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Treatment of seeds with cold atmospheric pressure plasma (CAPP) is in its proof-of-concept phase with regard to its effect on germination and plant growth. To increase the germination of hardseeded red clover ( Trifolium pratense L.), seeds are usually scarified, which is time-consuming and labour-intensive. The aim of this study was to compare the effect of different CAPP devices (indirect treatment: plasma processed air, direct treatment: corona discharge, argon and air dielectric barrier discharge) on germination and early growth of different long-term stored red clover accessions and to determine whether germination can be increased to meet seed management requirements. Sixty different red clover seed lots (diverse accessions and harvest years) with different initial germination percentages were divided into three batches of 20 lots each and the effect of the different plasma treatments on germination and development were examined in laboratory and greenhouse. The overall results indicate a plasma discharge- and accession-depended enhancement of germination speed which was detected in all batches but most pronounced in Batch 1. While direct treatments, especially with corona discharge-plasma, increased germination speed (up to 58% germination seven days after sowing vs. 44% in control in laboratory conditions), treatment with plasma processed air resulted partially in reduced germination speed (42%). Despite a small but significant increase in total germination of maximum five percentage points, no treatment led to an increase from 62% or 70% in control (depending on experiment) to at least 80% germination percentage to meet storage requirements for seed banks. Stimulating effects on biomass of young plants under greenhouse cultivation conditions were observed in Batch 1, but were absent in Batch 2 and 3 and therefore inconclusive. Future research is needed to elucidate influencing factors on plasma effects in red clover seed lots which include but are not limited to the effect of seed coat compounds and seed coat thickness.

Legumes have a high demand for phosphorus (P) due to energetically costly biological nitrogen fixation, but they also have effective physiological and morphological strategies for P mobilization. To evaluate the inter- and intraspecific P efficiency of small-grain legumes supplied with different P recycling fertilisers, eight accessions each of alfalfa (Medicago sativa L.) and red clover (Trifolium pratense L.) were cultivated in two pot experiments under greenhouse conditions until the flowering stage. To substantiate the results, some accessions were used in both experiments. Five treatments (no P, triple-superphosphate (TSP), sewage sludge ash (SSA), biowaste compost (compost), and struvite) were considered P sources. In addition to plant P uptake, the soil P pools were analysed in detail. Red clover showed higher yields and nutrient uptakes compared to alfalfa, but intraspecific effects were marginal. The addition of P resulted only partly in an increase in yield, despite the low P content in the soil. While struvite application clearly enhanced the P uptake of the plants in both experiments, SSA application had no effect compared to the control. The same treatment effect occurs with the bio-available soil P contents, which were on average 72.6 mg kg−1 after struvite and 44.3 mg kg−1 after SSA addition. Struvite as a P source was especially effective when applied to red clover. Our study aligns with previous field results and underscores the high potential of P mobilization of small-grain legumes without pronounced inter- or intraspecific differences. While struvite is suitable as a P fertiliser, the application of SSA to legumes is not recommended.

Background Seed retention is the basic prerequisite for seed harvest. However, only little breeding progress has been achieved for this trait in the major forage grasses. The aim of this study was to evaluate the potential of plant genetic resources of the important fodder grasses Festuca pratensis Huds. and Lolium perenne L. as source for seed retention in the breeding process. Furthermore, the morphology of the abscission zone, where shattering occurs, was studied on the cell tissue level in different developmental stages of contrasting accessions. Results 150 and 286 accessions of Festuca pratensis and Lolium perenne were screened for seed retention, respectively. Contrasting accessions were selected to be tested in a second year. We found a great variation in seed retention in Festuca pratensis and Lolium perenne , ranging from 13 to 71% (average: 35%) and 12 to 94% (average: 49%), respectively, in the first year. Seed retention was generally lower in the second year. Cultivars were within the accessions with highest seed retention in Festuca pratensis , but had lower seed retention than ecotypes in Lolium perenne . Field-shattered seeds had a lower thousand grain weight than retained seeds. Cell layers of the abscission zone appeared already in early seed stages and were nested within each other in accessions with high seed retention, while there were two to three superimposed layers in accessions with low seed retention. Conclusions Plant genetic resources of Lolium perenne might be a valuable source for breeding varieties with high seed retention. However, simultaneous selection for high seed weight is necessary for developing successful commercial cultivars.

Boron (B) is an essential micronutrient for seed plants. Information on B-efficiency mechanisms and B-efficient crop and model plant genotypes is very scarce. Studies evaluating the basis and consequences of B-deficiency and B-efficiency are limited by the facts that B occurs as a trace contaminant essentially everywhere, its bioavailability is difficult to control and soil-based B-deficiency growth systems allowing a high-throughput screening of plant populations have hitherto been lacking. The crop plant shows a very high sensitivity toward B-deficient conditions. To reduce B-deficiency-caused yield losses in a sustainable manner, the identification of B-efficient genotypes is indispensable. We developed a soil substrate-based cultivation system which is suitable to study plant growth in automated high-throughput phenotyping facilities under defined and repeatable soil B conditions. In a comprehensive screening, using this system with soil B concentrations below 0.1 mg B (kg soil) , we identified three highly B-deficiency tolerant cultivars ( , , and ) among a genetically diverse collection comprising 590 accessions from all over the world. The B-efficiency classification of cultivars was based on a detailed assessment of various physical and high-throughput imaging-based shoot and root growth parameters in soil substrate or in conditions, respectively. We identified cultivar-specific patterns of B-deficiency-responsive growth dynamics. Elemental analysis revealed striking differences only in B contents between contrasting genotypes when grown under B-deficient but not under standard conditions. Results indicate that B-deficiency tolerant cultivars can grow with a very limited amount of B which is clearly below previously described critical B-tissue concentration values. These results suggest a higher B utilization efficiency of , , and which would represent a unique trait among so far identified B-efficient cultivars which are characterized by a higher B-uptake capacity. Testing various other nutrient deficiency treatments, we demonstrated that the tolerance is specific for B-deficient conditions and is not conferred by a general growth vigor at the seedling stage. The identified B-deficiency tolerant cultivars will serve as genetic and physiological \"tools\" to further understand the mechanisms regulating the B nutritional status in rapeseed and to develop B-efficient elite genotypes.

With the ongoing cost decrease of genotyping and sequencing technologies, accurate and fast phenotyping remains the bottleneck in the utilizing of plant genetic resources for breeding and breeding research. Although cost-efficient high-throughput phenotyping platforms are emerging for specific traits and/or species, manual phenotyping is still widely used and is a time- and money-consuming step. Approaches that improve data recording, processing or handling are pivotal steps towards the efficient use of genetic resources and are demanded by the research community. Therefore, we developed PhenoApp, an open-source Android app for tablets and smartphones to facilitate the digital recording of phenotypical data in the field and in greenhouses. It is a versatile tool that offers the possibility to fully customize the descriptors/scales for any possible scenario, also in accordance with international information standards such as MIAPPE (Minimum Information About a Plant Phenotyping Experiment) and FAIR (Findable, Accessible, Interoperable, and Reusable) data principles. Furthermore, PhenoApp enables the use of pre-integrated ready-to-use BBCH (Biologische Bundesanstalt für Land- und Forstwirtschaft, Bundessortenamt und CHemische Industrie) scales for apple, cereals, grapevine, maize, potato, rapeseed and rice. Additional BBCH scales can easily be added. The simple and adaptable structure of input and output files enables an easy data handling by either spreadsheet software or even the integration in the workflow of laboratory information management systems (LIMS). PhenoApp is therefore a decisive contribution to increase efficiency of digital data acquisition in genebank management but also contributes to breeding and breeding research by accelerating the labour intensive and time-consuming acquisition of phenotyping data.

Legumes have a high demand for phosphorus (P) but also have effective physiological and morphological strategies of P mobilisation. In order to evaluate the inter- and intraspecific P efficiency of small-grain legumes under contrasting long-term P management, eight accessions each of alfalfa (Medicago sativa L.) and red clover (Trifolium pratense L.) were cultivated in two consecutive growing periods from 2020 to 2021 in a field trial established in 1998. Six treatments (no P, triple-superphosphate (TSP), biomass ash, cattle manure, biowaste compost, and biowaste compost + TSP) were considered as P sources. While the yield clearly varied between both growing seasons, the differences between alfalfa and red clover were relatively small (4.7 vs. 4.9 Mg ha−1 in 2020 and 12.0 vs. 10.5 Mg ha−1 in 2021, p < 0.05). Even after more than 20 years of P management, crop yields were hardly affected by mineral P sources (TSP and biomass ash) while organic fertilisers increased the yields and nutrient uptake of plants and also raised soil P pools and the activities of soil enzymes in comparison to the control. A relevant crop effect was only found for the nitrogen (N) leaching with higher mineral N contents in 60 to 90 cm soil depth measured for red clover compared to alfalfa (11.8 vs. 4.8 kg ha−1, p < 0.05). Our results emphasise the high P efficiency of small-grain legumes without pronounced inter- or intraspecific differences. The yield-enhancing effect of organic amendments was related to higher soil fertility rather than to P supply.

The natural genetic diversity of agricultural species is an essential genetic resource for breeding programs aiming to improve their ecosystem and production services. A large natural ecotype diversity is usually available for most grassland species. This could be used to recombine natural climatic adaptations and agronomic value to create improved populations of grassland species adapted to future regional climates. However describing natural genetic resources can be long and costly. Molecular markers may provide useful information to help this task. This opportunity was investigated for Lolium perenne L., using a set of 385 accessions from the natural diversity of this species collected right across Europe and provided by genebanks of several countries. For each of these populations, genotyping provided the allele frequencies of 189,781 SNP markers. GWAS were implemented for over 30 agronomic and/or putatively adaptive traits recorded in three climatically contrasted locations (France, Belgium, Germany). Significant associations were detected for hundreds of markers despite a strong confounding effect of the genetic background; most of them pertained to phenology traits. It is likely that genetic variability in these traits has had an important contribution to environmental adaptation and ecotype differentiation. Genomic prediction models calibrated using natural diversity were found to be highly effective to describe natural populations for almost all traits as well as commercial synthetic populations for some important traits such as disease resistance, spring growth or phenological traits. These results will certainly be valuable information to help the use of natural genetic resources of other species.

Although oilseed rape has become one of the most important oil crops in Europe, little is known regarding the viability of its seed under conditions of long-term storage. We report here an examination of oilseed rape seed longevity performed on a set of 42 accessions housed at the German ex situ genebank at IPK, Gatersleben. A comparison of germination between the accessions stored for 26 years showed that viability was in part genetically determined, since it ranged between 42 and 98%. An attempt was made to define the genetic basis of viability by subjecting a mapping population of doubled haploids to three artificial ageing treatments. Quantitative trait loci (QTL) were detected on six chromosomes: N6, N7, N8, N15, N16 and N18. The chromosomal locations of these QTL were compared with their syntenic regions in Arabidopsis thaliana in order to explore what genes might underlie genetic variation for longevity.

1. Climate extremes are expected to increase in frequency and magnitude as a consequence of global warming. 2. Managed permanent grasslands cover a large surface in Europe and contribute substantially to agricultural production. These managed plant communities are dominated by perennial clonal species. Their capacity to adapt to rapidly changing environmental conditions may be limited. 3. We hypothesize that those plant populations that have already been exposed to conditions that are expected to occur due to future climate change, particularly conditions that would be 'extreme' in the target area, are able to cope better with these conditions. 4. For a common-garden experiment we selected ecotypes (provenances as supported by accessions in seed banks) of important European grass species: Arrhenatherum elatius, Festuca pratensis, Holcus lanatus and Alopecurus pratensis. Southern target locations of ecotypes (populations) were identified based on climate model projections for the local site in Northern Bavaria, Germany. 5. In a controlled experiment, the plants were exposed to warming and extreme drought. Drought conditions(16—19) days, depending on the species) were imposed starting from the end of May in combination with and without an increase in the average temperature from May to September 2009 (+ 1.5 K compared with control; + 2.5 K compared with ambient conditions outside of the experimental units). 6. Ecotypes and drought manipulation had significant impacts on biomass production and tissue die-back. Significant interactions between ecotype and drought indicated a different drought tolerance of the ecotypes in some cases. The warming treatment yielded a less significant response. The local ecotype generally did not perform significantly worse than the presumably better-adapted southern ecotypes. 7. Synthesis. The selection of ecotypes that are adapted to more extreme climatic conditions could be an option for maintaining future ecosystem functioning in temperate managed grasslands, as was indicated by the clear differences between ecotypes in our experiment. Based on our data, however, performance cannot be predicted from climatic origin. Therefore, we recommend enhancing the genetic variability within populations of species in general.

The occurrence of race-specific resistance genes to the stem canker fungus, Leptosphaeria maculans, was analysed in 453 accessions of B. napus, mainly originating from the Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK) GeneBank. Major resistance genes Rlm1, Rlm2, Rlm4 and the putative RlmBBA gene were investigated using genetically improved strains of the fungus harbouring as few corresponding avirulence genes as possible. In addition, a screening with fully virulent isolates was used to uncover novel resistance sources. Major resistance genes were rarer in frequency and diversity in spring-type cultivars compared to winter types. In the former, 65.7% of the accessions were fully susceptible to all isolates, whereas only 12.2% of the winter types were devoid of at least one R gene. In spring cultivars, the most common R gene, Rlm4 was found in 26.6% of accessions, whereas the other R genes were rare. In winter cultivars, the most common R genes were Rlm2 (more than 45.9–54.0% of the accessions) and Rlm4 (26.4–27.7% of the genotypes). In winter types however, the improvement of the quality of oils, through the generation of single- and double-low genotypes improved the homogeneity of the cvs, whereas it impoverished R gene diversity, including the loss of complete resistance that was harboured by 18.4% of the less advanced accessions, and a reduction in the ratio of accessions harbouring Rlm1. Correlation between the R gene(s) present in the accessions and their field resistance is discussed.