Variations in chemical composition of common buckwheat (Fagopyrum esculentum Moench) as a result of different environmental conditions.

BACKGROUND: Common buckwheat (Fagopyrum esculentum Moench) is a pseudo cereal that is gaining interest in the world. The chemical profile of common buckwheat determines its high nutritional and health-promoting value. The accumulation of these valuable ingredients depends on many factors, such as: variety, location of cultivation and related weather and agrotechnical conditions. Due to the growing interest in common buckwheat as a natural plant material for food production, it is important to know the factors affecting the quantitative and qualitative composition of its grains. The aim of the research was to determine the effect of the genotype (G), environment (E) and G × E interaction on the content of nutrients (protein, starch, ash, lipids) and bioactive components [dietary fiber (DF), total phenolic content (TPC)] in the common buckwheat grains. The study covered four cultivars grown in three locations for three consecutive vegetation seasons (2016/2017, 2017/2018, 2018/2019). RESULTS: Based on the obtained results, a significant influence of the environment and G × E interaction on the content of the studied parameters was found. The greatest impact on the diversity of the content of nutrients had environmental conditions, which in the case of protein and ash determined these features in more than 80%, and in the case of starch, 70%. With regard to bioactive compounds, the greatest influence of the environment was observed for the amount of TPC (78%), lignin (51%) and the DF complex (56%). CONCLUSION: The obtained results are useful for breeders working on expanding the pool of common buckwheat genotypes, stable in changing environmental conditions. © 2023 Society of Chemical Industry.

Corrigendum: Glutathione and copper ions as critical factors of green plant regeneration efficiency of triticale in vitro anther culture.

[This corrects the article DOI: 10.3389/fpls.2022.926305.].

Glutathione and copper ions as critical factors of green plant regeneration efficiency of triticale in vitro anther culture.

Plant tissue culture techniques are handy tools for obtaining unique plant materials that are difficult to propagate or important for agriculture. Homozygous materials derived through in vitro cultures are invaluable and significantly accelerate the evaluation of new varieties, e.g., cereals. The induction of somatic embryogenesis/androgenesis and the regeneration and its efficiency can be influenced by the external conditions of tissue culture, such as the ingredients present in the induction or regeneration media. We have developed an approach based on biological system, molecular markers, Fourier Transform Infrared spectroscopy, and structural equation modeling technique to establish links between changes in sequence and DNA methylation at specific symmetric (CG, CHG) and asymmetric (CHH) sequences, glutathione, and green plant regeneration efficiency in the presence of variable supplementation of induction medium with copper ions. The methylation-sensitive Amplified Fragment Length Polymorphism was used to assess tissue culture-induced variation, Fourier Transform Infrared spectroscopy to describe the glutathione spectrum, and a structural equation model to develop the relationship between sequence variation, de novo DNA methylation within asymmetric sequence contexts, and copper ions in the induction medium, as well as, glutathione, and green plant efficiency. An essential aspect of the study is demonstrating the contribution of glutathione to green plant regeneration efficiency and indicating the critical role of copper ions in influencing tissue culture-induced variation, glutathione, and obtaining green regenerants. The model presented here also has practical implications, showing that manipulating the concentration of copper ions in the induction medium may influence cell function and increases green plant regeneration efficiency.

Structural Equation Modeling (SEM) Analysis of Sequence Variation and Green Plant Regeneration via Anther Culture in Barley.

The process of anther culture involves numerous abiotic stresses required for cellular reprogramming, microspore developmental switch, and plant regeneration. These stresses affect DNA methylation patterns, sequence variation, and the number of green plants regenerated. Recently, in barley (Hordeum vulgare L.), mediation analysis linked DNA methylation changes, copper (Cu2+) and silver (Ag+) ion concentrations, sequence variation, ß-glucans, green plants, and duration of anther culture (Time). Although several models were used to explain particular aspects of the relationships between these factors, a generalized complex model employing all these types of data was not established. In this study, we combined the previously described partial models into a single complex model using the structural equation modeling approach. Based on the evaluated model, we demonstrated that stress conditions (such as starvation and darkness) influence ß-glucans employed by cells for glycolysis and the tricarboxylic acid cycle. Additionally, Cu2+ and Ag+ ions affect DNA methylation and induce sequence variation. Moreover, these ions link DNA methylation with green plants. The structural equation model also showed the role of time in relationships between parameters included in the model and influencing plant regeneration via anther culture. Utilization of structural equation modeling may have both scientific and practical implications, as it demonstrates links between biological phenomena (e.g., culture-induced variation, green plant regeneration and biochemical pathways), and provides opportunities for regulating these phenomena for particular biotechnological purposes.

Functional Validation of cas9/guideRNA Constructs for Site-Directed Mutagenesis of Triticale ABA8'OH1 loci.

Cas endonuclease-mediated genome editing provides a long-awaited molecular biological approach to the modification of predefined genomic target sequences in living organisms. Although cas9/guide (g)RNA constructs are straightforward to assemble and can be customized to target virtually any site in the plant genome, the implementation of this technology can be cumbersome, especially in species like triticale that are difficult to transform, for which only limited genome information is available and/or which carry comparatively large genomes. To cope with these challenges, we have pre-validated cas9/gRNA constructs (1) by frameshift restitution of a reporter gene co-introduced by ballistic DNA transfer to barley epidermis cells, and (2) via transfection in triticale protoplasts followed by either a T7E1-based cleavage assay or by deep-sequencing of target-specific PCR amplicons. For exemplification, we addressed the triticale ABA 8'-hydroxylase 1 gene, one of the putative determinants of pre-harvest sprouting of grains. We further show that in-del induction frequency in triticalecan beincreased by TREX2 nuclease activity, which holds true for both well- and poorly performing gRNAs. The presented results constitute a sound basis for the targeted induction of heritable modifications in triticale genes.

Comparative Study of the Genetic and Biochemical Variability of Polyscias filicifolia (Araliaceae) Regenerants Obtained by Indirect and Direct Somatic Embryogenesis as a Source of Triterpenes.

Polyscias filicifolia (Araliaceae) is broadly used in traditional medicine in Southeast Asia due to its antimicrobial, immunomodulating and cytotoxic activities. The main groups of compounds responsible for pharmacological effects are believed to be oleanolic triterpene saponins. However, Polyscias plants demonstrate relatively slow growth in natural conditions, which led to applying a developing sustainable source of plant material via primary (PSE), secondary (DSE) and direct somatic embryogenesis from DSE (TSE). The AFLP and metAFLP genotyping resulted in 1277 markers, amplified by a total of 24 pairs of selective primers. Only 3.13% of the markers were polymorphic. The analysis of variance showed that the PSE and TSE regenerants differed only in terms of root number, while the DSE plantlets differed for all studied morphological characteristics. Further, the chemical analysis revealed that oleanolic acid (439.72 µg/g DW), ursolic acid (111.85 µg/g DW) and hederagenin (19.07 µg/g DW) were determined in TSE regenerants. Our results indicate that direct somatic embryogenesis ensures the production of homogeneous plant material, which can serve as a potential source of triterpene compounds. Plants obtained via somatic embryogenesis could also be reintroduced into the natural environment to protect and preserve its biodiversity.

Quantitative trait loci for starch-corrected chip color after harvest, cold storage and after reconditioning mapped in diploid potato.

The objective of this study was to map the quantitative trait loci (QTLs) for chip color after harvest (AH), cold storage (CS) and after reconditioning (RC) in diploid potato and compare them with QTLs for starch-corrected chip color. Chip color traits AH, CS, and RC significantly correlated with tuber starch content (TSC). To limit the effect of starch content, the chip color was corrected for TSC. The QTLs for chip color (AH, CS, and RC) and the starch-corrected chip color determined with the starch content after harvest (SCAH), after cold storage (SCCS) and after reconditioning (SCRC) were compared to assess the extent of the effect of starch and the location of genetic factors underlying this effect on chip color. We detected QTLs for the AH, CS, RC and starch-corrected traits on ten potato chromosomes, confirming the polygenic nature of the traits. The QTLs with the strongest effects were detected on chromosomes I (AH, 0 cM, 11.5% of variance explained), IV (CS, 43.9 cM, 12.7%) and I (RC, 49.7 cM, 14.1%). When starch correction was applied, the QTLs with the strongest effects were revealed on chromosomes VIII (SCAH, 39.3 cM, 10.8% of variance explained), XI (SCCS, 79.5 cM, 10.9%) and IV (SCRC, 43.9 cM, 10.8%). Applying the starch correction changed the landscape of QTLs for chip color, as some QTLs became statistically insignificant, shifted or were refined, and new QTLs were detected for SCAH. The QTLs on chromosomes I and IV were significant for all traits with and without starch correction.

Reconditioning and weather conditions affect black spot damage during storage of potato (Solanum tuberosum L.) tubers.

BACKGROUND: In potatoes (Solanum tuberosum L.), mechanical damage can cause the formation of black spots in the tuber flesh as the result of oxidation of phenolic compounds. This damage can result in substantial economic losses and degradation of quality. External factors contributing to the formation and the prevention of black spot damage (BSD) are not fully understood. The aim of this study was to determine the effect of weather conditions, using a hydrothermal coefficient, and of two potato tuber reconditioning methods on the formation of BSD. Five potato varieties were divided into high, moderate and low BSD susceptibility groups. The research was carried out over two growing seasons that differed greatly in precipitation and temperature. The black spot index was determined immediately after harvest and after 3 and 7 months of storage. Two reconditioning methods, applied for 7 days at 8 °C and for 7 days at 15 °C, were used for each variety. RESULTS: The incidence of BSD in susceptible varieties did not depend significantly on weather conditions, although statistically significant variation was observed. We found a correlation between the hydrothermal coefficient (dry conditions) and BSD in the Etiuda variety. There was no significant effect of storage time and temperature on BSD incidence in susceptible or moderately susceptible varieties. The reconditioning methods significantly reduced the formation of BSD in tubers after storage in all three groups. Regardless of the level of susceptibility of the variety, storage time and storage temperature, the most efficient treatment to limit BSD was reconditioning for 7 days at 15 °C. CONCLUSION: Identification of the significant effects of weather, and strong reduction of BSD in tubers that had been reconditioned, allows examination of the underlying mechanisms. The described reconditioning method can lead to satisfactory reduction of BSD in potato tubers. Data from this research will be of interest to potato breeders, particularly if valuable alleles that affect this phenomeon can be isolated. © 2019 Society of Chemical Industry.

Diversity Arrays Technology-based PCR markers for marker assisted selection of aluminum tolerance in triticale (x Triticosecale Wittmack).

The tolerance of triticale (x Triticosecale Wittmack) cultivars to aluminum (Al) stress observed in acid soils is an important agronomic trait affecting seed yield. Traditionally, breeding of Al-tolerant cultivars was selection based; for example, using a physiological test. However, such selection methods are relatively slow and require numerous plants for phenotype evaluation. Alternatively, DNA-based molecular marker systems could be applied to identify markers useful for selection purposes. Among many marker platforms available, Diversity Arrays Technology (DArT) is one of the most promising. DArT markers preselected for conversion to specific PCR assays were chosen based on association mapping studies using diverse materials. Forty-nine DArT markers were selected and tested for redundancy based on their segregation patterns and sequences, and 40 were successfully converted into specific PCR assays. However, only 24 of these proved to be polymorphic. Where possible, the chromosomal locations of the converted markers were verified. The markers assigned to chromosome 7R that were the most highly correlated with Al-tolerant and non-tolerant plants were chosen for marker assisted selection using genetically diverse triticale materials.