【Objective】This study aimed to establish a quintuple PCR method for rapid and simultaneous detection of Ralstonia solanacearum,Fusarium spp.,Pectobacterium spp.,Enterobacter spp.,and Pythium spp.,which provided tech...【Objective】This study aimed to establish a quintuple PCR method for rapid and simultaneous detection of Ralstonia solanacearum,Fusarium spp.,Pectobacterium spp.,Enterobacter spp.,and Pythium spp.,which provided technical support for early diagnosis of various soil-borne diseases on ginger.【Method】For five types of soil-borne pathogens causing ginger bacterial wilt and rhizome rot,specific primer combinations were designed and screened,the optimal quintuple reaction system was established by exploring optimal primer concentrations,annealing temperature,and sensitivity,and was applied to detect field plant samples to verify its utility.【Result】Specific primers pairs Rs1F/Rs1R,En1F/En1R,and Py1F/Py1R were designed according to flic gene of Ralstonia solanacearum,rpoB gene of Enterobacter spp.,and 18S rDNA of Pythium spp.,and combined with reported Fusarium spp.specific primers Fu3/Fu4 and specific primers 23SPecF/23SPecR of Pectobacterium spp.,a quintuple PCR reaction system for ginger soil-borne pathogens has been established(25.00μL):above primer dosage was 1.20,0.20,0.60,1.60,and 0.15μL respectively;2×PCR Mix 12.50μL;DNA templates of different pathogens were 1.00μL each;added ddH_(2)O to 25.00μL.Annealing temperature was optimized to 55.4℃.The specific fragments with sizes of 516,370,266,207,and 159 bp could be amplified simultaneously in the established quintuple PCR system,and the detection limit of this system for Ralstonia solanacearum,Enterobacter spp.and Pythium spp.reached 10^(-1)pg/μL,for Fusarium spp.and Pectobacterium spp.was 1 pg/μL,and for detecting five pathogens simultaneously was 10^(3)pg/μL.The multiplex PCR system established in this study could successfully detect the diseased plant samples from the field.【Conclusion】The quintuple PCR system established is able to rapid ly and accurately detect Ralstonia solanacearum,Fusarium spp.,Pectobacterium spp.,Enterobacter spp.,and Pythium spp.,which provides a useful tool for timely diagnosis and epidemic monitoring of various soil-borne diseases of ginger.展开更多
Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as s...Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as single embryo and easy rooting.However,Citron C-05 was found to be highly susceptible to root rot during cultivation,with the specific pathogens previously unknown.In this study,four candidate fungal species were isolated from Citron C-05 roots.Sequence analysis of ITS,EF-1a,RPB1,and RPB2 identified two Fusarium solani strains,Rr-2 and Rr-4,as the candidates causing root rot in Citron C-05.Resistance tests showed these two pathogens increased root damage rate from 10.30%to 35.69%in Citron C-05,sour orange(Citrus aurantium),sweet orange(Citrus sinensis)and pummelo(Citrus grandis).F.solani exhibited the weak pathogenicity towards trifoliate orange(Poncirus trifoliata).DAB staining revealed none of reddish-brown precipitation in the four susceptible citrus germplasm after infection with F.solani,while trifoliate orange exhibited significant H2O2 accumulation.Trypan blue staining indicated increased cell death in the four susceptible citrus germplasm following infection with these two pathogens but not in trifoliate orange.These findings provide a comprehensive understanding of citrus root rot and support future research on the mechanisms of root rot resistance in citrus.展开更多
Reductive soil disinfestation(RSD)is commonly employed for soil remediation in greenhouse cultivation.However,its influence on antibiotic resistance genes(ARGs)in soil remains uncertain.This study investigated the dyn...Reductive soil disinfestation(RSD)is commonly employed for soil remediation in greenhouse cultivation.However,its influence on antibiotic resistance genes(ARGs)in soil remains uncertain.This study investigated the dynamic changes in soil communities,potential bacterial pathogens,and ARG profiles under various organicmaterial treatments during RSD,including distillers’grains,potato peel,peanut vine,and peanut vine combined with charcoal.Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens(P<0.05).The relative abundance of high-risk ARGs decreased by 10.7%-30.6%after RSD treatments,the main decreased ARG subtypeswere AAC(3)_Via,dfrA1,ErmB,lnuB,aadA.Actinobacteria was the primary host of ARGs and was suppressed by RSD.Soil physicochemical properties,such as total nitrogen,soil pH,total carbon,were crucial factors affecting ARG profiles.Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.展开更多
In order to ensure food safety,controlling foodborne pathogen contamination is of utmost importance.Growing apprehensions regarding the safety of synthetic antimicrobials,due to their adverse health effects,have promp...In order to ensure food safety,controlling foodborne pathogen contamination is of utmost importance.Growing apprehensions regarding the safety of synthetic antimicrobials,due to their adverse health effects,have prompted a search for alternative options.Plant natural products(PNPs)with antimicrobial activity are being explored as a viable alternative.Among the various antimicrobial natural products studied,plant essential oils,plant flavonoids,plant polyphenols,plant polysaccharides,and plant antimicrobial peptides have been identified as potential candidates.PNPs demonstrate a diverse array of antimicrobial mechanisms,encompassing cell wall and membrane damage,interference with genetic replication,disruption of energy metabolism,and induction of oxidative stress at the single-cell level,as well as inhibition of biofilm formation and quorum sensing at the population level.Certain PNPs have been harnessed as natural antimicrobial agents for the food preservation.The utilization of encapsulation technology proves to be an effective strategy in protecting PNPs,thereby ensuring good antimicrobial efficacy,enhanced dispersibility,and controlled release within food products.The utilization of nanoemulsions,nanoliposomes,edible packaging,electrospun nanofibers,and microcapsules formed by encapsulation has enriched the ways in which PNPs can be applied in food preservation.Although PNPs have great potential in food preservation,their widespread application in the food industry is currently constrained by factors such as production costs,safety concerns,and legal considerations.Chemical synthesis and biosynthesis pathways offer viable strategies for reducing the cost of producing PNPs,and ongoing efforts to assess safety and improve regulatory frameworks are likely to facilitate the broader adoption of PNPs in food preservation practices.This article provides an overview of the main types of PNPs with antimicrobial activity and their properties,focusing on their mechanisms of action.Additionally,it summarizes the use of PNPs in food preservation and discusses the characteristics and applications of different encapsulation technologies.Lastly,the paper briefly analyzes current limitations and proposes potential future trends for this field.展开更多
Respiratory infections are associated with high morbidity and mortality and are a major global health problem[1].Acute respiratory infections are caused by multiple respiratory pathogens,including viruses and bacteria...Respiratory infections are associated with high morbidity and mortality and are a major global health problem[1].Acute respiratory infections are caused by multiple respiratory pathogens,including viruses and bacteria.Viral-bacterial co-infections,which have become increasingly common and a global concern,can lead to substantial complications,causing higher morbidity and adverse prognosis[2].Previous studies have reported low positive detection rates of targeted pathogens related to acute respiratory infections,owing to the limited number of detected pathogens and variations in the sensitivity of diagnostic methods[3-4].Low positive detection rates may impede our understanding of respiratory pathogen characteristics and hamper the development of precise treatment and prevention strategies.展开更多
The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency ...The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency and insufficient sensitivity.This article reviews the progress of rapid detection technology for aquatic pathogens based on molecular biology,immunology,biosensors,etc.,and analyzes the application value of innovative methods such as isothermal amplification and CRISPR.This technology injects core momentum into the new quality productivity of fisheries through early and precise identification of pathogens:reducing aquaculture losses to improve resource efficiency,promoting the transformation of aquaculture models to data-driven,ensuring the safety of aquatic products to enhance the competitiveness of the industry chain.The current technology has shortcomings such as lagging standardization,weak on-site anti-interference ability,and insufficient recognition of new pathogens.In the future,we need to focus on technological integration and innovation,intelligent upgrading,and standardization construction,promote technology from laboratories to industrial applications,and provide continuous support for the high-quality development of fisheries.展开更多
基金National Natural Science Foundation of China(32270237)Guangxi Key Research and Development Plan Project(Guike AB21238002)Basic Scientific Research Project of Guangxi Academy of Agricultural Sciences(Guinongke 2024YP082)。
文摘【Objective】This study aimed to establish a quintuple PCR method for rapid and simultaneous detection of Ralstonia solanacearum,Fusarium spp.,Pectobacterium spp.,Enterobacter spp.,and Pythium spp.,which provided technical support for early diagnosis of various soil-borne diseases on ginger.【Method】For five types of soil-borne pathogens causing ginger bacterial wilt and rhizome rot,specific primer combinations were designed and screened,the optimal quintuple reaction system was established by exploring optimal primer concentrations,annealing temperature,and sensitivity,and was applied to detect field plant samples to verify its utility.【Result】Specific primers pairs Rs1F/Rs1R,En1F/En1R,and Py1F/Py1R were designed according to flic gene of Ralstonia solanacearum,rpoB gene of Enterobacter spp.,and 18S rDNA of Pythium spp.,and combined with reported Fusarium spp.specific primers Fu3/Fu4 and specific primers 23SPecF/23SPecR of Pectobacterium spp.,a quintuple PCR reaction system for ginger soil-borne pathogens has been established(25.00μL):above primer dosage was 1.20,0.20,0.60,1.60,and 0.15μL respectively;2×PCR Mix 12.50μL;DNA templates of different pathogens were 1.00μL each;added ddH_(2)O to 25.00μL.Annealing temperature was optimized to 55.4℃.The specific fragments with sizes of 516,370,266,207,and 159 bp could be amplified simultaneously in the established quintuple PCR system,and the detection limit of this system for Ralstonia solanacearum,Enterobacter spp.and Pythium spp.reached 10^(-1)pg/μL,for Fusarium spp.and Pectobacterium spp.was 1 pg/μL,and for detecting five pathogens simultaneously was 10^(3)pg/μL.The multiplex PCR system established in this study could successfully detect the diseased plant samples from the field.【Conclusion】The quintuple PCR system established is able to rapid ly and accurately detect Ralstonia solanacearum,Fusarium spp.,Pectobacterium spp.,Enterobacter spp.,and Pythium spp.,which provides a useful tool for timely diagnosis and epidemic monitoring of various soil-borne diseases of ginger.
基金supported by Joint Funds of the National Natural Science Foundation of China(Grant No.U21A20228).
文摘Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as single embryo and easy rooting.However,Citron C-05 was found to be highly susceptible to root rot during cultivation,with the specific pathogens previously unknown.In this study,four candidate fungal species were isolated from Citron C-05 roots.Sequence analysis of ITS,EF-1a,RPB1,and RPB2 identified two Fusarium solani strains,Rr-2 and Rr-4,as the candidates causing root rot in Citron C-05.Resistance tests showed these two pathogens increased root damage rate from 10.30%to 35.69%in Citron C-05,sour orange(Citrus aurantium),sweet orange(Citrus sinensis)and pummelo(Citrus grandis).F.solani exhibited the weak pathogenicity towards trifoliate orange(Poncirus trifoliata).DAB staining revealed none of reddish-brown precipitation in the four susceptible citrus germplasm after infection with F.solani,while trifoliate orange exhibited significant H2O2 accumulation.Trypan blue staining indicated increased cell death in the four susceptible citrus germplasm following infection with these two pathogens but not in trifoliate orange.These findings provide a comprehensive understanding of citrus root rot and support future research on the mechanisms of root rot resistance in citrus.
基金supported by the Key Research and Development Program of Shandong Province,China(No 2021CXGC010803)Pan’an County Chinese Medicine Industry Project(No.PZYF202103).
文摘Reductive soil disinfestation(RSD)is commonly employed for soil remediation in greenhouse cultivation.However,its influence on antibiotic resistance genes(ARGs)in soil remains uncertain.This study investigated the dynamic changes in soil communities,potential bacterial pathogens,and ARG profiles under various organicmaterial treatments during RSD,including distillers’grains,potato peel,peanut vine,and peanut vine combined with charcoal.Results revealed that applying diverse organic materials in RSD significantly altered bacterial community composition and diminished the relative abundance of potential bacterial pathogens(P<0.05).The relative abundance of high-risk ARGs decreased by 10.7%-30.6%after RSD treatments,the main decreased ARG subtypeswere AAC(3)_Via,dfrA1,ErmB,lnuB,aadA.Actinobacteria was the primary host of ARGs and was suppressed by RSD.Soil physicochemical properties,such as total nitrogen,soil pH,total carbon,were crucial factors affecting ARG profiles.Our findings demonstrated that RSD treatment inhibited pathogenic bacteria and could be an option for reducing high-risk ARG proliferation in soil.
基金supported by the National Natural Science Foundation of China(32060520)Science and Technology Talents and Platform Program of Yunnan Province(202105AF150049)University Key Laboratory of Food Microbial Resources and Utilization in Yunnan Province(Yunjiaofa[2018]No.135).
文摘In order to ensure food safety,controlling foodborne pathogen contamination is of utmost importance.Growing apprehensions regarding the safety of synthetic antimicrobials,due to their adverse health effects,have prompted a search for alternative options.Plant natural products(PNPs)with antimicrobial activity are being explored as a viable alternative.Among the various antimicrobial natural products studied,plant essential oils,plant flavonoids,plant polyphenols,plant polysaccharides,and plant antimicrobial peptides have been identified as potential candidates.PNPs demonstrate a diverse array of antimicrobial mechanisms,encompassing cell wall and membrane damage,interference with genetic replication,disruption of energy metabolism,and induction of oxidative stress at the single-cell level,as well as inhibition of biofilm formation and quorum sensing at the population level.Certain PNPs have been harnessed as natural antimicrobial agents for the food preservation.The utilization of encapsulation technology proves to be an effective strategy in protecting PNPs,thereby ensuring good antimicrobial efficacy,enhanced dispersibility,and controlled release within food products.The utilization of nanoemulsions,nanoliposomes,edible packaging,electrospun nanofibers,and microcapsules formed by encapsulation has enriched the ways in which PNPs can be applied in food preservation.Although PNPs have great potential in food preservation,their widespread application in the food industry is currently constrained by factors such as production costs,safety concerns,and legal considerations.Chemical synthesis and biosynthesis pathways offer viable strategies for reducing the cost of producing PNPs,and ongoing efforts to assess safety and improve regulatory frameworks are likely to facilitate the broader adoption of PNPs in food preservation practices.This article provides an overview of the main types of PNPs with antimicrobial activity and their properties,focusing on their mechanisms of action.Additionally,it summarizes the use of PNPs in food preservation and discusses the characteristics and applications of different encapsulation technologies.Lastly,the paper briefly analyzes current limitations and proposes potential future trends for this field.
基金supported by the Beijing Science and Technology Planning Project of the Beijing Science and Technology Commission(Z241100009024047)the High-Level Public Health Technical Talent Training Plan(lingjunrencai-01-02).
文摘Respiratory infections are associated with high morbidity and mortality and are a major global health problem[1].Acute respiratory infections are caused by multiple respiratory pathogens,including viruses and bacteria.Viral-bacterial co-infections,which have become increasingly common and a global concern,can lead to substantial complications,causing higher morbidity and adverse prognosis[2].Previous studies have reported low positive detection rates of targeted pathogens related to acute respiratory infections,owing to the limited number of detected pathogens and variations in the sensitivity of diagnostic methods[3-4].Low positive detection rates may impede our understanding of respiratory pathogen characteristics and hamper the development of precise treatment and prevention strategies.
基金supported by the National Modern Agricultural Industry Technology System(CARS-45-33)Innovation Team of Tianjin Freshwater Aquaculture Industry Technology System(ITTFRS2021000-002,ITTFRS2021000-001)+1 种基金Tianjin Science and Technology Plan Project(24KPHDRC00280,24ZYCGSN00250,23YDTPJC00420)the Open Fund Project of Key Laboratory of Ocean Observation Technology,MNR(No.2023klootA03).
文摘The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency and insufficient sensitivity.This article reviews the progress of rapid detection technology for aquatic pathogens based on molecular biology,immunology,biosensors,etc.,and analyzes the application value of innovative methods such as isothermal amplification and CRISPR.This technology injects core momentum into the new quality productivity of fisheries through early and precise identification of pathogens:reducing aquaculture losses to improve resource efficiency,promoting the transformation of aquaculture models to data-driven,ensuring the safety of aquatic products to enhance the competitiveness of the industry chain.The current technology has shortcomings such as lagging standardization,weak on-site anti-interference ability,and insufficient recognition of new pathogens.In the future,we need to focus on technological integration and innovation,intelligent upgrading,and standardization construction,promote technology from laboratories to industrial applications,and provide continuous support for the high-quality development of fisheries.
