This research aimed to indicate mechanisms involved in protection against the imbalanced generation of reactive oxygen species (ROS) during heavy metals (HMs) exposition of Silene vulgaris ecotypes with different levels of metal tolerance. Specimens of non-metallicolous (NM), calamine (CAL), and serpentine (SER) ecotypes were treated in vitro with Zn, Pb, and Cd ions applied simultaneously in concentrations that reflected their contents in natural habitats of the CAL ecotype (1× HMs) and 2.5- or 5.0-times higher than the first one. Our findings confirmed the sensitivity of the NM ecotype and revealed that the SER ecotype was not fully adapted to the HM mixture, since intensified lipid peroxidation, ultrastructural alternations, and decline in photosynthetic pigments’ content were ascertained under HM treatment. These changes resulted from insufficient antioxidant defense mechanisms based only on ascorbate peroxidase (APX) activity assisted (depending on HMs concentration) by glutathione-S-transferase (GST) and peroxidase activity at pH 6.8 in the NM ecotype or by GST and guaiacol-type peroxidase in the SER one. In turn, CAL specimens showed a hormetic reaction to 1× HMs, which manifested by both increased accumulation of pigments and most non-enzymatic antioxidants and enhanced activity of catalase and enzymes from the peroxidase family (with the exception of APX). Interestingly, no changes in superoxide dismutase activity were noticed in metallicolous ecotypes. To sum up, the ROS scavenging pathways in S. vulgaris relied on antioxidants specific to the respective ecotypes, however the synthesis of polyphenols was proved to be a universal reaction to HMs.
Background Loss of genetic variation negatively impacts breeding efforts and food security. Genebanks house over 7 million accessions representing vast allelic diversity that is a resource for sustainable breeding. Discovery of DNA variations is an important step in the efficient use of these resources. While technologies have improved and costs dropped, it remains impractical to consider resequencing millions of accessions. Candidate genes are known for most agronomic traits, providing a list of high priority targets. Heterogeneity in seed stocks means that multiple samples from an accession need to be evaluated to recover available alleles. To address this we developed a pooled amplicon sequencing approach and applied it to the out-crossing cereal rye (Secale cereale L.). Results Using the amplicon sequencing approach 95 rye accessions of different improvement status and worldwide origin, each represented by a pooled sample comprising DNA of 96 individual plants, were evaluated for sequence variation in six candidate genes with significant functions on biotic and abiotic stress resistance, and seed quality. Seventy-four predicted deleterious variants were identified using multiple algorithms. Rare variants were recovered including those found only in a low percentage of seed. Conclusions We conclude that this approach provides a rapid and flexible method for evaluating stock heterogeneity, probing allele diversity, and recovering previously hidden variation. A large extent of within-population heterogeneity revealed in the study provides an important point for consideration during rye germplasm conservation and utilization efforts.
38Loss of genetic variation negatively impacts breeding efforts and food security. Genebanks house 39 over 7 million accessions representing vast allelic diversity that is a resource for sustainable 40 breeding. Discovery of DNA variations is an important step in the efficient use of these resources. 41While technologies have improved and costs dropped, it remains impractical to consider 42 resequencing millions of accessions. Candidate genes are known for most agronomic traits, 43 providing a list of high priority targets. Heterogeneity in seed stocks means that multiple samples 44 from an accession need to be evaluated to recover available alleles. To address this we developed a 45 pooled amplicon sequencing approach and applied it to the out-crossing cereal rye (Secale cereale). 46Ninety-six plants from an accession were pooled together, and 95 unique accessions evaluated for 47 sequence variation in six target genes involved in seed quality, biotic and abiotic stress resistance, 48 resulting in 74 predicted deleterious variants using multiple algorithms. Rare variants were 49 recovered including those found only in a low percentage of seed. We conclude that this approach 50 provides a rapid and flexible method for evaluating stock heterogeneity, probing allele diversity, 51 and recovering previously hidden variation in large germplasm collections. 52 53
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