【目的】探明四川地区小麦、玉米和大豆3种寄主禾谷镰孢菌复合种(Fusarium graminearum species complex,FGSC)的群体遗传多样性及相互侵染关系。【方法】采用多重PCR和PCR-VNTR分子标记技术分析不同寄主来源菌株的毒素类型和群体遗传...【目的】探明四川地区小麦、玉米和大豆3种寄主禾谷镰孢菌复合种(Fusarium graminearum species complex,FGSC)的群体遗传多样性及相互侵染关系。【方法】采用多重PCR和PCR-VNTR分子标记技术分析不同寄主来源菌株的毒素类型和群体遗传多样性,并通过高粱粒接种法检测菌株在不同寄主间的相互侵染作用。【结果】来源于3种寄主的禾谷镰孢菌复合种菌株被鉴定为禾谷镰孢菌(F. graminearum)和亚洲镰孢菌(F. asiaticum),其中禾谷镰孢菌的毒素化学型为3-乙酰基脱氧雪腐镰刀菌烯醇(3-ADON)和15-乙酰脱氧雪腐镰刀菌烯醇(15-ADON),而亚洲镰孢菌的毒素化学型为雪腐镰刀菌烯醇(NIV);各寄主菌株的群体遗传多样性存在差异,以小麦菌株群体遗传多样性最高,且与大豆菌株遗传距离较远;同时,各寄主菌株间能够相互侵染,但致病力存在差异。【结论】来源于四川小麦、玉米和大豆3种寄主的禾谷镰孢菌复合种遗传多样性存在差异,但可相互侵染致病,具有加重其所致病害在该地区发生的风险。展开更多
AIM: To characterize the CagA variable region of Helicobacter pylori isolates from Chinese patients.METHODS: DNA fragments in CagA variable region were amplified and sequenced respectively from genomic DNA of 19 isola...AIM: To characterize the CagA variable region of Helicobacter pylori isolates from Chinese patients.METHODS: DNA fragments in CagA variable region were amplified and sequenced respectively from genomic DNA of 19 isolates from patients with gastric cancer and 20isolates from patients with chronic gastritis. The tendency of phosphorylation in tyrosine(s) of CagA proteins was evaluated subsequently by phosphorylation assay in vivo and in vitro respectively.RESULTS: About 97.44% (38/39) H pylori isolates possessed CagA gene. CagA+ strains contained 2-4tandem five-amino-acid motifs EPIYA but only one EPIYA had repeated sequence in CagA variable region in different isolates. There was no significant difference between the number of EPIYA motifs in H pylori from patients with different diseases. However, only tyrosine site in EPIYA within repeated sequence could be phosphorylated by AGS cells in vivo although all tyrosine sites in EPIYA could be phosphorylated in vitro.CONCLUSION: CagA in Chinese has no functional difference in perturbing cellular signal pathway among different H pylori isolates.展开更多
To isolate the causal organism of soft rot of vegetables, diseased samples of potato, tomato, carrot, chilies, and bell pepper, were analyzed in the lab, using nutrient agar (NA) and/or the enrichment host (Bell peppe...To isolate the causal organism of soft rot of vegetables, diseased samples of potato, tomato, carrot, chilies, and bell pepper, were analyzed in the lab, using nutrient agar (NA) and/or the enrichment host (Bell pepper) technique. Successful isolations were purified by sub-culturing, identified as Erwinia carotovora subsp. carotovora through biochemical tests and their pathogenicity was confirmed through inoculation on green tomato fruits. The isolates were tested for their aggressiveness to find out the most aggressive one in term of producing maximum soft rot on tomatoes. CRD (completely randomized design), with four replication was used and data were analyzed using LSD (least significant test) test. Among the five isolates evaluated for aggressiveness on tomato fruits, chili isolate was found to be the most aggressive followed by tomato and potato isolates producing 22.3 mm, 7.9 mm, and 7.8 mm diameter soft rot lesions, respectively.展开更多
To find new genes involved in fungal pathogenicity, a mutant (B11 ) exhibiting enhanced pathogenicity was isolated from an Agrobacterium-mediated transformed Magnaporthe oryzae mutant library. Southern blotting anal...To find new genes involved in fungal pathogenicity, a mutant (B11 ) exhibiting enhanced pathogenicity was isolated from an Agrobacterium-mediated transformed Magnaporthe oryzae mutant library. Southern blotting analysis showed that T-DNA insertion in the B11 genome was a single copy. TAIL-PCR and sequence alignment analyses revealed that a putative gene locus MG01679 was interrupted by the T-DNA fragment. By using the PCR-based method, the DNA and cDNA of the mutant gene MG01679 was cloned and sequenced. The open reading frame of MG01679 includes one intron and two exons, and the coding sequence is 696 bp in length and encodes a 231 amino acid peptide. Protein similarity analysis indicated that the gene belongs to the ThiJ/Pfp I protein family, and the gene was thus designated MgThiJ1. MgThiJ1 showed 57% similarity to FOXG_09029 from Fusarium oxysporum and 54% similarity to FGSG_08979 from F. graminearum in protein sequence. MgThiJ1 gene might act as a negative regulator in vegetative growth and pathogenesis in filamentous fungi, and its specific mechanism needs to be studied further.展开更多
基金Supported by the National High Technology ResearchDevelopment Program of China, No. 2001AA227111
文摘AIM: To characterize the CagA variable region of Helicobacter pylori isolates from Chinese patients.METHODS: DNA fragments in CagA variable region were amplified and sequenced respectively from genomic DNA of 19 isolates from patients with gastric cancer and 20isolates from patients with chronic gastritis. The tendency of phosphorylation in tyrosine(s) of CagA proteins was evaluated subsequently by phosphorylation assay in vivo and in vitro respectively.RESULTS: About 97.44% (38/39) H pylori isolates possessed CagA gene. CagA+ strains contained 2-4tandem five-amino-acid motifs EPIYA but only one EPIYA had repeated sequence in CagA variable region in different isolates. There was no significant difference between the number of EPIYA motifs in H pylori from patients with different diseases. However, only tyrosine site in EPIYA within repeated sequence could be phosphorylated by AGS cells in vivo although all tyrosine sites in EPIYA could be phosphorylated in vitro.CONCLUSION: CagA in Chinese has no functional difference in perturbing cellular signal pathway among different H pylori isolates.
文摘To isolate the causal organism of soft rot of vegetables, diseased samples of potato, tomato, carrot, chilies, and bell pepper, were analyzed in the lab, using nutrient agar (NA) and/or the enrichment host (Bell pepper) technique. Successful isolations were purified by sub-culturing, identified as Erwinia carotovora subsp. carotovora through biochemical tests and their pathogenicity was confirmed through inoculation on green tomato fruits. The isolates were tested for their aggressiveness to find out the most aggressive one in term of producing maximum soft rot on tomatoes. CRD (completely randomized design), with four replication was used and data were analyzed using LSD (least significant test) test. Among the five isolates evaluated for aggressiveness on tomato fruits, chili isolate was found to be the most aggressive followed by tomato and potato isolates producing 22.3 mm, 7.9 mm, and 7.8 mm diameter soft rot lesions, respectively.
基金supported by the Natural Science Foundation of Zhejiang Province,China(Grant No.Y306638)the Project of Zhejiang Science and Technology,China(Grant No.2007C12905)the National Natural Science Foundation of China(GrantNos.30900933 and 30970082)
文摘To find new genes involved in fungal pathogenicity, a mutant (B11 ) exhibiting enhanced pathogenicity was isolated from an Agrobacterium-mediated transformed Magnaporthe oryzae mutant library. Southern blotting analysis showed that T-DNA insertion in the B11 genome was a single copy. TAIL-PCR and sequence alignment analyses revealed that a putative gene locus MG01679 was interrupted by the T-DNA fragment. By using the PCR-based method, the DNA and cDNA of the mutant gene MG01679 was cloned and sequenced. The open reading frame of MG01679 includes one intron and two exons, and the coding sequence is 696 bp in length and encodes a 231 amino acid peptide. Protein similarity analysis indicated that the gene belongs to the ThiJ/Pfp I protein family, and the gene was thus designated MgThiJ1. MgThiJ1 showed 57% similarity to FOXG_09029 from Fusarium oxysporum and 54% similarity to FGSG_08979 from F. graminearum in protein sequence. MgThiJ1 gene might act as a negative regulator in vegetative growth and pathogenesis in filamentous fungi, and its specific mechanism needs to be studied further.