Androgenic responsiveness in anther (AC) and isolated microspore cultures (MC) was analysed using 15 lines of Polish winter rye (Secale cereale L.). The effect of low temperature (LT) alone or in combination with osmotic stress induced by mannitol treatment (MAN) and/or with reduced glutathione (GSH) on the effectiveness of the process was studied. Interestingly, each treatment had a different effect on microspore (mcs) vitality and capability to divide symmetrically. The first criterion for successful embryogenesis was to exceed the threshold number of at least 25% dividing microspores, which determined ‘embryogenic suspension culture’. In some configurations a spectacular effect was achieved, especially in lines highly recalcitrant to androgenesis induction. Relatively high effectiveness of androgenesis induction (up to 4.58 AS per 10⁵ mcs per spike in MC and 21.29 AS per spike in AC) showed that the developed protocol with GSH and/or MAN tiller pre-treatments overcomes the genotypic barrier for androgenesis initiation in rye. Moreover, relatively high, spontaneous genome diploidization (55%) of regenerated plants demonstrated that the described protocols could be effectively integrated into conventional rye breeding programmes.

Gloeobacter violaceus is a cyanobacteria species with a lack of thylakoids, while photosynthetic antennas, i.e., phycobilisomes (PBSs), photosystem II (PSII), and I (PSI), are located in the cytoplasmic membrane. We verified the hypothesis that blue–red (BR) light supplemented with a far-red (FR), ultraviolet A (UVA), and green (G) light can affect the photosynthetic electron transport chain in PSII and explain the differences in the growth of the G. violaceus culture. The cyanobacteria were cultured under different light conditions. The largest increase in G. violaceus biomass was observed only under BR + FR and BR + G light. Moreover, the shape of the G. violaceus cells was modified by the spectrum with the addition of G light. Furthermore, it was found that both the spectral composition of light and age of the cyanobacterial culture affect the different content of phycobiliproteins in the photosynthetic antennas (PBS). Most likely, in cells grown under light conditions with the addition of FR and G light, the average antenna size increased due to the inactivation of some reaction centers in PSII. Moreover, the role of PSI and gloeorhodopsin as supplementary sources of metabolic energy in the G. violaceus growth is discussed.

Superoxide dismutases (SODs) belong to the group of metalloenzymes that remove superoxide anion radicals and they have been identified in three domains of life: Bacteria, Archaea and Eucarya. SODs in Synechocystis sp. PCC 6803, Gloeobacter violaceus CCALA 979, and Geitlerinema sp. ZHR1A were investigated. We hypothesized that iron (FeSOD) and/or manganese (MnSOD) dominate as active forms in these cyanobacteria. Activity staining and three different spectroscopic methods of SOD activity bands excised from the gels were used to identify a suitable metal in the separated samples. FeSODs or enzymes belonging to the Fe-MnSOD superfamily were detected. The spectroscopic analyses showed that only Fe is present in the SOD activity bands. We found FeSOD in Synechocystis sp. PCC 6803 while two forms in G. violaceus and Geitlerinema sp. ZHR1A: FeSOD1 and FeSOD2 were present. However, no active Cu/ZnSODs were identified in G. violaceus and Geitlerinema sp. ZHR1A. We have shown that selected spectroscopic techniques can be complementary to the commonly used method of staining for SOD activity in a gel. Furthermore, the occurrence of active SODs in the cyanobacteria studied is also discussed in the context of SOD evolution in oxyphotrophs.

DOI: 10.33963/KP.15397, Published online: May 28, 2020, Authors: Mariusz Kowalewski, Mirosław Gozdek, Kamil Zieliński, Giuseppe Maria Raffa, Piotr Suwalski, Roberto Lorusso