【目的】通过分析在逆境胁迫过程中绿僵菌菌株MAX-2的Hog1基因的转录表达,为揭示该菌株耐逆特性的分子机理提供理论依据。【方法】将具有不同耐逆能力的3个绿僵菌菌株MAX-2、BUM2495和BUM723的分生孢子分别用盐处理(0.4、0.8 mol/L Na ...【目的】通过分析在逆境胁迫过程中绿僵菌菌株MAX-2的Hog1基因的转录表达,为揭示该菌株耐逆特性的分子机理提供理论依据。【方法】将具有不同耐逆能力的3个绿僵菌菌株MAX-2、BUM2495和BUM723的分生孢子分别用盐处理(0.4、0.8 mol/L Na Cl)和紫外线照射(5、15 min),测定孢子萌发率,并提取RNA,通过半定量RT-PCR方法分析其Hog1基因的转录表达差异。【结果】从盐和紫外线逆境条件下菌株的孢子萌发率可以看出:这3个菌株的耐逆性能为MAX-2>BUM2495>BUM723,菌株间的差异都达到了极显著水平(P<0.01)。盐处理和紫外线辐射都能诱导绿僵菌Hog1基因的表达上调,不同菌株表达差异较大,MAX-2表达水平最高,BUM723表达水平最低。在较低程度的胁迫条件下(0.4 mol/L Na Cl和紫外线照射5 min),MAX-2的表达水平略高于BUM2495,而在较高程度的胁迫条件下(0.8 mol/L Na Cl和紫外线照射15 min)MAX-2的表达水平明显高于BUM2495,表明Hog1基因在较高程度压力条件下的高效表达与MAX-2的耐逆能力密切相关。【结论】绿僵菌菌株MAX-2较高的耐逆能力与其Hog1基因的高水平转录表达相关。展开更多
Arsenic is a highly toxic and carcinogenic metalloid widely dispersed in the environment, contaminating water and soil and accumulating in crops. Paradoxically, arsenic is also part of modern therapy and employed in t...Arsenic is a highly toxic and carcinogenic metalloid widely dispersed in the environment, contaminating water and soil and accumulating in crops. Paradoxically, arsenic is also part of modern therapy and employed in treating numerous ailments and diseases. Hence, inventing strategies to tune cellular arsenic uptake based on purpose is striking. Here, we describe an approach in which the arsenite uptake can be increased using a MAPK inhibitor. Employing microfluidic flow chambers in combination with optical tweezers and fluorescent microscopy, we elevated the influx of arsenite into the yeast Saccharomyces cerevisiae cells following short-term treatment with a Hog1 kinase inhibitor. The increase in arsenite uptake was followed on arsenite triggered redistribution of a reporter protein, Hsp104-GFP, which was imaged over time. The effect was even more pronounced when the yeast mother and daughter cells were analyzed disjointedly, an opportunity provided owing to single-cell analysis. Our data firstly provide a strategy to increase arsenite uptake and secondly show that arsenite triggered aggregates, previously shown to be sites of damaged proteins, are distributed asymmetrically and less accumulated in daughter cells. Inventing approaches to tune arsenite uptake has a great value for its use in environmental as well as medical applications.展开更多
In several filamentous fungi,incident light and environmental stress signaling share the mitogen-activated protein kinase(MAPK)HOG(SAK)pathway.It has been revealed that short-term illumination with blue light triggers...In several filamentous fungi,incident light and environmental stress signaling share the mitogen-activated protein kinase(MAPK)HOG(SAK)pathway.It has been revealed that short-term illumination with blue light triggers the activation of the HOG pathway in Trichoderma spp.In this study,we demonstrate the crucial role of the basic leucine zipper transcription factor ATF1 in blue light responses and signaling downstream of the MAPK HOG1 in Trichoderma guizhouense.The lack of ATF1 severely impaired photoconidiation and delayed vegetative growth and conidial germination.Upon blue light or H2O2 stimuli,HOG1 interacted with ATF1 in the nucleus.Genome-wide transcriptome analyses revealed that 61.8%(509 out of 824)and 85.2%(702 out of 824)of blue light-regulated genes depended on ATF1 and HOG1,respectively,of which 58.4%(481 out of 824)were regulated by both of them.Our results also show that blue light promoted conidial germination and HOG1 and ATF1 played opposite roles in controlling conidial germination in the dark.Additionally,the lack of ATF1 led to reduced oxidative stress resistance,probably because of the downregulation of catalase-encoding genes.Overall,our results demonstrate that ATF1 is the downstream component of HOG1 and is responsible for blue light responses,conidial germination,vegetative growth,and oxidative stress resistance in T.guizhouense.展开更多
文摘【目的】通过分析在逆境胁迫过程中绿僵菌菌株MAX-2的Hog1基因的转录表达,为揭示该菌株耐逆特性的分子机理提供理论依据。【方法】将具有不同耐逆能力的3个绿僵菌菌株MAX-2、BUM2495和BUM723的分生孢子分别用盐处理(0.4、0.8 mol/L Na Cl)和紫外线照射(5、15 min),测定孢子萌发率,并提取RNA,通过半定量RT-PCR方法分析其Hog1基因的转录表达差异。【结果】从盐和紫外线逆境条件下菌株的孢子萌发率可以看出:这3个菌株的耐逆性能为MAX-2>BUM2495>BUM723,菌株间的差异都达到了极显著水平(P<0.01)。盐处理和紫外线辐射都能诱导绿僵菌Hog1基因的表达上调,不同菌株表达差异较大,MAX-2表达水平最高,BUM723表达水平最低。在较低程度的胁迫条件下(0.4 mol/L Na Cl和紫外线照射5 min),MAX-2的表达水平略高于BUM2495,而在较高程度的胁迫条件下(0.8 mol/L Na Cl和紫外线照射15 min)MAX-2的表达水平明显高于BUM2495,表明Hog1基因在较高程度压力条件下的高效表达与MAX-2的耐逆能力密切相关。【结论】绿僵菌菌株MAX-2较高的耐逆能力与其Hog1基因的高水平转录表达相关。
文摘Arsenic is a highly toxic and carcinogenic metalloid widely dispersed in the environment, contaminating water and soil and accumulating in crops. Paradoxically, arsenic is also part of modern therapy and employed in treating numerous ailments and diseases. Hence, inventing strategies to tune cellular arsenic uptake based on purpose is striking. Here, we describe an approach in which the arsenite uptake can be increased using a MAPK inhibitor. Employing microfluidic flow chambers in combination with optical tweezers and fluorescent microscopy, we elevated the influx of arsenite into the yeast Saccharomyces cerevisiae cells following short-term treatment with a Hog1 kinase inhibitor. The increase in arsenite uptake was followed on arsenite triggered redistribution of a reporter protein, Hsp104-GFP, which was imaged over time. The effect was even more pronounced when the yeast mother and daughter cells were analyzed disjointedly, an opportunity provided owing to single-cell analysis. Our data firstly provide a strategy to increase arsenite uptake and secondly show that arsenite triggered aggregates, previously shown to be sites of damaged proteins, are distributed asymmetrically and less accumulated in daughter cells. Inventing approaches to tune arsenite uptake has a great value for its use in environmental as well as medical applications.
基金supported by the National Natural Science Foundation of China (NSFC) (Grant Nos.32070101 and 32270053)the Fundamental Research Funds for the Central Universities (Grant Nos.XUEKEN2023039,XUEKEN2023041,RENCAI2022005,and KYT2023001)the Jiangsu Agriculture Science and Technology Innovation fund (JASTIF) (Grant No.CX (21)2018).
文摘In several filamentous fungi,incident light and environmental stress signaling share the mitogen-activated protein kinase(MAPK)HOG(SAK)pathway.It has been revealed that short-term illumination with blue light triggers the activation of the HOG pathway in Trichoderma spp.In this study,we demonstrate the crucial role of the basic leucine zipper transcription factor ATF1 in blue light responses and signaling downstream of the MAPK HOG1 in Trichoderma guizhouense.The lack of ATF1 severely impaired photoconidiation and delayed vegetative growth and conidial germination.Upon blue light or H2O2 stimuli,HOG1 interacted with ATF1 in the nucleus.Genome-wide transcriptome analyses revealed that 61.8%(509 out of 824)and 85.2%(702 out of 824)of blue light-regulated genes depended on ATF1 and HOG1,respectively,of which 58.4%(481 out of 824)were regulated by both of them.Our results also show that blue light promoted conidial germination and HOG1 and ATF1 played opposite roles in controlling conidial germination in the dark.Additionally,the lack of ATF1 led to reduced oxidative stress resistance,probably because of the downregulation of catalase-encoding genes.Overall,our results demonstrate that ATF1 is the downstream component of HOG1 and is responsible for blue light responses,conidial germination,vegetative growth,and oxidative stress resistance in T.guizhouense.
