A new high-molecular-weight glutenin subunit from the slovak wheat (Triticum aestivum L.) cultivar ‘Trebišovská 76’

Historical and local varieties are an important source of genetic diversity in connection with the wheat baking quality. By analysis of DNA from the coding part of the genome of the Slovak regional wheat (Triticum aestivum L.) variety ‘Trebišovská 76’, differences in the Dy subunit composition coded by locus Glu-1D were found. Comparing the newly found sequence with a generally used 1Dy12 (GenBank accession no. X03041) sequence, existence of a new high-molecular-weight glutenin subunit, which was marked as 1Dy12* (GenBank accession no. EU266533) was confirmed. The molecular weight of probable protein coded by 1Dy12* subunit reaches 72.35 kDa comparing to the 1Dy12, which reaches 68.71 kDa. This difference is not likely to distinguish by SDS-PAGE. The easiest way for cereal genotyping according to baking quality is proteomic analysis. More exact and suitable method for genotype identification appears to be marker assisted selection using PCR.

     

Biosynthesis of Essential Polyunsaturated Fatty Acids in Wheat Triggered by Expression of Artificial Gene

The artificial gene D6D encoding the enzyme ∆⁶desaturase was designed and synthesized using the sequence of the same gene from the fungus Thamnidium elegans. The original start codon was replaced by the signal sequence derived from the wheat gene for high-molecular-weight glutenin subunit and the codon usage was completely changed for optimal expression in wheat. Synthesized artificial D6D gene was delivered into plants of the spring wheat line CY-45 and the gene itself, as well as transcribed D6D mRNA were confirmed in plants of T₀ and T₁ generations. The desired product of the wheat genetic modification by artificial D6D gene was the γ-linolenic acid. Its presence was confirmed in mature grains of transgenic wheat plants in the amount 0.04%–0.32% (v/v) of the total amount of fatty acids. Both newly synthesized γ-linolenic acid and stearidonic acid have been detected also in leaves, stems, roots, awns, paleas, rachillas, and immature grains of the T₁ generation as well as in immature and mature grains of the T₂ generation. Contents of γ-linolenic acid and stearidonic acid varied in range 0%–1.40% (v/v) and 0%–1.53% (v/v) from the total amount of fatty acids, respectively. This approach has opened the pathway of desaturation of fatty acids and production of essential polyunsaturated fatty acids in wheat.     

Establishment of Stem Cell-like Cells of Sida hermaphrodita (L.) Rusby from Explants Containing Cambial Meristems

The in vitro cultures of plant stem cells and stem cell-like cells can be established from tissues containing meristematic cells. Chemical compounds—as well as their production potential—is among the emerging topics of plant biotechnology. We induced the callus cell biomass growth and characterized the parameters indicating the presence of stem cells or stem cell-like cells. Four types of explants (stem, petiole, leaf, root) from Sida hermaphrodita (L.) Rusby and various combinations of auxins and cytokinins were tested for initiation of callus, growth of sub-cultivated callus biomass, and establishment of stem cells or stem cell-like cells. Induction of callus and its growth parameters were significantly affected both by the explant type and the combination of used plant growth hormones and regulators. The responsibility for callus initiation and growth was the highest in stem-derived explants containing cambial meristematic cells. Growth parameters of callus biomass and specific characteristics of vacuoles confirmed the presence of stem cells or stem cell-like cells in sub-cultivated callus cell biomass. Establishment of in vitro stem cell or stem cell-like cell cultures in S. hermaphrodita can lead to the development of various applications of in vitro cultivation systems as well as alternative applications of this crop.     

Biodiversity of Legume Health‐promoting Starch

The content of retrograded resistant starch (RS₃) and RS₃/total starch ratio was determined in 74 cultivars of six legumes currently found in human food. There were significant differences in individual crops as well as their cultivars. All analysed legumes (except soybean) are a suitable natural source of health‐promoting RS₃. The highest levels of RS₃ were found in peas. There were extensive differences in RS₃ content and RS₃/total starch ratio in the latter crop types. Higher resistant and lower total starch amounts were found in wrinkle seeded peas. Maximal RS₃ content was detected in Ctirad, Dinara, Royal Salute and Elkan cultivars. In smooth seeded peas, lower resistant and higher total starch were estimated. High RS₃ levels were also observed in lentils, especially in the LCMB12/58 and LCMB1941 cultivars. Similar RS₃ values were found in the chickpea, faba bean and kidney bean. The highest RS₃ levels were demonstrated in the kidney bean Ultima, Fabia, 545/97472 and 545/974532; chickpea Alfa, CM7185, BG004233, PK51814 and 88199; and faba bean Alfréd and Omar cultivars. No soybean cultivar was found to be a suitable source of RS₃.