Abstract
We used soybean (Glycine max) cDNA microarrays to identify candidate genes for a stable mutation at the Wp locus in soybean, which changed a purple-flowered phenotype to pink, and found that flavanone 3-hydroxylase cDNAs were overexpressed in purple flower buds relative to the pink. Restriction fragment length polymorphism analysis and RNA gel blots of purple and pink flower isolines, as well as the presence of a 5.7-kb transposon insertion in the wp mutant allele, have unequivocally shown that flavanone 3-hydroxylase gene 1 is the Wp locus. Moreover, the 5.7-kb insertion in wp represents a novel transposable element (termed Tgm-Express1) with inverted repeats closely related to those of other Tgms (transposable-like elements, G. max) but distinct in several characteristics, including the lack of subterminal inverted repeats. More significantly, Tgm-Express1 contains four truncated cellular genes from the soybean genome, resembling the Pack-MULEs (Mutator-like transposable elements) found in maize (Zea mays), rice (Oryza sativa), and Arabidopsis thaliana and the Helitrons of maize. The presence of the Tgm-Express1 element causing the wp mutation, as well as a second Tgm-Express2 element elsewhere in the soybean genome, extends the ability to acquire and transport host DNA segments to the CACTA family of elements, which includes both Tgm and the prototypical maize Spm/En.
MeSH Terms
DNA Transposable Elements/genetics
DNA, Complementary/analysis,genetics
Flowers/genetics
Gene Expression Profiling
Gene Expression Regulation, Enzymologic/genetics
Gene Expression Regulation, Plant/genetics
Genes, Plant/genetics
Genome, Plant/genetics
Mixed Function Oxygenases/genetics
Molecular Sequence Data
Mutation/genetics
Oligonucleotide Array Sequence Analysis
Phenotype
Pigmentation/genetics
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Soybeans/enzymology,genetics
Chemicals
DNA Transposable Elements
DNA, Complementary
Mixed Function Oxygenases
flavanone 3-dioxygenase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zabala Gracia
Department of Crop Sciences, University of Illinois, Urbana, IL 61801, USA.
Vodkin Lila O
References (32)
32 references, click to expand
-
Characterization of a highly conserved sequence related to mutator transposable elements in maize.
Mol Biol Evol. 1988 Sep;5(5):519-29
PMID: 2848175
-
Patterning of virus-infected Glycine max seed coat is associated with suppression of endogenous silencing of chalcone synthase genes.
Plant Cell. 2004 Apr;16(4):807-18
PMID: 15037735
-
Molecular characterization of flavanone 3 beta-hydroxylases. Consensus sequence, comparison with related enzymes and the role of conserved histidine residues.
Eur J Biochem. 1993 Oct 15;217(2):745-54
PMID: 8223617
-
Highly structured sequence homology between an insertion element and the gene in which it resides.
Proc Natl Acad Sci U S A. 1985 Jan;82(2):493-7
PMID: 16593538
-
The maize genome contains a helitron insertion.
Plant Cell. 2003 Feb;15(2):381-91
PMID: 12566579
-
Pack-MULE transposable elements mediate gene evolution in plants.
Nature. 2004 Sep 30;431(7008):569-73
PMID: 15457261
-
cA lectin gene insertion has the structural features of a transposable element.
Cell. 1983 Oct;34(3):1023-31
PMID: 6313203
-
Transposon diversity in Arabidopsis thaliana.
Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7376-81
PMID: 10861007
-
A single-base deletion in soybean flavonoid 3'-hydroxylase gene is associated with gray pubescence color.
Plant Mol Biol. 2002 Sep;50(2):187-96
PMID: 12175012
-
Tissue-specific gene silencing mediated by a naturally occurring chalcone synthase gene cluster in Glycine max.
Plant Cell. 2004 Apr;16(4):819-35
PMID: 15064367
-
Capture of a genomic HMG domain sequence by the En/Spm-related transposable element Tpn1 in the Japanese morning glory.
Mol Gen Genet. 1999 Apr;261(3):447-51
PMID: 10323224
-
Microarrays for global expression constructed with a low redundancy set of 27,500 sequenced cDNAs representing an array of developmental stages and physiological conditions of the soybean plant.
BMC Genomics. 2004 Sep 29;5:73
PMID: 15453914
-
Analysis of flavanone 3-hydroxylase in Arabidopsis seedlings. Coordinate regulation with chalcone synthase and chalcone isomerase.
Plant Physiol. 1996 May;111(1):339-45
PMID: 8685272
-
Bottlenecks for metabolic engineering of isoflavone glycoconjugates in Arabidopsis.
Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14578-83
PMID: 12384577
-
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
Anal Biochem. 1983 Jul 1;132(1):6-13
PMID: 6312838
-
Two functional soybean genes encoding p34cdc2 protein kinases are regulated by different plant developmental pathways.
Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):943-7
PMID: 8430109
-
Molecular characterization and expression of alfalfa (Medicago sativa L.) flavanone-3-hydroxylase and dihydroflavonol-4-reductase encoding genes.
Plant Mol Biol. 1995 Nov;29(4):773-86
PMID: 8541503
-
A catalogue of splice junction and putative branch point sequences from plant introns.
Nucleic Acids Res. 1986 Dec 22;14(24):9549-59
PMID: 3808952
-
A novel class of Helitron-related transposable elements in maize contain portions of multiple pseudogenes.
Plant Mol Biol. 2005 Jan;57(1):115-27
PMID: 15821872
-
Phenotypic diversity mediated by the maize transposable elements Ac and Spm.
Science. 1988 Oct 21;242(4877):399-405
PMID: 2845581
-
Maize transposable elements.
Annu Rev Genet. 1989;23:71-85
PMID: 2559653
-
Survey of transposable elements from rice genomic sequences.
Plant J. 2001 Jan;25(2):169-79
PMID: 11169193
-
Mutator-like elements in Arabidopsis thaliana. Structure, diversity and evolution.
Genetics. 2000 Dec;156(4):2019-31
PMID: 11102392
-
Purification and characterization of (2S)-flavanone 3-hydroxylase from Petunia hybrida.
Eur J Biochem. 1986 May 2;156(3):569-77
PMID: 3699024
-
Cloning of the pleiotropic T locus in soybean and two recessive alleles that differentially affect structure and expression of the encoded flavonoid 3' hydroxylase.
Genetics. 2003 Jan;163(1):295-309
PMID: 12586717
-
Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.).
Plant Mol Biol. 1994 Mar;24(5):743-55
PMID: 8193299
-
Chalcone synthase mRNA and activity are reduced in yellow soybean seed coats with dominant I alleles.
Plant Physiol. 1994 Jun;105(2):739-48
PMID: 8066134
-
Duplications That Suppress and Deletions That Restore Expression from a Chalcone Synthase Multigene Family.
Plant Cell. 1996 Apr;8(4):687-699
PMID: 12239396
-
Clustering of microarray data reveals transcript patterns associated with somatic embryogenesis in soybean.
Plant Physiol. 2003 May;132(1):118-36
PMID: 12746518
-
Organization of the Tgm family of transposable elements in soybean.
Genetics. 1988 Oct;120(2):597-604
PMID: 2848748
-
Multiple sequence alignment with hierarchical clustering.
Nucleic Acids Res. 1988 Nov 25;16(22):10881-90
PMID: 2849754
-
Site-directed mutagenesis of the active site serine290 in flavanone 3beta-hydroxylase from Petunia hybrida.
Eur J Biochem. 2000 Feb;267(3):853-60
PMID: 10651823