The High-Amylose Maize Line GEMS-0067 Is Associated with an Intron 8 Insertion in Sbe1a, Altered Native PAGE Zymogram Patterns, and Development of PCR Markers for Breeding Applications.
4 affiliations
- 1Department of Agricultural Research and Development Program, Central State University, Wilberforce, OH 45384, USA.
- 2Syngenta Seeds, DeKalb, IL 60150, USA.
- 3Department of Agronomy, Iowa State University, Ames, IA 50011, USA.
- 4United States Department of Agriculture-Agricultural Research Service (USDA-ARS), Iowa State University, Ames, IA 50011, USA.
Abstract
The high-amylose maize (Zea mays L.) germplasm line GEMS-0067 carries the recessive amylose-extender (ae/sbe2b) allele together with modifier loci that elevate starch amylose content from approximately 55% to more than 70%. Previous quantitative trait locus (QTL) studies associated a major component of this modifier effect with the Sbe1a region on chromosome 5S; however, the underlying DNA polymorphism and practical breeding markers remained unresolved. To identify candidate variation, a targeted genomic amplicon-scanning strategy was applied across coding and non-coding regions of Sbe1a. Screening with 23 gene-specific primer pairs identified a unique fragment-length polymorphism spanning the intron 8-exon 9 boundary. Sequencing revealed a 6 bp TACATA insertion together with a downstream 2 bp GA insertion within intron 8. The polymorphism was present in GEMS-0067 and its ancestral source Guatemala 209 but absent from 33 surveyed USDA Germplasm Enhancement of Maize (GEM) releases. Native PAGE zymogram analysis identified a distinct starch-metabolizing enzyme activity band in the putative SBEI region that displayed altered migration in GEMS-0067 relative to reference genotypes. A PCR-CAPS marker based on the insertion was developed using MslI digestion and enabled clear codominant discrimination of homozygous and heterozygous individuals. These results establish a novel gene-based marker associated with the GEMS-0067 high-amylose modifier background and provide a practical tool for marker-assisted breeding.
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BibTeX
@article{campbell2026highamylose,
title = {{The High-Amylose Maize Line GEMS-0067 Is Associated with an Intron 8 Insertion in Sbe1a, Altered Native PAGE Zymogram Patterns, and Development of PCR Markers for Breeding Applications.}},
author = {Mark R Campbell and Avinash Karn and Marvin Paul Scott and Luke A Farno},
journal = {Plants (Basel, Switzerland)},
year = {2026},
month = oct,
doi = {10.3390/plants15203157},
url = {https://europepmc.org/article/MED/42943110},
note = {CC BY},
}
RIS
TY - JOUR
TI - The High-Amylose Maize Line GEMS-0067 Is Associated with an Intron 8 Insertion in Sbe1a, Altered Native PAGE Zymogram Patterns, and Development of PCR Markers for Breeding Applications.
AU - Mark R Campbell
AU - Avinash Karn
AU - Marvin Paul Scott
AU - Luke A Farno
PY - 2026
DA - 2026/10/15
JO - Plants (Basel, Switzerland)
DO - 10.3390/plants15203157
UR - https://europepmc.org/article/MED/42943110
AB - The high-amylose maize (Zea mays L.) germplasm line GEMS-0067 carries the recessive amylose-extender (ae/sbe2b) allele together with modifier loci that elevate starch amylose content from approximately 55% to more than 70%. Previous quantitative trait locus (QTL) studies associated a major component of this modifier effect with the Sbe1a region on chromosome 5S; however, the underlying DNA polymorphism and practical breeding markers remained unresolved. To identify candidate variation, a targeted genomic amplicon-scanning strategy was applied across coding and non-coding regions of Sbe1a. Screening with 23 gene-specific primer pairs identified a unique fragment-length polymorphism spanning the intron 8-exon 9 boundary. Sequencing revealed a 6 bp TACATA insertion together with a downstream 2 bp GA insertion within intron 8. The polymorphism was present in GEMS-0067 and its ancestral source Guatemala 209 but absent from 33 surveyed USDA Germplasm Enhancement of Maize (GEM) releases. Native PAGE zymogram analysis identified a distinct starch-metabolizing enzyme activity band in the putative SBEI region that displayed altered migration in GEMS-0067 relative to reference genotypes. A PCR-CAPS marker based on the insertion was developed using MslI digestion and enabled clear codominant discrimination of homozygous and heterozygous individuals. These results establish a novel gene-based marker associated with the GEMS-0067 high-amylose modifier background and provide a practical tool for marker-assisted breeding.
SN - 2223-7747
ER -