We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a...We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a strain of A. niger by adopting a variant Czapek medium （using NazHPO4 as a substitute for KzHPO4） in two groups, Group A （containing silicate minerals bearing potassium but without KC1） and Group B （with KCI） . We extracted the mRNAs of CA and CysM from these two groups and performed real-time quantitative polymerase chain reactions （RT-qPCR）. We constructed relative standard curves by using glyceraldehyde-3-phosphate dehydrogenase （GAPDH） as the reference to confirm a consistent amplification effi- ciency of the target genes （CA and CysM） and the reference gene and quantified the gene expression of the targets in a relative manner. Our results showed that CA and CysM in Group A were upregulated for 1.7 times and 11.7 times, respectively, com- pared with those in Group B. Furthermore, we also analyzed some metabolic pathways and functions of the A. niger-induced weathering of potassium-bearing minerals, which involved the synthesizing of these two enzymes. Thus our work provides materials for further study of the roles of A. niger in the metabolic regulation during the weathering process of potassi- um-beating minerals.展开更多
Both activated carbon and magnetite have been reported to promote the syntrophic growth of Geobacter metallireducens and Geobacter sulfurreducens co-cultures, the first model to show direct interspecies electron trans...Both activated carbon and magnetite have been reported to promote the syntrophic growth of Geobacter metallireducens and Geobacter sulfurreducens co-cultures, the first model to show direct interspecies electron transfer (DIET); however, differential transcriptomics of the promotion on co-cultures with these two conductive materials are unknown. Here, the comparative transcriptomic analysis of G. metallireducens and G. sulfurreducens co-cultures with granular activated carbon (GAC) and magnetite was reported. More than 2.6-fold reduced transcript abundances were determined for the uptake hydrogenase genes of G. sulfurreducens as well as other hydrogenases in those co-cultures to which conductive materials had been added. This is consistent with electron transfer in G. metallireducens-G. sulfurreducens co-cultures as evinced by direct interspecies electron transfer (DIET). Transcript abundance for the structural component of electrically conductive pili (e-pili), PilA, was 2.2-fold higher in G. metallireducens, and, in contrast, was 14.9-fold lower in G. sulfurreducens in co-cultures with GAC than in Geobacters co-cultures without GAC. However, it was 9.3-fold higher in G. sulfurreducens in co-cultures with magnetite than in Geobacters co-cultures. Mutation results showed that GAC can be substituted for the e-pili of both strains but magnetite can only compensate for that of G. sulfurreducens, indicating that the e-pili is a more important electron acceptor for the electron donor strain of G. metallireducens than for G. sulfurreducens. Transcript abundance for G. metallireducens c-type cytochrome gene GMET_RS14535, a homologue to c-type cytochrome gene omcE of G. sulfurreducens was 9.8-fold lower in co-cultures with GAC addition, while that for OmcS of G. sulfurreducens was 25.1-fold higher in co-cultures with magnetite, than in that without magnetite. Gene deletion studies showed that neither GAC nor magnetite can completely substitute the cytochrome (OmcE homologous) of G. metallireducens but compensate for the cytochrome (OmcS) of G. sulfurreducens. Moreover, some genes associated with central metabolism were up-regulated in the presence of both GAC and magnetite; however, tricarboxylic acid cycle gene transcripts in G. sulfurreducens were not highly-expressed in each of these amended co-cultures, suggesting that there was considerable redundancy in the pathways utilised by G. sulfurreducens for electron transfer to reduce fumarate with the amendment of GAC or magnetite. These results support the DIET model of G. metallireducens and G. sulfurreducens and suggest that e-pili and cytochromes of the electron donor strain are more important than that of the electron acceptor strain, indicating that comparative transcriptomics may be a promising route by which to reveal different responses of electron donor and acceptor during DIET in co-cultures.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.41173091)National Key Basic Research Program of China(Grant No.2013CB956702)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘We investigated the differences in the gene expression of carbonic anhydrase （CA） and cysteine synthase （CysM） between two weathering conditions, with either soluble potassium or insoluble potassium. We cultured a strain of A. niger by adopting a variant Czapek medium （using NazHPO4 as a substitute for KzHPO4） in two groups, Group A （containing silicate minerals bearing potassium but without KC1） and Group B （with KCI） . We extracted the mRNAs of CA and CysM from these two groups and performed real-time quantitative polymerase chain reactions （RT-qPCR）. We constructed relative standard curves by using glyceraldehyde-3-phosphate dehydrogenase （GAPDH） as the reference to confirm a consistent amplification effi- ciency of the target genes （CA and CysM） and the reference gene and quantified the gene expression of the targets in a relative manner. Our results showed that CA and CysM in Group A were upregulated for 1.7 times and 11.7 times, respectively, com- pared with those in Group B. Furthermore, we also analyzed some metabolic pathways and functions of the A. niger-induced weathering of potassium-bearing minerals, which involved the synthesizing of these two enzymes. Thus our work provides materials for further study of the roles of A. niger in the metabolic regulation during the weathering process of potassi- um-beating minerals.
基金supported by the Major Research plan(91751112)the General Programme(41371257,41573071)of the National Natural Science Foundation of China+2 种基金Shandong Natural Science Fund for Distinguished Young Scholars(JQ201608)the Young Taishan Scholars Programme of Shandong Province(tsqn20161054)the Key Research Project for Frontier Science of the Chinese Academy of Sciences(QYZDJ-SSW-DQC015)
文摘Both activated carbon and magnetite have been reported to promote the syntrophic growth of Geobacter metallireducens and Geobacter sulfurreducens co-cultures, the first model to show direct interspecies electron transfer (DIET); however, differential transcriptomics of the promotion on co-cultures with these two conductive materials are unknown. Here, the comparative transcriptomic analysis of G. metallireducens and G. sulfurreducens co-cultures with granular activated carbon (GAC) and magnetite was reported. More than 2.6-fold reduced transcript abundances were determined for the uptake hydrogenase genes of G. sulfurreducens as well as other hydrogenases in those co-cultures to which conductive materials had been added. This is consistent with electron transfer in G. metallireducens-G. sulfurreducens co-cultures as evinced by direct interspecies electron transfer (DIET). Transcript abundance for the structural component of electrically conductive pili (e-pili), PilA, was 2.2-fold higher in G. metallireducens, and, in contrast, was 14.9-fold lower in G. sulfurreducens in co-cultures with GAC than in Geobacters co-cultures without GAC. However, it was 9.3-fold higher in G. sulfurreducens in co-cultures with magnetite than in Geobacters co-cultures. Mutation results showed that GAC can be substituted for the e-pili of both strains but magnetite can only compensate for that of G. sulfurreducens, indicating that the e-pili is a more important electron acceptor for the electron donor strain of G. metallireducens than for G. sulfurreducens. Transcript abundance for G. metallireducens c-type cytochrome gene GMET_RS14535, a homologue to c-type cytochrome gene omcE of G. sulfurreducens was 9.8-fold lower in co-cultures with GAC addition, while that for OmcS of G. sulfurreducens was 25.1-fold higher in co-cultures with magnetite, than in that without magnetite. Gene deletion studies showed that neither GAC nor magnetite can completely substitute the cytochrome (OmcE homologous) of G. metallireducens but compensate for the cytochrome (OmcS) of G. sulfurreducens. Moreover, some genes associated with central metabolism were up-regulated in the presence of both GAC and magnetite; however, tricarboxylic acid cycle gene transcripts in G. sulfurreducens were not highly-expressed in each of these amended co-cultures, suggesting that there was considerable redundancy in the pathways utilised by G. sulfurreducens for electron transfer to reduce fumarate with the amendment of GAC or magnetite. These results support the DIET model of G. metallireducens and G. sulfurreducens and suggest that e-pili and cytochromes of the electron donor strain are more important than that of the electron acceptor strain, indicating that comparative transcriptomics may be a promising route by which to reveal different responses of electron donor and acceptor during DIET in co-cultures.
