Molecular selection of soybean towards adaptation to Central European agroclimatic conditions.

Europe is highly dependent on soybean meal imports and anticipates an increase of domestic plant protein production. Ongoing climate change resulted in northward shift of plant hardiness zones, enabling spring-sowing of freezing-sensitive crops, including soybean. However, it requires efficient reselection of germplasm adapted to relatively short growing season and long-day photoperiod. In the present study, a PCR array has been implemented, targeting early maturity (E1-E4, E7, E9, and E10), pod shattering (qPHD1), and growth determination (Dt1) genes. This array was optimized for routine screening of soybean diversity panel (204 accessions), subjected to the 2018-2020 survey of phenology, morphology, and yield-related traits in a potential cultivation region in Poland. High broad-sense heritability (0.84-0.88) was observed for plant height, thousand grain weight, maturity date, and the first pod height. Significant positive correlations were identified between the number of seeds and pods per plant, between these two traits and seed yield per plant as well as between flowering, maturity, plant height, and first pod height. PCR array genotyping revealed high genetic diversity, yielding 98 allelic combinations. The most remarkable correlations were identified between flowering and E7 or E1, between maturity and E4 or E7 and between plant height and Dt1 or E4. The study demonstrated high applicability of this PCR array for molecular selection of soybean towards adaptation to Central Europe, designating recessive qPHD1 and dominant Dt1, E3, and E4 alleles as major targets to align soybean growth season requirements with the length of the frost-free period, improve plant performance, and increase yield.

Biochemical Properties of Bioactive Compounds in the Oil from Polish Varieties of Camelina sativa Cultivated in 2019-2022.

Cold-pressed Camelina oil is a traditional oil registered as a traditional food in Poland. Camelina oil has health-promoting properties and high oxidative stability. This may be due to the presence of various bioactive antioxidant compounds such as carotenoids, sterols and polyphenols. Bioactive compounds content in Camelina oil depends mainly on the varieties and on the conditions under which the crop was grown therefore the aim of the research was to analyse antioxidant bioactive compounds in oil from different cultivars of Camelina sativa seeds and to determine their relationship with oil parameters.

Novel Tetraploid Triticale (Einkorn Wheat × Rye)-A Source of Stem Rust Resistance.

Among cereals, triticale (×Trititcoseale Wittmack ex A. Camus) represents a number of advantages such as high grain yield even in marginal environments, tolerance to drought, cold and acid soils, as well as lower production costs. Together with high biomass of grain and straw, triticale is also considered as an industrial energy crop. As an artificial hybrid, it has not evolved naturally, which is reflected in narrow genetic diversity causing a resistance collapse in recent years. Here, we describe a novel, synthetic tetraploid triticale, which was developed by the crossing of rye (Secale cereale L.) with einkorn wheat (Triticum monococcum spp. monococcum), which possess Sr35 stem rust resistance gene. Three subsequent generations of alloploids were obtained by chromosome doubling followed by self-pollination. The cytogenetic analyses revealed that the amphiploids possess a set of 28 chromosomes (14 of Am-genome and 14 of R-genome). The values of the most important yield-shaping traits for these tetraploid triticale form, including thousand-grain weight, plant height and stem length were higher compared to parental genotypes, as well as standard hexaploid triticale cultivars. This study shows that this tetraploid triticale genetic stock can be an interesting pre-breeding germplasm for triticale improvement or can be developed as a new alternative crop.

An Analysis of Variability in the Content of Phenolic Acids and Flavonoids in Camelina Seeds Depending on Weather Conditions, Functional Form, and Genotypes.

Camelina oil obtained from the seeds of Camelina sativa exhibits strong antioxidative properties. This study was based on four years of field experiments conducted on 63 genotypes of spring and 11 genotypes of winter camelina. The aim of the study was to determine the variability in the content of the selected bioactive compounds, depending on the weather conditions during the cultivation, the functional form, and genotype. The cultivation form of the genotypes analysed in our study did not exhibit significant differences in the quantitative profiles of the tested phenolic acids and flavonoids. Sinapic acid was the most abundant of all phenolic acids under analysis (617-668 mg/kg), while quercetin was the main flavonoid (91-161 mg/kg). Camelina has great potential not only for the food industry but also for researchers attempting to breed an oil plant with the stable biosynthesis of bioactive compounds to ensure oxidative protection of obtained fat.

Assessment of Antimicrobial Properties of Phenolic Acid Extracts from Grain Infected with Fungi from the Genus Fusarium.

Problems related with biological contamination of plant origin raw materials have a considerable effect on prevention systems at each stage of food production. Concerning the antimicrobial action of phenolic acids, studies were undertaken to investigate antibacterial properties against bacterial strains of Escherichia coli (EC), Pseudomonas fluorescence (PF), Micrococcus luteus (ML) and Proteus mirabilis (PM), as well as antifungal properties targeting microscopic fungi Fusarium spp., extracts of phenolic compounds coming from inoculated grain from various genotypes of cereals. This study evaluated the antimicrobial action of phenolic acids extracts obtained from both naturally infested and inoculated with microorganisms. For this purpose a total of 24 cereal cultivars were selected, including 9 winter and 15 spring cultivars. The analyses showed a bactericidal effect in the case of 4 extracts against Micrococcus luteus (ML), 14 extracts against Pseudomonas fluorescence (PF), 17 extracts against Escherichia coli (EC) as well as 16 extracts against Proteus mirabilis (PM). It was found that 3 out of the 24 extracts showed no antibacterial activity. In turn, fungicidal action was observed in the case of 17 extracts against Fusarium culmorum (FC) (NIV), 16 extracts against FC (3AcDON), 12 extracts against Fusarium graminearum (FG) (3AcDON), while 12 other extracts showed antifungal action against FG (NIV) and 19 extracts against Fusarium langsethiae (FL). Based on the conducted analyses it was found that grain of small-grained cereals exposed to fungal infection is a source of bioactive compounds exhibiting antimicrobial properties. It was observed that the qualitative and quantitative profiles of polyphenols vary depending on the cereal cultivar. This extracts may be used to develop an antimicrobial preparation applicable in organic farming.

Identification of leaf rust resistance genes Lr34 and Lr46 in common wheat (Triticum aestivum L. ssp. aestivum) lines of different origin using multiplex PCR.

Leaf rust caused by the fungus Puccinia recondita f. sp. tritici is one of the most dangerous diseases of common wheat. Infections caused by fungal pathogens reduce the quantity and quality of yields of many cereal species. The most effective method to limit plant infection is to use cultivars that show rust resistance. Genetically conditioned horizontal-type resistance (racial-nonspecific) is a desirable trait because it is characterized by more stable expression compared to major (R) genes that induce racially specific resistance, often overcome by pathogens. Horizontal resistance is conditioned by the presence of slow rust genes, which include genes Lr34 and Lr46. This study aimed to identify markers linked to both genes in 64 common wheat lines and to develop multiplex PCR reaction conditions that were applied to identify both genes simultaneously. The degree of infestation of the analyzed lines was also assessed in field conditions during the growing season of 2017 and 2018. Simple sequence repeat anchored-polymerase chain reaction (SSR-PCR) marker csLV was identified during analysis in line PHR 4947. The presence of a specific sequence has also been confirmed in multiplex PCR analyses. In addition to gene Lr34, gene Lr46 was identified in this genotype. Lines PHR 4947 and PHR 4819 were characterized by the highest leaf rust resistance in field conditions. During STS-PCR analyses, the marker wmc44 of gene Lr46 was identified in most of the analyzed lines. This marker was not present in the following genotypes: PHR 4670, PHR 4800, PHR 4859, PHR 4907, PHR 4922, PHR 4949, PHR 4957, PHR 4995, and PHR 4997. The presence of a specific sequence has also been confirmed in multiplex PCR analyses. Genotypes carrying the markers of the analyzed gene showed good resistance to leaf rust in field conditions in both 2017 and 2018. Research has demonstrated that marker assisted selection (MAS) and multiplex PCR techniques are excellent tools for selecting genotypes resistant to leaf rust.

Cytomolecular analysis of mutants, breeding lines, and varieties of camelina (Camelina sativa L. Crantz).

Camelina sativa L. Crantz (Brassicaceae family), known as camelina, has gained new attention as a re-emerging oil seed crop. With a unique seed oil profile, with the majority of the fatty acids consisting of linolenic (C18:3), oleic (C18:1), linoleic (C18:2), and eicosenoic (C20:1), camelina oil is reported to be useful as a food oil and biofuel. However, there are still many unknown factors about the structure and genetic variability of this crop. Chromosomal localization of ribosomal DNA was performed using fluorescence in situ hybridization (FISH) with 5S rDNA and 25S rDNA sequences as molecular probes on mitotic chromosomes of enzymatically digested root-tip meristematic cells. Here, we present for the first time a comparative analysis of selected genotypes (cultivars, breeding lines and mutants) of C. sativa with the use of cytogenetic techniques. The main aim of the study was to determine the intraspecific and interspecific polymorphisms in the structure of chromosomes of selected accessions using conserved 5S and 25S rDNA repetitive sequences as molecular probes. The results were compared with C. microcarpa (closely related to C. sativa) rDNA gene loci distribution. The presence of minor rDNA sites was discussed and compared with other Brassicaceae species. In addition, demonstration karyograms of C. sativa and C. microcarpa mapped with rDNA probes were prepared based on the cv. "Przybrodzka" and GE2011-02 genotype, respectively. The use of 5S and 25S rDNA probes provided an insight on the genome structure of C. sativa at the cytogenetic level and can help to understand the genome organization of this crop. The putative role of cytogenetic markers in phylogenetic analyses of camelina was discussed, as well.

Winter camelina seeds as a raw material for the production of erucic acid-free oil.

Camelina oil is increasingly popular as consumption as oil. Erucic acid is an unwanted fatty acid in oil. First studies on several genotypes have shown that this oil contains varying amounts of eriuc acid. The aim of the study was to analyses content of eriuc acid in all genotypes camelina. Hypothesis was that the content of erucic acid in winter forms is lower than in spring ones. A field experiment with 65 spring genotypes and 9 winter genotypes of camelina was conducted in Poland from 2016 to 2018. The analyses based on two chromatographic methods, i.e. UPLC-DAD and GC-MS, showed no differences in the results for the camelina samples. The average percentage content of the erucic acid in the spring genotypes was 3.432%, and in the winter genotypes was 0.1%. Our three-year research shows that some winter varieties can be used as low erucic acid forms.

Effect of Zearalenone and Hormone Regulators on Microspore Embryogenesis in Anther Culture of Wheat.

The purpose of this work was to assess the impact of zearalenone (ZEN) and selected hormone regulators on the effectiveness of microspore embryogenesis in anther culture of wheat. The plant material comprised F1 hybrids of winter and spring wheat. Six combinations of media inducing microspore proliferation and formation of embryogenic structures were investigated: two combinations of growth regulators (D - 2,4-D + dicamba, K - 2,4-D + kinetin), each with three ZEN concentrations (0 mL/L, 0.1 mL/L, 0.2 mL/L). A significant increase in microspore embryogenesis effectiveness on media with the addition of ZEN was observed both at the stages of its induction and the formation of green plants in some genotypes. In case of both combinations of growth regulators, an increased concentration of ZEN resulted in more effective induction of microspore embryogenesis. The most effective induction medium was the D medium supplemented with 0.2 mL/L ZEN. As a result of the use of zearalenone together with two combinations of growth regulators, all genotypes tested produced androgenic structures, which indicates the breakdown of genotypic recalcitrant in the analysed hybrids. In addition, green plants were obtained from 18 out of 19 tested hybrids. The addition of ZEN to the medium did not affect the number of regenerated albino plants nor the number of spontaneous genome doublings proportion.

Cytological markers used for identification and transfer of Aegilops spp. chromatin carrying valuable genes into cultivated forms of Triticum.

There are many reports describing chromosome structure, organization and evolution within goatgrasses (Aegilops spp.). Chromosome banding and fluorescence in situ hybridization techniques are main methods used to identify Aegilops Linnaeus, 1753 chromosomes. These data have essential value considering the close genetic and genomic relationship of goatgrasses with wheat (Triticumaestivum Linnaeus, 1753) and triticale (× Triticosecale Wittmack, 1899). A key question is whether those protocols are useful and effective for tracking Aegilops chromosomes or chromosome segments in genetic background of cultivated cereals. This article is a review of scientific reports describing chromosome identification methods, which were applied for development of prebreeding plant material and for transfer of desirable traits into Triticum Linnaeus, 1753 cultivated species. Moreover, this paper is a resume of the most efficient cytomolecular markers, which can be used to follow the introgression of Aegilops chromatin during the breeding process.

Identification of Leaf Rust Resistance Genes in Selected Wheat Cultivars and Development of Multiplex PCR.

Ten leading wheat cultivars originating from the Plant Breeding and Acclimatization Institute (IHAR) - National Research Institute (Poland) and the Department of Gene Bank (Czech Republic) were used to establish a field experiment in 2017 and 2018 at the Dlon Experimental Farm. The analyzed wheat genotypes were characterized by diversified field resistance to leaf rust. Jubilatka, Thatcher and Sparta were the most resistant cultivars in field conditions in both 2017 and 2018. The aim of the work was to identify the Lr11, L13, Lr16 and Lr26 genes encoding resistance to leaf rust using molecular SSR markers (wmc24, wmc261, Xgwm630, Xwmc764 and P6M12) and to develop multiplex PCR conditions to accelerate identification of these genes. Markers of three leaf rust resistance genes have been identified simultaneously in these cultivars. Jubilatka, Thatcher and Sparta cultivars may serve as a good source of the analyzed leaf rust resistance genes. In addition, multiplex PCR conditions have been developed for the simultaneous identification of the Lr11 and Lr16 and Lr11 and Lr26 gene pairs.

Ammonium bio-ionic liquids based on camelina oil as potential novel agrochemicals.

Third generation bio-ionic liquids (bio-ILs) were synthesized based on cheap and increasingly available camelina oil. The ionic liquids were obtained with high yield based on the reaction between camelina oil, which contained the following carboxylic acids: C18:3ω-3 linolenic >30%, C20:1 eicosenoic 28%, C18:2ω-6 linoleic 13%, C18:1 oleic 13%, C16:0 palmitic 4.5%, C22:1 erucic 4.5% and C18:0 stearic 2.5%, and quaternary ammonium hydroxides comprising cations such as: choline, di(hydrogenated tallow)dimethylammonium, oleylmethylbis(2-hydroxyethyl)ammonium, benzalkonium, tetradecyltrimethylammonium, tetramethylammonium and didecyldimethylammonium. The synthesized bio-ILs were characterized as high viscosity liquids which are thermally stable and their solubility in water and organic solvents depended on the type of cation. Two extreme examples of bio-ILs include the water soluble one comprising choline as the cation and the one comprising the di(hydrogenated tallow)dimethylammonium cation, which is soluble in hexane. The presented results show the importance of ammonium bio-ILs as antifeedants with a wide spectrum of activity. The tested beetles (Insecta: Coleoptera) of storage pests: grain weevil (Sitophilus granarius (L.)), confused flour beetle (Tribolium confusum Duv.) and khapra beetle (Trogoderma granarium Ev.) presented notable differences in terms of susceptibility to the synthesized ILs. The synthesized bio-ILs are effective adjuvants for herbicides belonging to the sylfonylurea group. They exhibited high activity despite the fact they were applied at a dose almost half that used for commercial adjuvants, which opens the era of adjuvant ILs. Aside from its use in the production of biodiesel, renewable diesel and renewable jet fuel, camelina oil is starting to become a potential resource for the production of novel agrochemicals.