Linkage mapping, molecular cloning and functional analysis of soybean gene Fg2 encoding flavonol 3-O-glucoside (1 → 6) rhamnosyltransferase
| Autor: | Ryoji Takahashi, Junichi Kitajima, Kazuki Saito, Tito O. Rodriguez, Tsukasa Iwashina, Makoto Suzuki, Satoko Sugawara, Ryo Nakabayashi, Felipe Rojas Rodas, Kyoko Toda, Keiko Yonekura-Sakakibara, Yoshinori Murai |
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| Rok vydání: | 2013 |
| Předmět: |
Candidate gene
DNA Complementary Rhamnose Molecular Sequence Data Plant Science Molecular cloning Biology Genes Plant chemistry.chemical_compound Gene Expression Regulation Plant Genetic linkage Genetics Coding region Amino Acid Sequence Cloning Molecular Gene Chromatography High Pressure Liquid DNA Primers chemistry.chemical_classification Base Sequence Sequence Homology Amino Acid Chromosome Mapping General Medicine Recombinant Proteins Amino acid Hexosyltransferases chemistry Biochemistry Soybean Proteins Soybeans Kaempferol Agronomy and Crop Science |
| Zdroj: | Plant Molecular Biology. 84:287-300 |
| ISSN: | 1573-5028 0167-4412 |
| DOI: | 10.1007/s11103-013-0133-1 |
| Popis: | There are substantial genotypic differences in the levels of flavonol glycosides (FGs) in soybean leaves. The first objective of this study was to identify and locate genes responsible for FG biosynthesis in the soybean genome. The second objective was to clone and verify the function of these candidate genes. Recombinant inbred lines (RILs) were developed by crossing the Kitakomachi and Koganejiro cultivars. The FGs were separated by high performance liquid chromatography (HPLC) and identified. The FGs of Koganejiro had rhamnose at the 6″-position of the glucose or galactose bound to the 3-position of kaempferol, whereas FGs of Kitakomachi were devoid of rhamnose. Among the 94 RILs, 53 RILs had HPLC peaks classified as Koganejiro type, and 41 RILs had peaks classified as Kitakomachi type. The segregation fitted a 1:1 ratio, suggesting that a single gene controls FG composition. SSR analysis, linkage mapping and genome database survey revealed a candidate gene in the molecular linkage group O (chromosome 10). The coding region of the gene from Koganejiro, designated as GmF3G6″Rt-a, is 1,392 bp long and encodes 464 amino acids, whereas the gene of Kitakomachi, GmF3G6″Rt-b, has a two-base deletion resulting in a truncated polypeptide consisting of 314 amino acids. The recombinant GmF3G6″Rt-a protein converted kaempferol 3-O-glucoside to kaempferol 3-O-rutinoside and utilized 3-O-glucosylated/galactosylated flavonols and UDP-rhamnose as substrates. GmF3G6″Rt-b protein had no activity. These results indicate that GmF3G6″Rt encodes a flavonol 3-O-glucoside (1 → 6) rhamnosyltransferase and it probably corresponds to the Fg2 gene. GmF3G6″Rt was designated as UGT79A6 by the UGT Nomenclature Committee. |
| Databáze: | OpenAIRE |
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