从西藏搭格架铯硅华矿床区热泉中分离培养高温菌T4-1,并进行了格兰氏染色、显微镜观察、室内温度实验、16SrRNA基因分析等。结果表明,T4-1为杆状菌,格兰氏染色阳性,其生长范围为45-80℃,最适生长温度70℃。16SrRNA基因分析结果表明,该...从西藏搭格架铯硅华矿床区热泉中分离培养高温菌T4-1,并进行了格兰氏染色、显微镜观察、室内温度实验、16SrRNA基因分析等。结果表明,T4-1为杆状菌,格兰氏染色阳性,其生长范围为45-80℃,最适生长温度70℃。16SrRNA基因分析结果表明,该菌株属于地芽孢杆菌属（Geobacillus）,在发育树上,T4-1菌株与高温烷烃地芽孢杆菌（Geobacillus the rmoleovorans）非常近。本研究为进一步开展西藏高温微生物资源以及微生物参与成矿作用的研究提供了首例。展开更多
The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6V...The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.展开更多
文摘从西藏搭格架铯硅华矿床区热泉中分离培养高温菌T4-1,并进行了格兰氏染色、显微镜观察、室内温度实验、16SrRNA基因分析等。结果表明,T4-1为杆状菌,格兰氏染色阳性,其生长范围为45-80℃,最适生长温度70℃。16SrRNA基因分析结果表明,该菌株属于地芽孢杆菌属（Geobacillus）,在发育树上,T4-1菌株与高温烷烃地芽孢杆菌（Geobacillus the rmoleovorans）非常近。本研究为进一步开展西藏高温微生物资源以及微生物参与成矿作用的研究提供了首例。
基金国家"8 6 3"计划资助项目 (Z 17 0 4 0 1) 国家转基因植物研究与产业化资助项目 (J0 0 A 0 0 2 )~~
文摘The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.
