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ARS Home » Southeast Area » Florence, South Carolina » Coastal Plain Soil, Water and Plant Conservation Research » Research » Publications at this Location » Publication #71864
Title: A COMBINED APPROACH OF PCR AND RFLP TO IDENTIFY DNA MARKERS IN G. HIRSUTUM

Author
item FENG, XIANG - ALABAMA A&M UNIV
item SAHA, S - ALABAMA A&M UNIV
item SOLIMAN, K - ALABAMA A&M UNIV
item May Iii, Oscar

Submitted to: International Crop Science Congress Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 12/15/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: An integrated approach of PCR and RFLP was used to develop DNA markers in cotton. The PCR generated amplification products of the ACC synthase gene, a microsatellite fragment, and a cDNA probe. These PCR products were then used as probes in a RFLP mapping experiment. Interspecific crosses between G. hirsutum/G. barbadense, aneuploid, and chromosome substitution lines of G. hirsutum were used to map markers to chromosomes. Also, 96 F2 plants resulting from the intraspecific (G. hirsutum) cross between germplasm PD-3-14 and Chinese cultivar Zhongmian 12 were screened for polymorphic markers as part of an effort to develop a DNA marker linkage map in a G. hirsutum background. Monosomic F1 plants indicated that an 1140 bp RFLP marker of microsatellite probe (HindIII) is located on the short arm of chromosome 6. Additionally, a RFLP marker of cDNA probe (HindIII) is located on chromosome 20. RFLP screening of the F2 population revealed two polymorphic bands (1180 and 1240 bp with ACC synthase probe and HindIII) that are allelic. A unique band was found in the F2 population using the microsatellite probe. None of the RFLP markers are linked. A band in some F2 samples not found in the F1 or parents may be due to contamination from a seed mix or possibly an outcross. Alternatively, DNA sequence rearrangement resulting from the cross of genetically divergent parents could explain the novel band.