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‘Speed breeding’ (SB) shortens the breeding cycle and accelerates crop research through rapid generation advancement. SB can be carried out in numerous ways, one of which involves extending the duration of plants’ daily exposure to light, combined with early seed harvest, to cycle quickly from seed to seed, thereby reducing the generation times for some long-day (LD) or day-neutral crops. In this protocol, we present glasshouse and growth chamber–based SB approaches with supporting data from experimentation with several crops. We describe the conditions that promote the rapid growth of bread wheat, durum wheat, barley, oat, various Brassica species, chickpea, pea, grass pea, quinoa and Brachypodium distachyon. Points of flexibility within the protocols are highlighted, including how plant density can be increased to efficiently scale up plant numbers for single-seed descent (SSD). In addition, instructions are provided on how to perform SB on a small scale in a benchtop growth cabinet, enabling optimization of parameters at a low cost.

Melatonin (MT; N-acetyl-5-methoxytryptamine) is a pleiotropic signaling molecule that has been demonstrated to play an important role in plant growth, development, and regulation of environmental stress responses. Studies have been conducted on the role of the exogenous application of MT in a few species, but the potential mechanisms of MT-mediated stress tolerance under salt stress are still largely unknown. In this study, naked oat seedlings under salt stress (150 mM NaCl) were pretreated with two different concentrations of MT (50 and 100 μM), and the effects of MT on the growth and antioxidant capacity of naked oat seedlings were analyzed to explore the regulatory effect of MT on salt tolerance. The results showed that pretreating with different concentrations of MT promoted the growth of seedlings in response to 150 mM NaCl. Different concentrations of MT reduced hydrogen peroxide, superoxide anion, and malondialdehyde contents. The exogenous application of MT also increased superoxide dismutase, peroxidase, catalase, and ascorbate peroxide activities. Chlorophyll content, leaf area, leaf volume, and proline increased in the leaves of naked oat seedlings under 150 mM NaCl stress. MT upregulated the expression levels of the lipid peroxidase genes lipoxygenase and peroxygenase, a chlorophyll biosynthase gene (ChlG), the mitogen-activated protein kinase genes Asmap1 and Aspk11, and the transcription factor genes (except DREB2), NAC, WRKY1, WRKY3, and MYB in salt-exposed MT-pretreated seedlings when compared with seedlings exposed to salt stress alone. These results demonstrate an important role of MT in the relief of salt stress and, therefore, provide a reference for managing salinity in naked oat.

Plants are subjected to various biotic and abiotic stresses that significantly impact their growth and productivity. To achieve balanced crop growth and yield, including for leafy vegetables, the continuous application of micronutrient is crucial. This study investigates the effects of different concentrations of copper sulphate (0, 75, 125, and 175 ppm) on the morphological and biochemical features of Spinacia oleracea and Avena sativa . Morphological parameters such as plant height, leaf area, root length, and fresh and dry weights were optimized at a concentration of 75 ppm copper sulfate. At this concentration, chlorophyll a & b levels increased significantly in Spinacia oleracea (462.9 and 249.8 𝜇𝑔/𝑔), and Avena sativa (404.7 and 437.63𝜇𝑔/𝑔). However, carotenoid content and sugar levels in Spinacia oleracea were negatively affected, while sugar content in Avena sativa increased at 125 ppm (941.6 µg/ml). Protein content increased in Spinacia oleracea (75 ppm, 180.3 µg/ml) but decreased in Avena sativa . Phenol content peaked in both plants at 75 ppm (362.2 and 244.5 µg/ml). Higher concentrations (175 ppm) of copper sulfate reduced plant productivity and health. Plants exposed to control and optimal concentrations (75 and 125 ppm) of copper sulpate exhibited the best health and growth compared to those subjected to higher concentrations. Maximum plant height, leaf area, root length, fresh and dry weights were observed at lower concentrations (75 and 125 ppm) of copper sulfate, while higher concentrations caused toxicity. Optimal copper sulfate levels enhanced chlorophyll a, chlorophyll b, total chlorophyll, protein, and phenol contents but inhibited sugar and carotenoid contents in both Spinacia oleracea and Avena sativa . Overall, increased copper sulfate treatment adversely affected the growth parameters and biochemical profiles of these plants.

Melatonin (N-acetyl-5-methoxytryptamine, MT) is a molecule with pleiotropic effects including antioxidant activity, regulated plant growth, development, and reduced environmental stress in plants. However, only a few studies have analyzed the effect of exogenous MT on drought stress in naked oat seedlings. Therefore, in this study, we studied the effects of exogenous MT on the antioxidant capacity of naked oat under drought stress to understand the possible antioxidant mechanism. The results showed that a pretreatment of 100 μM MT reduced the hydrogen peroxide (H2O2) and superoxide anion (O2−•) contents. MT also enhanced superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) activities in the leaves of naked oat seedlings under 20% PEG-6000 drought stress. MT upregulated the expression levels of the mitogen-activated protein kinases (MAPKs) Asmap1 and Aspk11, and the transcription factor (TF) genes (except for NAC), WRKY1, DREB2, and MYB increased in drought with MT pretreatment seedlings when compared with seedlings exposed to drought stress alone. These data indicated that the MT-mediated induction of the antioxidant response may require the activation of reactive oxygen species (ROS) and MAPK, followed by triggering a downstream MAPK cascade such as Asmap1 and Aspk11, to regulate the expression of antioxidant-related genes. This study demonstrated that MT could induce the expression of MAPKs and TFs and regulate the expression of downstream stress-responsive genes, thereby increasing the plant’s tolerance. This may provide a new idea for MT modulation in the regulation of plant antioxidant defenses. These results provide a theoretical basis for MT to alleviate drought stress in naked oat.

Avena species, especially A . fatua , A . sterilis and A . ludoviciana , are among the most problematic weed species in many crops worldwide. The growth cycles of these three species could be helpful in understanding their growth cycle and their implications for agriculture and weed management. The growth cycles of these species were studied altogether or in combination with any single or double combinations of the other species in cereal fields in Türkiye, using two populations of each in a common garden experiment in Bornova district, Izmir, Türkiye. Germination and growth experiments were conducted in the laboratory and screen house, respectively. Various phenological parameters were recorded during the experiment and data were analyzed using R software. There were no significant differences in germination, emergence, SPAD values, leaf width, plant height, or plant dry weight among the species or populations. The SPAD values and width of the flag leaf and the leaf before the flag leaf were strongly correlated. Plant weight increased with increasing tiller number. The length of the ligule in a population of A . sterilis was significantly greater than that in populations of two other species, and it was concluded that the species is not A . fatua or A . ludoviciana if the length of the ligule is greater than 10 mm. The length of the spikelets of A . sterilis was greater than 65 mm with awn and greater than 35 mm without awn; these values were significantly greater than those of two other species that were shorter than 55 with awn for A . fatua and 30 mm without awn for A . ludoviciana , respectively. Avena ludoviciana had fewer tillers than the other two species. The plants emerged at 37.58 GDD at the soil surface temperature, which corresponds to 7 days after sowing. The growing cycles of the species differed: 196 days for A . sterilis , 201 days for A . fatua , and 209 days for A . ludoviciana after emergence, although there were no clear differences in earlier growth stages. This study provides initial basic information about the Avena spp., and it is concluded that even if a field has mixed Avena populations, herbicides can be applied simultaneously because the early development stages of the three species are very similar. In future prospects, there is a need for proper studies about the management of all Avena spp. on the basis of growth stages and growing degree days in regional context.

Avena fatua and A . ludoviciana (commonly known as wild oats) are the most problematic winter grass species in fallows and winter crops in the northeast region of Australia. A series of experiments were conducted to evaluate the performance of glyphosate and alternative post-emergence herbicides on A . fatua and A . ludoviciana . This study reports the world’s first glyphosate-resistant (GR) biotypes of A . fatua and A . ludoviciana . The glyphosate dose required to kill 50% of the plants (LD 50 ) and to reduce 50% of the biomass (GR 50 ) for the GR biotype of A . fatua was 556 g a.e./ha and 351 g a.e./ha, respectively. These values for A . ludoviciana were 848 g a.e./ha and 289 g a.e./ha. Regardless of the growth stage (3–4 or 6–7 leaf stages), clethodim (120 g a.i./ha), haloxyfop (78 g a.i./ha), pinoxaden (20 g a.i./ha), and propaquizafop (30 g a.i./ha) were the best alternative herbicide options for the control of A . fatua and A . ludoviciana . The efficacy of butroxydim (45 g a.i./ha), clodinafop (120 g a.i./ha), imazamox + imazapyr (36 g a.i./ha), and paraquat (600 g a.i./ha) reduced at the advanced growth stage. Glufosinate (750 g a.i./ha), flamprop (225 g a.i./ha), and pyroxsulam + halauxifen (20 g a.i./ha) did not provide effective control of Avena species. This study identified alternative herbicide options to manage GR biotypes of A . fatua and A . ludoviciana .

Background The high fat content in oat seeds makes them susceptible to aging during storage, leading to reduced seed vigor, delayed germination, and even seed death. Much evidence suggests that lipid remodeling is closely associated with successful seed germination. However, the dynamic behavior and response mechanisms of lipids during the germination of aged oat seeds remain unclear. In this study, ‘Monida’ (aging-tolerant) and ‘Haywire’ (aging-sensitive), were used to investigate the lipid profiles in the embryo and endosperm and the dynamic transcriptomic differences in the embryo during the germination. Results The results demonstrate that phospholipid alterations during the germination of aged seeds are more significant compared to unaged seeds, indicating that aging affects lipid remodeling during germination, particularly in the ‘Haywire’. Further analysis revealed that the most critical lipid response events occurred at the end of germination stage II (32 h) in embryo, primarily regulated through the PLC-DGK pathway to modulate phospholipid and glycerolipid molecules. Specifically, transcripts of PLC, DGK, and DGAT were upregulated, promoting the generation of diacylglycerol (DG) from various phospholipids, which further increased the monogalactosyldiacylglycerol/digalactosyldiacylglycerol (MGDG/DGDG) ratio, thereby influencing membrane repair. Additionally, at 6 h of germination in aged seeds, PC(3:0/0:0) levels significantly decreased. Compared to ‘Monida,’ the aging-sensitive ‘Haywire’ seeds exhibited substantial production of PE(19:0/0:0) and PC(15:0/0:0) at 32 h of germination, which may be key factors contributing to the seed's sensitivity to aging and the significant reduction in germination percentage after aging. Therefore, PC(3:0/0:0), PE(19:0/0:0), and PC(15:0/0:0) could serve as important lipid metabolic markers in future studies on the mechanisms of oat seed vigor. Conclusions The findings of this study provide insights into the specificity of lipid remodeling and its response mechanisms during the germination of aged oat seeds, providing a theoretical foundation for the safe preservation of oat germplasm and the development of aging-tolerant varieties.

A critical step in the production of doubled haploids is a conversion of the haploid embryos into plants. Our study aimed to recognize the reasons for the low germination rate of Avena sativa haploid embryos obtained by distant crossing with maize. Oat cultivars of ‘Krezus’ and ‘Akt’ were investigated regarding embryo anatomy, the endogenous phytohormone profiles, and antioxidant capacity. The zygotic embryos of oat were used as a reference. It was found that twenty-one days old haploid embryos were smaller and had a less advanced structure than zygotic ones. Morphology and anatomy modifications of haploid embryos were accompanied by extremely low levels of endogenous auxins. Higher levels of cytokinins, as well as tenfold higher cytokinin to auxin ratio in haploid than in zygotic embryos, may suggest an earlier stage of development of these former. Individual gibberellins reached higher values in ‘Akt’ haploid embryos than in the respective zygotic ones, while the differences in both types of ‘Krezus’ embryos were not noticed. Additionally to the hormonal regulation of haploid embryogenesis, the poor germination of oat haploid embryos can be a result of the overproduction of reactive oxygen species, and therefore higher levels of low molecular weight antioxidants and stress hormones.

Background Seed aging, a natural and inevitable process occurring during storage. Oats, an annual herb belonging to the Gramineae family and pooideae. In addition to being a healthy food, oats serve as ecological pastures, combating soil salinization and desertification. They also play a role in promoting grassland agriculture and supplementing winter livestock feed. However, the high lipid and fat derivatives contents of oat seeds make them susceptible to deterioration, as fat derivatives are prone to rancidity, affecting oat seed production, storage, development, and germplasm resource utilization. Comparative studies on the effects of aging on physiology and cytological structure in covered and naked oat seeds are limited. Thus, our study aimed to determine the mechanism underlying seed deterioration in artificially aged ‘LongYan No. 3’ ( A. sativa ) and ‘BaiYan No. 2’ ( A. nuda ) seeds, providing a basis for the physiological evaluation of oat seed aging and serving as a reference for scientifically safe storage and efficient utilization of oats. Results In both oat varieties, superoxide dismutase and catalase activities in seeds showed increasing and decreasing trends, respectively. Variance analysis revealed significant differences and interaction in all measured indicators of oat seeds between the two varieties at different aging times. ‘LongYan No. 3’ seeds, aged for 24–96 h, exhibited a germination rate of < 30%, Conductivity, malondialdehyde, soluble sugar, and soluble protein levels increased more significantly than the ‘BaiYan No. 2’. With prolonged aging leading to cell membrane degradation, reactive oxygen species accumulation, disrupted antioxidant enzyme system, evident embryo cell swelling, and disordered cell arrangement, blocking the nutrient supply route. Simultaneously, severely concentrated chromatin in the nucleus, damaged mitochondrial structure, and impaired energy metabolism were noted, resulting in the loss of ‘LongYan No. 3’ seed vitality and value. Conversely, ‘BaiYan No. 2’ seeds showed a germination rate of 73.33% after 96 h of aging, consistently higher antioxidant enzyme activity during aging, normal embryonic cell shape, and existence of the endoplasmic reticulum. Conclusions ROS accumulation and antioxidant enzyme system damage in aged oat seeds, nuclear chromatin condensation, mitochondrial structure damage, nucleic acid metabolism and respiration weakened, oat seed vigor decreased. ‘LongYan No. 3’ seeds were more severely damaged under artificial aging than ‘BaiYan No. 2’ seeds, highlighting their heightened susceptibility to aging effects.

Naked oats ( Avena nuda ) are a key cereal crop that are valued for their high nutritional content and desirable agronomic traits. In this study, 80 developed strains and three bred varieties were evaluated in Kelan County, Shanxi Province, China, from 2022 to 2023. Six agronomic, eight physical, and five chemical traits were assessed. Significant phenotypic variations were observed across the 83 strains, with marked improvements in agronomic traits in 2023, particularly in grain shape, thousand grain weight, and spike length. The grain shape showed the most substantial improvement, whereas the thousand grain weight remained relatively stable. Cluster analysis identified three agronomic groups, with Group III comprising high-yield, stable strains, such as YM49. Regarding the physical traits, a general reduction in grain size and fullness was observed in 2023, with hardness exhibiting the greatest variation. Principal component analysis classified the strains into three morphological groups, with YM23 representing a large-grain, soft-textured type. Chemical quality traits indicated an increase in β -glucan and water content in 2023, whereas fat content decreased while exhibiting high variability. Nutritional profiling clustered the strains into three groups, with YM39 leading in protein, β -glucan, and fat content. Correlation analysis across the selected 19 traits highlighted the key tradeoffs between grain morphology, yield, and nutritional quality. Effective tillering was identified as a crucial trait that influences the overall performance. Strain YM49 demonstrated a consistent high-ranking performance across multiple traits, indicating its potential for breeding high-quality oats with high yields.