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| 烟草单染色体的分离与扩增及其SSR的检测 | |
| 引用本文： | 焦天雷,谢小芳,童治军,肖炳光,吴为人.烟草单染色体的分离与扩增及其SSR的检测[J].福建农林大学学报(自然科学版),2012,41(1):58-62. |
| 作者姓名： | 焦天雷 谢小芳 童治军 肖炳光 吴为人 |
| 作者单位： | 1. 浙江大学农业与生物技术学院,浙江杭州,310029 2. 福建农林大学生命科学学院,福建福州,350002 3. 云南省烟草农业科学研究院,云南玉溪,653100 4. 浙江大学农业与生物技术学院,浙江杭州310029;福建农林大学生命科学学院,福建福州350002 |
| 基金项目： | 国家烟草专卖局和云南省烟草公司项目(110200701023、08A0) |
| 摘 要： | 以普通烟草根尖为材料,在显微操作仪下分离单染色体,并用LAM-PCR方法进行扩增.对扩增产物用点杂交进行鉴定,用Southern杂交确定目的扩增片段的大小,并用本实验室开发的SSR特异引物检测SSR位点.结果表明:获得的单染色体扩增产物均存在来自烟草基因组的序列,扩增产物大小为250-3000 bp;利用SSR特异引物可以从单染色体扩增产物中检测到目标SSR位点. |
| 关 键 词： | 烟草 显微分离 LAM-PCR SSR |
Isolation and amplification of single chromosomes and detection of their SSRs in tobacco |
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| JIAO Tian-lei , XIE Xiao-fang , TONG Zhi-jun , XIAO Bing-guang , WU Wei-ren.Isolation and amplification of single chromosomes and detection of their SSRs in tobacco[J].Journal of Fujian Agricultural and Forestry University,2012,41(1):58-62. | |
| Authors: | JIAO Tian-lei XIE Xiao-fang TONG Zhi-jun XIAO Bing-guang WU Wei-ren |
| Institution: | 1,2(1.College of Agriculture and Biotechnology,Zhejiang University,Hangzhou,Zhejiang 310029,China;2.College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China; 3.Yunnan Academy of Tobacco Agricultural Sciences,Yuxi,Yunnan 653100,China) |
| Abstract: | With root tips of tobacco(Nicotana tobacum L.) as material,single chromosomes were isolated under the microscope operating instrument and amplified using LAM-PCR(linker adaptor-mediated PCR) method.The amplified products were examined by dot hybridization and their sizes were determined by Southern hybridization.Meanwhile,SSRs in the amplified fragments were detected with specific SSR primers developed by ourselves.The results showed that the amplified products of single chromosomes contained sequences from the tobacco genome and the sizes of amplified products approximately ranged from 250 to 3000 bp;SSRs could be detected from the amplified products of single chromosomes using the specific SSR primers.The results of this study will facilitate further application of the isolation and amplification technology in tobacco. |
| Keywords: | tobacco microdissection LAM-PCR SSR |
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