Phenotypic, molecular and biochemical evaluation of somatic hybrids between Solanum tuberosum and S. bulbocastanum.

Somatic hybridization has been frequently used to overcome sexual incompatibility between potato and its secondary germplasm. The primary objective of this study was to produce and evaluate somatic hybrids of Solanum tuberosum (Stub) and S. bulbocastanum (Sblb) for breeding purposes. In 2007, 23 somatic hybrids were produced using an electrofusion of mesophyll protoplasts of diploid (2n = 2x = 24) potato line StubDH165 and S. bulbocastanum PI24351 (Sblb66). Phenotype of somatic hybrids in field conditions were evaluated, together with constitution and stability of 30 nuclear (ncSSR) and 27 cytoplasmic (cpSSR) microsatellite markers and content of main glycoalkaloids. All somatic hybrids had very high field resistance against late blight, but the plants were infertile: the viability of pollen grains insignificantly varied between 0.58 and 8.97%. A significant somaclonal variation was observed in terms of the morphology of plants, the date of emergence, the quantity of harvested tubers, the content of glycoalkaloids in foliage, and nuclear microsatellite markers (ncSSR). The analysis of ncSSR identified five distinct genotypes of hybrids partly associated with phenotype variations. The process of somatic hybridization with regeneration of shoots was identified as the most likely source of somaclonal variation because the ncSSR genotypes of hybrids, which were maintained in vitro, remained stable for more than 10 years. The infertile somatic hybrids have no practical breeding potential, but they are considered very suitable for advanced studies of the differential expression of genes in the pathways linked to dormancy of tubers and synthesis of glycoalkaloids.

Carotenoid changes of colored-grain wheat flours during bun-making.

Colored-grain wheat genotypes were used in the preparation of flour, dough, buns, and buns stored for a short period of time. The main carotenoid in all genotypes was lutein, followed by its esters, zeaxanthin, and ß-carotene, while antheraxanthin and α-carotene occurred only at negligible levels. The highest carotenoid contents were observed in yellow- and purple-grained genotypes. After the preparation of dough, total carotenoid content (TCC) decreased significantly by an average of 61.5%. Zeaxanthin was shown to be stable, whereas α-carotene was destroyed. In baked buns, the average decrease of TCC and all-E-lutein was lower than in unbaked dough. Greater decreases were recorded for esters, antheraxanthin, and ß-carotene. After storing buns for 24 h at room temperature, approximately one-quarter of TCC observed in the original flour was preserved. Z-Isomers of lutein occurred in minor concentrations, but the degradation of this component, and that of zeaxanthin, was low, suggesting E- to Z-isomerization.

Free and esterified carotenoids in pigmented wheat, tritordeum and barley grains.

Carotenoids are important phytonutrients responsible for the yellow endosperm color in cereal grains. Five carotenoids, namely lutein, zeaxanthin, antheraxanthin, α- and ß-carotene, were quantified by HPLC-DAD-MS in fourteen genotypes of wheat, barley and tritordeum harvested in Czechia in 2014 and 2015. The highest carotenoid contents were found in yellow-grained tritordeum HT 439 (12.16µg/gDW), followed by blue-grained wheat V1-131-15 (7.46µg/gDW), and yellow-grained wheat TA 4024 (7.04µg/gDW). Comparing carotenoid contents, blue varieties had lower whereas purple ones had the same or higher levels than conventional bread wheat. Lutein was the main carotenoid found in wheat and tritordeum while zeaxanthin dominated in barley. The majority of cereals contained considerable levels of esterified forms (up to 61%) of which lutein esters prevailed. It was assessed that cereal genotype determines the proportion of free and esterified forms. High temperatures and drought during the growing season promoted carotenoid biosynthesis.

Tocotrienols and tocopherols in colored-grain wheat, tritordeum and barley.

Colored-grain spring and winter wheat, spring tritordeum and barley (blue aleurone, purple pericarp, and yellow endosperm) from the harvests 2014 and 2015 were evaluated for tocol contents by HPLC-FD. Higher content of total tocols was found in spring wheat varieties compared with winter varieties. Four tocols (ß-tocotrienol, α-tocotrienol, ß-tocopherol, and α-tocopherol) were identified in wheat and tritordeum varieties. Dominant tocols in purple- and blue-grained wheat and yellow-grained tritordeum were α-tocopherol and ß-tocotrienol, whereas spring barley varieties differed from wheat and tritordeum by high α-tocotrienol content. Tocol content was significantly affected by genotype and in a lesser extent in some varieties and lines also by rainfall and temperatures during crop year. Higher rainfall and lower temperatures caused in most varieties higher tocol contents. Purple- and blue-grained wheat lines with higher tocol, anthocyanin and phenolic acids with health benefits may be useful for breeding new varieties.

Carotenoid profile and retention in yellow-, purple- and red-fleshed potatoes after thermal processing.

This research aimed to investigate the effect of thermal processing on carotenoid profile, quantity and stability in 22 colour-fleshed potato cultivars grown in the Czech Republic. The total of nine carotenoids was analysed by HPLC using a C30 column and PDA detection. The total carotenoid content for all cultivars ranged from 1.44 to 40.13 µg/g DM. Yellow cultivars showed a much higher average total carotenoid content (26.22 µg/g DM) when compared to red/purple-fleshed potatoes (5.69 µg/g DM). Yellow cultivars were dominated by antheraxanthin, whereas neoxanthin was the main carotenoid in red/purple cultivars. Thermal processing significantly impacted all potato cultivars. Boiling decreased the total carotenoids by 92% compared to baking (88%). Lutein was the most stable carotenoid against thermal processing (decreased by 24-43%) followed by ß-carotene (decreased by 78-83%); other carotenoids were degraded nearly completely. Increased formation of (Z)-isomers by thermal processing has not been confirmed.

Endogenously produced hydrogen sulfide is involved in porcine oocyte maturation in vitro.

Hydrogen sulfide, one of three known gasotransmitters, is involved in physiological processes, including reproductive functions. Oocyte maturation and surrounding cumulus cell expansion play an essential role in female reproduction and subsequent embryonic development. Although the positive effects of exogenous hydrogen sulfide on maturing oocytes are well known, the role of endogenous hydrogen sulfide, which is physiologically released by enzymes, has not yet been described in oocytes. In this study, we observed the presence of Cystathionine ß-Synthase (CBS), Cystathionine γ-Lyase (CTH) and 3-Mercaptopyruvate Sulfurtransferase (3-MPST), hydrogen sulfide-releasing enzymes, in porcine oocytes. Endogenous hydrogen sulfide production was detected in immature and matured oocytes as well as its requirement for meiotic maturation. Individual hydrogen sulfide-releasing enzymes seem to be capable of substituting for each other in hydrogen sulfide production. However, meiosis suppression by inhibition of all hydrogen sulfide-releasing enzymes is not irreversible and this effect is a result of M-Phase/Maturation Promoting Factor (MPF) and Mitogen-Activated Protein Kinase (MAPK) activity inhibition. Futhermore, cumulus expansion expressed by hyaluronic acid (HA) production is affected by the inhibition of hydrogen sulfide production. Moreover, quality changes of the expanded cumuli are indicated. These results demonstrate hydrogen sulfide involvement in oocyte maturation as well as cumulus expansion. As such, hydrogen sulfide appears to be an important cell messenger during mammalian oocyte meiosis and adequate cumulus expansion.