Biochemical genetics of black pigmentation of soybean seed

Buzzell, R. I.; Buttery, B. R.; MacTavish, D. C.

doi:10.1093/oxfordjournals.jhered.a110309

Cited by 46 publications

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“…7A, lane 11). We have confirmed that the pattern (Buzzell et al, 1987). Brown (irT) and buff (irt) do not contain anthocyanins but do contain polymeric procyanidin and propelargonidin, respectively (Todd and Vodkin, 1993).…”

Section: Chs Genomic Dna Polymorphisms In Near-lsogenic Linessupporting

confidence: 64%

“…Neither the I , R, or T genes have been cloned and only the T gene is defined at the biochemical level. The T gene is thought to encode a microsomal 3' flavonoid hydroxylase (Buzzell et al, 1987;Todd and Vodkin, 1993) because the major anthocyanins (cyanidin-3-monoglucoside) and proanthocyanidins (procyanidin) that are present in T soybean seed coats have hydroxyl groups at the 3' position of the flavonoid B ring. The R gene is likely to act at a step specific to the anthocyanin pathway because procyanidins are present in both black (iRT) and brown (irT) seed coats (Todd and Vodkin, 1993), but anthocyanins are absent in brown (irT) and buff (irt) seed coats (Todd and Vodkin, 1993).…”

Section: Chs Genomic Dna Polymorphisms In Near-lsogenic Linesmentioning

confidence: 99%

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Chalcone Synthase mRNA and Activity Are Reduced in Yellow Soybean Seed Coats with Dominant I Alleles

Wang¹,

Todd²,

Vodkin³

1994

Plant Physiol.

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The seed of all wild Clycine accessions have black or brown pigments because of the homozygous recessive i allele in combination with alleles at the R and T loci. In contrast, nearly all commercial soybean (GIycine max) varieties are yellow due to the presence of a dominant allele of the I locus (either I or rl) that inhibits pigmentation in the seed coats. Spontaneous mutations to the recessive i allele occur in these varieties and result in pigmented seed coats. We have isolated a clone for a soybean dihydroflavonol reductase (DFR) gene using polymerase chain reaction. We examined expression of DFR and two other genes of the flavonoid pathway during soybean seed coat development in a series of nearisogenic isolines that vary in pigmentation as specified by combinations of alleles of the I , R, and T loci. The expression of phenylalanine ammonia-lyase and DFR mRNAs was similar in all of the gene combinations at each stage of seed coat development. In contrast, chalcone synthase (CHS) mRNA was barely detectable at all stages of development in seed coats that carry the dominant I allele that results in yellow seed coats. CHS activity in yellow seed coats (I) was also 7-to 10-fold less than in the pigmented seed coats that have the homozygous recessive i allele. It appears that the dominant I allele results in reduction of CHS mRNA, leading to reduction of CHS activity as the basis for inhibition of anthocyanin and proanthocyanin synthesis in soybean seed coats. A further connection between CHS and the I locus is indicated by the occurrence of multiple restriction site polymorphisms in genomic DNA blots of the CHS gene family in near-isogenic lines containing alleles of the I locus.Pigmentation of soybean (Glycine max) seed coats is influenced by at least three genes, I, R, and T. The I gene (for inhibitor) controls synthesis and spatial occurrence of pigments in the epidermal layer of the seed coat, whereas R and T are responsible for the specific color (reviewed in Bemard and Weiss, 1973;Palmer and Kilen, 1987). The dominant I allele inhibits pigment accumulation, resulting in a yellow seed coat color at maturity, whereas the homozygous, recessive i allele specifies full pigmentation across the entire seedWe are grateful for a graduate fellowship award from the. Na- Recently, we showed that polymeric proanthocyanidins are also synthesized in pigmented seed coats with the homozygous recessive i genotype (Todd and Vodkin, 1993). Proanthocyanidins are not precursors of anthocyanins but are synthesized as polymeric condensation products of leucoanthocyanins, the precursors of anthocyanins. Brown (irT) and buff (irt) seed coats do not contain anthocyanins but they do synthesize proanthocyanidins. The mechanism of inhibition of anthocyanin and proanthocyanidin synthesis by the I allele is unknown.Spontaneous mutations of the dominant I alleles ( I or 2%) to the recessive i allele have occurred in many soybean varieties, resulting in isogenic lines with pigmented seed coats. A Prorich cell wall protein, PRP1, and its ...

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Section: Chs Genomic Dna Polymorphisms In Near-lsogenic Linessupporting

confidence: 64%

Section: Chs Genomic Dna Polymorphisms In Near-lsogenic Linesmentioning

confidence: 99%

Chalcone Synthase mRNA and Activity Are Reduced in Yellow Soybean Seed Coats with Dominant I Alleles

Wang¹,

Todd²,

Vodkin³

1994

Plant Physiol.

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“…The anthocyanins (anthocyanidin glycosides) of the mature, black seed coat (i,R,T) have been identified as cyanidin-3-monoglucoside and delphinidin-3-monoglucoside (Buzzell et al, 1987), which have 3',4' and 3',4',5' B-ring hydroxylation pattems, respectively. In mature, imperfect-black seed coats (i,R,t), delphinidin is the primary anthocyanin, whereas cyanidin-3-monoglucoside is drastically reduced in quantity (Buzzell et al, 1987).…”

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confidence: 99%

“…In mature, imperfect-black seed coats (i,R,t), delphinidin is the primary anthocyanin, whereas cyanidin-3-monoglucoside is drastically reduced in quantity (Buzzell et al, 1987). Thus, the T locus is thought to encode a 3'-flavonoid hydroxylase that results in conversion of pelargonidin or its dihydroflavonol precursors to cyanidin.…”

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confidence: 99%

Pigmented Soybean (Glycine max) Seed Coats Accumulate Proanthocyanidins during Development

1993

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The dominant I gene inhibits accumulation of anthocyanin pigments in the epidermal layer of soybean (Crycine max) seed coats.Seed-coat color i s also influenced by the R locus and by the pubescence color alleles (T, tawny; f, gray). Protein and RNA from cultivars with black (i,R,T) and brown (i,r,T) seed coats are difficult to extract. To determine the nature of the interfering plant products, we examined seed-coat extracts from Clark isogenic lines for flavonoids, anthocyanins, and possible proanthocyanidins by thinlayer chromatography. We show that yellow seed-coat varieties ( I ) do not accumulate anthocyanins (anthocyanidin glycosides) or proanthocyanidins (polymeric anthocyanidins). Mature, black (i,R,T) and imperfect-black (i,R,f) seed coats contained anthocyanins, whereas mature, brown (i,r,r) and buff (i,r,f) seed coats did not contain anthocyanins. In contrast, all colored ( i ) genotypes tested positive for the presence of proanthocyanidins by butanol/ HCI and 0.5% vanillin assays. Immature, black (/R,T) and brown (&r,T) seed coats contained significant amounts of procyanidin, a 3',4'-hydroxylated proanthocyanidin. Immature, black (i,R,T) or brown (i,r,r) seed-coat extracts also tested positive for the ability to precipitate proteins in a radial diffusion assay and to bind RNA in vitro. Imperfect-black (i,R,f) or buff (i,r,f) seed coats contained lesser amounts of propelargonidin, a 4'-hydroxylated proanthocyanidin. Seed-coat extracts from these genotypes did not have the ability to precipitate protein or bind to RNA. In summary, the dominant I gene controls inhibition of not only anthocyanins but also proanthocyanidins in soybean seed coats. In homozygous recessive i genotypes, the T-f gene pair determines the types of proanthocyanidins present, which is consistent with the hypothesis that the T locus encodes a microsomal 3'-flavonoid hydroxylase.Commercially grown soybean (Glycine max) varieties have yellow seed. However, some soybean varieties are selfcolored and accumulate anthocyanins within the epidermal layer of the seed coat. The I locus and the R locus along with the pleiotropic T locus have major effects on seed-coat color (reviewed by Bemard and Weiss, 1973;Palmer and Kilen, 1987). The 1 locus (inhibitor) has four alleles that determine the absence or presence of pigment along with the spatial distribution of the pigments. Unlike other species in which the presence of pigment is usually the dominant phenotype, the absence of anthocyanin pigments in soybean seed coats is dominant because of the 1 allele, which inhibits pigment The anthocyanins (anthocyanidin glycosides) of the mature, black seed coat (i,R,T) have been identified as cyanidin-

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“…Thus, various combinations of flavonol glycoside genes need to be evaluated in different genetic backgrounds. The Wm/wm isolines used in our research carried Wl/wl as markers which could have contributed a nonisogenic effect; however, WI is a phenolase gene for anthocyanidins and not for flavonols (6). On the average in the work of Ciha and Brun (7), purple-flowered cultivars had a similar adaxial stomatal frequency as white-flowered cultivars.…”

Section: Discussionmentioning

confidence: 88%

Leaf Traits Associated with Flavonol Glycoside Genes in Soybean

Buttery¹,

Buzzell²

1987

Plant Physiol.

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In the soybea (Glciw sx (IL) Experimental plants were grown in the field (a fertile sandy loam) at Harrow, Ontario, machine planted in rows 3.7 m long, 0.6 m apart. Photosynthetic rate (PA) was measured by a '4CO2 method (4) on the youngest fully mature leaves in August 1985 and 1986. Stomatal numbers were counted in leaf epidermal strips secured to glass slides by a cyanoacrylate adhesive (9) at a magnification of 400x using five random fields of 0.184 mm2 for each leaf. Chl was determined in 80% acetone extracts (1). Flavonol glycosides were routinely monitored by cellulose TLC. For hydrolysis of these glycosides, an equal volume of 2 N HCI was added to the acetone extracts used for Chl determinations and the mixture heated for 30 min on a boiling water bath. Kaempferol was determined quantitatively in the hydrolysate using a Hewlett Packard 1090 HPLC equipped with a Hypersil MOSS am column (100 x 2.1 mm). Kaempferol had a retention time of 17 min at a flow rate of 0.5 ml min-', an oven temperature of 38C, and a methanol gradient between 30 and 50% MeOH in 10% HCOOH in H20. Elution was monitored at 365 nm. RESULTSThe 3t OX941 and the St OX943 contained several hundred nanograms of kaempferol (after hydrolysis) per mg of leaf tissue, while OX942 and (OX944 with the wm gene contained very

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Biochemical genetics of black pigmentation of soybean seed

Cited by 46 publications

References 0 publications

Chalcone Synthase mRNA and Activity Are Reduced in Yellow Soybean Seed Coats with Dominant I Alleles

Chalcone Synthase mRNA and Activity Are Reduced in Yellow Soybean Seed Coats with Dominant I Alleles

Pigmented Soybean (Glycine max) Seed Coats Accumulate Proanthocyanidins during Development

Leaf Traits Associated with Flavonol Glycoside Genes in Soybean

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