A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phyl...A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phylogenetic relationships.Based on structures of N-terminal motif and leucine-rich repeat domains motif,we found 381 NBS-coding sequences with 122 non-regular NBS genes and 259 regular NBS genes that were further classified into 13 types such as TNL,CNL,NL,XNL,TN and other minor types.Meanwhile 81.9% of the NBS genes were distributed in cluster,and 81.8% of the cluster genes had duplicates.The results showed that there were many duplicate phenomenon occurred in the evolution of disease resistance genes of P.trichocarpa.After analysis of NBS standard phylogenetic tree in the genome of P.trichocarpa,the structure of tree exhibited a star topology,and the regular NBS genes were classified into 68 groups by less than 30% amino acid sequence diversity in each domain.展开更多
目的探讨白头翁汤正丁醇提取物(Butyl alcohol extract of Bai Tou Weng decoction,BAEB)对外阴阴道念珠菌病(Vulvovaginal Candidiasis,VVC)白念珠菌临床分离株黏附作用的影响。方法平板法检测与BAEB共培养2 h、4 h时白念珠菌的CFU;XT...目的探讨白头翁汤正丁醇提取物(Butyl alcohol extract of Bai Tou Weng decoction,BAEB)对外阴阴道念珠菌病(Vulvovaginal Candidiasis,VVC)白念珠菌临床分离株黏附作用的影响。方法平板法检测与BAEB共培养2 h、4 h时白念珠菌的CFU;XTT还原法检测与BAEB共培养2 h、4 h的白念珠菌代谢活性;试管法检测白念珠菌絮凝能力;水-烃两相法检测白念珠菌细胞表面疏水性(cell surface hydrophobicity,CSH);荧光显微镜观察菌体活力;扫描电镜观察白念珠菌细胞形态;qRT-PCR法检测黏附相关基因ALS1、CSH1、HWP1、ALS5、ALA1、INT1、MNT2与ALS3的表达。结果 512、1 024μg/m L BAEB可显著抑制黏附2 h、4 h时的白念珠菌的CFU及代谢活性;肉眼和倒置显微镜下观察发现512、1 024μg/m L BAEB可明显延缓絮凝产生的时间;256、512、1 024μg/m L的BAEB均能够显著减少白念珠菌的细胞表面疏水性;qRT-PCR检测表明1 024μg/m L的BAEB作用后,ALS1、CSH1、ALA1、ALS3、INT1、HWP1分别下调18.7%、36%、37.8%、29.6%、19.9%、21.6%,ALS5与MNT2未产生明显变化。结论 BAEB可能通过调节黏附相关基因抑制白念珠菌VVC临床株的黏附作用。展开更多
目的探讨白头翁汤正丁醇提取物（Butyl alcohol extract of Bai Tou Weng decoction,BAEB）对分离自外阴阴道念珠菌病（vulvovaginal candidiasis,VVC）的白念珠菌临床分离株（以下简称VVC临床株）生物膜形成的影响。方法采用微量稀释...目的探讨白头翁汤正丁醇提取物（Butyl alcohol extract of Bai Tou Weng decoction,BAEB）对分离自外阴阴道念珠菌病（vulvovaginal candidiasis,VVC）的白念珠菌临床分离株（以下简称VVC临床株）生物膜形成的影响。方法采用微量稀释法测定BAEB对白念珠菌的最低抑菌浓度（Minimal Inhibitory Concentration,MIC）;甲基四氮盐（XTT）还原法测定BAEB对白念珠菌生物膜代谢活性的影响,Time-kill法检测BAEB对白念珠菌活菌数的影响;结晶紫染色法测定BAEB对白念珠菌生物膜生物量（Biomass）的影响;扫描电镜（SEM）观察BAEB对白念珠菌生物膜形态结构的影响;激光共聚焦显微镜（CLSM）检测BAEB对白念珠菌生物膜荧光信号强度的影响;实时荧光定量PCR（qRT-PCR）检测生物膜相关基因UME6、PES1和HSP90的转录水平变化。结果 BAEB对12株白念珠菌的MIC在64~256μg/mL之间,对白念珠菌生物膜的SMIC80（抑制80%生物膜形成的最低药物浓度）为1 024μg/mL或以上;Time-Kill曲线显示在12h之后,512、1 024μg/mL浓度的BAEB对白念珠菌均具良好的杀伤作用;结晶紫染色法表明512、1 024μg/mLBAEB能够减少其生物膜生物量;SEM观察到1 024μg/mL BAEB能够有效抑制白念珠菌在不同黏附介质上生物膜的完整度;CLSM显示512、1 024μg/mL的BAEB可以明显降低生物膜荧光信号强度;qRT-PCR检测显示在256、512、1 024μg/mL的BAEB作用下,UME6转录水平分别下调了72%、71%、77%,在512、1 024μg/mL的BAEB下HSP90转录水平上调了2.23和3.31倍,而PES1未有明显变化。结论 BAEB可以抑制白念珠菌VVC临床株体外生物膜的形成。展开更多
文摘A full set of disease resistance(R) candidate genes encoding nucleotide-binding sites(NBS) in a complete genome of Populus trichocarpa was identified and characterized by structural diversities,physical positions,phylogenetic relationships.Based on structures of N-terminal motif and leucine-rich repeat domains motif,we found 381 NBS-coding sequences with 122 non-regular NBS genes and 259 regular NBS genes that were further classified into 13 types such as TNL,CNL,NL,XNL,TN and other minor types.Meanwhile 81.9% of the NBS genes were distributed in cluster,and 81.8% of the cluster genes had duplicates.The results showed that there were many duplicate phenomenon occurred in the evolution of disease resistance genes of P.trichocarpa.After analysis of NBS standard phylogenetic tree in the genome of P.trichocarpa,the structure of tree exhibited a star topology,and the regular NBS genes were classified into 68 groups by less than 30% amino acid sequence diversity in each domain.
基金Supported by the National High Technology Research and Development Program(863Program of China)(NO.2006AA10Z1B4 and NO.2008AZ10A408)the Scientific and Technological Key Program of Ministry of Education(206065)
