Effect of partial cotyledon damage on soybean seed viability and sample suitability PCR assay

Abstract

Marker-assisted selection is a key tool in modern plant breeding but its application towards soybean is limited by the destructive nature of standard PCR analysis which leads to the loss of valuable genotypes. The research goal was to evaluate the feasibility of non-destructive cotyledon tissue sampling for simultaneous PCR genotyping and preservation of seed viability. The research targets were soybean seeds of the variety Sentyabrinka with a total sample size of 200. In the experimental group, a wedge-shaped section of the lateral part of the cotyledon weighing 8 to 12 milligrams was made followed by DNA extraction and viability assessment under laboratory conditions. It was found that partial tissue removal did not critically affect seed viability. Laboratory germination rate in the experimental group was 94 percent compared to 96 percent in the control group, while germination energy was 89 percent compared to 91 percent in the control. Root and hypocotyl lengths showed no statistically significant differences from the control. DNA concentration in 50 experimental samples ranged from 336 to 625 nanograms per microliter, with an average value of 473.6 nanograms per microliter. DNA purity measured by the A260 slash 280 ratio was within the range of 1.8 to 1.92 which met the requirements for real-time PCR and SNP genotyping. The proposed approach allows genetic screening at the seed stage without material loss, enables the exclusion of samples lacking target alleles, and concentrates resources on promising lines. The method is recommended for work with limited collections and unique soybean genetic material, as it virtually fully combines molecular genetic analysis with the preservation of valuable genotypes for further use in the breeding process and significantly accelerates the development of new varieties.