以甘蔗茎组织总DNA为模板,以16S rRNA基因赖氏细菌属(L eif son ia)通用引物为第一轮引物、甘蔗宿根矮化病菌(L eif son ia xy li subsp.xy li,Lxx)亚种特异引物为第二轮引物建立了Lxx巢式PCR检测技术。根据已报道的Lxx巴西分离物基因...以甘蔗茎组织总DNA为模板,以16S rRNA基因赖氏细菌属(L eif son ia)通用引物为第一轮引物、甘蔗宿根矮化病菌(L eif son ia xy li subsp.xy li,Lxx)亚种特异引物为第二轮引物建立了Lxx巢式PCR检测技术。根据已报道的Lxx巴西分离物基因组全序列(G enB ank登录号AE 016822.1)设计了扩增致病相关基因片段的3个引物对,经多种组合进行了RCR检验,筛选出两个特异性好、灵敏高的引物对,建立了Lxx的多重PCR检测技术。克隆测序表明,PCR产物与巴西分离物基因组相应区段同一率为99%以上,从而证实了上述PCR技术的正确性。检测结果表明,广东样品的阳性率为90%,海南样品的阳性率为60%。展开更多
Based on the specific primer from the nucleotide sequences between 16S-23S rDNA,Polymerase chain reaction(PCR) approach for detecting Leifsonia xyli subsp.xyli(Lxx) was established.The approach was applied to detect L...Based on the specific primer from the nucleotide sequences between 16S-23S rDNA,Polymerase chain reaction(PCR) approach for detecting Leifsonia xyli subsp.xyli(Lxx) was established.The approach was applied to detect Lxx from 30 cultivars collected from two main sugar cane producing areas in Yunnan and 10 hot-water treatment samples.The results showed that 80% of the cultivars were infected by Lxx and hot-water treatment was proved to be an efficient measure to control but not completely eliminate Lxx in sugar cane tissues.The PCR products amplified from six infected cultivars were cloned and sequenced,six sequences were acquired and analyzed.It indicated that the six sequences were identical and showed 100% similarity with the isolates from Brazil,Australia and Fujian,and 99.54% with the isolate from Louisiana.展开更多
文摘以甘蔗茎组织总DNA为模板,以16S rRNA基因赖氏细菌属(L eif son ia)通用引物为第一轮引物、甘蔗宿根矮化病菌(L eif son ia xy li subsp.xy li,Lxx)亚种特异引物为第二轮引物建立了Lxx巢式PCR检测技术。根据已报道的Lxx巴西分离物基因组全序列(G enB ank登录号AE 016822.1)设计了扩增致病相关基因片段的3个引物对,经多种组合进行了RCR检验,筛选出两个特异性好、灵敏高的引物对,建立了Lxx的多重PCR检测技术。克隆测序表明,PCR产物与巴西分离物基因组相应区段同一率为99%以上,从而证实了上述PCR技术的正确性。检测结果表明,广东样品的阳性率为90%,海南样品的阳性率为60%。
文摘Based on the specific primer from the nucleotide sequences between 16S-23S rDNA,Polymerase chain reaction(PCR) approach for detecting Leifsonia xyli subsp.xyli(Lxx) was established.The approach was applied to detect Lxx from 30 cultivars collected from two main sugar cane producing areas in Yunnan and 10 hot-water treatment samples.The results showed that 80% of the cultivars were infected by Lxx and hot-water treatment was proved to be an efficient measure to control but not completely eliminate Lxx in sugar cane tissues.The PCR products amplified from six infected cultivars were cloned and sequenced,six sequences were acquired and analyzed.It indicated that the six sequences were identical and showed 100% similarity with the isolates from Brazil,Australia and Fujian,and 99.54% with the isolate from Louisiana.
