Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy ...Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast. Thus, it is a good recipient cell for exogenous gene expression. In this study, the expression plasmid pAN7-1 （containing promoter gpd-An derived from Aspergillus nidulans and selectable marker gene hph conferring resistance to hygromycin B） and plasmid pLg-hph （containing promoter gpd-Le derived from Lentinula edodes and selectable marker gene hph） were transformed into the yeast-like conidia of T. fuciformis by PEG-mediated protoplast transformation, respectively. The primary putative transformants were selected by the sandwich screening method with the selective medium containing 50 μg mL^-1 hygromycin. The putative transformants were obtained from the primary putative transformants transferred on PDSA plates containing 100 μg mL^-1 hygromycin for second round selection. Experimental results showed that the optimal concentration of PEG 4000 for mediating protoplast transformation was 25%. PCR and Southern blotting confirmed that the selectable marker gene hph was integrated effectively into the genome of the yeast-like conidia of T. fuciformis with plasmid pLg-hph transformation. Its transformation efficiency was 110 transformants per μg DNA, and the hph gene was integrated into the genome of some yeast-like conidia with plasmid pAN7-1 transformation. However, its transformation efficiency was only 9 transformants per μg DNA. The presence of hph gene in the genome of transformants after 5 generations of sub- culturing on PDSB medium was confirmed by PCR, suggesting that the foreign gene hph was stable during subculture.展开更多
The newly-hatched nymphs of the small brown planthopper (SBPH), Laodelphax striatellus, including field and sensitive populations, were subjected to the high-temperature (35°C) treatment. The number of yeast-like...The newly-hatched nymphs of the small brown planthopper (SBPH), Laodelphax striatellus, including field and sensitive populations, were subjected to the high-temperature (35°C) treatment. The number of yeast-like endosymbiotes in SBPH reduced by 23.47%–34.23%, 57.86%–61.51% and 88.96%–90.71% after the high-temperature treatment for 1 d, 2 d, and 3 d, respectively. However, the size of yeast-like endosymbiotes was not obviously affected. Resistance of SBPH to three insecticides (imidacloprid, chlorpyrifos and fipronil) decreased with the increase of treatment time.展开更多
To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the libra...To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the library was 20 kb. This library was screened for antibiotic activity using paper disc assaying. Two clones displayed the antibacterial activity against Micrococcus tetragenus. The metabolites of these two clones were analyzed through HPLC. The result showed that their metabolites were quite different from those of the host E. coli DNA and the host containing vector pHZ132. This study may present a new approach to exploring bioactive metabolites of sponge symbionts.展开更多
Marine biofouling has been regarded as a serious problem in the marine environment. The application of TBT and other heavy metal-based antifoulants has created another environmental problem. The present study explored...Marine biofouling has been regarded as a serious problem in the marine environment. The application of TBT and other heavy metal-based antifoulants has created another environmental problem. The present study explored the possible role of baterial symbionts of seagrasses Thalassia hemprichii, and Enhalus acoroides, which were successfully screened for antifouling activity against marine biofilm-forming bacteria isolated from the surrounding colonies of seagrasses. Bacterial symbionts were isolated and tested against biofilm-forming bacteria resulted in 4 bacterial symbionts capable of inhibiting the growth biofilm-forming isolates. Molecular identification based on 16S rRNA gene sequences revealed that the active bacterial symbionts belonged to the members of the genera Bacillus and Virgibacillus. Further tests of the crude extracts of the active bacterial symbionts supported the potential of these symbionts as the alternative source of environmentally friendly marine antifoulants.展开更多
The objective of this research was to improve the growth and biomass of Pterocarpus santalinus L.f.(an endangered leguminous tree)using native microbial symbionts such as arbuscular mycorrhizal fungi and Rhizobium ass...The objective of this research was to improve the growth and biomass of Pterocarpus santalinus L.f.(an endangered leguminous tree)using native microbial symbionts such as arbuscular mycorrhizal fungi and Rhizobium associated with native populations of P.santalinus.The native arbuscular mycorrhizal fungi isolated from P.santalinus soils were identifi ed as(1)Glomus fasciculatum;(2)Glomus geosporum;and Glomus aggregatum.A nitrogenfi xing microbial symbiont was isolated from the root nodules of P.santalinus and identifi ed as Rhizobium aegyptiacum by 16s rRNA gene sequencing.These microbial symbionts were inoculated individually and in combination into P.santalinus seedling roots.After 90 days,growth and biomass had improved compared with uninoculated controls.Shoot and root lengths,number of leaves,stem circumference,number of root nodules,biomass,nutrient uptake and seedling quality index were signifi cantly increased by a combined inoculation of arbuscular mycorrhizal fungi+Rhizobium aegyptiacum.It was concluded that native microbial symbionts positively infl uenced P.santalinus seedling growth which will be helpful for successful fi eld establishment.展开更多
The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen fixation.However,the whole nitrogen-cycling network has rema...The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen fixation.However,the whole nitrogen-cycling network has remained incomplete in any individual sponge holobiont.In this study,454 pyrosequencing of the 16S rRNA genes revealed that the sponge Spheciospongia vesparium from the South China Sea has a unique bacterial community(including 12 bacterial phyla),dominated particularly by the genus Shewanella(order Alteromonadales).A total of 10 functional genes,nifH,amoA,narG,napA,nirK,norB,nosZ,ureC,nrfA,and gltB,were detected in the microbiome of the sponge S.vesparium by gene-targeted analysis,revealing an almost complete nitrogen-cycling network in this sponge.Particularly,bacterial urea utilization and the whole denitrification pathway were highlighted.MEGAN analysis suggests that Proteobacteria(e.g.,Shewanella)and Bacteroidetes(e.g.,Bizionia)are probably involved in the nitrogen cycle in the sponge S.vesparium.展开更多
基金funded by the National Hi-Tech Research and Development Program of China (863 Program,2006AA10Z301)the National Natural Science Foundation of China (30371000, 30671457)
文摘Tremellafuciformis is one of higher basidiomycetes. Its basidiospore can reproduce yeast-like conidia, which is also called the blastospore by budding. The yeast-like conidia of T. fuciformis is monokaryotic and easy to culture by submerged fermentation similar to yeast. Thus, it is a good recipient cell for exogenous gene expression. In this study, the expression plasmid pAN7-1 （containing promoter gpd-An derived from Aspergillus nidulans and selectable marker gene hph conferring resistance to hygromycin B） and plasmid pLg-hph （containing promoter gpd-Le derived from Lentinula edodes and selectable marker gene hph） were transformed into the yeast-like conidia of T. fuciformis by PEG-mediated protoplast transformation, respectively. The primary putative transformants were selected by the sandwich screening method with the selective medium containing 50 μg mL^-1 hygromycin. The putative transformants were obtained from the primary putative transformants transferred on PDSA plates containing 100 μg mL^-1 hygromycin for second round selection. Experimental results showed that the optimal concentration of PEG 4000 for mediating protoplast transformation was 25%. PCR and Southern blotting confirmed that the selectable marker gene hph was integrated effectively into the genome of the yeast-like conidia of T. fuciformis with plasmid pLg-hph transformation. Its transformation efficiency was 110 transformants per μg DNA, and the hph gene was integrated into the genome of some yeast-like conidia with plasmid pAN7-1 transformation. However, its transformation efficiency was only 9 transformants per μg DNA. The presence of hph gene in the genome of transformants after 5 generations of sub- culturing on PDSB medium was confirmed by PCR, suggesting that the foreign gene hph was stable during subculture.
文摘The newly-hatched nymphs of the small brown planthopper (SBPH), Laodelphax striatellus, including field and sensitive populations, were subjected to the high-temperature (35°C) treatment. The number of yeast-like endosymbiotes in SBPH reduced by 23.47%–34.23%, 57.86%–61.51% and 88.96%–90.71% after the high-temperature treatment for 1 d, 2 d, and 3 d, respectively. However, the size of yeast-like endosymbiotes was not obviously affected. Resistance of SBPH to three insecticides (imidacloprid, chlorpyrifos and fipronil) decreased with the increase of treatment time.
基金This work was supported by High Tech R&D Program of China(Grant Nos.2002AA628130 and 2003AA624020)the National Natural Science Foundation of China(30171102)+2 种基金the Fund for Cheung Kong Scholar from the Cheung Kong Scholar Program of Ministry of Education of Chinathe Fund from the Natural Science Foundation of Shandong Province(No.Z2001C01)the High Tech R&D Program of Shandong Province(No.0121100107).The authors would like to thank Professor Li Jinhe of Institute of 0ceanology,Chinese Academy of Sciences,for the identification of the sponge.
文摘To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the library was 20 kb. This library was screened for antibiotic activity using paper disc assaying. Two clones displayed the antibacterial activity against Micrococcus tetragenus. The metabolites of these two clones were analyzed through HPLC. The result showed that their metabolites were quite different from those of the host E. coli DNA and the host containing vector pHZ132. This study may present a new approach to exploring bioactive metabolites of sponge symbionts.
文摘Marine biofouling has been regarded as a serious problem in the marine environment. The application of TBT and other heavy metal-based antifoulants has created another environmental problem. The present study explored the possible role of baterial symbionts of seagrasses Thalassia hemprichii, and Enhalus acoroides, which were successfully screened for antifouling activity against marine biofilm-forming bacteria isolated from the surrounding colonies of seagrasses. Bacterial symbionts were isolated and tested against biofilm-forming bacteria resulted in 4 bacterial symbionts capable of inhibiting the growth biofilm-forming isolates. Molecular identification based on 16S rRNA gene sequences revealed that the active bacterial symbionts belonged to the members of the genera Bacillus and Virgibacillus. Further tests of the crude extracts of the active bacterial symbionts supported the potential of these symbionts as the alternative source of environmentally friendly marine antifoulants.
基金a research project(No.IFGTB/NFRP 168)of the Indian Council of Forestry Research and Education,Dehra Dun.
文摘The objective of this research was to improve the growth and biomass of Pterocarpus santalinus L.f.(an endangered leguminous tree)using native microbial symbionts such as arbuscular mycorrhizal fungi and Rhizobium associated with native populations of P.santalinus.The native arbuscular mycorrhizal fungi isolated from P.santalinus soils were identifi ed as(1)Glomus fasciculatum;(2)Glomus geosporum;and Glomus aggregatum.A nitrogenfi xing microbial symbiont was isolated from the root nodules of P.santalinus and identifi ed as Rhizobium aegyptiacum by 16s rRNA gene sequencing.These microbial symbionts were inoculated individually and in combination into P.santalinus seedling roots.After 90 days,growth and biomass had improved compared with uninoculated controls.Shoot and root lengths,number of leaves,stem circumference,number of root nodules,biomass,nutrient uptake and seedling quality index were signifi cantly increased by a combined inoculation of arbuscular mycorrhizal fungi+Rhizobium aegyptiacum.It was concluded that native microbial symbionts positively infl uenced P.santalinus seedling growth which will be helpful for successful fi eld establishment.
基金Financial support from the National Natural Science Foundation of China(NSFC)(Nos.31861143020,41776138)was used to conduct this research and is greatly appreciated.
文摘The microbes associated with sponges play important roles in the nitrogen cycle of the coral reefs ecosystem,e.g.,nitrification,denitrification,and nitrogen fixation.However,the whole nitrogen-cycling network has remained incomplete in any individual sponge holobiont.In this study,454 pyrosequencing of the 16S rRNA genes revealed that the sponge Spheciospongia vesparium from the South China Sea has a unique bacterial community(including 12 bacterial phyla),dominated particularly by the genus Shewanella(order Alteromonadales).A total of 10 functional genes,nifH,amoA,narG,napA,nirK,norB,nosZ,ureC,nrfA,and gltB,were detected in the microbiome of the sponge S.vesparium by gene-targeted analysis,revealing an almost complete nitrogen-cycling network in this sponge.Particularly,bacterial urea utilization and the whole denitrification pathway were highlighted.MEGAN analysis suggests that Proteobacteria(e.g.,Shewanella)and Bacteroidetes(e.g.,Bizionia)are probably involved in the nitrogen cycle in the sponge S.vesparium.
