Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds
Soybean (<i>Glycine max</i> (L.) Merrill) isoflavones are among the most important secondary metabolites, with functional benefits for human health. Soybeans accumulate three aglycone forms of isoflavones: genistein, daidzein, and glycitein. Soybean landrace Kumachi-1 does not accumulate...
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MDPI AG
2024-01-01
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| Online Access: | https://www.mdpi.com/2223-7747/13/2/156 |
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| author | Masaki Horitani Risa Yamada Kanami Taroura Akari Maeda Toyoaki Anai Satoshi Watanabe |
| author_facet | Masaki Horitani Risa Yamada Kanami Taroura Akari Maeda Toyoaki Anai Satoshi Watanabe |
| author_sort | Masaki Horitani |
| collection | DOAJ |
| description | Soybean (<i>Glycine max</i> (L.) Merrill) isoflavones are among the most important secondary metabolites, with functional benefits for human health. Soybeans accumulate three aglycone forms of isoflavones: genistein, daidzein, and glycitein. Soybean landrace Kumachi-1 does not accumulate malonylglycitin at all. Gene structure analysis indicated that <i>Glyma.11G108300</i> (<i>F6H4</i>) of Kumachi-1 has a 3.8-kbp insertion, resulting in a truncated flavonoid 6-hydroxylase (<i>F6H</i>) sequence compared to the wild-type sequence in Fukuyutaka. Mapping experiments using a mutant line (MUT1246) with a phenotype similar to that of Kumachi-1, with a single-nucleotide polymorphism (SNP) in <i>F6H4</i>, revealed co-segregation of this mutation and the absence of glycitein isoflavones. We also identified a mutant line (K01) that exhibited a change in the HPLC retention time of glycitein isoflavones, accumulating glycoside and malonylglycoside forms of 6-hydroxydaidzein. K01 contains an SNP that produces a premature stop codon in <i>Glyma.01G004200</i> (<i>IOMT3</i>), a novel soybean isoflavone O-methyltransferase (<i>IOMT</i>) gene. We further analyzed transgenic hairy roots of soybeans expressing <i>Glyma.11G108300</i> (<i>F6H4</i>) and <i>Glyma.01G004200</i> (<i>IOMT3</i>). Those overexpressing <i>F6H4</i> accumulated malonylglycoside forms of 6-hydroxydaidzein (M_6HD), and co-expression of <i>F6H4</i> and <i>IOMT3</i> increased the level of malonylglycitin but not of M_6HD. These results indicate that <i>F6H4</i> and <i>IOMT3</i> are responsible for glycitein biosynthesis in soybean seed hypocotyl. |
| first_indexed | 2024-03-08T09:48:29Z |
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| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
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| spelling | doaj.art-2741ea557d994d8089f585d4e2693c682024-01-29T14:10:36ZengMDPI AGPlants2223-77472024-01-0113215610.3390/plants13020156Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean SeedsMasaki Horitani0Risa Yamada1Kanami Taroura2Akari Maeda3Toyoaki Anai4Satoshi Watanabe5Faculty of Agriculture, Saga University, 1 Honjo-machi, Saga 840-8502, JapanFaculty of Agriculture, Saga University, 1 Honjo-machi, Saga 840-8502, JapanFaculty of Agriculture, Saga University, 1 Honjo-machi, Saga 840-8502, JapanFaculty of Agriculture, Saga University, 1 Honjo-machi, Saga 840-8502, JapanFaculty of Agriculture, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, JapanFaculty of Agriculture, Saga University, 1 Honjo-machi, Saga 840-8502, JapanSoybean (<i>Glycine max</i> (L.) Merrill) isoflavones are among the most important secondary metabolites, with functional benefits for human health. Soybeans accumulate three aglycone forms of isoflavones: genistein, daidzein, and glycitein. Soybean landrace Kumachi-1 does not accumulate malonylglycitin at all. Gene structure analysis indicated that <i>Glyma.11G108300</i> (<i>F6H4</i>) of Kumachi-1 has a 3.8-kbp insertion, resulting in a truncated flavonoid 6-hydroxylase (<i>F6H</i>) sequence compared to the wild-type sequence in Fukuyutaka. Mapping experiments using a mutant line (MUT1246) with a phenotype similar to that of Kumachi-1, with a single-nucleotide polymorphism (SNP) in <i>F6H4</i>, revealed co-segregation of this mutation and the absence of glycitein isoflavones. We also identified a mutant line (K01) that exhibited a change in the HPLC retention time of glycitein isoflavones, accumulating glycoside and malonylglycoside forms of 6-hydroxydaidzein. K01 contains an SNP that produces a premature stop codon in <i>Glyma.01G004200</i> (<i>IOMT3</i>), a novel soybean isoflavone O-methyltransferase (<i>IOMT</i>) gene. We further analyzed transgenic hairy roots of soybeans expressing <i>Glyma.11G108300</i> (<i>F6H4</i>) and <i>Glyma.01G004200</i> (<i>IOMT3</i>). Those overexpressing <i>F6H4</i> accumulated malonylglycoside forms of 6-hydroxydaidzein (M_6HD), and co-expression of <i>F6H4</i> and <i>IOMT3</i> increased the level of malonylglycitin but not of M_6HD. These results indicate that <i>F6H4</i> and <i>IOMT3</i> are responsible for glycitein biosynthesis in soybean seed hypocotyl.https://www.mdpi.com/2223-7747/13/2/156<i>Glycine max</i>isoflavoneQTLglycitein6-hydroxydaidzeinflavonoid 6-hydroxylase |
| spellingShingle | Masaki Horitani Risa Yamada Kanami Taroura Akari Maeda Toyoaki Anai Satoshi Watanabe Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds Plants <i>Glycine max</i> isoflavone QTL glycitein 6-hydroxydaidzein flavonoid 6-hydroxylase |
| title | Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds |
| title_full | Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds |
| title_fullStr | Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds |
| title_full_unstemmed | Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds |
| title_short | Identification of Genes Responsible for the Synthesis of Glycitein Isoflavones in Soybean Seeds |
| title_sort | identification of genes responsible for the synthesis of glycitein isoflavones in soybean seeds |
| topic | <i>Glycine max</i> isoflavone QTL glycitein 6-hydroxydaidzein flavonoid 6-hydroxylase |
| url | https://www.mdpi.com/2223-7747/13/2/156 |
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