Well-established doubled haploid (DH) plant production methods enhance the efficiency of modern plant breeding programs. In vitro anther culture is one biotechnological tool that can produce homozygous lines in one generation, while conventional breeding programs require more generations to develop genetically pure lines. In this chapter, an efficient protocol for DH rice production via anther culture is described after a cool pretreatment for 3-5 days at 10°C. We combined the beneficial effects of the "N6NDK" induction medium and the "MSNBK1" regeneration medium to obtain healthy green DH plantlets from japonica and indica rice genotypes. These plantlets can be introduced directly into research or breeding projects after a successful acclimatization in glasshouses and under field conditions.

Gastrointestinal diseases, including irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD), which encompasses Crohn's disease (CD) and ulcerative colitis (UC), significantly impact the quality of life of affected individuals. While IBS is a functional disorder without structural changes in the digestive tract, IBD is manifested in chronic and immune-mediated intestinal alterations. Globally, IBS is 10 times more prevalent than IBD, with IBS accounting for 11% of the population and contributing to a substantial proportion of gastroenterology consultations. In Hungary, approximately 15–20% of the adult population experiences gastrointestinal symptoms. Dietary approaches, such as the low-FODMAP diet, have been validated to alleviate symptoms in IBS patients and offer potential benefits for IBD patients with IBS-like symptoms. This randomised, single-blind, crossover study evaluated the effects of a low-FODMAP long-fermented yeast spelt bread compared to a gluten-free bread on symptoms and quality of life in individuals with IBS and IBD. The study highlights the potential for low-FODMAP long-fermented yeast spelt bread to serve as a practical and palatable alternative to gluten-free bread, emphasising the importance of preparation methods and ingredient selection. While promising, further research with larger sample sizes is needed to establish the broader applicability of these findings.

Drought stress is a major environmental constraint that significantly reduces wheat productivity worldwide. In this study, seventeen wheat genotypes were evaluated under well-watered and drought-stressed conditions across two consecutive years (2023–2024) in a controlled greenhouse experiment. Twenty morphological and agronomic traits were recorded, and their responses to prolonged water limitation were assessed using multivariate statistical methods, including three-way ANOVA, principal component analysis (PCA), and cluster analysis. Drought stress significantly decreased all traits except the harvest index (HI), with the most severe reductions observed in traits related to secondary spikes (e.g., grain weight reduced by 95%). The ANOVA results confirmed significant genotype × treatment (G × T) interactions for key agronomic traits, with the strongest effect observed for total grain weight (F = 7064.30, p < 0.001). A PCA reduced the 20 original variables to five principal components, explaining 87.2% of the total variance. These components reflected distinct trait groups associated with productivity, spike architecture, and development in phenology. Cluster analysis based on PCA scores grouped genotypes into three clusters with contrasting drought response profiles. A yield-based evaluation confirmed the cluster structure, distinguishing genotypes with a stable performance (average yield loss ~58%) from highly sensitive ones (~70% loss). Overall, the findings demonstrate that drought tolerance in wheat is governed by complex trait interactions. Integrating a trait-based multivariate analysis with a yield stability assessment enables the identification of genotypes with superior adaptation to water-limited environments, providing an excellent genotype background for future breeding efforts.

Remote sensing is the process of detecting and monitoring a plant's characteristics by measuring its reflected and emitted radiation at a distance, typically from a satellite or aircraft. The handheld leaf spectrometers help validate these images at the field scale. This dataset was captured by the CI-710s SpectraVue Leaf Spectrometer (Cid-Bioscience, Camas, WA, U.S.A.). The absorbance, reflectance, and transmittance of albino plants were measured under natural cold stress in a temperate rice-growing area [1]. The experiment was carried out in field conditions at the seedling stage. The chlorophyll degradation takes place, starting with the yellowing of the leaf until plant death. Albinos and different level of leaf colour mutants are very useful for research and as well as breeding [2]. The symptoms of cool-temperature-induced chlorosis (CTIC) are widely examined in higher plants, especially in rice [3]. Beside laboratory induction, CTIC is appearing natural low temperature in early spring, especially cold-sensitive genotypes, such as indica rice cultivars (e.g ‘Dular’) [4]. The dataset contains raw data from 400 nm to 1100nm with the wavelength data increment of 0.6nm [5]. These data may provide reliable support to researcher and breeder to make a simple comparison of the extent of chlorophyll degradation.

The efficient doubled haploid (DH) plant production methods play a key role in accelerating the breeding of new varieties and hybrids in cultivated plants. Consequently, DH plant production methods are continuously improving for barley (Hordeum vulgare L.) breeding and research programs. Two plant regeneration (FHGR and K4NB) and three rooting media (MSr, N6I and ½N6I + Ca) were compared with four F1 barley cross-combinations to clarify the effect of medium on the regeneration of green and albino plantlets and acclimatization. The plant regeneration efficiency was higher using K4NB medium (74.53 green plantlets/100 anthers and 30.85 albino/100 anthers) compared to FHGR (55.77 green plantlets/100anthers and 21.32 albino/100 anthers). The percentage of acclimatization was highest when the K4NB regeneration medium was combined with the MSr rooting medium. Altogether, 61.83% of the anther culture-derived plantlets of 8 cross-combinations acclimatized to the greenhouse conditions, and 1403 acclimatized plantlets were produced from the F1 cross-combinations. Haploid (22.52%), diploid (69.37%) and tetraploid (8.11%) plantlets were identified among the 111 tested green plantlets by flow cytometric analyses. The tetraploid lines can be explored to offer new scopes for future barley research and breeding directions. Nearly one thousand DH plants have been integrated into our barley breeding program.

In this study, we examined over 200-year-old Ginkgo biloba L. specimens under different environmental conditions. The overall aim was to explore which factors influence their vitality and general fitness in urban environments and thus their ability to tolerate stressful habitats. In order to determine this, we used a number of different methods, including histological examinations (stomatal density and size) and physiological measurements (peroxidase enzyme activity), as well as assessing the air pollution tolerance index (APTI). The investigation of the genetic relationships between individuals was performed using flow cytometry and miRNA marker methods. The genetic tests revealed that all individuals are diploid, whereas the lus-miR168 and lus-miR408 markers indicated a kinship relation between them. These results show that the effect of different habitat characteristics can be detected through morphological and physiological responses, thus indicating relatively higher stress values for all studied individuals. A significant correlation can be found between the level of adaptability and the relatedness of the examined individuals. These results suggest that Ginkgo biloba L. is well adapted to an environment with increased stress factors and therefore suitable for use in urban areas.

Changes of water-soluble carbohydrate (WSC) content such as fructose, glucose, sucrose, maltose, nystose, raffinose, stachyose and fructan were analyzed in wheat kernels in Fusarium epidemic and non-epidemic seasons. In both season types, eight commercial fungicides were applied and three wheat varieties with differing Fusarium resistance were tested. In the epidemic year, the average total amount of WSC was above 1.6% which was 2 times higher than in the non-epidemic year (0.7%). Sucrose, maltose, raffinose and fructan components determined the increased WSC value, but the most substantial change was observed in maltose content where its average amount was 28 times higher in the epidemic year. Fungicide application also significantly increased all the carbohydrate components except maltose, where significant reduction was observed. WSC components had strong correlation with several farinograph or extensograph parameters, but only the maltose content showed positive strong correlation (r = 0.9) with deoxynivalenol (DON) toxin that was highly affected by the applied fungicide. The changes of WSC indicate altered carbohydrate synthesis along with abnormal degradation processes and thus have impaction on the baking features. It seems that the sugar metabolism interacts with DON synthesis and the results give important additional information to the altered metabolism of the attacked plant.

Anther culture is an efficient biotechnological tool in modern plant breeding programs to produce new varieties and parental lines in hybrid seed productions. However, some bottlenecks—low induction rate, genotype dependency, albinism—restrict the widespread utilization of in vitro anther culture in rice breeding, especially in Oryza sativa ssp. indica (indica) genotypes, while an improved efficient protocol can shorten the process of breeding. Three different induction media (N6NDK, N6NDZ, Ali-1) and four plant regeneration media (mMSNBK1, MSNBK3, MSNBKZ1, MSNBKZ2) were tested with five indica rice genotypes to increase the efficiency of in vitro androgenesis (number of calli and regenerated green plantlets). The production of calli was more efficient on the N6NDK medium with an average 88.26 calli/100 anthers and N6NDZ medium with an average of 103.88 calli/100 anthers as compared to Ali-1 with an average of 6.96 calli/100 anthers. The production of green plantlets was greater when calli was produced on N6NDK medium (2.15 green plantlets/100 anthers) compared when produced on to N6NDZ medium (1.18 green plantlets/100 anthers). Highest green plantlets production (4.7 green plantlets/100 anthers) was achieved when mMSNBK1 plant regeneration medium was used on calli produced utilizing N6NDK induction medium. In the best overall treatment (N6NDK induction medium and mMSNBK1 plant regeneration medium), four tested genotypes produced green plantlets. However, the genotype influenced the efficiency, and the green plantlets production ranged from 0.4 green plantlets/100 anthers to 8.4 green plantlets/100 anthers. The ploidy level of 106 acclimatized indica rice plantlets were characterized with flow cytometric analyses to calculate the percentage of spontaneous chromosome doubling. Altogether, 48 haploid-, 55 diploid-, 2 tetraploid- and 1 mixoploid plantlets were identified among the regenerant plantlets, and the spontaneous chromosome doubling percentage was 51.89%. Utilization of DH plants have been integrated as a routine method in the Hungarian rice breeding program. The tetraploid lines can be explored for their potential to offer new scopes for rice research and breeding directions in the future.

Nowadays, hyperspectral remote sensing data are widely used in nutrient management, crop yield forecasting and stress monitoring. These data can be acquired with satellites, drones and handheld spectrometers. In this research, handheld spectrometer data were validated by chlorophyll-a fluorescence measurements under natural cold stress. The performance of 16 rice cultivars with different origins and tolerances was monitored in the seedling stage. The studies were carried out under field conditions across two seasons to simulate different temperature regimes. Twenty-four spectral indices and eleven rapid chlorophyll-a fluorescence parameters were compared with albino plants. We identified which wavelengths are affected by low temperatures. Furthermore, the differences between genotypes were characterized by certain well-known and two newly developed (AAR and RAR) indices based on the spectral difference between the genotype and albino plant. The absorbance, reflectance and transmittance differences from the control are suitable for the discrimination of tolerant-sensitive varieties, especially based on their shape, peak and shifting distance. The following wavelengths are capable of determining the tolerant varieties, namely 548-553 nm, 667-670 nm, 687-688 nm and 800-950 nm in case of absorbance; above 700 nm for reflectance; and the whole spectrum (400-1100 nm) for transmittance.

In cereal breeding, in vitro androgenesis methods are frequently applied to achieve doubled haploid (DH) plants. The aim of this study was to determine the effects of genotype (three registered varieties and eight F1 crossing combinations) and induction medium (W14mf and P4mf) on anther cultures (ACs) of triticale (×Triticosecale Wittmack). Androgenesis was induced in the treatment of each tested genotype, and the genotype significantly influenced the efficiency of AC, including in embryo-like structures (ELSs), albinos, green plantlets, and transplanted plantlets. The utilized medium also had a significant effect on the number of ELSs, albinos, and transplanted plantlets. Both media were suitable for AC in triticale DH plant production. The efficiency of AC was higher when using the P4mf medium (103.7 ELS/100 anthers, 19.7 green plantlets/100 anthers) than when using the W14mf medium (90.0 ELS/100 anthers, 17.0 green plantlets/100 anthers). However, the green plantlet regeneration efficiency of microspore-derived structures was 18.0% when using the W14mf medium, while this value was 15.9% in the case of ELSs induced with the P4mf medium. After nursery seed evaluation and propagation (DH1), the genetic homogeneity of the offspring generation (DH2) was tested using a molecular genetic method. Most of the tested DH lines showed homogeneity and were progressed into a breeding program after agronomic selection. Some DH lines showed inhomogeneity, which could be explained by the outcross inclination of triticale. We would like to call breeders’ attention to the outcross character of triticale and emphasize the vigilant propagation and maintenance of the triticale DH lines in breeding programs. Due to the outcross nature of triticale, even in self-pollinated genotypes, breeders should focus on careful maintenance, along with isolation in the case of line propagations, in triticale breeding programs.

Rice production is affected by several environmental factors, such as cold, salinity and drought stress. These unfavourable factors could have a serious impact on germination as well as on later growth, causing many types of damage. Recently, polyploid breeding can offer an alternative opportunity to enhance the yield and abiotic stress tolerance in rice breeding. This article describes some germination parameters of 11 different autotetraploid breeding lines and their parental lines under different environmental stresses. Each genotype was grown in a climate chamber under controlled conditions: 13 °C for 4 weeks in the cold test and 30/25 °C for 5 days in control, salinity (150 mM NaCl) and drought (15% PEG 6000) treatments, respectively. The germination process was monitored throughout the experiment. The average data were calculated using three replicates. This dataset contains germination raw data and three calculated germination parameters, such as median germination time (MGT), final germination percentage (FGP), and germination index (GI). These data may provide reliable support to clarify whether the tetraploid lines can exceed the performance of their diploid parental lines under germination phase or not.

Einkorn ( Triticum monococcum L.) can be applied as a model species for cereal genomic studies due to its small genome size and high level of polymorphism. The in vitro somatic tissue culture protocol in einkorn was significantly improved recently, however the in vitro androgenesis remained an unresolved research topic. Five different pre-treatments were compared to study the effects of stress pre-treatments on the efficiency of androgenesis in two einkorn genotypes. The long cold pre-treatment (2 weeks, 4 °C) of donor tillers increased significantly the number of microspore derived embryo-like structures (ELS). Green and albino plantlets were regenerated from these structures. The ploidy level of microspore-derived green plantlet was determined as haploid by flow cytometric analyses. This is the first report published on the successful androgenesis induction (ELS production) and green- and albino plantlet regeneration in in vitro anther culture of the recalcitrant einkorn wheat ( Triticum monococcum L.).

Detailed experiments were performed to investigate the effects of the marginal cultivation of temperate japonica rice on salinity tolerance in different developmental stages and to detect new genetic resources for further research and breeding projects. Eight rice varieties were screened for salinity tolerance in a greenhouse at two developmental stages. During seedling-stage screening, stress was induced with a 12 dS m−1 NaCl solution. For reproductive-stage analysis, we applied continuous salinity stress (8.39 dS m−1 of soil extract at 25 °C (EC25)). We compared the marginal rice genotypes (japonica_1) to widely grown controls (those being Mediterranean japonica genotypes, referred as the japonica_2 group, which also included one indica genotype). Without stress, marginally cultivated genotypes had significantly higher potassium storage capacity (406 µmol g−1) in flag leaves than did japonica_2 (347) or indica (267). Moreover, the seeds contained twice as much sodium (5.27) as did the others (2.19 and 2.00). Based on root-to-grain allocation, we identified two sodium excluders (‘Dular’ and ‘Dáma’) and six includers under constant salinity stress. Among the includers, we found that ‘M488’ had extremely high sodium concentration in both the flag leaves (250.54 µmol g−1) and the grains (13.57 µmol g−1). Rice varieties bred for the edge of their habitat show special physiological patterns. The differences are mainly manifested as lower levels of antioxidant pigments, higher amounts of potassium in flag leaves, and higher sodium levels in grains under normal conditions. With the onset of stress, higher tillering ability, enhanced antioxidant pigment synthesis, and rapid potassium translocation from the root are the key mechanisms which help the plants to avoid serious damage caused by salinity.

Doubled haploid (DH) plant production, such as anther culture (AC), is an effective tool used in modern rice breeding programs. The improved efficient protocols applied can shorten the process of breeding. The effect of combinations of plant growth regulators (2.5 mg/L NAA, 1 mg/L 2,4-D and 0.5 mg/L kinetin; 2 mg/L 2,4-D and 0.5 mg/L BAP) in the induction medium were compared in AC for five rice breeding materials and combinations. Induction of calli ranged from 264.6 ± 67.07 to 468.8 ± 123.2 calli/100 anthers in AC of rice genotypes. Two basal media (MS and N6) and two combinations of growth regulators (1 mg/L NAA, 1 mg/L BAP and 1 mg/L kinetin; 1.5 mg/L BAP, 0.5 mg/L NAA and 0.5 mg/L kinetin) were used as regeneration media. The in vitro green plant production was the highest with the application of the N6NDK induction medium (NAA, 2,4-D and kinetin) and the MS-based regeneration medium (1 mg/L NAA, 1 mg/BAP and 1 mg/L kinetin) in anther culture of the ‘1009’ genotype (95.2 green plantlets/100 anthers). The mean of five genotypes was 24.48 green plantlets/100 anthers for the best treatment. Flow cytometric analyses conducted identified the microspore origin of the haploid calli produced in AC, while the uniformity of spontaneous DH plants was checked in the DH1 and DH2 generations. Spontaneous chromosome doubling ranged from 38.1% to 57.9% (mean 42.1%), depending on the breeding source. The generated and selected DH lines were tested in micro- and small-plot field experiments to identify promising lines for a pedigree breeding program. The improved AC method was integrated in a Hungarian temperate rice pedigree breeding program.

The Pannonian Plain, as the most productive region of Southeast Europe, has a long tradition of agronomic production as well as agronomic research and plant breeding. Many research institutions from the agri-food sector of this region have a significant impact on agriculture. Their well-developed and fruitful breeding programmes resulted in productive crop varieties highly adapted to the specific regional environmental conditions. Rapid climatic changes that occurred during the last decades led to even more investigations of complex interactions between plants and their environments and the creation of climate-smart and resilient crops. Plant phenotyping is an essential part of botanical, biological, agronomic, physiological, biochemical, genetic, and other omics approaches. Phenotyping tools and applied methods differ among these disciplines, but all of them are used to evaluate and measure complex traits related to growth, yield, quality, and adaptation to different environmental stresses (biotic and abiotic). During almost a century-long period of plant breeding in the Pannonian region, plant phenotyping methods have changed, from simple measurements in the field to modern plant phenotyping and high-throughput non-invasive and digital technologies. In this review, we present a short historical background and the most recent developments in the field of plant phenotyping, as well as the results accomplished so far in Croatia, Hungary, and Serbia. Current status and perspectives for further simultaneous regional development and modernization of plant phenotyping are also discussed.

Hungary is northernmost temperate rice growing country in Europe. One of the main limiting factors is low temperature, especially at germination and seedling developmental stages. In early developmental stages, low temperature can impair and delay germination, as well as have negative impacts on seedling growth, causing poor stand establishment and non-uniform crop maturation [1]. Temperatures lower than 15 °C are generally detrimental for germination [2] under filed conditions for establishment of the crop. This article describes some key germination parameters of 165 rice accessions including breeding lines and varieties. Each genotype was grown in three replicates in a controlled cabinet under 13 °C for 4 weeks’ duration. Growth was measured every 7th day. Growth traits such as coleoptile and radicle length were measured at the end of the experiment. The average data were calculated for three replicates. This dataset contains germination raw data and five germination parameters such as median germination time (MGT), final germination percentage (FGP), germination index (GI), coleoptile length (CL) and radicle length (RL). These data may provide reliable support for researchers and breeders to select the right rice genotypes for low temperature conditions.

Aerobic rice production is an alternative growing method to reduce water consumption of rice and thus increase the water productivity of the system without a significant reduction of yield and quality. Evapotranspiration (ETc) of a Hungarian rice variety, ‘SZV Tünde’ under aerobic conditions was measured in large weighing lysimeter during the growing season in 2020. In our experiment, 506.7 g/m2 grain yield and a total above-ground biomass of 1140.4 g/m2 were produced with the application of 315.6 mm of irrigation. Water use-efficiency (WUE) based on the water input and the grain yield was 0.65 g/L. Total ETc for the whole season was measured as 648.3 mm. However, ETc values were ranged 2.04-3.86 mm/day, 3.57-7.90 mm/day and 0.90-4.26 mm/day at the initial, mid and end stages, respectively. Crop coefficients for the different periods of the season were calculated as Kcini=0.82, Kcmid=1.40 and Kcend=0.77. Negative effects of drought can seriously damage rice crop; therefore irrigation scheduling has significant role in successful aerobic rice cultivation. Reliable estimation of evapotranspiration rate in different crop developmental stages can promote this goal.

The efficiency of in vitro anther culture (AC) method on six common wheat (Triticum aestivum L.) F3-6 bulk combinations was investigated for the AC-induced androgenic traits, such as embryo-like structures (ELSs), regenerated plantlets, green plantlets, albinos, transplanted plantlets, and acclimatized plantlets. Moreover, genotype dependency was tracked among the tested combinations in the AC method. The results showed that all androgenic traits were produced in each tested combination and non-significant differences were found among combinations for all androgenic traits except albinos. The number of green plantlets ranged between 8.00 and 19.00 green plantlets/100 anthers based on the combination, with an overall mean of 14.37 green plantlets/100 anthers. The total number of acclimatized plantlets was 496 plantlets out of the 604 transplanted plantlets, with a rate of 82.12%. This investigation confirms the efficiency and utility of in vitro AC method in winter wheat to produce doubled haploid (DH) lines based on androgenesis for the breeding program and genetic studies. AC-induced DH lines produced subsequently from acclimatized plantlet individuals in the nursery will be inserted in breeding programs for wheat drought tolerance evaluation and in crossing programs for more improvement in green plants production in the nursery. Albinism and genotype dependency, limiting-factors for wheat AC-induced DH production, were mitigated by the application of some modifications of the wheat AC protocol. The modifications included a 2-week cold pre-treatment of donor tiller at 4? in constant dim light and the use of the W14 mf induction medium instead of P4 mf.

In vitro anther culture is the most commonly-used technology for doubled haploid production in wheat (Triticum aestivum L.) and various cereals. The technology application allows the genetic attainment of homozygous pure lines from heterozygous breeding material in a single generation, and has become a rapid alternative to the conventional breeding methods. This technology also contributes to more accurately assess QTL × environmental interactions and has been applied in: genetic researches for marker-trait association study, genomics and as a target for transformation, genetic engineering, gene mapping, and mapping of quantitative trait loci (QTLs). This paper reviews the most important factors that limit the efficiency of winter wheat (Triticum aestivum L.) anther culture, such as genotype dependency and albinism, in addition to other affecting factors including the collection time of tillers and physiological growth environments of plants. Factors that improve the efficiency of wheat anther culture in mitigating both genotypic dependency and albinism have been included in this review, too, for example, genetic improvements by following a strategy to realize this purpose through using responsive breeding material for crossing, application of convenient stress pre-treatments (cold pre-treatments, heat shock treatment, colchicine, hormones, and chemicals) to induce androgenesis, and improvement of the composition of anther culture media and culture conditions. We have also highlighted the methods of chromosome doubling for the haploid wheat produced by anther culture androgenesis, such as spontaneous chromosome doubling or use of colchicine and anti-mitotic chemicals at an early stage of anther culture or as root immersion treatment.