Bakanae disease,caused by several species ofFusarium section Liseola is an important dis-ease of rice(Oryza sativa L.).It is unclearwhether the isolates with different morpholog-ical features in anamorphs were genetic...Bakanae disease,caused by several species ofFusarium section Liseola is an important dis-ease of rice(Oryza sativa L.).It is unclearwhether the isolates with different morpholog-ical features in anamorphs were genetic differ-ent.Gibberella fujikuroi is the teleomorph ofspecies of Fusarium section Liseola,and en-compasses at least seven mating populations(MPs).Vegetative compatibility group(VCG)has been used to study the relation- ships between the strains within a fungalspecies.We studied the genetic diversity ofpathogenes of rice bakanae disease by usingthe MP and VCG techniques.展开更多
Gibberellic acid(GA3)is a vital plant growth hormone widely used in agriculture.Currently,GA3 production relies on liquid fermentation by the filamentous fungus Fusarium fujikuroi.However,the lack of an effective sele...Gibberellic acid(GA3)is a vital plant growth hormone widely used in agriculture.Currently,GA3 production relies on liquid fermentation by the filamentous fungus Fusarium fujikuroi.However,the lack of an effective selection marker recycling system hampers the application of metabolic engineering technology in F.fujikuroi,as multiple-gene editing and positive-strain screening still rely on a limited number of antibiotics.In this study,we developed a strategy using pyr4-blaster and CRISPR/Cas9 tools for recycling orotidine-5'-phosphate decarboxylase(Pyr4)selection markers.We demonstrated the effectiveness of this method for iterative gene integration and large gene-cluster deletion.We also successfully improved GA3 titers by overexpressing geranylgeranyl pyrophosphate synthase and truncated 3-hydroxy-3-methyl glutaryl coenzyme A reductase,which rewired the GA3 biosynthesis pathway.These results highlight the efficiency of our established system in recycling selection markers during iterative gene editing events.Moreover,the selection marker recycling system lays the foundation for further research on metabolic engineering for GA3 industrial production.展开更多
Rice(Oryza sativa L.)is one of the most important cereal crops in the world.Bakanae disease is a significant rice disease widely distributed in rice-growing regions worldwide.Therefore,the present investigation aimed ...Rice(Oryza sativa L.)is one of the most important cereal crops in the world.Bakanae disease is a significant rice disease widely distributed in rice-growing regions worldwide.Therefore,the present investigation aimed to assess the optimal concentrations of paclobutrazol(PBZ)as a treatment for rice grains(cv.Sakha 108)to control bakanae disease,also evaluating its impact on grain germination,seedling growth parameters as well as disease index.Paclobutrazol concentrations had no significant impact on seed germination,regardless of whether the seeds were incubated with Fusarium fujikuroi or not.Application of PBZ,either alone or in combination with fungal pathogens,negatively impacted the rice seedlings’height.Paclobutrazol at 25,50 and 100 mg/L,combined with the fungal pathogen positively impacted root length.Paclobutrazol at 3 and 6 mg/L mitigated the adverse impact on chlorophyll pigments content in infected seedlings.The highest proline contents were achieved by 100 mg/L PBZ alone or in combination with fungal pathogens.It has been observed that the application of PBZ,either alone or in combination with a fungal pathogen,leads to the enhancement of catalase,peroxidase,and polyphenol oxidase activities.The median lethal concentration of PBZ was 0.874 mg/L;applying low concentrations of paclobutrazol effectively increased the percentage of fungal growth suppression.Application of PBZ,at higher concentrations(50 and 100 mg/L),decreased infection percentage and disease severity index(DSI)significantly.These findings suggest that PBZ can be an effective treatment for controlling bakanae disease and enhancing resistance in rice plants.展开更多
The regio- and stereo-selective hydroxylations of two ingenane diterpenoids, 20-deoxyingenol(1) and 13-oxyingenol dodecanoat(2), by the filamentous fungi Mortierella ramanniana and Gibberella fujikuroi were investigat...The regio- and stereo-selective hydroxylations of two ingenane diterpenoids, 20-deoxyingenol(1) and 13-oxyingenol dodecanoat(2), by the filamentous fungi Mortierella ramanniana and Gibberella fujikuroi were investigated in the present study. Four undescribed metabolites(3–6) of substrate 1 and two undescribed metabolites(7 and 8) of substrate 2 were isolated. All the lites were identified as hydroxylated ingenane derivatives by extensive NMR and HR-ESI-MS data analyses. All the biotransformed compounds and the substrates were evaluated for their cytotoxicities against three human cancer cell lines, including human colon cancer Caco-2, breast cancer MCF-7, and adriamycin(ADM)-resistant MCF-7/ADM cell lines. All ingenane alcohols(1, and 3–6) displayed no significant cytotoxic activities. The substrate 13-oxyingenol dodecanoat(2) showed moderate cytotoxicity with IC50 values being 35.59 ± 5.37 μmol·L^(-1)(Caco-2), 24.04 ± 4.70 μmol·L^(-1)(MCF-7), and 22.24 ± 5.19 μmol·L^(-1)(MCF-7/ADM). However, metabolites 7 and 8 displayed no significant cytotoxicity. These results indicated that the hydroxylation at the C^(-1)3 aliphatic acid ester of substrate 2 can significantly reduce the cytotoxic activity.展开更多
文摘Bakanae disease,caused by several species ofFusarium section Liseola is an important dis-ease of rice(Oryza sativa L.).It is unclearwhether the isolates with different morpholog-ical features in anamorphs were genetic differ-ent.Gibberella fujikuroi is the teleomorph ofspecies of Fusarium section Liseola,and en-compasses at least seven mating populations(MPs).Vegetative compatibility group(VCG)has been used to study the relation- ships between the strains within a fungalspecies.We studied the genetic diversity ofpathogenes of rice bakanae disease by usingthe MP and VCG techniques.
基金supported by National key research and development program of China(2021YFC2104300)Natural Science Foundation of Jiangsu Province(BK20210573,20230381)+2 种基金Project of Leading Innovative Talents in Short-term Jiangxi Province(jxsq2023102172)Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project TSBICIP-PTJS-003-04Postgraduate Research&Practice Innovation Program of Jiangsu Province KYCX23_1788.
文摘Gibberellic acid(GA3)is a vital plant growth hormone widely used in agriculture.Currently,GA3 production relies on liquid fermentation by the filamentous fungus Fusarium fujikuroi.However,the lack of an effective selection marker recycling system hampers the application of metabolic engineering technology in F.fujikuroi,as multiple-gene editing and positive-strain screening still rely on a limited number of antibiotics.In this study,we developed a strategy using pyr4-blaster and CRISPR/Cas9 tools for recycling orotidine-5'-phosphate decarboxylase(Pyr4)selection markers.We demonstrated the effectiveness of this method for iterative gene integration and large gene-cluster deletion.We also successfully improved GA3 titers by overexpressing geranylgeranyl pyrophosphate synthase and truncated 3-hydroxy-3-methyl glutaryl coenzyme A reductase,which rewired the GA3 biosynthesis pathway.These results highlight the efficiency of our established system in recycling selection markers during iterative gene editing events.Moreover,the selection marker recycling system lays the foundation for further research on metabolic engineering for GA3 industrial production.
基金supported and funded by Deanship of Scientific Research,Vice Presidency for Graduate Studies and Scientific Research,King Faisal University,Saudi Arabia(KFU241897).
文摘Rice(Oryza sativa L.)is one of the most important cereal crops in the world.Bakanae disease is a significant rice disease widely distributed in rice-growing regions worldwide.Therefore,the present investigation aimed to assess the optimal concentrations of paclobutrazol(PBZ)as a treatment for rice grains(cv.Sakha 108)to control bakanae disease,also evaluating its impact on grain germination,seedling growth parameters as well as disease index.Paclobutrazol concentrations had no significant impact on seed germination,regardless of whether the seeds were incubated with Fusarium fujikuroi or not.Application of PBZ,either alone or in combination with fungal pathogens,negatively impacted the rice seedlings’height.Paclobutrazol at 25,50 and 100 mg/L,combined with the fungal pathogen positively impacted root length.Paclobutrazol at 3 and 6 mg/L mitigated the adverse impact on chlorophyll pigments content in infected seedlings.The highest proline contents were achieved by 100 mg/L PBZ alone or in combination with fungal pathogens.It has been observed that the application of PBZ,either alone or in combination with a fungal pathogen,leads to the enhancement of catalase,peroxidase,and polyphenol oxidase activities.The median lethal concentration of PBZ was 0.874 mg/L;applying low concentrations of paclobutrazol effectively increased the percentage of fungal growth suppression.Application of PBZ,at higher concentrations(50 and 100 mg/L),decreased infection percentage and disease severity index(DSI)significantly.These findings suggest that PBZ can be an effective treatment for controlling bakanae disease and enhancing resistance in rice plants.
文摘The regio- and stereo-selective hydroxylations of two ingenane diterpenoids, 20-deoxyingenol(1) and 13-oxyingenol dodecanoat(2), by the filamentous fungi Mortierella ramanniana and Gibberella fujikuroi were investigated in the present study. Four undescribed metabolites(3–6) of substrate 1 and two undescribed metabolites(7 and 8) of substrate 2 were isolated. All the lites were identified as hydroxylated ingenane derivatives by extensive NMR and HR-ESI-MS data analyses. All the biotransformed compounds and the substrates were evaluated for their cytotoxicities against three human cancer cell lines, including human colon cancer Caco-2, breast cancer MCF-7, and adriamycin(ADM)-resistant MCF-7/ADM cell lines. All ingenane alcohols(1, and 3–6) displayed no significant cytotoxic activities. The substrate 13-oxyingenol dodecanoat(2) showed moderate cytotoxicity with IC50 values being 35.59 ± 5.37 μmol·L^(-1)(Caco-2), 24.04 ± 4.70 μmol·L^(-1)(MCF-7), and 22.24 ± 5.19 μmol·L^(-1)(MCF-7/ADM). However, metabolites 7 and 8 displayed no significant cytotoxicity. These results indicated that the hydroxylation at the C^(-1)3 aliphatic acid ester of substrate 2 can significantly reduce the cytotoxic activity.
