| 基于ISSR和AFLP标记开发甜菜 SSR 引物的研究 |
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| 引用本文： | 牛泽如 杨文柱 庞,磊 田自华 邵金旺 史树德.基于ISSR和AFLP标记开发甜菜 SSR 引物的研究[J].中国农学通报,2010,26(21):147-151. |
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| 作者姓名： | 牛泽如 杨文柱 庞 磊 田自华 邵金旺 史树德 |
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| 作者单位： | 内蒙古农业大学农学院,呼和浩特,010019 |
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| 基金项目： | 内蒙古自治区高等学校科学研究项目，内蒙古农业大学博士科研启动基金，现代农业产业技术体系建设专项资金 |
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| 摘 要： | 本研究以ISSR-PCR和AFLP标记原理为基础，介绍一种新的分离甜菜基因组微卫星引物的方法。首先对甜菜基因组DNA进行酶切并连接已知序列的接头，构建基因组DNA酶切文库，同时用一个或两个ISSR引物，扩增文库中两端含微卫星序列片段并进行克隆测序，根据测序结果设计微卫星序列间的IP1引物和IP1与微卫星序列间IP2 引物；再根据侧翼序列克隆原理，采用巢式PCR进行基因组步移，扩增IP2引物下游序列，根据巢式PCR产物测序结果，设计微卫星序列另一侧的引物IP3 ，IP2和IP3即为SSR标记引物，对获得的SSR引物进行PCR验证，结果表明SSR引物产率为16%，本研究获得的SSR引物具有较高的多态性，对于后续的遗传多样性检测和遗传连锁图构建具有重要意义。
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| 关 键 词： | 烤烟 烘烤 密集烤房 散叶堆积式 烤烟 烘烤 密集烤房 散叶堆积式 |
| 收稿时间： | 2010/4/12 0:00:00 |
| 修稿时间： | 6/1/2010 12:00:00 AM |
A Method for Developing Sugar Beet (Beta vulgaris L.) Microsatellite Markers |
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| Niu Zeru,Yang Wenzhu,Pang Lei,Tian Zihua,Shao Jinwang,Shi Shude.A Method for Developing Sugar Beet (Beta vulgaris L.) Microsatellite Markers[J].Chinese Agricultural Science Bulletin,2010,26(21):147-151. |
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| Authors: | Niu Zeru Yang Wenzhu Pang Lei Tian Zihua Shao Jinwang Shi Shude |
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| Institution: | (College of Agronomy,Inner Mongolia Agricultural University,Hohhot 010019) |
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| Abstract: | Based on the principle and characteristics of the molecular marker techniques of amplified AFLP and ISSR,used genome walking technology,a reliable and valid method was established for isolating microsatellites from genomic DNA of sugar beet （Beta vulgaris L.）.Fragments flanked by one or two microsatellites were amplified using a microsatellite primer,（AG） 10 or （AT） 10 etc..A primer IP l designed from the sequenced region at one end of the microsatellite,and for nested PCR another primer IP 2 based on the sequence between IP 1 and the microsatellite were prepared.Then primer IP 3 of the other side of the microsatellite DNA was designed by nested PCR.IP2/IP3 was examined as a potential SSR marker.The results showed that the success rate was 16%,which was significantly higher than that of traditional methods.The SSR primers which were got from this study showing higher polymorphic,it is great significance for the following testing of the genetic diversity and genetic linkage map construction. |
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| Keywords: | ISSR-PCR ALFP-PCR |
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