Major facilitator superfamily(MFS)transporters are secondary active membrane transporters that play an important role in solute interchange and energy metabolism.Peronophythora litchii causes the most destructive dise...Major facilitator superfamily(MFS)transporters are secondary active membrane transporters that play an important role in solute interchange and energy metabolism.Peronophythora litchii causes the most destructive disease on lichi,litchi downy blight.PlM90 was reported as a key oosporogenesis regulator.Here,we identified an MFS transporter gene PlMFS1,which is up-regulated during oospore formation at the late infection stage,while down-regulated in the PlM90 mutant.To investigate PlMFS1 function,we generated PlMFS1knockout mutants using CRISPR/Cas9-mediated genome editing technology.Compared with the wild-type strain SHS3,PlMFS1 deletion impaired mycelium growth,zoospore release,oospore production and pathogenicity.Furthermore,PlMFS1 deletion significantly affected P.litchii utilization of fructose,lactose and maltose,and may be the PlMFS1 mechanism involved in mycelial growth.PlMFS1 gene deletion also led to deceased laccase activity,laccase-encoding gene downregulation and impaired P.litchii pathogenicity.To our knowledge,this is the first report of an MFS transporter involved in sugar utilization,sexual reproduction,asexual reproduction and pathogenesis in oomycetes.展开更多
Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the develo...Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the development and pathogenicity of the filamentous fungi.However,the function of autophagy in oomycetes remain elusive.Here,an autophagy-related protein Atg3 homolog PlAtg3 was identified and characterized in P.litchii.The absence of PlATG3 through the CRISPR/Cas9 gene replacement strategy compromised vegetative growth and sexual/asexual development.Cytological analyses revealed that the deletion of PlATG3 impaired autophagosome formation with monodansylcadaverine(MDC)staining and significantly disrupted zoospore release due to defects of sporangial cleavage with FM4-64 staining.Atg8 is considered to be an autophagy marker protein in various species.Western blot analysis indicated that PlAtg3 is involved in degradation of PlAtg8-PE.Interestingly,PlAtg3 was unable to interact with PlAtg8 in yeast two hybrid(Y2H)assays,possibly due to the absence of the Atg8 family interacting motif(AIM)in PlAtg3.Furthermore,pathogenicity assays revealed that the deletion of PlATG3 considerably reduced the virulence of P.litchii.Taken together,our data reveal that PlAtg3 plays an important role in radial growth,asexual/sexual development,sporangial cleavage and zoospore release,autophagosome formation,and pathogenicity in P.litchii.This study contributes to a better understanding of the pathogenicity mechanisms of P.litchii and provides insights for the development of more effective strategies to control oomycete diseases.展开更多
为寻找果树病害防治的植物源抑菌物质,以乙醇为溶剂,采用超声波辅助提取法获得17种植物的提取物,并用提取物对荔枝霜疫霉菌(Peronophythora litchii Chen ex Ko et al.)进行抑菌试验,结果显示,丁香(Syringa aramaticum)、黄连(Coptischi...为寻找果树病害防治的植物源抑菌物质,以乙醇为溶剂,采用超声波辅助提取法获得17种植物的提取物,并用提取物对荔枝霜疫霉菌(Peronophythora litchii Chen ex Ko et al.)进行抑菌试验,结果显示,丁香(Syringa aramaticum)、黄连(Coptischinensis Franch)、细辛(Asarum sieboldii)提取物5 mg/mL浓度处理药后7 d抑菌率均为100%,1 mg/mL浓度处理抑菌率则分别为60.68%、51.88%、15.64%,黄柏(Cortex Phellodendri)、苦参(Sophora flavescens)、花椒(Pricklyash Peel)提取物5 mg/mL浓度处理药后7 d抑菌率均达60%以上;5 mg/mL浓度处理花椒、黄连、黄柏、细辛提取物对荔枝霜疫霉菌孢子囊萌发抑制率都在60%以上,而花椒的抑制率最高,达73.08%。不同植物提取物抑菌效果受温度和pH的影响明显。展开更多
测定了64种植物的甲醇提取物对荔枝霜疫霉菌(Peronophythora litchii Chen ex Ko et al.)和香蕉炭疽病菌(Colletotrichummusae(Berk.et Curt.)Arx)的离体抗菌活性。试验结果显示,在提取物质量浓度为0.01g/mL 时,对荔枝霜疫霉菌菌丝生长...测定了64种植物的甲醇提取物对荔枝霜疫霉菌(Peronophythora litchii Chen ex Ko et al.)和香蕉炭疽病菌(Colletotrichummusae(Berk.et Curt.)Arx)的离体抗菌活性。试验结果显示,在提取物质量浓度为0.01g/mL 时,对荔枝霜疫霉菌菌丝生长的抑制率高于50%的植物共有30种.对霜疫霉菌孢子囊萌发的抑制率高于50%的植物共有26种;对香蕉炭疽病菌菌丝生长的抑抑制率高于50%的植物共有18种,对香蕉炭疽病孢子萌发的抑制率高于50%的植物共有6种;对两种病原菌的菌丝生长与孢子萌发抑制率均大于50%的植物有开口箭(Tupistra chinensis Bak.)和苍耳(Xanthinum sibiricum Patrin)。展开更多
基金funded by the Natural Science Foundation of Guangdong Province,China(Grant Nos.2023A1515012617,2022A1515010458 and 2023A1515030267)Guangzhou Science&Technology Program(Grant No.202201010410)the earmarked fund for CARS-32。
文摘Major facilitator superfamily(MFS)transporters are secondary active membrane transporters that play an important role in solute interchange and energy metabolism.Peronophythora litchii causes the most destructive disease on lichi,litchi downy blight.PlM90 was reported as a key oosporogenesis regulator.Here,we identified an MFS transporter gene PlMFS1,which is up-regulated during oospore formation at the late infection stage,while down-regulated in the PlM90 mutant.To investigate PlMFS1 function,we generated PlMFS1knockout mutants using CRISPR/Cas9-mediated genome editing technology.Compared with the wild-type strain SHS3,PlMFS1 deletion impaired mycelium growth,zoospore release,oospore production and pathogenicity.Furthermore,PlMFS1 deletion significantly affected P.litchii utilization of fructose,lactose and maltose,and may be the PlMFS1 mechanism involved in mycelial growth.PlMFS1 gene deletion also led to deceased laccase activity,laccase-encoding gene downregulation and impaired P.litchii pathogenicity.To our knowledge,this is the first report of an MFS transporter involved in sugar utilization,sexual reproduction,asexual reproduction and pathogenesis in oomycetes.
基金supported by the grants from the Hainan Provincial Natural Science Foundation,China(321QN190 and 321CXTD437)the National Natural Science Foundation of China(32202246 and 32160614)+1 种基金the Open Project Program of Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests,China(MIMCP-202102)the Scientific Research Foundation of Hainan University,China(KYQD(ZR)-21042 and KYQD(ZR)-20080)。
文摘Litchi downy blight,caused by the plant pathogenic oomycete Peronophythora litchii,is one of the most devastating diseases on litchi and resulted in huge economic losses.Autophagy plays an essential role in the development and pathogenicity of the filamentous fungi.However,the function of autophagy in oomycetes remain elusive.Here,an autophagy-related protein Atg3 homolog PlAtg3 was identified and characterized in P.litchii.The absence of PlATG3 through the CRISPR/Cas9 gene replacement strategy compromised vegetative growth and sexual/asexual development.Cytological analyses revealed that the deletion of PlATG3 impaired autophagosome formation with monodansylcadaverine(MDC)staining and significantly disrupted zoospore release due to defects of sporangial cleavage with FM4-64 staining.Atg8 is considered to be an autophagy marker protein in various species.Western blot analysis indicated that PlAtg3 is involved in degradation of PlAtg8-PE.Interestingly,PlAtg3 was unable to interact with PlAtg8 in yeast two hybrid(Y2H)assays,possibly due to the absence of the Atg8 family interacting motif(AIM)in PlAtg3.Furthermore,pathogenicity assays revealed that the deletion of PlATG3 considerably reduced the virulence of P.litchii.Taken together,our data reveal that PlAtg3 plays an important role in radial growth,asexual/sexual development,sporangial cleavage and zoospore release,autophagosome formation,and pathogenicity in P.litchii.This study contributes to a better understanding of the pathogenicity mechanisms of P.litchii and provides insights for the development of more effective strategies to control oomycete diseases.
文摘测定了64种植物的甲醇提取物对荔枝霜疫霉菌(Peronophythora litchii Chen ex Ko et al.)和香蕉炭疽病菌(Colletotrichummusae(Berk.et Curt.)Arx)的离体抗菌活性。试验结果显示,在提取物质量浓度为0.01g/mL 时,对荔枝霜疫霉菌菌丝生长的抑制率高于50%的植物共有30种.对霜疫霉菌孢子囊萌发的抑制率高于50%的植物共有26种;对香蕉炭疽病菌菌丝生长的抑抑制率高于50%的植物共有18种,对香蕉炭疽病孢子萌发的抑制率高于50%的植物共有6种;对两种病原菌的菌丝生长与孢子萌发抑制率均大于50%的植物有开口箭(Tupistra chinensis Bak.)和苍耳(Xanthinum sibiricum Patrin)。
