Long Wenbo, Li Ye, Zhou Wenjuan, Ling Hong-Qing, Zheng Shusong
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
PLoS One. 2013 Dec 5;8(12):e81091. doi: 10.1371/journal.pone.0081091. eCollection 2013.
The introgression lines (ILs) from cv. M82 (Solanum lycopersicum) × LA0716 (S. pennellii) have been proven to be exceptionally useful for genetic analysis and gene cloning. The introgressions were originally defined by RFLP markers at their development. The objectives of this study are to develop polymorphic SSR markers, and to re-define the DNA introgression from LA0716 in the ILs. Tomato sequence data was scanned by software to generate SSR markers. In total, 829 SSRs, which could be robustly amplified by PCR, were developed. Among them, 658 SSRs were dinucleotide repeats, 162 were trinucleotide repeats, and nine were tetranucleotide repeats. The 829 SSRs together with 96 published RFLPs were integrated into the physical linkage map of S. lycopersicum. Introgressions of DNA fragments from LA0716 were re-defined among the 75 ILs using the newly developed SSRs. A specific introgression of DNA fragment from LA0716 was identified in 72 ILs as described previously by RFLP, whereas the specific DNA introgression described previously were not detected in the ILs LA4035, LA4059 and LA4091. The physical location of each investigated DNA introgression was finely determined by SSR mapping. Among the 72 ILs, eight ILs showed a shorter and three ILs (IL3-2, IL12-3 and IL12-3-1) revealed a longer DNA introgression than that framed by RFLPs. Furthermore, 54 previously undefined segments were found in 21 ILs, ranging from 1 to 11 DNA introgressions per IL. Generally, the newly developed SSRs provide additional markers for genetic studies of tomatoes, and the fine definition of DNA introgressions from LA0716 would facilitate the use of the ILs for genetic analysis and gene cloning.
栽培品种M82（番茄）×LA0716（潘那利番茄）的渐渗系已被证明对遗传分析和基因克隆极为有用。这些渐渗系在最初开发时是由限制性片段长度多态性（RFLP）标记定义的。本研究的目的是开发多态性简单序列重复（SSR）标记，并重新定义渐渗系中来自LA0716的DNA渐渗。通过软件扫描番茄序列数据以生成SSR标记。总共开发了829个可通过聚合酶链反应（PCR）进行有效扩增的SSR标记。其中，658个SSR为二核苷酸重复序列，162个为三核苷酸重复序列，9个为四核苷酸重复序列。这829个SSR与96个已发表的RFLP一起被整合到番茄的物理连锁图谱中。利用新开发的SSR在75个渐渗系中重新定义了来自LA0716的DNA片段渐渗。如先前通过RFLP所描述的那样，在72个渐渗系中鉴定出了来自LA0716的特定DNA片段渐渗，而在渐渗系LA4035、LA4059和LA4091中未检测到先前描述的特定DNA渐渗。通过SSR图谱精细确定了每个研究的DNA渐渗的物理位置。在72个渐渗系中，8个渐渗系显示出比RFLP界定的更短的DNA渐渗，3个渐渗系（IL3 - 2、IL12 - 3和IL12 - 3 - 1）显示出更长的DNA渐渗。此外，在21个渐渗系中发现了54个先前未定义的片段，每个渐渗系中有1至11个DNA渐渗。总体而言，新开发的SSR为番茄的遗传研究提供了额外的标记，对来自LA0716的DNA渐渗的精细定义将有助于利用渐渗系进行遗传分析和基因克隆。
