| 利用重组自交系群体构建番茄AFLP 遗传连锁图谱 |
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| 引用本文： | 陈丽静,王利,王玉坤,陶承光,李君明,王晓武,李天来.利用重组自交系群体构建番茄AFLP 遗传连锁图谱[J].园艺学报,2012,39(12):2377-2384. |
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| 作者姓名： | 陈丽静 王利 王玉坤 陶承光 李君明 王晓武 李天来 |
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| 作者单位： | （1 沈阳农业大学，辽宁省农业生物技术重点实验室，设施园艺省部共建教育部重点实验室，沈阳 110866；2 中国农业科学院蔬菜花卉研究所，北京 100081；3 辽宁省农业科学院，沈阳 110866） |
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| 基金项目： | 辽宁省自然科学基金项目，辽宁省自然科学基金博士启动基金项目 |
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| 摘 要： | 以普通栽培番茄（Solanum lycopersicum）99165-30为母本，野生多毛番茄（Solanum habrochaites）LA1777为父本进行杂交，通过单粒传得到了含有80个F5︰6家系的重组自交系分离群体，利用荧光AFLP分子标记技术构建番茄分子遗传连锁图谱。AFLP标记采用MseⅠ和EcoRⅠ两种内切酶及荧光标记（IRD-700或IRD-800）的E + 3和非荧光标记的M + 3引物组合进行选择性扩增，扩增结果经95 ℃预变性后在6%变性聚丙烯酰胺凝胶上电泳2.5 h，运用LICOR 公司的NEN Global Edition IR2 DNA Analyzer（Model 5200 LI-COR Biosciences，Lincoln，NE）荧光扫描检测DNA多态性。对RILs群体中产生分离的274个AFLP标记运用JoinMap 3.0软件分析，得到一张番茄分子遗传连锁图谱，图谱总长度为662 cM，共包括18个主要连锁群，125个多态性分子标记。每条连锁群上的标记数在3 ~ 22个之间，连锁群的长度在14.0 ~ 58.0 cM的范围内，平均图距在 2.27 ~ 13.3 cM。总平均距离5.3 cM，本研究中构建的番茄永久遗传图谱，为番茄分子辅助育种及重要农艺性状的定位奠定了基础。
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| 关 键 词： | 番茄 RILs 群体 AFLP 标记 荧光标记引物 遗传连锁图谱 |
Construction AFLP Genetic Linkage Map of Tomato Using Recombinant Inbred Lines (RILs) Population |
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| CHEN Li-jing,WANG Li,WANG Yu-kun,TAO Cheng-guang,LI Jun-ming,WANG Xiao-wu,and LI Tian-lai.Construction AFLP Genetic Linkage Map of Tomato Using Recombinant Inbred Lines (RILs) Population[J].Acta Horticulturae Sinica,2012,39(12):2377-2384. |
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| Authors: | CHEN Li-jing WANG Li WANG Yu-kun TAO Cheng-guang LI Jun-ming WANG Xiao-wu and LI Tian-lai |
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| Institution: | （1Bioscience and Technology Institute of Shenyang Agricultural University，Key Laboratory of Agriculture Biotechnology Liaoning Province，Key Laboratory of Protected Horticulture，Shenyang Agricultural University，Ministry of Education，Shenyang 110866，China； 2 Institute of Vegetables and Flowers，Chinese Academy of Agricultural Sciences，Beijing 100081，China；3The Liaoning Academy of Agricultural Sciences，Shenyang 110866，China） |
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| Abstract: | A genetic linkage map of tomatowas constructed using a RILs（recombinant inbred lines）population of 80 individuals which was developed by crossing Solanum lycopersicum 99165-30 andSolanum habrochaites LA1777 through single-seed descent（SSD）. AFLPs were generated by the use of restriction enzymes EcoRⅠin combination with either MseⅠ. Pre-amplification was carried out using primers corresponding to EcoRⅠ and MseⅠ adaptors with no selective base. Selective amplifications were performed using IRD700 or IRD800 labeled EcoRⅠ primers and non-labeled MseⅠ primers. The resulting products were denatured in formamide at 95 ℃ and separated by electrophoresis 2.5 h on 6% polyacrylamide gel using IR2 DNA Analyzer（Model 5200 LI-COR Biosciences，Lincoln，NE）. The segregation of each marker and linkage analysis was done using the program JoinMap version 3.0. With 22 primer pairs，a total of 247 parental polymorphic bands were detected and 125 used for mapping，the total map length was 662 cM，consisted of 18 linkage groups，number of markers in the linkage groups varied from 3 to 22，the length of linkage groups were from 14.0 cM to 58.0 cM and mean marker interval distance from 2.27 cM to 13.3 cM individually，and a mean marker interval distance of 5.3 cM between markers. The map developed in the present study could be used for genetic mapping and molecular marker assisted breeding and quantitative trait locus mapping of tomato. |
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| Keywords: | tomato recombinant inbred line AFLP IRD700 or IRD800 labeled EcoR I primers genetic linkage map |
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