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Linkage mapping, molecular cloning and functional analysis of soybean gene Fg3 encoding flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase.

Di S, Yan F, Rodas FR, Rodriguez TO, Murai Y, Iwashina T, Sugawara S, Mori T, Nakabayashi R, Yonekura-Sakakibara K, Saito K, Takahashi R - BMC Plant Biol. (2015)

Bottom Line: GmF3G2″Gt of Nezumisaya showed a broad activity for kaempferol/quercetin 3-O-glucoside/galactoside derivatives but it did not glucosylate kaempferol 3-O-rhamnosyl-(1 → 4)-[rhamnosyl-(1 → 6)-glucoside] and 3-O-rhamnosyl-(1 → 4)-[glucosyl-(1 → 6)-glucoside].GmF3G2″Gt was designated as UGT79B30 by the UGT Nomenclature Committee.Based on substrate specificity of GmF3G2″Gt, 2″-glucosylation of flavonol 3-O-glycoside may be irreconcilable with 4″-glycosylation in soybean leaves.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8518, Japan. dishaokang@affrc.go.jp.

ABSTRACT

Background: Flavonol glycosides (FGs) are major components of soybean leaves and there are substantial differences in FG composition among genotypes. The first objective of this study was to identify genes responsible for FG biosynthesis and to locate them in the soybean genome. The second objective was to clone the candidate genes and to verify their function. Recombinant inbred lines (RILs) were developed from a cross between cultivars Nezumisaya and Harosoy.

Results: HPLC comparison with authentic samples suggested that FGs having glucose at the 2″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Nezumisaya, whereas FGs of Harosoy were devoid of 2″-glucose. Conversely, FGs having glucose at the 6″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Harosoy, whereas these FGs were absent in Nezumisaya. Genetic analysis suggested that two genes control the pattern of attachment of these sugar moieties in FGs. One of the genes may be responsible for attachment of glucose to the 2″-position, probably encoding for a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase. Nezumisaya may have a dominant whereas Harosoy may have a recessive allele of the gene. Based on SSR analysis, linkage mapping and genome database survey, we cloned a candidate gene designated as GmF3G2″Gt in the molecular linkage group C2 (chromosome 6). The open reading frame of GmF3G2″Gt is 1380 bp long encoding 459 amino acids with four amino acid substitutions among the cultivars. The GmF3G2″Gt recombinant protein converted kaempferol 3-O-glucoside to kaempferol 3-O-sophoroside. GmF3G2″Gt of Nezumisaya showed a broad activity for kaempferol/quercetin 3-O-glucoside/galactoside derivatives but it did not glucosylate kaempferol 3-O-rhamnosyl-(1 → 4)-[rhamnosyl-(1 → 6)-glucoside] and 3-O-rhamnosyl-(1 → 4)-[glucosyl-(1 → 6)-glucoside].

Conclusion: GmF3G2″Gt encodes a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase and corresponds to the Fg3 gene. GmF3G2″Gt was designated as UGT79B30 by the UGT Nomenclature Committee. Based on substrate specificity of GmF3G2″Gt, 2″-glucosylation of flavonol 3-O-glycoside may be irreconcilable with 4″-glycosylation in soybean leaves.

No MeSH data available.


Related in: MedlinePlus

Linkage mapping of F3G2″Gt using recombinant inbred lines derived from a cross between soybean cultivars Nezumisaya and Harosoy. The name of the linkage group is indicated at the top followed by the chromosome number in parenthesis. Distances (cM) of markers from the top of the linkage group are shown on the left.
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Fig2: Linkage mapping of F3G2″Gt using recombinant inbred lines derived from a cross between soybean cultivars Nezumisaya and Harosoy. The name of the linkage group is indicated at the top followed by the chromosome number in parenthesis. Distances (cM) of markers from the top of the linkage group are shown on the left.

Mentions: Among the 465 SSR markers tested, 185 markers that exhibited polymorphism between the parents and distinctly segregated in the RILs were used for linkage mapping. A total of 99 markers were classified into 28 linkage groups spanning 2,172 cM. For mapping of the gene responsible for attachment of glucose to the 2″-position, RILs having FG composition of the Nezumisaya type and ‘type 4’ were considered to have the genotype of Nezumisaya, whereas RILs having FG composition of the Harosoy type and ‘type 3’ were considered to have the genotype of Harosoy. Linkage mapping revealed that the gene responsible for attachment of glucose to the 2″-position was located in the molecular linkage group C2 (chromosome 6) between Satt307 and Sat_202 (Figure 2).Figure 2


Linkage mapping, molecular cloning and functional analysis of soybean gene Fg3 encoding flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase.

Di S, Yan F, Rodas FR, Rodriguez TO, Murai Y, Iwashina T, Sugawara S, Mori T, Nakabayashi R, Yonekura-Sakakibara K, Saito K, Takahashi R - BMC Plant Biol. (2015)

Linkage mapping of F3G2″Gt using recombinant inbred lines derived from a cross between soybean cultivars Nezumisaya and Harosoy. The name of the linkage group is indicated at the top followed by the chromosome number in parenthesis. Distances (cM) of markers from the top of the linkage group are shown on the left.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4494776&req=5

Fig2: Linkage mapping of F3G2″Gt using recombinant inbred lines derived from a cross between soybean cultivars Nezumisaya and Harosoy. The name of the linkage group is indicated at the top followed by the chromosome number in parenthesis. Distances (cM) of markers from the top of the linkage group are shown on the left.
Mentions: Among the 465 SSR markers tested, 185 markers that exhibited polymorphism between the parents and distinctly segregated in the RILs were used for linkage mapping. A total of 99 markers were classified into 28 linkage groups spanning 2,172 cM. For mapping of the gene responsible for attachment of glucose to the 2″-position, RILs having FG composition of the Nezumisaya type and ‘type 4’ were considered to have the genotype of Nezumisaya, whereas RILs having FG composition of the Harosoy type and ‘type 3’ were considered to have the genotype of Harosoy. Linkage mapping revealed that the gene responsible for attachment of glucose to the 2″-position was located in the molecular linkage group C2 (chromosome 6) between Satt307 and Sat_202 (Figure 2).Figure 2

Bottom Line: GmF3G2″Gt of Nezumisaya showed a broad activity for kaempferol/quercetin 3-O-glucoside/galactoside derivatives but it did not glucosylate kaempferol 3-O-rhamnosyl-(1 → 4)-[rhamnosyl-(1 → 6)-glucoside] and 3-O-rhamnosyl-(1 → 4)-[glucosyl-(1 → 6)-glucoside].GmF3G2″Gt was designated as UGT79B30 by the UGT Nomenclature Committee.Based on substrate specificity of GmF3G2″Gt, 2″-glucosylation of flavonol 3-O-glycoside may be irreconcilable with 4″-glycosylation in soybean leaves.

View Article: PubMed Central - PubMed

Affiliation: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305-8518, Japan. dishaokang@affrc.go.jp.

ABSTRACT

Background: Flavonol glycosides (FGs) are major components of soybean leaves and there are substantial differences in FG composition among genotypes. The first objective of this study was to identify genes responsible for FG biosynthesis and to locate them in the soybean genome. The second objective was to clone the candidate genes and to verify their function. Recombinant inbred lines (RILs) were developed from a cross between cultivars Nezumisaya and Harosoy.

Results: HPLC comparison with authentic samples suggested that FGs having glucose at the 2″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Nezumisaya, whereas FGs of Harosoy were devoid of 2″-glucose. Conversely, FGs having glucose at the 6″-position of glucose or galactose that is bound to the 3-position of kaempferol were present in Harosoy, whereas these FGs were absent in Nezumisaya. Genetic analysis suggested that two genes control the pattern of attachment of these sugar moieties in FGs. One of the genes may be responsible for attachment of glucose to the 2″-position, probably encoding for a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase. Nezumisaya may have a dominant whereas Harosoy may have a recessive allele of the gene. Based on SSR analysis, linkage mapping and genome database survey, we cloned a candidate gene designated as GmF3G2″Gt in the molecular linkage group C2 (chromosome 6). The open reading frame of GmF3G2″Gt is 1380 bp long encoding 459 amino acids with four amino acid substitutions among the cultivars. The GmF3G2″Gt recombinant protein converted kaempferol 3-O-glucoside to kaempferol 3-O-sophoroside. GmF3G2″Gt of Nezumisaya showed a broad activity for kaempferol/quercetin 3-O-glucoside/galactoside derivatives but it did not glucosylate kaempferol 3-O-rhamnosyl-(1 → 4)-[rhamnosyl-(1 → 6)-glucoside] and 3-O-rhamnosyl-(1 → 4)-[glucosyl-(1 → 6)-glucoside].

Conclusion: GmF3G2″Gt encodes a flavonol 3-O-glucoside/galactoside (1 → 2) glucosyltransferase and corresponds to the Fg3 gene. GmF3G2″Gt was designated as UGT79B30 by the UGT Nomenclature Committee. Based on substrate specificity of GmF3G2″Gt, 2″-glucosylation of flavonol 3-O-glycoside may be irreconcilable with 4″-glycosylation in soybean leaves.

No MeSH data available.


Related in: MedlinePlus