利用26S r DNA D1/D2区序列分析法鉴定了内蒙古牧区传统开菲尔发酵液中的酵母菌,为筛选可发酵特色乳酒的酵母菌奠定基础。对所分离纯化得的酵母菌进行菌落特征和细胞显微形态区分,在形态鉴定基础上,选择代表菌进行26S r DNA D1/D2区序...利用26S r DNA D1/D2区序列分析法鉴定了内蒙古牧区传统开菲尔发酵液中的酵母菌,为筛选可发酵特色乳酒的酵母菌奠定基础。对所分离纯化得的酵母菌进行菌落特征和细胞显微形态区分,在形态鉴定基础上,选择代表菌进行26S r DNA D1/D2区序列分子鉴定。结果表明,共分离到36株酵母菌,形态聚类为5类,经DNA提取、26S r DNA D1/D2区PCR扩增、酶切、基因序列分析和同源性比对,鉴定为5种分子类型的酵母菌,分别为胶红酵母菌(Rhodotorula mucilaginosa)、诞沫假丝酵母菌(Candida zeylanoides)、发酵毕赤酵母菌(Pichia fermentans)、酿酒酵母菌(Saccharomyces cerevisiae)、有孢圆酵母菌(Torulaspora delbrueckii)。传统开菲尔发酵液中分离到的酿酒酵母,具有可发酵特色乳酒的潜质。展开更多
Periwinkle(Catharanthus roseus) yellows is a common disease in Hainan. Periwinkle′s leaf tissue with symptoms was assayed for phytoplasma infection by using PCR assay employing phytoplasma universal 16S rRNA gene pri...Periwinkle(Catharanthus roseus) yellows is a common disease in Hainan. Periwinkle′s leaf tissue with symptoms was assayed for phytoplasma infection by using PCR assay employing phytoplasma universal 16S rRNA gene primers (Rl6mF2/Rl6mR1). A PCR product (about 1.4 kb) was amplified from periwinkle showed yellows. Nucleotide sequencing and phylogenetic tree analysis showed that the amplified 16S rDNA contained 1 432 nucleotides, the most homology was 98.1% with the members of elm yellows group (16S r Ⅴ) and clustered in the same clade, while it was under 96.1% with other phytoplasma groups. Our results suggested that the phytoplasma sample belonged to 16S rⅤgroup and was tentatively named as Hainan periwinkle yellows phytoplasma (PY-Hn). This is the first report of existence of 16S r Ⅴ group phytoplasma in naturally infected periwinkle.展开更多
Mulberry yellow dwarf(MYD)disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer,P1/P7 and Rm16F2/Rm16R1,based on the 16S-23S rDNA sequence of phytoplasma and to...Mulberry yellow dwarf(MYD)disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer,P1/P7 and Rm16F2/Rm16R1,based on the 16S-23S rDNA sequence of phytoplasma and total DNA extracted from infected mulberry tissues were employed for PCR and nested-PCR detection.The results revealed that a phytoplasma-specific 1 830 bp fragment with a G+C content of 46.01% was sequenced(GenBank accession No.GQ249410).The sequence shared 99.7% and 99.8% identity with aster yellows,the representatiive phytoplasma in 16SrI group,and mulberry dwarf phytoplasma classified into subgroup B in 16SrI group and named as the MYD phytoplasma strain Anhui(MYD-Anh).A phylogenetic tree based on 16S rDNA sequences was constructed and showed that MYD-Anh was clustered into 16SrI group.Identity of 16S rDNA sequence between MYD-Anh and mulberry yellow dwarf phytoplasma strain Zhenjiang(MD-zj) was nearly 100%,and they might belong to the same strain.Nested-PCR was used to detect the pathogenic phytoplasma from the differential tissues of mulberry infected with MYD-Anh.The results showed that a phytoplasma-specific 1.4 kb fragment was amplified with total DNA extracted from bark and vein.Nested-PCR was more sensitive than PCR for detecting MYD phytoplasma.展开更多
目的通过三种假丝酵母菌鉴定方法的比较,为临床选择假丝酵母菌的鉴定方法提供参考。方法用科玛嘉显色培养基、API 20C AUX生化系统和rRNA-ITS序列测定分析鉴定住院患者口腔培养出的198株假丝酵母菌,比较3种鉴定方法的结果。结果 API 20C...目的通过三种假丝酵母菌鉴定方法的比较,为临床选择假丝酵母菌的鉴定方法提供参考。方法用科玛嘉显色培养基、API 20C AUX生化系统和rRNA-ITS序列测定分析鉴定住院患者口腔培养出的198株假丝酵母菌,比较3种鉴定方法的结果。结果 API 20C AUX生化系统与rDNA-ITS序列测定分析鉴定结果一致,科玛嘉显色培养基与API 20C AUX生化系统和rDNA-ITS序列测定分析鉴定的符合率白假丝酵母菌为97.84%,热带假丝酵母菌的符合率为93.33%,光滑假丝酵母菌和克柔假丝酵母菌的符合率分别为90.91%、88.89%,其他假丝酵母菌科玛嘉不能鉴定。结论 API 20C AUX生化系统与rDNA-ITS序列测定分析鉴定假丝酵母菌的结果有较好的一致性,rDNA-ITS序列测定分析鉴定假丝酵母菌有望成为假丝酵母菌鉴定较好的方法。科玛嘉显色培养基方法可以简便有效的鉴定最常见的4种假丝酵母菌。展开更多
文摘利用26S r DNA D1/D2区序列分析法鉴定了内蒙古牧区传统开菲尔发酵液中的酵母菌,为筛选可发酵特色乳酒的酵母菌奠定基础。对所分离纯化得的酵母菌进行菌落特征和细胞显微形态区分,在形态鉴定基础上,选择代表菌进行26S r DNA D1/D2区序列分子鉴定。结果表明,共分离到36株酵母菌,形态聚类为5类,经DNA提取、26S r DNA D1/D2区PCR扩增、酶切、基因序列分析和同源性比对,鉴定为5种分子类型的酵母菌,分别为胶红酵母菌(Rhodotorula mucilaginosa)、诞沫假丝酵母菌(Candida zeylanoides)、发酵毕赤酵母菌(Pichia fermentans)、酿酒酵母菌(Saccharomyces cerevisiae)、有孢圆酵母菌(Torulaspora delbrueckii)。传统开菲尔发酵液中分离到的酿酒酵母,具有可发酵特色乳酒的潜质。
文摘Periwinkle(Catharanthus roseus) yellows is a common disease in Hainan. Periwinkle′s leaf tissue with symptoms was assayed for phytoplasma infection by using PCR assay employing phytoplasma universal 16S rRNA gene primers (Rl6mF2/Rl6mR1). A PCR product (about 1.4 kb) was amplified from periwinkle showed yellows. Nucleotide sequencing and phylogenetic tree analysis showed that the amplified 16S rDNA contained 1 432 nucleotides, the most homology was 98.1% with the members of elm yellows group (16S r Ⅴ) and clustered in the same clade, while it was under 96.1% with other phytoplasma groups. Our results suggested that the phytoplasma sample belonged to 16S rⅤgroup and was tentatively named as Hainan periwinkle yellows phytoplasma (PY-Hn). This is the first report of existence of 16S r Ⅴ group phytoplasma in naturally infected periwinkle.
文摘Mulberry yellow dwarf(MYD)disease is an quarantine disease and the causal agent is a phytoplasma.Two pairs of published universal primer,P1/P7 and Rm16F2/Rm16R1,based on the 16S-23S rDNA sequence of phytoplasma and total DNA extracted from infected mulberry tissues were employed for PCR and nested-PCR detection.The results revealed that a phytoplasma-specific 1 830 bp fragment with a G+C content of 46.01% was sequenced(GenBank accession No.GQ249410).The sequence shared 99.7% and 99.8% identity with aster yellows,the representatiive phytoplasma in 16SrI group,and mulberry dwarf phytoplasma classified into subgroup B in 16SrI group and named as the MYD phytoplasma strain Anhui(MYD-Anh).A phylogenetic tree based on 16S rDNA sequences was constructed and showed that MYD-Anh was clustered into 16SrI group.Identity of 16S rDNA sequence between MYD-Anh and mulberry yellow dwarf phytoplasma strain Zhenjiang(MD-zj) was nearly 100%,and they might belong to the same strain.Nested-PCR was used to detect the pathogenic phytoplasma from the differential tissues of mulberry infected with MYD-Anh.The results showed that a phytoplasma-specific 1.4 kb fragment was amplified with total DNA extracted from bark and vein.Nested-PCR was more sensitive than PCR for detecting MYD phytoplasma.
基金Co-constructed Program of Beijing Education Committee for Scientific Research BaseResearch Fund for the Doctoral Program of Higher Education (20030022015)Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT0607)~~
