Gedil M A, Wye C, Berry S, Segers B, Peleman J, Jones R, Leon A, Slabaugh M B, Knapp S J
Department of Crop and Soil Science, Oregon State University, Corvallis 97331-3002, USA.
Genome. 2001 Apr;44(2):213-21.
Restriction fragment length polymorphism (RFLP) maps have been constructed for cultivated sunflower (Helianthus annuus L.) using three independent sets of RFLP probes. The aim of this research was to integrate RFLP markers from two sets with RFLP markers for resistance gene candidate (RGC) and amplified fragment length polymorphism (AFLP) markers. Genomic DNA samples of HA370 and HA372, the parents of the F2 population used to build the map, were screened for AFLPs using 42 primer combinations and RFLPs using 136 cDNA probes (RFLP analyses were performed on DNA digested with EcoRI, HindIII, EcoRV, or DraI). The AFLP primers produced 446 polymorphic and 1101 monomorphic bands between HA370 and HA372. The integrated map was built by genotyping 296 AFLP and 104 RFLP markers on 180 HA370 x HA372 F2 progeny (the AFLP marker assays were performed using 18 primer combinations). The HA370 x HA372 map comprised 17 linkage groups, presumably corresponding to the 17 haploid chromosomes of sunflower, had a mean density of 3.3 cM, and was 1326 cM long. Six RGC RFLP loci were polymorphic and mapped to three linkage groups (LG8, LG13, and LG15). AFLP markers were densely clustered on several linkage groups, and presumably reside in centromeric regions where recombination is reduced and the ratio of genetic to physical distance is low. Strategies for targeting markers to euchromatic DNA need to be tested in sunflower. The HA370 x HA372 map integrated 14 of 17 linkage groups from two independent RFLP maps. Three linkage groups were devoid of RFLP markers from one of the two maps.
利用三套独立的限制性片段长度多态性(RFLP)探针构建了栽培向日葵(Helianthus annuus L.)的RFLP图谱。本研究的目的是将两组RFLP标记与抗性基因候选(RGC)的RFLP标记及扩增片段长度多态性(AFLP)标记整合起来。使用42对引物组合对用于构建图谱的F2群体亲本HA370和HA372的基因组DNA样本进行AFLP筛选,并使用136个cDNA探针进行RFLP分析(RFLP分析在经EcoRI、HindIII、EcoRV或DraI消化的DNA上进行)。AFLP引物在HA370和HA372之间产生了446条多态性条带和1101条单态性条带。通过对180个HA370×HA372 F2后代进行296个AFLP和104个RFLP标记的基因分型构建了整合图谱(AFLP标记分析使用18对引物组合)。HA370×HA372图谱包含17个连锁群,大概对应向日葵的17条单倍体染色体,平均密度为3.3 cM,长度为1326 cM。六个RGC RFLP位点具有多态性,并定位到三个连锁群(LG8、LG13和LG15)。AFLP标记在几个连锁群上密集聚类,大概位于着丝粒区域,那里重组减少,遗传距离与物理距离的比率较低。需要在向日葵中测试将标记靶向常染色质DNA的策略。HA370×HA372图谱整合了来自两个独立RFLP图谱的17个连锁群中的14个。三个连锁群没有来自两个图谱之一的RFLP标记。