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a Dep. of Agronomy and Horticulture, New Mexico State Univ., Las Cruces, NM 88003
b Dep. of Crop, Soil, and Environmental Sci., Univ. of Arkansas, Fayetteville, AR 72701
* Corresponding author (jinzhang{at}nmsu.edu).
The identification of molecular markers closely linked to restorer genes of the cytoplasmic male sterile (CMS) D8 system could facilitate the development of parental lines for hybrid cotton (Gossypium hirsutum L.). Our objective was to develop molecular markers closely linked to the two independent dominant restorer genes, Rf1 from the D2 restorer line transferred from G. harknessii Brandegee (D2 genome) and Rf2 from the D8 restorer line. Bulked segregant analysis was used to identify random amplified polymorphic DNA (RAPD) markers that were linked to the restorer genes. Three testcrosses were used for mapping Rf2 and two for Rf1 relative to the markers. Two RAPD markers, UBC1113000 and UBC188500, were associated with Rf2 in coupling phase. UBC188500 was closely linked to Rf2 with an average genetic distance of 2.9 cM. A survey of cotton species and cultivars revealed that UBC188500 was absent in normal cotton cultivars and most Gossypium spp. However, it was present in the D8 restorer line, G. raimondii Ulbr. (D5), G. trilobum (DC.) Skovst. (D8), and selected wild species from Australia, indicating that Rf2 and the DNA fragment yielding the RAPD marker were both introgressed into the tetraploid cottons from the D8 genome. A new RAPD marker, UBC169700, and a previously identified RAPD marker, UBC6591500, cosegregated with Rf1 in the two populations examined. The three RAPD markers, UBC188500, UC169700, and UBC6591500 were converted into reliable and genome-specific sequence tagged site (STS) markers on the basis of their sequence information. These markers are restorer-specific and should be useful in marker-based selection for developing restorer parental lines and constructing a high-resolution linkage map containing Rf1 and Rf2.
Abbreviations: AFLP, amplified fragment length polymorphism CMS, cytoplasmic male sterile/sterility EB, ethidium bromide F, fertile MAS, marker-assisted selection PCR, polymerase chain reaction RAPD, random amplified polymorphic DNA RFLP, restriction fragment length polymorphism S, sterile SSR, simple sequence repeat STS, sequence tagged site
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