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Stress was recognized as a major factor changing microspore development from a gamethophytic to a sporophytic pathway. Stress factors effectively inducing androgenesis in a wide spectrum of genotypes include high temperature, carbohydrate/nitrogen starvation and high-osmotic shocks. A low temperature treatment usually improve plant responsiveness, however in some cases, is not sufficient for embryogenesis initiation. Thus low temperature is not consider by some authors to be a true „stress factor”. In the experiment the effect of prolonged cold pretreatment of tillers on the effectiveness of androgenesis induction in isolated microspore cultures of hexaploid triticale (X Triticisecale Wittm.) was tested. In the study, two spring cultivars (cv. ‘Mieszko’ and cv. ‘Wanad’) varying in androgenic responsiveness were used. Prolongation of tillers cold pretreatment from 3 to 5 weeks, increased the number of microspores of 'Mieszko' isolated per spike from 5.8·10⁴ to 9.9·10⁴. It also promoted ELS development and its regeneration ability. ELS production increased more then 5-fold, when the number of regenerating ELS increased from 1.4% to 29%. At the same time, prolonged cold pretreatment of ‘Wanad’ tillers had no effect on the microspores yield received from one spike. Moreover, it reduced the number of produced ELS about 3-fold. Due to great variation among replicates the increase of ELS regeneration ability (from 2.8% to 24.5%) was not statistically significant. For both cultivars, the highest green plant regeneration percentage (24% and 8.2% for ‘Wanad’ and ‘Mieszko’, respectively) was noted after the longest cold pretreatment of tillers. However, the great variation among replicates made the comparison among treatments difficult for statistical analyses. Proper interpretation of the received results needs a further and more detailed examination.
The effect of various stress treatments (cold, heat, starvation) and its combinations on androgenesis induction in isolated microspore cultures of spring wheat (Triticum aestivum L.) was under the study. On the basis of our preliminary experiments, spring wheat cultivar Minaret recognized as responsive for androgenic development was selected and used in the study. The anthers were aseptically isolated from freshly cut or cold-pretreated (9-14 days at 4°C) tillers. The anthers were precultured in starvation B medium or rich AT3 medium at 5, 26 or 33°C. After 4 days, microspores were mechanically isolated and cultured in AT3 medium at a density 1 x 10⁵ microspores per cm³, at 26°C, in the dark. Well-developed embryos were transferred to solid 190-2 medium and cultured at 26°C, in the dim light. Cytological observations were done in 2-weeks intervals with inverted light microscope. The analysis of the number of spontaneously shed microspores and its cytological characterization conducted after 4-day anther pre-culture was a good indicator of culture androgenic potential. The effectiveness of androgenesis induction was depended on the type and sequence of stress treatments. The highest intensity of stress treatment (combination of cold pretreatment of tillers with heat and carbohydrate/nitrogen starvation stresses applied to anthers) induced the most effective androgenic development and as result a great number (about 100 per dish) of multicellular androgenic structures were formed. However, the final effect (the number of globular embryos big enough to be transferred onto regeneration medium) was not satisfying (max 50 embryos per dish). Cold shock applied both to tillers or anthers enhanced androgenic development and plant regeneration. The percentage of regenerating embryos ranging from 0 to 50%. However all regenerates were chlorophyll-lacking (albino) plants. In the control cultures (no stress treatment) microspores continued their normal gametophytic development.
Plants growing under natural conditions are constantly exposed to ultraviolet (UV), primarily UV-A, radiation. Grass pea (Lathyrus sativus L.) is a legume species resistant to harsh growing conditions, such as drought, salinity or periodic flooding. Due to the advantageous composition of seeds, it is used for consumption in such regions as South Asia or East Africa where high intensity of UV radiation occurs. Absorption of this spectral range causes changes in the photosynthetic apparatus of plants, including damage to the photosystem II (PSII) reaction centres. The aim of the work was to examine whether the use of the combination UV-A: red light as a source of radiation would enable quick acclimatization of the photosynthetic apparatus of grass pea to the negative effect of UV-A radiation. 14-day-old plants were exposed to UV-A:red radiation for 48 h. The plants exposed to UV-A:red radiation showed enhanced effective efficiency of PSII and increased total electron carriers, which enabled more effective photosynthesis at higher values of radiation intensity in comparison with control plants, kept under white LED light. At the same time, there were no statistically significant differences in both the photosynthetic pigment contents and the level of lipid peroxidation. The obtained results indicate that the observed increase in the efficiency of CO₂ carboxylation after short-term UV-A:red radiation has resulted from the efficient linear electron transport due to maintaining the effective oxygen evolving complex (OEC) and increased total electron carriers.
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