Trehalose metabolism is related to the sclerotial development of Rhizoctonia solani AG-1 IA,the causal agent of rice sheath blight(RSB).Here,we further elucidated the functions of three genes Rstre,Rstps1 and Rstpp th...Trehalose metabolism is related to the sclerotial development of Rhizoctonia solani AG-1 IA,the causal agent of rice sheath blight(RSB).Here,we further elucidated the functions of three genes Rstre,Rstps1 and Rstpp that encode three key enzymes trehalase(TRE),alpha,alpha-trehalosephosphate synthase(TPS1)and trehalose 6-phosphate phosphatase(TPP)in the sclerotial development of R.solani AG-1 IA.Due to the lack of a stable genetic transformation system for R.solani,the heterologous expression of these three genes in Pichia pastoris GS115 was performed.The results showed that reactive oxygen species(ROS)contents and enzyme activities in R.solani decreased significantly in the treatments of the fermentation broths of Rstps1 and Rstpp transformants,and that in the treatment of the fermentation broth of Rstre transformant visibly increased.Furthermore,the fermentation broths of the transformants of all the three genes were added to potato dextrose agar(PDA)medium for the cultivation of R.solani,as a result,the dry weight of sclerotia in each PDA plate containing the fermentation broths of Rstps1 and Rstpp transformants significantly increased compared with the control,and that of Rstre transformant obviously decreased.Finally,178 proteins were found to interact with RSTPS1,and 16 of them were associated with ROS.Taken together,the findings suggest that all these three genes related to trehalose metabolism play important roles in the sclerotial development of R.solani AG-1 IA,and can be used as new targets for the development of novel high-efficiency fungicides for the controlling of RSB.展开更多
A field under rice-wheat rotation was selected near Chengdu, China, to study thepopulation of Rhizoctonia solani anastomosis group 1 (AG-1), pathogen causing ricesheath blight disease, in natural soil ecosystem. Inocu...A field under rice-wheat rotation was selected near Chengdu, China, to study thepopulation of Rhizoctonia solani anastomosis group 1 (AG-1), pathogen causing ricesheath blight disease, in natural soil ecosystem. Inocula of the fungus recovered fromthe field were divided into three types, i.e., sclerotia, free mycelium retained in thesoil passed through a 0.355mm sieve, and colonized plant debris which was subdividedinto small colonized debris retained between 2.00 and 0.355mm sieves and large colonizeddebris retained on 2.00mm sieve after wet screening. Quantitative estimation of thethree types of inocula in one year indicated that small colonized debris was the dominantinoculum type for most of the time. The population peaked in March and September at 1210and 480 colonized debris 100g-1 air-dry soil respectively, and fell down in December andAugust to 0 and 177 colonized debris 100g-1 air-dry soil respectively. Free mycelium wasonly detectable in March, September and October with 1209, 7.9 and 14.5g fresh wtmyceliumg-1 air-dry soil respectively, which corresponded to the two peaks and the secondhighest level of small debris density in the year. Viable sclerotia and large colonizeddebris were rare with populations ranging from 0 to 3 for sclerotia and 0 to 14 for largecolonized debris 100g-1 air-dry soil, but were the main structures to survive overwinter. It was expected that soil temperature was the main factor determining populationdynamics of R.solani AG-1 in natural soil. Optimum temperature for population increasingis predicted to be around 15℃, with a range from 10 to 25℃. Viability tests indicatedthat 60.9% sclerotia could survive after 265d being buried in natural sandy loam in fieldconditions in Beijing, while colonized rice straw debris (0.5-1.0cm long) could notyield the fungus on medium plates after 88d of being buried under the same conditions.展开更多
【目的】为明确水稻纹枯病菌(Rhizoctonia solani AG 1-IA)黑色素形成前体物质酪氨酸等氨基酸成份在不同培养处理中的差异。【方法】利用氨基酸分析仪对水稻纹枯病菌在3种培养处理(PDB对照,添加了300μg/mL质量浓度莨菪碱的PDB和50μg/m...【目的】为明确水稻纹枯病菌(Rhizoctonia solani AG 1-IA)黑色素形成前体物质酪氨酸等氨基酸成份在不同培养处理中的差异。【方法】利用氨基酸分析仪对水稻纹枯病菌在3种培养处理(PDB对照,添加了300μg/mL质量浓度莨菪碱的PDB和50μg/mL质量浓度儿茶酚的PDB)后菌丝体和发酵液的氨基酸成份进行了分析。【结果】水稻纹枯病菌菌丝体中谷氨酸、天冬氨酸和丙氨酸3种氨基酸含量相对比较丰富,而谷氨酸的含量最高,达到总氨基酸的15.49%;水稻纹枯病菌发酵液中天冬氨酸和谷氨酸含量最为丰富,分别达到了总氨基酸的17.83%和58.21%;在3种处理中,不管是菌丝体还是发酵液中,都是谷氨酸和天冬氨酸最为丰富。影响水稻纹枯病菌菌丝体黑色素形成的前体物质酪氨酸的含量在3种处理的菌丝体中没有显著差异;但300μg/mL质量浓度的莨菪碱处理组与对照组和儿茶酚处理组中的苯丙氨酸含量都存在显著差异,而对照组和50μg/mL质量浓度的儿茶酚处理组的苯丙氨酸含量差异不显著。水稻纹枯病菌发酵液中对照组和50μg/mL质量浓度的儿茶酚处理组酪氨酸含量没有显著差异,但是300μg/mL质量浓度的莨菪碱处理组与对照组和儿茶酚处理组酪氨酸含量都存在显著差异,而苯丙氨酸在发酵液中差异不显著。【结论】影响水稻纹枯病菌菌丝体和发酵液黑色素形成的氨基酸不同,苯丙氨酸对水稻纹枯病菌菌丝体黑色素形成更加重要,而酪氨酸对水稻纹枯病菌发酵液黑色素形成更加重要,以上结果为氨基酸对水稻纹枯病菌黑色素形成的影响提供了参考依据。展开更多
为了探明影响水稻纹枯病菌营养体亲和性分化的因子,将水稻纹枯病菌菌株cx-2在不同水稻品种继代接种,并在含不同杀菌剂、具不同p H及不同温度条件的PDA平板上继代培养,用对峙法测定继代菌株与原始接种菌株的营养体亲和性。此外,对营养体...为了探明影响水稻纹枯病菌营养体亲和性分化的因子,将水稻纹枯病菌菌株cx-2在不同水稻品种继代接种,并在含不同杀菌剂、具不同p H及不同温度条件的PDA平板上继代培养,用对峙法测定继代菌株与原始接种菌株的营养体亲和性。此外,对营养体亲和性分化菌株与原始菌株的AFLP指纹图谱进行比较。结果显示,在供试的30个水稻品种上连续接种4次后,从9个水稻品种中分离出与原始菌株营养体不亲和的菌株。菌株在不同p H值的PDA平板继代培养4次后,在p H偏碱性端(p H 10、p H 11)开始出现营养体亲和性分化的菌株。在不同农药和温度条件下继代培养10次的菌株中没有分离到营养体亲和性分化的菌株。营养体亲和性分化菌株与原始菌株的AFLP指纹图谱没有差异。展开更多
基金supported by the Major Program of Guangdong Basic and Applied Basic Research(Grant No.2019B030302006)the National Natural Science Foundation of China(Grant No.31801677)。
文摘Trehalose metabolism is related to the sclerotial development of Rhizoctonia solani AG-1 IA,the causal agent of rice sheath blight(RSB).Here,we further elucidated the functions of three genes Rstre,Rstps1 and Rstpp that encode three key enzymes trehalase(TRE),alpha,alpha-trehalosephosphate synthase(TPS1)and trehalose 6-phosphate phosphatase(TPP)in the sclerotial development of R.solani AG-1 IA.Due to the lack of a stable genetic transformation system for R.solani,the heterologous expression of these three genes in Pichia pastoris GS115 was performed.The results showed that reactive oxygen species(ROS)contents and enzyme activities in R.solani decreased significantly in the treatments of the fermentation broths of Rstps1 and Rstpp transformants,and that in the treatment of the fermentation broth of Rstre transformant visibly increased.Furthermore,the fermentation broths of the transformants of all the three genes were added to potato dextrose agar(PDA)medium for the cultivation of R.solani,as a result,the dry weight of sclerotia in each PDA plate containing the fermentation broths of Rstps1 and Rstpp transformants significantly increased compared with the control,and that of Rstre transformant obviously decreased.Finally,178 proteins were found to interact with RSTPS1,and 16 of them were associated with ROS.Taken together,the findings suggest that all these three genes related to trehalose metabolism play important roles in the sclerotial development of R.solani AG-1 IA,and can be used as new targets for the development of novel high-efficiency fungicides for the controlling of RSB.
文摘A field under rice-wheat rotation was selected near Chengdu, China, to study thepopulation of Rhizoctonia solani anastomosis group 1 (AG-1), pathogen causing ricesheath blight disease, in natural soil ecosystem. Inocula of the fungus recovered fromthe field were divided into three types, i.e., sclerotia, free mycelium retained in thesoil passed through a 0.355mm sieve, and colonized plant debris which was subdividedinto small colonized debris retained between 2.00 and 0.355mm sieves and large colonizeddebris retained on 2.00mm sieve after wet screening. Quantitative estimation of thethree types of inocula in one year indicated that small colonized debris was the dominantinoculum type for most of the time. The population peaked in March and September at 1210and 480 colonized debris 100g-1 air-dry soil respectively, and fell down in December andAugust to 0 and 177 colonized debris 100g-1 air-dry soil respectively. Free mycelium wasonly detectable in March, September and October with 1209, 7.9 and 14.5g fresh wtmyceliumg-1 air-dry soil respectively, which corresponded to the two peaks and the secondhighest level of small debris density in the year. Viable sclerotia and large colonizeddebris were rare with populations ranging from 0 to 3 for sclerotia and 0 to 14 for largecolonized debris 100g-1 air-dry soil, but were the main structures to survive overwinter. It was expected that soil temperature was the main factor determining populationdynamics of R.solani AG-1 in natural soil. Optimum temperature for population increasingis predicted to be around 15℃, with a range from 10 to 25℃. Viability tests indicatedthat 60.9% sclerotia could survive after 265d being buried in natural sandy loam in fieldconditions in Beijing, while colonized rice straw debris (0.5-1.0cm long) could notyield the fungus on medium plates after 88d of being buried under the same conditions.
文摘【目的】为明确水稻纹枯病菌(Rhizoctonia solani AG 1-IA)黑色素形成前体物质酪氨酸等氨基酸成份在不同培养处理中的差异。【方法】利用氨基酸分析仪对水稻纹枯病菌在3种培养处理(PDB对照,添加了300μg/mL质量浓度莨菪碱的PDB和50μg/mL质量浓度儿茶酚的PDB)后菌丝体和发酵液的氨基酸成份进行了分析。【结果】水稻纹枯病菌菌丝体中谷氨酸、天冬氨酸和丙氨酸3种氨基酸含量相对比较丰富,而谷氨酸的含量最高,达到总氨基酸的15.49%;水稻纹枯病菌发酵液中天冬氨酸和谷氨酸含量最为丰富,分别达到了总氨基酸的17.83%和58.21%;在3种处理中,不管是菌丝体还是发酵液中,都是谷氨酸和天冬氨酸最为丰富。影响水稻纹枯病菌菌丝体黑色素形成的前体物质酪氨酸的含量在3种处理的菌丝体中没有显著差异;但300μg/mL质量浓度的莨菪碱处理组与对照组和儿茶酚处理组中的苯丙氨酸含量都存在显著差异,而对照组和50μg/mL质量浓度的儿茶酚处理组的苯丙氨酸含量差异不显著。水稻纹枯病菌发酵液中对照组和50μg/mL质量浓度的儿茶酚处理组酪氨酸含量没有显著差异,但是300μg/mL质量浓度的莨菪碱处理组与对照组和儿茶酚处理组酪氨酸含量都存在显著差异,而苯丙氨酸在发酵液中差异不显著。【结论】影响水稻纹枯病菌菌丝体和发酵液黑色素形成的氨基酸不同,苯丙氨酸对水稻纹枯病菌菌丝体黑色素形成更加重要,而酪氨酸对水稻纹枯病菌发酵液黑色素形成更加重要,以上结果为氨基酸对水稻纹枯病菌黑色素形成的影响提供了参考依据。
文摘为了探明影响水稻纹枯病菌营养体亲和性分化的因子,将水稻纹枯病菌菌株cx-2在不同水稻品种继代接种,并在含不同杀菌剂、具不同p H及不同温度条件的PDA平板上继代培养,用对峙法测定继代菌株与原始接种菌株的营养体亲和性。此外,对营养体亲和性分化菌株与原始菌株的AFLP指纹图谱进行比较。结果显示,在供试的30个水稻品种上连续接种4次后,从9个水稻品种中分离出与原始菌株营养体不亲和的菌株。菌株在不同p H值的PDA平板继代培养4次后,在p H偏碱性端(p H 10、p H 11)开始出现营养体亲和性分化的菌株。在不同农药和温度条件下继代培养10次的菌株中没有分离到营养体亲和性分化的菌株。营养体亲和性分化菌株与原始菌株的AFLP指纹图谱没有差异。
