures. Moreover, as capillary electrophoresis (CE) is an ideal way to differentiate positive and false-positive PCR products, a CE system was built to achieve efficient separation of the DNA fragments. This paper describes the process of amplification of Escherichia coli (E. coli) by the CF-PCR system built in-house and the detection of the PCR products by CE. The results demonstrate that the target gene of E. coli was successfully amplified within 10 min, indicating that these two systems can be used for the rapid amplification and detection of nucleic acids.
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2018
·Vol. 19
·期 8
·2403
·
IF 3.687
Yukun Chen,Xiaoping Xu,Xiaohui Chen,Yan Chen,Zihao Zhang,Xu Xuhan,Yuling Lin,Zhongxiong Lai
Mother of FT and TFL1(MFT) belongs to phosphatidylethanolamine-binding protein (PEBP) family, which plays an important role in flowering time regulation, seed development, and germination. To gain insight into the molecular function ofDlMFTinDimocarpus longanLour., we isolatedDlMFTand its promoter sequence from longan embryogenic callus (EC). Bioinformatic analysis indicated that the promoter contained multiphytohormones and light responsive regulatory elements. Subcellular localization showed that the given the DlMFT signal localized in the nucleus, expression profiling implied thatDlMFTshowed significant upregulation during somatic embryogenesis (SE) and zygotic embryogenesis (ZE), and particular highly expressed in late or maturation stages. The accumulation ofDlMFTwas mainly detected in mature fruit and seed, while it was undetected in abortive seeds, and notably decreased during seed germination.DlMFTresponded differentially to exogenous hormones in longan EC. Auxins, salicylic acid (SA) and methyl jasmonate (MeJa) suppressed its expression, however, abscisic acid (ABA), brassinosteroids (BR) showed the opposite function. Meanwhile,DlMFTdifferentially responded to various abiotic stresses. Our study revealed thatDlMFTmight be a key regulator of longan somatic and zygotic embryo development, and in seed germination, it is involved in complex plant hormones and abiotic stress signaling pathways.Keywords:mother of FT and TFL1;longan;embryogenesis;phytohormones;stress response;quantitative real-time PCR
研究领域:
植物生物学
·
分子遗传学
·
发育生物学
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PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2026
·
IF 7.1
Juping Zheng,Tiantian Huang,Chao Li,Yichen Tian,Yuzhu Song
A broad-spectrum bacterial enrichment strategy was developed using antimicrobial peptide-modified magnetic beads (MBs@AMP). • The method combined with mPCR simultaneously detects five foodborne pathogens in fruit juice samples. • This approach eliminates food matrix interference and improves detection sensitivity compared to conventional PCR. Rapid detection of foodborne pathogens in food samples is essential to mitigate threats to public health and ensure food safety. Due to the complexity of food matrix and low concentration of pathogens, traditional detection methods have certain limitations in actual detection. The effective enrichment of foodborne pathogens during sample pretreatment will greatly improve the sensitivity and accuracy of subsequent detection. In this study, an antimicrobial peptide (AMP) was designed as a capture element capable of binding multiple bacterial species, including both Gram-negative and Gram-positive ones. The AMP was loaded on the surface of the magnetic beads (MBs@AMP) for magnetic separation. The MBs@AMP were then combined with multiplex PCR (MBs@AMP-mPCR) to rapidly and sensitively detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Shigella spp. , and Staphylococcus aureus in food samples. The detection limit of MBs@AMP-mPCR was 100-fold lower than that of conventional PCR, which is of great significance for the rapid detection of foodborne pathogens.
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高影响因子
MICROCHEMICAL JOURNAL
·
2022
·
IF 5.304
Xuekun Bai,Zhengzheng Wang,Weiqiang Li,Fangbin Xiao,Jin Huang,Qian Xu,Hengyi Xu
Listeria monocytogenes ( L . monocytogenes ) is a typical food-borne pathogen which poses a serious threat to global public health. Achieving rapid and sensitive detection of pathogens in food is critical to solve food safety problems. In this work, ampicillin functionalized magnetic beads (Amp-MBs) were synthesized and used as a capture probe for detection of L . monocytogenes in milk sample combined with quantitative real-time polymerase chain reaction (qPCR) with high sensitivity and specificity. Amp, as a broad spectrum antibiotic was selected to identify bacteria of the Listeria spp. powerfully, and the hly as a specific gene of L . monocytogenes was selected to establish qPCR for accurate detection. At optimal conditions, the capture efficiency of Amp-MBs for L . monocytogenes (10 1 -10 6 CFU/mL) and other bacteria of Listeria spp. (10 5 CFU/mL) was higher than 90 %. The limit of detection of Amp-MBs-qPCR strategy for L . monocytogenes was 10 1 and 10 2 CFU/mL in PBS and artificially contaminated milk within 2.5 h, respectively. And this strategy could detect L. monocytogenes from high concentration Listeria spp. with ascendant specificity. These results demonstrated that the Amp-MBs-qPCR strategy was successfully established and applied to the specific detection of L. monocytogenes in milk samples.
研究领域:
食品科学
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微生物学
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分子生物学
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DOI
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2021
·
IF 3.963
Meng-Wen Yan,Xiao-Rong Xing,Fu-An Wu,Jun Wang,Sheng Sheng
Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae) is a predominant endoparasitoid of lepidopteran pests in mulberry fields. Extensive application of insecticides puts natural enemies under threat. UDP-glycosyltransferases (UGTs), as important detoxification enzymes , potentially contribute to the detoxification of pesticides in insects. To investigate the roles of UGTs in the process of tolerance towards commonly used insecticides in M. pulchricornis , ten UGT genes were identified from the transcriptome database of M. pulchricornis . Seven UGT genes contained full-length ORFs and shared 47.12–78.28% identity with other homologous hymenopteran insects. qRT-PCR validation revealed that UGT genes can be induced by treatment of sublethal doses of phoxim, cypermethrin and chlorfenapyr, respectively, and these upregulations were depending on the time post insecticide treatments. To further explore the functions of UGT genes, three MpulUGT genes were singly knocked down, which resulted in the decline of UGT expression and significantly increased mortality of parasitoids under sublethal doses of insecticides exposure. This study revealed that UGTs in M. pulchricornis contributed to the tolerance towards insecticides and provided basic insight into the insecticide detoxification mechanism in parasitoid wasps .
研究领域:
昆虫学
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分子生物学
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害虫管理
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PubMed
高影响因子
ANALYST
·
2023
·Vol. 148
·期 7
·1492-1499
·
IF 4.2
Jiahai Wang,Lanfang Chen,Cenlin Gui,Jianji Zhu,Baian Zhu,Zhuobin Zhu,Yunhui Li,Daqi Chen
DNA methylation has been considered an essential epigenetic biomarker for diagnosing various diseases, such as cancer. A simple and sensitive way for DNA methylation level detection is necessary. Inspired by the label-free and ultra-high sensitivity of solid-state nanopores to double-stranded DNA (dsDNA), we proposed a nanopore counter for evaluating DNA methylation by integrating a dual-restriction endonuclease digestion strategy coupled with polymerase chain reaction (PCR) amplification. Simultaneous application of BstUI/HhaI endonucleases can ensure the full digestion of the unmethylated target DNA but shows no effect on the methylated ones. Therefore, only the methylated DNA remains intact and can trigger the subsequent PCR reaction, producing a large quantity of fixed-length PCR amplicons, which can be directly detected through glassy nanopores. By simply counting the event rate of the translocation signals, the concentration of methylated DNA can be determined to range from 1 aM to 0.1 nM, with the detection limit as low as 0.61 aM. Moreover, a 0.01% DNA methylation level was successfully distinguished. The strategy of using the nanopore counter for highly sensitive DNA methylation evaluation would be a low-cost but reliable alternative in the analysis of DNA methylation.
研究领域:
生物化学
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分子生物学
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医学诊断
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PubMed
高影响因子
Frontiers in Microbiology
·
2025
·
IF 4.5
Zibei Wu,Xufeng Zhang,Lili Wang,Jinlei Sui,Mingqi Yu,Bingzhangke Bao,Jianing Qian,Guohui Yi,Yinzheng Ma,Xiaowen He
Streptococcus mutans is one of the most significant pathogens leading to dental caries by producing highly cariogenic biofilms within the oral cavity. However, the increasing antimicrobial resistance brings great difficulties to the prevention and treatment of dental caries. Therefore, it is very important to develop new and effective antimicrobial agents from natural sources. Clausena lansium (Lour.) Skeels, a plant commonly cultivated in Southern China, is rich in various active constituents. In this study, the main constituents of the essential oils extracted from Clausena lansium leaves (EOL), seeds (EOS), peels (EOP) were precisely analyzed by gas chromatography–mass spectrometry. The antimicrobial and anti-biofilm activities of EOL, EOS, and EOP, along with their main chemical constituents, against Streptococcus mutans were comprehensively evaluated. The primary constituent in EOL was β-caryophyllene, with a content of 112.29 mg/mL. EOS was found to be abundant in sabinene and 4-terpineol, with constituents of 147.21 mg/mL and 54.36 mg/mL, respectively. In EOP, the most predominant constituent was β-phellandrene, with a content of 173.95 mg/mL. The minimum inhibitory concentrations of EOL, EOS, and EOP were 0.12, 2.00, and 1.00 mg/mL, respectively. The minimum concentrations of inhibiting at least 90% biofilm formation of EOL, EOS, and EOP was 0.12 mg/mL, 2.00, and 2.00 mg/mL, respectively. Intriguingly, the antimicrobial and anti-biofilm efficacies of the main constituents were significantly lower than those of the essential oils. By observing the dynamic changes of the biofilm formation of S. mutans treated with essential oils, it was found that the process of biofilm formation was greatly delayed. In some cases, there was even no biofilm formation. The results of real-time quantitative PCR indicated that the expression levels of brpA, gtfB, gtfC, gtfD, and luxS genes were down-regulated after treatment with the essential oils. This study reinforces the potential of Clausena lansium as a promising source for developing new antimicrobial and anti-biofilm agents. They could inhibit the proliferation and virulence of S. mutans, thereby contributing to the control of dental caries.
研究领域:
植物学
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微生物学
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药理学
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分子生物学
·
牙科医学
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高影响因子
危险品化学品经营许可证（带存储）