Ilona Czyczyło-Mysza

Alleviation of Osmotic Stress Effects by Exogenous Application of Salicylic or Abscisic Acid on Wheat Seedlings

The aim of the study was to assess the role of salicylic acid (SA) and abscisic acid (ABA) in osmotic stress tolerance of wheat seedlings. This was accomplished by determining the impact of the acids applied exogenously on seedlings grown under osmotic stress in hydroponics. The investigation was unique in its comprehensiveness, examining changes under osmotic stress and other conditions, and testing a number of parameters simultaneously. In both drought susceptible (SQ1) and drought resistant (CS) wheat cultivars, significant physiological and biochemical changes were observed upon the addition of SA (0.05 mM) or ABA (0.1 μM) to solutions containing half-strength Hoagland medium and PEG 6000 (−0.75 MPa). The most noticeable result of supplementing SA or ABA to the medium (PEG + SA and PEG + ABA) was a decrease in the length of leaves and roots in both cultivars. While PEG treatment reduced gas exchange parameters, chlorophyll content in CS, and osmotic potential, and conversely, increased lipid peroxidation, soluble carbohydrates in SQ1, proline content in both cultivars and total antioxidants activity in SQ1, PEG + SA or PEG + ABA did not change the values of these parameters. Furthermore, PEG caused a two-fold increase of endogenous ABA content in SQ1 and a four-fold increase in CS. PEG + ABA increased endogenous ABA only in SQ1, whereas PEG + SA caused a greater increase of ABA content in both cultivars compared to PEG. In PEG-treated plants growing until the harvest, a greater decrease of yield components was observed in SQ1 than in CS. PEG + SA, and particularly PEG + ABA, caused a greater increase of these yield parameters in CS compared to SQ1. In conclusion, SA and ABA ameliorate, particularly in the tolerant wheat cultivar, the harmful effects and after effects of osmotic stress induced by PEG in hydroponics through better osmotic adjustment achieved by an increase in proline and carbohydrate content as well as by an increase in antioxidant activity.

Production of double haploids in oat (Avena sativa L.) by pollination with maize (Zea mays L.)

The aim of the study was to optimize the method of oat haploid production by pollination with maize. Seventeen oat genotypes were used in the experiment. Various factors influencing the growth and development of ovaries and embryo production were investigated: genotype, time of pollination, growth regulators and time of their application. Emasculated before anthesis, oat florets were pollinated with maize pollen after 0, 1 or 2 days. Next, one of two auxins analogues (2,4-D or dicamba) were applied to oat pistils. These auxins had no significant influence on the number of enlarged ovaries and embryos. The time of application of these growth regulators had a significant influence on embryo production. Haploid embryos were obtained from all used genotypes, although the frequency of enlarged ovaries and obtained embryos did not differ markedly between the genotypes. On average, 85% of ovaries were enlarged and 11.7% of them produced haploid embryos. Depending on the regeneration medium, 24–41% of embryos were germinated, of which 12% had developed into green plants. A strong significant difference in the number of germinating embryos and haploid plants was observed between the kind of regenerating medium used. There were no albino plants and all the obtained plants were haploid.

QTL mapping for germination of seeds obtained from previous wheat generation under drought

The QTLs controlling germination and early seedling growth were mapped using seeds acquired from mapping population and parental lines of Chinese Spring and SQ1 grown under water-limited conditions, severe drought (SDr) and well-watered plants (C). Germination ability was determined by performing a standard germination test based on the quantification of the germination percentage (GP24) of seeds incubated for 24 h at 25°C in the dark. Early seedling growth was evaluated on the basis of the length of the root and leaf at the 6th day of the experiment. QTLs were identified by composite interval mapping method using Windows QTLCartographer 2.5 software. For the traits studied, a total of thirty eight additive QTLs were identified. Seventeen QTLs were mapped in C on chromosomes: 1A, 2A, 7A, 1B, 2B, 3B, 4B, 5B, 6B, 7B, 2D, 3D, 4D and 6D, while twenty one QTLs were identified in SDr on chromosomes: 1A, 2A, 5A, 2B, 3B, 4B, 5B, 6B, 7B, 3D, 5D and 6D. Most of the QTLs for GP and early leaf growth parameters were clustered on chromosome 4B (associated with the Rht-B1 marker) both in C and SDr plants. The results indicate the complex and polygenic nature of germination.

Application of chosen factors in the wide crossingmethod for the production of oat doubled haploids

Abstract Oat (Avena sativa L.) has recently gained importance due to the discovery of a variety of health benefits and new opportunities of use. There is no efficient protocol for the production of oat doubled haploid (DH) lines. The aim of this study was to increase the efficiency of obtaining DHs of oat by the wide crossing method. The study was performed on five oat genotypes. We have compared the induction of embryos after pollination with maize, sorghum and millet pollen as well as the development of haploid embryos isolated 2, 3 and 4 weeks after pollination and cultivated on media with different sugar content. Haploid plants were treated with colchicine after or before acclimation to natural conditions. Of the three types of pollen used, the largest number of haploid embryos was obtained using maize pollen. Three weeks after pollination was the most suitable time for the isolation and cultivation of the embryos. The most efficient medium enabling the development of embryos and conversion to plants was 190-2 containing 9% of maltose. Colchicine treatment of acclimated plants provided high survival rate.

The effect of light intensity on the production of oat (Avena sativa L.) doubled haploids through oat × maize crosses

Oat haploid embryos were obtained by wide crossing with maize. The effect of light intensity during the growing period of donor plants (450 and 800 μmol m−2 s−1) and in vitro cultures (20, 40, 70 and 110 μmol m−2 s−1) was examined for the induction and development of oat DH lines. Oat florets (26008) from 32 genotypes were pollinated with maize and treated with 2,4-dichlorophenoxyacetic acid. All the tested genotypes formed more haploid embryos when donor plants were grown in a greenhouse (9.4%) compared to a growth chamber (6.1%). The light intensity of 110 μmol m−2 s−1 during in vitro culture resulted in the highest percentage of embryo germination (38.9%), conversion into plants (36.4%) and DH line production (9.2%) when compared with lower light intensities (20, 40 and 70 μmol m−2 s−1). The results show that the growth conditions of the donor plant and light intensity during in vitro culture can affect the development of haploid embryos. This fact may have an impact on oat breeding programs using oat × maize crosses.

Application of PV Powered High Intensity LEDs for Supplementary Irradiation of Horticultural Plants

The work presents a comprehensive study on problems of supplementary photosynthesis in horticultural plants realised by artificial light sources of optimised wavelength spectrum. A purely photovoltaic method of powering these sources is provided in order to avoid mains energy consumption. Three goals of the project are specified i.e. scientific, applique and usable ones. The first one deals with elaborating of algorithms for controlling, measuring and supplementing the photosynthesis process. The second - with laboratory engineering and equipment, while the third goal deals with utilization of elaborated methods and means for improving the quality and harvesting of the irradiated plants. Apart from the natural sunlight portions the entire supplementary photosynthesis bases on sun energy PV conversion to the red and blue light energy, additionally emitted by groups of red and blue LEDs, each of 1 to 3 W power. Their light intensity and mutual light proportions are controlled and optimised by a computer programme and by the electronic PWM and MPPT technique. Such an irradiation process, if necessary, may be carried on also in night hours or in cloudy periods by means of PV energy stored in batteries

Genetic analysis of water loss of excised leaves associated with drought tolerance in wheat

Background Wheat is widely affected by drought. Low excised-leaf water loss (ELWL) has frequently been associated with improved grain yield under drought. This study dissected the genetic control of ELWL in wheat, associated physiological, morphological and anatomical leaf traits, and compared these with yield QTLs. Methods Ninety-four hexaploid wheat (Triticum aestivum L.) doubled haploids, mapped with over 700 markers, were tested for three years for ELWL from detached leaf 4 of glasshouse-grown plants. In one experiment, stomata per unit area and leaf thickness parameters from leaf cross-sections were measured. QTLs were identified using QTLCartographer. Results ELWL was significantly negatively correlated with leaf length, width, area and thickness. Major QTLs for ELWL during 0–3 h and 3–6 h were coincident across trials on 3A, 3B, 4B, 5B, 5D, 6B, 7A, 7B, 7D and frequently coincident (inversely) with leaf size QTLs. Yield in other trials was sometimes associated with ELWL and leaf size phenotypically and genotypically, but more frequently under non-droughted than droughted conditions. QTL coincidence showed only ELWL to be associated with drought/control yield ratio. Discussion Our results demonstrated that measures of ELWL and leaf size were equally effective predictors of yield, and both were more useful for selecting under favourable than stressed conditions.

Complex characterization of oat (Avena sativa L.) lines obtained by wide crossing with maize (Zea mays L.)

Background The oat × maize addition (OMA) lines are used for mapping of the maize genome, the studies of centromere-specific histone (CENH3), gene expression, meiotic chromosome behavior and also for introducing maize C4 photosynthetic system to oat. The aim of our study was the identification and molecular-cytogenetic characterization of oat × maize hybrids. Methods Oat DH lines and oat × maize hybrids were obtained using the wide crossing of Avena sativa L. with Zea mays L. The plants identified as having a Grande-1 retrotransposon fragment, which produced seeds, were used for genomic in situ hybridization (GISH). Results A total of 138 oat lines obtained by crossing of 2,314 oat plants from 80 genotypes with maize cv. Waza were tested for the presence of maize chromosomes. The presence of maize chromatin was indicated in 66 lines by amplification of the PCR product (500 bp) generated using primers specific for the maize retrotransposon Grande-1. Genomic in situ hybridization (GISH) detected whole maize chromosomes in eight lines (40%). All of the analyzed plants possessed full complement of oat chromosomes. The number of maize chromosomes differed between the OMA lines. Four OMA lines possessed two maize chromosomes similar in size, three OMA—one maize chromosome, and one OMA—four maize chromosomes. In most of the lines, the detected chromosomes were labeled uniformly. The presence of six 45S rDNA loci was detected in oat chromosomes, but none of the added maize chromosomes in any of the lines carried 45S rDNA locus. Twenty of the analyzed lines did not possess whole maize chromosomes, but the introgression of maize chromatin in the oat chromosomes. Five of 66 hybrids were shorter in height, grassy type without panicles. Twenty-seven OMA lines were fertile and produced seeds ranging in number from 1–102 (in total 613). Sixty-three fertile DH lines, out of 72 which did not have an addition of maize chromosomes or chromatin, produced seeds in the range of 1–343 (in total 3,758). Obtained DH and OMA lines were fertile and produced seeds. Discussion In wide hybridization of oat with maize, the complete or incomplete chromosomes elimination of maize occur. Hybrids of oat and maize had a complete set of oat chromosomes without maize chromosomes, and a complete set of oat chromosomes with one to four retained maize chromosomes.

Obtaining of winter rye (Secale cereale L. ssp. cereale) haploid embryos through hybridization with maize (Zea Mays L.)

The aim of this study was to determine the effect of selected factors on rye (Secale cereale L.) haploid embryo production by the wide crossing method. The study was performed on fifteen winter rye genotypes. This is the first time for rye when besides the genotype, on the enlargement of ovaries and haploid embryo production, such factors as: type of auxin analogues 2,4-dichlorophenoxyacetic acid (2,4-D), 3,6-dichloro-2-methoxybenzoic acid (dicamba) and 4-amino-3,5,6-trichloropyridine-2-carboxylic acid (picloram), and the time between florets emasculation and pollination were investigated. All factors had a significant impact on rye ovary enlargement, however the haploid embryo formation depended only on rye genotype, not on kind of auxin and days between emasculation to pollination. In total, twenty one haploid embryos were formed by six genotypes of fifteen tested. On average, 13.86% (after 2,4-D treatment) to 20.05% (after dicamba treatment) enlarged ovaries per emasculated florets were obtained. Most of the ovaries enlarged when florets were pollinated 4 and 6 days after emasculation. Most of the haploid embryos formed when florets were pollinated 6 days after emasculation. The obtained haploid embryos did not germinate.

Putative candidate genes responsible for leaf rolling in rye ( Secale cereale L.)

BackgroundRolling of leaves (RL) is a phenomenon commonly found in grasses. Morphology of the leaf is an important agronomic trait in field crops especially in rice; therefore, majority of the rice breeders are interested in RL. There are only few studies with respect to RL of wheat and barley; however, the information regarding the genetic base of RL with respect to the shape of leaf in rye is lacking. To the best of our knowledge, this is the first study on the localization of loci controlling RL on high density consensus genetic map of rye.ResultsGenotypic analysis led to the identification of 43 quantitative trait loci (QTLs) for RL, grouped into 28 intervals, which confirms the multigenic base of the trait stated for wheat and rice. Four stable QTLs were located on chromosomes 3R, 5R, and 7R.Co-localization of QTL for RL and for different morphological, biochemical and physiological traits may suggests pleiotropic effects of some QTLs. QTLs for RL were associated with QTLs for such morphological traits as: grain number and weight, spike number per plant, compactness of spike, and plant height. Two QTLs for RL were found to coincide with QTLs for drought tolerance (4R, 7R), two with QTLs for heading earliness (2R, 7R), one with α-amylase activity QTL (7R) and three for pre-harvest sprouting QTL (1R, 4R, 7R).The set of molecular markers strongly linked to RL was selected, and the putative candidate genes controlling the process of RL were identified. Twelve QTLs are considered as linked to candidate genes on the base of DArT sequences alignment, which is a new information for rye.ConclusionsOur results expand the knowledge about the network of QTLs for different morphological, biochemical and physiological traits and can be a starting point to studies on particular genes controlling RL and other important agronomic traits (yield, earliness, pre-harvest sprouting, reaction to water deficit) and to appoint markers useful in marker assisted selection (MAS). A better knowledge of the rye genome and genes could both facilitate rye improvement itself and increase the efficiency of utilizing rye genes in wheat breeding.