We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （...We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （ITS） sequences of Verticillium spp., a pair of species-specific primers, Vaal/Vaa2, was synthesized. After screening 17 isolates of V. alboatrum, 121 isolates from the Ascomycota, B asidiomycota, Deuteromycota, and Oomycota, the Vaal/Vaa2 primers amplified only a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaal/Vaa2 was 10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedures were developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogens was 100-conidiag^-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.展开更多
Polysaccharides from genus Ganoderma have prominent anti-aging effect,but their mechanisms are incompletely clarified.In our previous experiments,Ganoderma atrum polysaccharide(PSG)was exhibited to significantly allev...Polysaccharides from genus Ganoderma have prominent anti-aging effect,but their mechanisms are incompletely clarified.In our previous experiments,Ganoderma atrum polysaccharide(PSG)was exhibited to significantly alleviate senescence and DNA damage of A375 cells.To investigate its underlying mechanism,we conducted RNA-Seq analysis and found that PSG upregulated autophagy and mitochondria pathways compared to the model group.Further experiments showed that PSG relieved mitochondrial dysfunction via inhibiting the accumulation of reactive oxygen species(ROS)and loss of the mitochondrial membrane potential(MMP).Meanwhile,PSG activated autophagy and enhanced lysosomal function through transcription factor EB(TFEB).Interestingly,the pretreatment of autophagy inhibitors bafilomycin A1(Baf A1)reversed the ROS decline induced by PSG,which subsequently increased cellular senescence of A375.In addition,PSG had a similar anti-aging effect on normal fibroblast WI-38.Taken together,PSG might reduce ROS levels through activating autophagy,which in turn suppressed mitochondrial dysfunction and cellular senescence.展开更多
基金We thank Dr Chen Qinghe,Dr Ko Wenhong,Dr Ho Hanhin,Dr Hu Baishi,Dr Peng Jinghuo and China General Microbiological Culture Collection Center(CGMCC)for providing some isolates.This work was supported by the China National 863 Program(2003AA249020).
文摘We developed one species-specific PCR assays for rapid and accurate detection of the pathogenic fungi Verticillium albo-atrum in diseased plant tissues and soil. Based on differences in internal transcribed spacer （ITS） sequences of Verticillium spp., a pair of species-specific primers, Vaal/Vaa2, was synthesized. After screening 17 isolates of V. alboatrum, 121 isolates from the Ascomycota, B asidiomycota, Deuteromycota, and Oomycota, the Vaal/Vaa2 primers amplified only a single PCR band of approximately 330 bp from V. albo-atrum. The detection sensitivity with primers Vaal/Vaa2 was 10 fg of genomic DNA. Using ITS1/ITS4 as the first-round primers, combined with Vaa1/Vaa2, the nested PCR procedures were developed, and the detection sensitivity increased 1 000-fold to 10 ag. The detection sensitivity for the soil pathogens was 100-conidiag^-1 soil. The PCR-based methods developed here could simplify both plant disease diagnosis and pathogen monitoring as well as guide plant disease management.
基金This work was supported by the National Science Fund for Distinguished Young Scholars of China(31825020)the Technology Innovation Guidance Program of Jiangxi Province(20203AEI91008)Key laboratory of Bioactive Polysaccharides of Jiangxi Province(20212BCD42016).
文摘Polysaccharides from genus Ganoderma have prominent anti-aging effect,but their mechanisms are incompletely clarified.In our previous experiments,Ganoderma atrum polysaccharide(PSG)was exhibited to significantly alleviate senescence and DNA damage of A375 cells.To investigate its underlying mechanism,we conducted RNA-Seq analysis and found that PSG upregulated autophagy and mitochondria pathways compared to the model group.Further experiments showed that PSG relieved mitochondrial dysfunction via inhibiting the accumulation of reactive oxygen species(ROS)and loss of the mitochondrial membrane potential(MMP).Meanwhile,PSG activated autophagy and enhanced lysosomal function through transcription factor EB(TFEB).Interestingly,the pretreatment of autophagy inhibitors bafilomycin A1(Baf A1)reversed the ROS decline induced by PSG,which subsequently increased cellular senescence of A375.In addition,PSG had a similar anti-aging effect on normal fibroblast WI-38.Taken together,PSG might reduce ROS levels through activating autophagy,which in turn suppressed mitochondrial dysfunction and cellular senescence.
