A single-base deletion in soybean flavonoid 3′-hydroxylase gene is associated with gray pubescence color

A single-base deletion in soybean flavonoid 3′-hydroxylase gene is associated with gray... The T locus of soybean (Glycine max (L.) Merr.) controls pubescence and seed coat color and is presumed to encode flavonoid 3′-hydroxylase (F3′H). The dominant T and the recessive t allele of the locus produce brown and gray pubescence, respectively. PCR primers were constructed based on the sequence of a soybean EST clone homologous to the F3′H gene. A putative full-length cDNA, sf3′h1 was isolated by 3′ and 5′ RACE. Sequence analysis revealed that sf3′h1 consists of 1690 nucleotides encoding 513 amino acids. It had 68% and 66% homology with corresponding F3′H protein sequences of petunia and Arabidopsis, respectively. A conserved amino acid sequence of F3′H proteins, GGEK, was found in the deduced polypeptide. Sequence analysis of the gene from a pair of near-isogenic lines for T, To7B (TT, brown) and To7G (tt, gray) revealed that they differed by a single C deletion in the coding region of To7G. The deletion changed the subsequent reading frame resulting in a truncated polypeptide lacking the GGEK consensus sequence and the heme-binding domain. Genomic Southern analysis probed by sf3′h1 revealed restriction fragment length polymorphisms between cultivars with different pubescence color. Further, sf3′h1 was mapped at the same position with T locus on LG3(c2). PCR-RFLP analysis was performed to detect the single-base deletion. To7B and three cultivars with brown pubescence exhibited shorter fragments, while To7G and three cultivars with gray pubescence had longer fragments due to the single-base deletion. The PCR-RFLP marker co-segregated with genotypes at the Tlocus in a F2 population segregating for the T locus. The above results strongly suggest that sf3′h1 represents the T gene of soybean responsible for pubescence color and that the single-base deletion may be responsible for gray pubescence color. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

A single-base deletion in soybean flavonoid 3′-hydroxylase gene is associated with gray pubescence color

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Publisher
Springer Journals
Copyright
Copyright © 2002 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1016087221334
Publisher site
See Article on Publisher Site

Abstract

The T locus of soybean (Glycine max (L.) Merr.) controls pubescence and seed coat color and is presumed to encode flavonoid 3′-hydroxylase (F3′H). The dominant T and the recessive t allele of the locus produce brown and gray pubescence, respectively. PCR primers were constructed based on the sequence of a soybean EST clone homologous to the F3′H gene. A putative full-length cDNA, sf3′h1 was isolated by 3′ and 5′ RACE. Sequence analysis revealed that sf3′h1 consists of 1690 nucleotides encoding 513 amino acids. It had 68% and 66% homology with corresponding F3′H protein sequences of petunia and Arabidopsis, respectively. A conserved amino acid sequence of F3′H proteins, GGEK, was found in the deduced polypeptide. Sequence analysis of the gene from a pair of near-isogenic lines for T, To7B (TT, brown) and To7G (tt, gray) revealed that they differed by a single C deletion in the coding region of To7G. The deletion changed the subsequent reading frame resulting in a truncated polypeptide lacking the GGEK consensus sequence and the heme-binding domain. Genomic Southern analysis probed by sf3′h1 revealed restriction fragment length polymorphisms between cultivars with different pubescence color. Further, sf3′h1 was mapped at the same position with T locus on LG3(c2). PCR-RFLP analysis was performed to detect the single-base deletion. To7B and three cultivars with brown pubescence exhibited shorter fragments, while To7G and three cultivars with gray pubescence had longer fragments due to the single-base deletion. The PCR-RFLP marker co-segregated with genotypes at the Tlocus in a F2 population segregating for the T locus. The above results strongly suggest that sf3′h1 represents the T gene of soybean responsible for pubescence color and that the single-base deletion may be responsible for gray pubescence color.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 13, 2004

References

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