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Novel genetic variants of GA-insensitive Rht-1 genes in hexaploid wheat and their potential agronomic value.

Tan MK, Koval J, Ghalayini A - PLoS ONE (2013)

Bottom Line: In addition, novel mutants which differ from reported alleles of Rht-B1b, Rht-B1e and Rht-D1b have been found in Quarrion, Carnamah and Whistler.Mutations in the conserved C-terminal GRAS domain of Rht-A1 alleles with possible effects on expression have been found in WW1842, Quarrion and Drysdale.Genetic variants with putative spliceosomal introns in the GRAS domain have been found in all accessions except Spica.

View Article: PubMed Central - PubMed

Affiliation: Elizabeth Macarthur Agricultural Institute, New South Wales (NSW) Department of Primary Industries, Menangle, New South Wales, Australia. mui-keng.tan@dpi.nsw.gov.au

ABSTRACT
This study has found numerous novel genetic variants of GA-insensitive dwarfing genes with potential agricultural value for crop improvement. The cultivar, Spica is a tall genotype and possesses the wild-type genes of Rht-A1a, Rht-B1a and Rht-D1a. The cultivar Quarrion possesses a mutant in the DELLA motif in each of the 3 genomes. This is a first report of a mutant of Rht-A1. In addition, novel mutants which differ from reported alleles of Rht-B1b, Rht-B1e and Rht-D1b have been found in Quarrion, Carnamah and Whistler. The accession, Aus1408 has an allele of Rht-B1 with a mutation in the conserved 'TVHYNP' N-terminal signal binding domain with possible implications on its sensitivity to GA. Mutations in the conserved C-terminal GRAS domain of Rht-A1 alleles with possible effects on expression have been found in WW1842, Quarrion and Drysdale. Genetic variants with putative spliceosomal introns in the GRAS domain have been found in all accessions except Spica. Genome-specific cis-sequences about 124 bp upstream of the start codon of the Rht-1 gene have been identified for each of the three genomes.

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HRM analysis of Rht-1 variants at the C-terminal GRAS domain in WW1842 X Whistler DH population.Normalized HRM curve for genotyping genetic variation in the Rht-1 gene bounded by the primers Rht-F6R6_F1a, Rht-F6R6_F1b and Rht-F6R6_FR1 (Table 1). The results are shown for parents, WW1842 and Whistler and eight progeny lines of the DH population (180 lines). Genotype data enabled the mapping of the molecular variation on chromosome 4A.
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pone-0069690-g004: HRM analysis of Rht-1 variants at the C-terminal GRAS domain in WW1842 X Whistler DH population.Normalized HRM curve for genotyping genetic variation in the Rht-1 gene bounded by the primers Rht-F6R6_F1a, Rht-F6R6_F1b and Rht-F6R6_FR1 (Table 1). The results are shown for parents, WW1842 and Whistler and eight progeny lines of the DH population (180 lines). Genotype data enabled the mapping of the molecular variation on chromosome 4A.

Mentions: A distinctive variant characterized by deletions, substitutions and an insertion ‘YG’ and occurring in low relative percentages of 3 to 5% was found for the accessions Drysdale, Quarrion and WW1842 (Fig. 3). The pedigree of Drysdale is Hartog*3/Quarrion and this allele must thus be inherited from Quarrion. The location of this ‘rare’ genetic variation has been determined in this work by mapping this molecular locus on the molecular marker map of WW1842 X Whistler doubled haploid population [10] using HRM (Fig. 4). Mapping the genetic variation involved the design of an allele-specific forward primer for cultivar Whistler (Rht-F6R6_F1a), an allele-specific forward primer for accession, WW1842 (Rht-F6R6_F1b) and a common reverse primer, Rht-F6R6_R1 (Table 1). The two forward primers overlap by 15 nucleotides. All the three primers were incorporated in equi-concentrations in each HRM assay, which gave unequivocal differentiation of the 2 parental alleles in the DH population (Fig. 4). This enabled the high stringency (p = 0.001) mapping of the genetic locus on chromosome 4A at a distance of about 2.4 cM from the DarT marker wPt4424 [10].


Novel genetic variants of GA-insensitive Rht-1 genes in hexaploid wheat and their potential agronomic value.

Tan MK, Koval J, Ghalayini A - PLoS ONE (2013)

HRM analysis of Rht-1 variants at the C-terminal GRAS domain in WW1842 X Whistler DH population.Normalized HRM curve for genotyping genetic variation in the Rht-1 gene bounded by the primers Rht-F6R6_F1a, Rht-F6R6_F1b and Rht-F6R6_FR1 (Table 1). The results are shown for parents, WW1842 and Whistler and eight progeny lines of the DH population (180 lines). Genotype data enabled the mapping of the molecular variation on chromosome 4A.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3716649&req=5

pone-0069690-g004: HRM analysis of Rht-1 variants at the C-terminal GRAS domain in WW1842 X Whistler DH population.Normalized HRM curve for genotyping genetic variation in the Rht-1 gene bounded by the primers Rht-F6R6_F1a, Rht-F6R6_F1b and Rht-F6R6_FR1 (Table 1). The results are shown for parents, WW1842 and Whistler and eight progeny lines of the DH population (180 lines). Genotype data enabled the mapping of the molecular variation on chromosome 4A.
Mentions: A distinctive variant characterized by deletions, substitutions and an insertion ‘YG’ and occurring in low relative percentages of 3 to 5% was found for the accessions Drysdale, Quarrion and WW1842 (Fig. 3). The pedigree of Drysdale is Hartog*3/Quarrion and this allele must thus be inherited from Quarrion. The location of this ‘rare’ genetic variation has been determined in this work by mapping this molecular locus on the molecular marker map of WW1842 X Whistler doubled haploid population [10] using HRM (Fig. 4). Mapping the genetic variation involved the design of an allele-specific forward primer for cultivar Whistler (Rht-F6R6_F1a), an allele-specific forward primer for accession, WW1842 (Rht-F6R6_F1b) and a common reverse primer, Rht-F6R6_R1 (Table 1). The two forward primers overlap by 15 nucleotides. All the three primers were incorporated in equi-concentrations in each HRM assay, which gave unequivocal differentiation of the 2 parental alleles in the DH population (Fig. 4). This enabled the high stringency (p = 0.001) mapping of the genetic locus on chromosome 4A at a distance of about 2.4 cM from the DarT marker wPt4424 [10].

Bottom Line: In addition, novel mutants which differ from reported alleles of Rht-B1b, Rht-B1e and Rht-D1b have been found in Quarrion, Carnamah and Whistler.Mutations in the conserved C-terminal GRAS domain of Rht-A1 alleles with possible effects on expression have been found in WW1842, Quarrion and Drysdale.Genetic variants with putative spliceosomal introns in the GRAS domain have been found in all accessions except Spica.

View Article: PubMed Central - PubMed

Affiliation: Elizabeth Macarthur Agricultural Institute, New South Wales (NSW) Department of Primary Industries, Menangle, New South Wales, Australia. mui-keng.tan@dpi.nsw.gov.au

ABSTRACT
This study has found numerous novel genetic variants of GA-insensitive dwarfing genes with potential agricultural value for crop improvement. The cultivar, Spica is a tall genotype and possesses the wild-type genes of Rht-A1a, Rht-B1a and Rht-D1a. The cultivar Quarrion possesses a mutant in the DELLA motif in each of the 3 genomes. This is a first report of a mutant of Rht-A1. In addition, novel mutants which differ from reported alleles of Rht-B1b, Rht-B1e and Rht-D1b have been found in Quarrion, Carnamah and Whistler. The accession, Aus1408 has an allele of Rht-B1 with a mutation in the conserved 'TVHYNP' N-terminal signal binding domain with possible implications on its sensitivity to GA. Mutations in the conserved C-terminal GRAS domain of Rht-A1 alleles with possible effects on expression have been found in WW1842, Quarrion and Drysdale. Genetic variants with putative spliceosomal introns in the GRAS domain have been found in all accessions except Spica. Genome-specific cis-sequences about 124 bp upstream of the start codon of the Rht-1 gene have been identified for each of the three genomes.

Show MeSH
Related in: MedlinePlus