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Mendel's dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins.

ABSTRACT: The major gibberellin (GA) controlling stem elongation in pea (Pisum sativum L.) is GA1, which is formed from GA20 by 3beta-hydroxylation. This step, which limits GA1 biosynthesis in pea, is controlled by the Le locus, one of the original Mendelian loci. Mutations in this locus result in dwarfism. We have isolated cDNAs encoding a GA 3beta-hydroxylase from lines of pea carrying the Le, le, le-3, and led alleles. The cDNA sequences from le and le-3 each contain a base substitution resulting in single amino acid changes relative to the sequence from Le. The cDNA sequence from led, a mutant derived from an le line, contains both the le "mutation" and a single-base deletion, which causes a shift in reading frame and presumably a null mutation. cDNAs from each line were expressed in Escherichia coli. The expression product for the clone from Le converted GA9 to GA4, and GA20 to GA1, with Km values of 1.5 microM and 13 microM, respectively. The amino acid substitution in the clone from le increased Km for GA9 100-fold and reduced conversion of GA20 to almost nil. Expression products from le and le-3 possessed similar levels of 3beta-hydroxylase activity, and the expression product from led was inactive. Our results suggest that the 3beta-hydroxylase cDNA is encoded by Le. Le transcript is expressed in roots, shoots, and cotyledons of germinating pea seedlings, in internodes and leaves of established seedlings, and in developing seeds.

SUBMITTER: Martin DN

PROVIDER: S-EPMC23192 | biostudies-literature | 1997 Aug

REPOSITORIES: biostudies-literature

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Mendel's dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins.

Martin D N DN Proebsting W M WM Hedden P P

Proceedings of the National Academy of Sciences of the United States of America 19970801 16

The major gibberellin (GA) controlling stem elongation in pea (Pisum sativum L.) is GA1, which is formed from GA20 by 3beta-hydroxylation. This step, which limits GA1 biosynthesis in pea, is controlled by the Le locus, one of the original Mendelian loci. Mutations in this locus result in dwarfism. We have isolated cDNAs encoding a GA 3beta-hydroxylase from lines of pea carrying the Le, le, le-3, and led alleles. The cDNA sequences from le and le-3 each contain a base substitution resulting in si ...[more]

PMID: 9238076

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Project description:Optimizing wheat height to maximize yield has been an important aspect which is evident from a successful example of green revolution. Dwarfing genes (Rht) are known for yield gains due to lodging resistance and partitioning of assimilates into ear. The available and commercially exploited sources of dwarfism in Indian spring wheat are Rht1 and Rht2 genes inspite of availability of over 20 dwarfing genes. Rht8 a Gibberellic acid sensitive dwarfing gene is another reduced height gene commercially exploited in some Mediterranean countries. Two F2 populations segregating for Rht1 and Rht8 genes with each comprising 398 and 379 plants were developed by crossing European winter wheat cultivars Beauchamp and Capitole with Indian spring wheat cultivar PBW 621. Different genotypic combinations for Rht1 and Rht8 genes were selected from these populations through linked molecular markers and selected F3:4 lines were evaluated for various agronomic traits in a replicated trial. Reduction in plant height with Rht8 and Rht1 averaged 2.86% and 13.3% respectively as compared to the group of lines lacking dwarfing gene. Reduction was spread along all the internodes of wheat culm and reduction was lower as progress towards the lower internode. Grain number per spike and highest yield was observed in lines carrying only Rht1 gene. Reduction in plant biomass was observed with either of the dwarfing gene. Longest coleoptile length and seedling shoot length averaged 4.4 ± 0.09 cm and 19.5 ± 0.48, respectively was observed in lines lacking any of the dwarfing gene. Negligible reduction of 6.75% and 2.84% in coleoptile length and seedling shoot length, respectively was observed in lines carrying only Rht8 gene whereas F3:4 lines with Rht1 gene showed 21.64% and 23.35% reduction in coleoptile length and seedling shoot length, respectively. Additive effect of genes was observed as double dwarfs showed 43.31% and 43.34% reduction in coleoptile length and seedling shoot length.

Dataset Information

Mendel's dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins.

Publications

Mendel's dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins.

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