Fine mapping and cloning of the major seed protein quantitative trait loci on soybean chromosome 20

Christina E. Fliege, Russell A. Ward, Pamela Vogel, Hanh Nguyen, Truyen Quach, Ming Guo, João Paulo Gomes Viana, Lucas Borges dos Santos, James E. Specht, Tom E. Clemente, Matthew E. Hudson, Brian W. Diers

Research output: Contribution to journalArticlepeer-review


Soybean [Glycine max (L.) Merr.] is a unique crop species because it has high levels of both protein and oil in its seed. Of the many quantitative trait loci (QTL) controlling soybean seed protein content, alleles of the cqSeed protein-003 QTL on chromosome 20 exert the greatest additive effect. The high-protein allele exists in both cultivated and wild soybean (Glycine soja Siebold & Zucc.) germplasm. Our objective was to fine map this QTL to enable positional-based cloning of its underlying causative gene(s). Fine mapping was achieved by developing and testing a series of populations in which the chromosomal region surrounding the segregating high- versus low-protein alleles was gradually narrowed, using marker-based detection of recombinant events. The resultant 77.8 kb interval was directly sequenced from a G. soja source and compared with the reference genome to identify structural and sequence polymorphisms. An insertion/deletion variant detected in Glyma.20G85100 was found to have near-perfect +/- concordance with high/low-protein allele genotypes inferred for this QTL in parents of published mapping populations. The indel structure was concordant with an evolutionarily recent insertion of a TIR transposon into the gene in the low-protein lineage. Seed protein was significantly greater in soybean expressing an RNAi hairpin downregulation element in two independent events relative to control null segregant lineages. We conclude that a transposon insertion within the CCT domain protein encoded by the Glyma.20G85100 gene accounts for the high/low seed protein alleles of the cqSeed protein-003 QTL.

Original languageEnglish (US)
Pages (from-to)114-128
Number of pages15
JournalPlant Journal
Issue number1
StatePublished - Apr 2022


  • Glycine max (L.) Merr.
  • QTL
  • fine mapping
  • gene cloning
  • seed protein
  • soybean

ASJC Scopus subject areas

  • Genetics
  • Plant Science
  • Cell Biology


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