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Lolium perenne (L.) is one of the most important species in temperate grasslands. Temperature is a major factor controlling plant development. Breeding L. perenne cultivars adapted to new ranges of temperature could be necessary under most climate change scenarios. However, before any breeding effort in such direction, knowing the intraspecific variability of responses to temperature is essential. Our objective was to analyze a sample of accessions of L. perenne for their response to constant temperature during germination and initial heterotrophic growth. Eight accessions were taken out from a genebank containing 160 accessions. Six accessions were wild populations collected in different places in France and the other two were populations from a selection program. For germination, four replicates of 100 seeds per accession were tested in Petri dishes in the dark at eight constant temperatures, from 5 to 40°C with increases of 5°C. Germination countings were carried out at variable time intervals and durations that depended on treatments. For initial heterotrophic growth analysis, seeds were germinated at 25°C. Sets of 30 seedlings per accession were placed in polypropylene boxes on blotter blue paper and transferred to each one of the eight treatments in the dark. They were pictured at variable time intervals for root and shoot growth measurement by image analysis. Neither seed germination nor heterotrophic growth was observed at 40°C, for any accession. Temperature and time course response surfaces were markedly different between accessions. Equally, maximum germinability and the shape of the response curves to temperature were significantly different between accessions. This means that limited similarities between responses were observed. Furthermore, germination rates followed the non-linear beta function with significant differences between some accessions. These also showed significant differences in their root and shoot growth rate in response to temperature. In general, the relative growth rates of roots and shoots were slow at 5°C, peaked between 25 and 30°C, and showed a sharp reduction afterward. These results reveal, for the first time, high genetic variability within L. perenne germplasm for the response to temperature in the initial life phases. This discovered variability should serve breeders to create perennial ryegrass varieties for the future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ahmed and Escobar-Gutiérrez.)

Various adaptive mechanisms can ensure that seedlings are established at the most favourable time and place. These mechanisms include seed dormancy i.e., incapacity to germinate in any environment without a specific environmental trigger and inhibition i.e., incapacity to germinate in an unfavourable environment (water availability, temperature: thermoinhibition and light). The objective of this research was to study in the temperate range for germination of forage and turf grass species perennial ryegrass, if the thermal requirements for germination are under genetic controlled and could be selectively bred. Two divergent selections of three cycles were realized on a natural population: one to select for the capacity to germinate at 10°C vs. the impossibility to germinate at 10°C, and one to select for the capacity to germinate at 32°C vs. the impossibility to germinate at 32°C. Seeds of all the lots obtained from the two divergent selections were then germinated at constant temperatures from 5 to 35°C to evaluate their germination ability. Concerning the positive selection, the first cycle of positive selection at 10°C was highly efficient with a very strong increase in the germination percentage. However, afterward no selection effect was observed during the next two cycles of positive selection. By contrast, the positive selection at 32°C was efficient during all cycles with a linear increase of the percentage of germination at 32°C. Concerning the negative selection, we observed only a large positive effect of the first cycle of selection at 10°C. These findings demonstrate that seed thermoinhibition at 10 and 32°C observed in a natural population of perennial ryegrass has a genetic basis and a single recessive gene seems to be involved at 10°C., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ghaleb, Barre, Teulat, Ahmed and Escobar-Gutiérrez.)

Perennial ryegrass is an important forage crop in dairy farming, either for grazing or haying purposes. To further optimise the forage use, this study focused on understanding forage digestibility in the two most important cuts of perennial ryegrass, the spring cut at heading and the autumn cut. In a highly diverse collection of 592 Lolium perenne genotypes, the organic matter digestibility (OMD) and underlying traits such as cell wall digestibility (NDFD) and cell wall components (cellulose, hemicellulose, and lignin) were investigated for 2 years. A high genotype × season interaction was found for OMD and NDFD, indicating differences in genetic control of these forage quality traits in spring versus autumn. OMD could be explained by both the quantity of cell wall content (NDF) and the quality of the cell wall content (NDFD). The variability in NDFD in spring was mainly explained by differences in hemicellulose. A 1% increase of the hemicellulose content in the cell wall (HC.NDF) resulted in an increase of 0.81% of NDFD. In autumn, it was mainly explained by the lignin content in the cell wall (ADL.NDF). A 0.1% decrease of ADL.NDF resulted in an increase of 0.41% of NDFD. The seasonal traits were highly heritable and showed a higher variation in autumn versus spring, indicating the potential to select for forage quality in the autumn cut. In a candidate gene association mapping approach, in which 503 genes involved in cell wall biogenesis, plant architecture, and phytohormone biosynthesis and signalling, identified significant quantitative trait loci (QTLs) which could explain from 29 to 52% of the phenotypic variance in the forage quality traits OMD and NDFD, with small effects of each marker taken individually (ranging from 1 to 7%). No identical QTLs were identified between seasons, but within a season, some QTLs were in common between digestibility traits and cell wall composition traits confirming the importance of hemicellulose concentration for spring digestibility and lignin concentration in NDF for autumn digestibility., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Colas, Barre, van Parijs, Wolters, Quitté, Ruttink, Roldán-Ruiz, Escobar Gutiérrez and Muylle.)

Lucerne ( Medicago sativa ), a major perennial pasture legume, belongs to a species complex that includes several subspecies with wild and cultivated populations. Stand establishment may be compromised by poor germination. Seed scarification, deterioration and temperature have an impact on germination. The objective of this study was to analyse the genetic diversity of lucerne germination in response to three factors: (1) temperature, with seven constant temperatures ranging from 5 to 40°C, was tested on 38 accessions, (2) seed scarification was tested on the same accessions at 5 and 22°C, (3) seed deterioration was tested on two accessions and two seed lots at the seven temperatures. The germination dynamics of seed lots over time was modelled and three parameters were analysed: germinability (germination capacity), maximum germination rate (maximum% of seeds germinating per time unit), and lag time before the first seed germinates. Seed scarification enhanced germinability at both temperatures and its effect was much higher on falcata and wild sativa accessions. Incomplete loss of the hardseededness trait during domestication and selection is hypothesised, indicating that the introduction of wild material in breeding programmes should be followed by the selection for germinability without scarification. Seed lots with altered germinability had low germination at extreme temperatures, both cold and hot, suggesting that mild temperatures are required to promote germination of damaged seed lots. A large genetic diversity was revealed for germination (both capacity and rate) in response to temperature. All accessions had an optimal germination at 15 or 22°C and a poor germination at 40°C. The sativa varieties and landraces had a high germination from 5 to 34°C while the germination of falcata and the wild sativa accessions were weakened at 5 or 34°C, respectively. These differences are interpreted in terms of adaptation to the climate of their geographical origin regions in order to escape frost or heat/drought risks. These new findings give insights on adaptation and domestication of lucerne in its wide geographic area. They suggest further improvement of germination is needed, especially when introducing wild material in breeding pools to remove scarification requirements and to limit differences in response to temperature., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ghaleb, Ahmed, Escobar-Gutiérrez and Julier.)

A great variety of legume species are used for forage production and grown in multi-species grasslands. Despite their close phylogenetic relationship, they display a broad range of morphologies that markedly affect their competitive abilities and persistence in mixtures. Little is yet known about the component traits that control the deployment of plant architecture in most of these species. During the present study, we compared the patterns of shoot organogenesis and shoot organ growth in contrasting forage species belonging to the four morphogenetic groups previously identified in herbaceous legumes (i.e., stolon-formers, rhizome-formers, crown-formers tolerant to defoliation and crown-formers intolerant to defoliation). To achieve this, three greenhouse experiments were carried out using plant species from each group (namely alfalfa, birdsfoot trefoil, sainfoin, kura clover, red clover, and white clover) which were grown at low density under non-limiting water and soil nutrient availability. The potential morphogenesis of shoots characterized under these conditions showed that all the species shared a number of common morphogenetic features. All complied with a generalized classification of shoot axes into three types (main axis, primary and secondary axes). A common quantitative framework for vegetative growth and development involved: (i) the regular development of all shoot axes in thermal time and a deterministic branching pattern in the absence of stress; (ii) a temporal coordination of organ growth at the phytomer level that was highly conserved irrespective of phytomer position, and (iii) an identical allometry determining the surface area of all the leaves. The species differed in their architecture as a consequence of the values taken by component traits of morphogenesis. Assessing the relationships between the traits studied showed that these species were distinct from each other along two main PCA axes which explained 68% of total variance: the first axis captured a trade-off between maximum leaf size and the ability to produce numerous phytomers, while the second distinguished morphogenetic strategies reliant on either petiole or internode expansion to achieve space colonization. The consequences of this quantitative framework are discussed, along with its possible applications regarding plant phenotyping and modeling.