Linkage mapping, molecular cloning and functional analysis of soybean gene Fg2 encoding flavonol 3-O-glucoside (1→6) rhamnosyltransferase

Linkage mapping, molecular cloning and functional analysis of soybean gene Fg2 encoding flavonol... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Linkage mapping, molecular cloning and functional analysis of soybean gene Fg2 encoding flavonol 3-O-glucoside (1→6) rhamnosyltransferase

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Publisher
Springer Journals
Copyright
Copyright © 2013 by Springer Science+Business Media Dordrecht
Subject
Life Sciences; Plant Sciences; Biochemistry, general; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1007/s11103-013-0133-1
Publisher site
See Article on Publisher Site

Abstract

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.

Journal

Plant Molecular BiologySpringer Journals

Published: Sep 27, 2013

References

  • Flavonol glycoside genes in soybeans
    Buzzell, RI; Buttery, BR
  • The nonsense-mediated decay RNA surveillance pathway
    Chang, YF; Imam, JS; Wilkinson, MF
  • QTL analysis of low temperature-induced browning in soybean seed coats
    Githiri, SM; Yang, D; Khan, NA; Xu, D; Komatsuda, T; Takahashi, R
  • Analysis of flavonoids in flower petals of soybean near-isogenic lines for flower and pubescence color genes
    Iwashina, T; Githiri, SM; Benitez, ER; Takemura, T; Kitajima, J; Takahashi, R
  • Cloning and structural analysis of the anthocyanin pigmentation locus Rt of Petunia hybrida: characterization of the insertion sequences in two mutant alleles
    Kroon, J; Souer, E; Graaff, A; Xue, Y; Mol, J; Koes, R
  • Glycosyltransferases: structures, functions, and mechanisms
    Lairson, LL; Henrissat, B; Davies, GJ; Withers, SG
  • A new integrated genetic linkage map of the soybean
    Song, QJ; Marek, LF; Shoemaker, RC; Lark, KG; Concibido, VC; Delannay, X; Specht, JE; Cregan, PB
  • A single-base deletion in soybean flavonol synthase gene is associated with magenta flower color
    Takahashi, R; Githiri, SM; Hatayama, K; Dubouzet, EG; Shimada, N; Aoki, T; Ayabe, S; Toda, K; Matsumura, H

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