Alfalfa （Medicago sativa L.） is an important forage crop and is also a target of many fungal diseases including Fusarium spp. As of today, very little information is available about molecular mechanisms that contrib...Alfalfa （Medicago sativa L.） is an important forage crop and is also a target of many fungal diseases including Fusarium spp. As of today, very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against Fusarium spp. and specifically against Fusarium proliferatum, the causal agent of alfalfa root rot. In this study, we used a proteomic approach to identify inducible proteins in alfalfa during a compatible interaction with F. proliferatum strain YQC-LI. Samples used for the two-dimensional gel electrophoresis （2-DE） and MALDI-TOF/TOF mass spectrometry were from roots and leaves of alfalfa cultivar AmeriGraze 401＋Z and WL656HQ. Plants were grown in hydroponic conditions and at 4 days post inoculation with YQC-LI. Our disease symptom assays indicated thatAmeriGraze 401＋Z was tolerant to YQC-L1 infection while WL656HQ was highly susceptible. Analysis of differentially expressed proteins found in the 2-DE was further characterized using the MASCOT MS/MS ion search software and associated databases to identify multiple proteins that might be involved in F. proliferatum resistance. A total of 66 and 27 differentially expressed proteins were found in the roots and leaves of the plants inoculated with YQC-L1, respectively. These identified proteins were placed in various categories including defense and stress response related metabolism, photosynthesis and protein synthesis. Thirteen identified proteins were validated for their expressions by quantitative reverse transcription （qRT）-PCR. Our results suggested that some of the identified proteins might play important roles in alfalfa resistance against Fusarium spp. These finding could facilitate further dissections of molecular mechanisms controlling root rot disease in alfalfa and potentially other legume crops.展开更多
Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the ...Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the filaments, and infected toward ovary. Both the parenchyma cells and the epidermal cells were wrinkled, and the filament was deformed. The result showed that F. proliferatum could infect filaments directly and cause maize ear rot through filament channel.展开更多
[Objectives]To find out the suitable inoculation method for Fusarium proliferatum.[Methods]Longmu 801 and TG4 were inoculated with rice grain-based inoculum at the bud and seedling stages and with root irrigation at t...[Objectives]To find out the suitable inoculation method for Fusarium proliferatum.[Methods]Longmu 801 and TG4 were inoculated with rice grain-based inoculum at the bud and seedling stages and with root irrigation at the seedling stage,respectively,and then,the plant height,aboveground biomass,underground biomass,incidence rate and disease index of the alfalfa were determined to conduct comprehensive resistance evaluation.[Results]There was obvious difference in disease resistance between the two alfalfa varieties.Among the three inoculation methods,the relative plant height,relative aboveground biomass and relative underground biomass of Longmu 801 differed insignificantly(P>0.05),all around 100.There were significant differences in TG4 among different inoculation methods(P<0.05).The incidence rates and disease indices of Longmu 801 and TG4 inoculated with rice grain-based inoculum at the bud stage were the highest,and their incidence rates and disease indices were 73.3%and 100%,and 40 and 60,respectively,significantly higher than those under the other two inoculation methods.The incidence rates and disease indices of Longmu 801 and TG4 inoculated with rice grain-based inoculum at the seedling stage were the lowest,and their incidence rates and disease indices were 0%and 13.30%,and 0 and 5.33,respectively,significantly lower than those under the other two inoculation methods.[Conclusions]The inoculation time and inoculation method of the pathogen both affect the inoculation effect.Among the three inoculation methods,the inoculation with rice grain-based inoculum at the bud stage is the most pathogenic to alfalfa seedlings,and can be popularized in the resistance evaluation of alfalfa to root diseases.展开更多
Rice spikelet rot disease(RSRD), caused by Fusarium proliferatum, is an emerging disease. So far, the effects of diseased rice floral organs as well as the primary infection sites and stages of this pathogen are not d...Rice spikelet rot disease(RSRD), caused by Fusarium proliferatum, is an emerging disease. So far, the effects of diseased rice floral organs as well as the primary infection sites and stages of this pathogen are not determined. We investigated changes in the floral organs, along with the infection processes of the pathogen in plants inoculated with F. proliferatum and labelled with a green fluorescent protein during different growth stages of rice. The results showed that RSRD is not a systemic infectious disease, which has negative effects on the fertility of the infected rice. F. proliferatum caused brown colored anthers, crinkled pistils and ovaries, pollen grain deformities and anther indehiscence. The number of pollen grains on the stigmas decreased significantly in the infected spikelets, and the anther dehiscence and seed-setting rate successively declined by 69% and 73%, respectively, as a result of the infection. The initial infection stage occurred at the pollen cell maturity stage, and the primary invasion sites were determined to be the anthers of rice. It was noted that the pathogen mainly damaged the pollen cells, and with the exception of the filaments, proceeded to colonize the pistils and endosperm.展开更多
Beauvericin, a cyclohexadepsipeptide-possessing natural product with synergistic antifungal, insecticidal, and cytotoxic activities. We isolated and characterized the fpBeas gene cluster, devoted to beauvericin biosyn...Beauvericin, a cyclohexadepsipeptide-possessing natural product with synergistic antifungal, insecticidal, and cytotoxic activities. We isolated and characterized the fpBeas gene cluster, devoted to beauvericin biosynthesis, from the filamentous fungus Fusarium proliferatum LF061. Targeted inactivation of the F. proliferatum genomic copy of fpBeas abolished the production of beauvericin. Comparative sequence analysis of the FpBEAS showed 74% similarity with the BbBEAS that synthesizes the cyclic trimeric ester beauvericin in Beauveria bassiana, which assembles N-methyl-dipeptidol monomer intermediates by the programmed iterative use of the nonribosomal peptide synthetase modules. Differences between the organization of the beauvericin loci in F. proliferaturm and B. bassiana revealed the mechanism for high production of beauvericin in F. proliferatum. Our work provides new insights into beauvericin biosynthesis, and may lead to beauvericin overproduction and creation of new analogs via synthetic biology approaches.展开更多
The effects of solarisation using clear, UV stabilized, 25μm low density polyethylene mulching combined with soil amendment of chicken manures 12 th^-1, mixed fungicides of Metalaxyl 2 g-Benlate 1.5 g L^-l, Biocontro...The effects of solarisation using clear, UV stabilized, 25μm low density polyethylene mulching combined with soil amendment of chicken manures 12 th^-1, mixed fungicides of Metalaxyl 2 g-Benlate 1.5 g L^-l, Biocontrol agent of Trichoderma harzianum （T.h.） and NPK fertilizer 180 Kg h^-1, on the survived micro and macrocondia of Fusarium proliferatum （Matsushita） Nirenberg were ascertained during summer 2008. Mulched treatments within 45 and 60 days significantly reduced viable propagules to 6 and 3.4 × l03 cfu1 g soil, respectively at 5 cm depth followed by 15 and 30 cm soil depth. Natural heating of dry soil reduced an initial population of 76 × 10^3 to 46.73, 49 and 49.13 × 103 cfu＇lg soil at 5, 15 and 30 cm depth, respectively. Therefore, Fusarium proliferatum reduced by 85.29 and 89.22% within 45 and 60 days compared to 38% in control aired soil. Application of mulching with T. harzianum caused a worthwhile reduction 84.37% in viable propagules, solar with low doses of Metalaxyl-Benlate further reduced Fusarium propagules at all depths 95.5%. Combining chicken manures proved its reduction of fungi cfu at 5 cm depth only, whereas NPK amendments failed in their reduction effects at various depths. However, almost 95% reduction in Fusarium propagules was achieved at various soil depths when combining Met.-Ben., with soil mulching. Combining T.h. was also proved to be significant in reducing fungus inoculum by 88.58, 96.06% at 5 and 15 cm depth. Generally, manures and NPK fertilizers amended soil found equally effective in reducing cfu of Fusarium at 5 and 15 cm depth similar to polyethylene mulching for both solar duration.展开更多
Disease suppression of Fusarium crown rot and the changes in free amino acid contents in mycorrhizal asparagus (Asparagus officinalis L., cv. “Welcome”) plants were investigated. Sixteen weeks after arbuscular mycor...Disease suppression of Fusarium crown rot and the changes in free amino acid contents in mycorrhizal asparagus (Asparagus officinalis L., cv. “Welcome”) plants were investigated. Sixteen weeks after arbuscular mycorrhizal fungus (AMF;Glomus intraradices) inoculation, mycorrhizal plants showed higher dry weight of shoots than non-mycorrhizal plants, and AMF colonization level in a root system reached up to 73.3%. Ten weeks after Fusarium proliferatum (Fp;N1-31, SUF1207) inoculation, control plants showed 100% incidence and high severity in the 2 Fp isolates. However, AMF plants showed lower severity than non-AMF plants in the 2 Fp isolates. Ten weeks after Fp (N1-31) inoculation, the increase in 7 constituents of amino acids (glutamine, arginine, aspartic acid, alanine, citrulline, GABA, glycine) in shoots, and 9 in roots (asparagine, arginine, threonine, serine, glutamine, citrulline, valine, GABA, histidine) occurred in AMF plants. From these findings, plant growth promotion and suppression of Fusarium crown rot occurred in mycorrhizal asparagus plants, and the disease tolerance was supposed to be associated with the symbiosis-specific increase in free amino acids.展开更多
基金supported by the earmarked fund for China Agriculture Research System(CARS-35-04)the Chinese Academy of Agricultural Science and Technology Innovation Project(ASTIP-IAS14)
文摘Alfalfa （Medicago sativa L.） is an important forage crop and is also a target of many fungal diseases including Fusarium spp. As of today, very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against Fusarium spp. and specifically against Fusarium proliferatum, the causal agent of alfalfa root rot. In this study, we used a proteomic approach to identify inducible proteins in alfalfa during a compatible interaction with F. proliferatum strain YQC-LI. Samples used for the two-dimensional gel electrophoresis （2-DE） and MALDI-TOF/TOF mass spectrometry were from roots and leaves of alfalfa cultivar AmeriGraze 401＋Z and WL656HQ. Plants were grown in hydroponic conditions and at 4 days post inoculation with YQC-LI. Our disease symptom assays indicated thatAmeriGraze 401＋Z was tolerant to YQC-L1 infection while WL656HQ was highly susceptible. Analysis of differentially expressed proteins found in the 2-DE was further characterized using the MASCOT MS/MS ion search software and associated databases to identify multiple proteins that might be involved in F. proliferatum resistance. A total of 66 and 27 differentially expressed proteins were found in the roots and leaves of the plants inoculated with YQC-L1, respectively. These identified proteins were placed in various categories including defense and stress response related metabolism, photosynthesis and protein synthesis. Thirteen identified proteins were validated for their expressions by quantitative reverse transcription （qRT）-PCR. Our results suggested that some of the identified proteins might play important roles in alfalfa resistance against Fusarium spp. These finding could facilitate further dissections of molecular mechanisms controlling root rot disease in alfalfa and potentially other legume crops.
基金Supported by Science and Technology Innovation Team Support Program for Colleges and Universities in Henan Province(2010JRTSTHN012)Henan Foundation and Frontier Technology Research Program of Henan Science and Technology Department(162300410137)+1 种基金Henan Educational Committee Key Research Projects of Henan Higher Education(16A210034)Nanyang Normal University Special Project(ZX2016002)
文摘Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the filaments, and infected toward ovary. Both the parenchyma cells and the epidermal cells were wrinkled, and the filament was deformed. The result showed that F. proliferatum could infect filaments directly and cause maize ear rot through filament channel.
基金National Natural Science Foundation of China(31702169)Innovative Talent Cultivation Project of Heilongjiang Bayi Agricultural University(CXRC2017006)Scientific Research Innovation Fund for Postgraduates of Heilongjiang Bayi Agricultural University(YJSCX2018-Y39)。
文摘[Objectives]To find out the suitable inoculation method for Fusarium proliferatum.[Methods]Longmu 801 and TG4 were inoculated with rice grain-based inoculum at the bud and seedling stages and with root irrigation at the seedling stage,respectively,and then,the plant height,aboveground biomass,underground biomass,incidence rate and disease index of the alfalfa were determined to conduct comprehensive resistance evaluation.[Results]There was obvious difference in disease resistance between the two alfalfa varieties.Among the three inoculation methods,the relative plant height,relative aboveground biomass and relative underground biomass of Longmu 801 differed insignificantly(P>0.05),all around 100.There were significant differences in TG4 among different inoculation methods(P<0.05).The incidence rates and disease indices of Longmu 801 and TG4 inoculated with rice grain-based inoculum at the bud stage were the highest,and their incidence rates and disease indices were 73.3%and 100%,and 40 and 60,respectively,significantly higher than those under the other two inoculation methods.The incidence rates and disease indices of Longmu 801 and TG4 inoculated with rice grain-based inoculum at the seedling stage were the lowest,and their incidence rates and disease indices were 0%and 13.30%,and 0 and 5.33,respectively,significantly lower than those under the other two inoculation methods.[Conclusions]The inoculation time and inoculation method of the pathogen both affect the inoculation effect.Among the three inoculation methods,the inoculation with rice grain-based inoculum at the bud stage is the most pathogenic to alfalfa seedlings,and can be popularized in the resistance evaluation of alfalfa to root diseases.
基金supported by the National Key Research and Development Program of China(Grant Nos.2016YFD0200801,2017YFD0300409 and 2018YFD 020030405)National Natural Science Foundation of China(Grant No.31800133)+5 种基金Zhejiang Provincial Natural Science Foundation of China(Grant No.LQ18C140005)Agriculture,Rural Areas,and Farmers Six-Party Science and Technology Cooperation Projects of Zhejiang Province(Grant No.CTZB-F160728AWZ-SNY1-4)the Innovation Project of the Chinese Academy of Agricultural Sciences(CAAS)the Collaborative Innovation Project of the CAAS(Grant No.CAAS-XTCX2016012)the Core Research Budget of Nonprofit Governmental Research Insititute of China(Grant No.2014RG005-2)Zhejiang Key Research and Development Program of China(Grant No.2019C02018)
文摘Rice spikelet rot disease(RSRD), caused by Fusarium proliferatum, is an emerging disease. So far, the effects of diseased rice floral organs as well as the primary infection sites and stages of this pathogen are not determined. We investigated changes in the floral organs, along with the infection processes of the pathogen in plants inoculated with F. proliferatum and labelled with a green fluorescent protein during different growth stages of rice. The results showed that RSRD is not a systemic infectious disease, which has negative effects on the fertility of the infected rice. F. proliferatum caused brown colored anthers, crinkled pistils and ovaries, pollen grain deformities and anther indehiscence. The number of pollen grains on the stigmas decreased significantly in the infected spikelets, and the anther dehiscence and seed-setting rate successively declined by 69% and 73%, respectively, as a result of the infection. The initial infection stage occurred at the pollen cell maturity stage, and the primary invasion sites were determined to be the anthers of rice. It was noted that the pathogen mainly damaged the pollen cells, and with the exception of the filaments, proceeded to colonize the pistils and endosperm.
基金supported by the National Basic Research Program of China (2013CB734000, 2012CB725200)the CAS Pillar Program (XDA04074000)+1 种基金Ministry of Science and Technology of China (2011ZX11102-011-11)ZHANG LiXin is an Awardee for the National Distinguished Young Scholar Program in China
文摘Beauvericin, a cyclohexadepsipeptide-possessing natural product with synergistic antifungal, insecticidal, and cytotoxic activities. We isolated and characterized the fpBeas gene cluster, devoted to beauvericin biosynthesis, from the filamentous fungus Fusarium proliferatum LF061. Targeted inactivation of the F. proliferatum genomic copy of fpBeas abolished the production of beauvericin. Comparative sequence analysis of the FpBEAS showed 74% similarity with the BbBEAS that synthesizes the cyclic trimeric ester beauvericin in Beauveria bassiana, which assembles N-methyl-dipeptidol monomer intermediates by the programmed iterative use of the nonribosomal peptide synthetase modules. Differences between the organization of the beauvericin loci in F. proliferaturm and B. bassiana revealed the mechanism for high production of beauvericin in F. proliferatum. Our work provides new insights into beauvericin biosynthesis, and may lead to beauvericin overproduction and creation of new analogs via synthetic biology approaches.
文摘The effects of solarisation using clear, UV stabilized, 25μm low density polyethylene mulching combined with soil amendment of chicken manures 12 th^-1, mixed fungicides of Metalaxyl 2 g-Benlate 1.5 g L^-l, Biocontrol agent of Trichoderma harzianum （T.h.） and NPK fertilizer 180 Kg h^-1, on the survived micro and macrocondia of Fusarium proliferatum （Matsushita） Nirenberg were ascertained during summer 2008. Mulched treatments within 45 and 60 days significantly reduced viable propagules to 6 and 3.4 × l03 cfu1 g soil, respectively at 5 cm depth followed by 15 and 30 cm soil depth. Natural heating of dry soil reduced an initial population of 76 × 10^3 to 46.73, 49 and 49.13 × 103 cfu＇lg soil at 5, 15 and 30 cm depth, respectively. Therefore, Fusarium proliferatum reduced by 85.29 and 89.22% within 45 and 60 days compared to 38% in control aired soil. Application of mulching with T. harzianum caused a worthwhile reduction 84.37% in viable propagules, solar with low doses of Metalaxyl-Benlate further reduced Fusarium propagules at all depths 95.5%. Combining chicken manures proved its reduction of fungi cfu at 5 cm depth only, whereas NPK amendments failed in their reduction effects at various depths. However, almost 95% reduction in Fusarium propagules was achieved at various soil depths when combining Met.-Ben., with soil mulching. Combining T.h. was also proved to be significant in reducing fungus inoculum by 88.58, 96.06% at 5 and 15 cm depth. Generally, manures and NPK fertilizers amended soil found equally effective in reducing cfu of Fusarium at 5 and 15 cm depth similar to polyethylene mulching for both solar duration.
文摘Disease suppression of Fusarium crown rot and the changes in free amino acid contents in mycorrhizal asparagus (Asparagus officinalis L., cv. “Welcome”) plants were investigated. Sixteen weeks after arbuscular mycorrhizal fungus (AMF;Glomus intraradices) inoculation, mycorrhizal plants showed higher dry weight of shoots than non-mycorrhizal plants, and AMF colonization level in a root system reached up to 73.3%. Ten weeks after Fusarium proliferatum (Fp;N1-31, SUF1207) inoculation, control plants showed 100% incidence and high severity in the 2 Fp isolates. However, AMF plants showed lower severity than non-AMF plants in the 2 Fp isolates. Ten weeks after Fp (N1-31) inoculation, the increase in 7 constituents of amino acids (glutamine, arginine, aspartic acid, alanine, citrulline, GABA, glycine) in shoots, and 9 in roots (asparagine, arginine, threonine, serine, glutamine, citrulline, valine, GABA, histidine) occurred in AMF plants. From these findings, plant growth promotion and suppression of Fusarium crown rot occurred in mycorrhizal asparagus plants, and the disease tolerance was supposed to be associated with the symbiosis-specific increase in free amino acids.
