Early Detection of Phenotypic Diversity of Alfalfa (Medicago sativa L.) in Response to Temperature.

Climate change may have important consequences on plant distribution because local environments could change faster than the pace of natural selection and adaptation of wild populations and cultivars of perennial forages. Temperature is a primary factor affecting seed germination and primary heterotrophic growth processes. Medicago sativa (L.) is the most important forage legumes globally. The accelerated breeding of alfalfa cultivars adapted to new ranges of temperature could be necessary under most future climate scenarios. This work aims to explore the genetic diversity of a sample of accessions for responses to temperature during seed germination and seedling heterotrophic growth. Seeds or seedlings were placed in the dark under eight constant temperatures in the range of 5 °C to 40 °C. Germinated seeds were manually counted, while hypocotyl and radicle growth were estimated by using image analysis and curve fitting. Multivariate analyses highlighted links between responses and the origin of accessions. Variability was high, within and between accessions, for all the response variables. Accessions showed significant differences in their non-linear response curves in terms of germinability, germination rates and relative elongation rates. Nevertheless, differences were more noticeable in germination rations and rates compared to seedling heterotrophic growth. Consequently, these could be easier to use as early markers for alfalfa selection and breeding for the future.

Unexpected Intraspecific Variability of Perennial Ryegrass (Lolium perenne L.) in Response to Constant Temperature During Germination and Initial Heterotrophic Growth.

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.

Interactions Between Environment and Genetic Diversity in Perennial Grass Phenology: A Review of Processes at Plant Scale and Modeling.

In perennial grasses, the reproductive development consists of major phenological stages which highly determine the seasonal variations of grassland biomass production in terms of quantity and quality. The reproductive development is regulated by climatic conditions through complex interactions subjected to high genetic diversity. Understanding these interactions and their impact on plant development and growth is essential to optimize grassland management and identify the potential consequences of climate change. Here, we review the main stages of reproductive development, from floral induction to heading, i.e., spike emergence, considering the effect of the environmental conditions and the genetic diversity observed in perennial grasses. We first describe the determinants and consequences of reproductive development at individual tiller scale before examining the interactions between plant tillers and their impact on grassland perenniality. Then, we review the available grassland models through their ability to account for the complexity of reproductive development and genetic × environmental interactions. This review shows that (1) The reproductive development of perennial grasses is characterized by a large intraspecific diversity which has the same order of magnitude as the diversity observed between species or environmental conditions. (2) The reproductive development is determined by complex interactions between the processes of floral induction and morphogenesis of the tiller. (3) The perenniality of a plant is dependent on the reproductive behavior of each tiller. (4) Published models only partly explain the complex interactions between morphogenesis and climate on reproductive development. (5) Introducing more explicitly the underlying processes involved in reproductive development in models would improve our ability to anticipate grassland behavior in future growth conditions.

Leaf elongation response to blue light is mediated by stomatal-induced variations in transpiration in Festuca arundinacea.

Reduced blue light irradiance is known to enhance leaf elongation rate (LER) in grasses, but the mechanisms involved have not yet been elucidated. We investigated whether leaf elongation response to reduced blue light could be mediated by stomata-induced variations of plant transpiration. Two experiments were carried out on tall fescue in order to monitor LER and transpiration under reduced blue light irradiance. Additionally, LER dynamics were compared with those observed in the response to vapour pressure deficit (VPD)-induced variations of transpiration. Finally, we developed a model of water flow within a tiller to simulate the observed short-term response of LER to various transpiration regimes. LER dramatically increased in response to blue light reduction and then reached new steady states, which remained higher than the control. Reduced blue light triggered a simultaneous stomatal closure which induced an immediate decrease of leaf transpiration. The hydraulic model of leaf elongation accurately predicted the LER response to blue light and VPD, resulting from an increase in the growth-induced water potential gradient in the leaf growth zone. Our results suggest that the blue light signal is sensed by stomata of expanded leaves and transduced to the leaf growth zone through the hydraulic architecture of the tiller.

Divergent Selection for Seed Ability to Germinate at Extreme Temperatures in Perennial Ryegrass (Lolium perenne L.).

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.

Seasonal Differences in Structural and Genetic Control of Digestibility in Perennial Ryegrass.

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.

Canonical correlations reveal adaptive loci and phenotypic responses to climate in perennial ryegrass.

Germplasm from perennial ryegrass (Lolium perenne L.) natural populations is useful for breeding because of its adaptation to a wide range of climates. Climate-adaptive genes can be detected from associations between genotype, phenotype and climate but an integrated framework for the analysis of these three sources of information is lacking. We used two approaches to identify adaptive loci in perennial ryegrass and their effect on phenotypic traits. First, we combined Genome-Environment Association (GEA) and GWAS analyses. Then, we implemented a new test based on a Canonical Correlation Analysis (CANCOR) to detect adaptive loci. Furthermore, we improved the previous perennial ryegrass gene set by de novo gene prediction and functional annotation of 39,967 genes. GEA-GWAS revealed eight outlier loci associated with both environmental variables and phenotypic traits. CANCOR retrieved 633 outlier loci associated with two climatic gradients, characterized by cold-dry winter versus mild-wet winter and long rainy season versus long summer, and pointed out traits putatively conferring adaptation at the extremes of these gradients. Our CANCOR test also revealed the presence of both polygenic and oligogenic climatic adaptations. Our gene annotation revealed that 374 of the CANCOR outlier loci were positioned within or close to a gene. Co-association networks of outlier loci revealed a potential utility of CANCOR for investigating the interaction of genes involved in polygenic adaptations. The CANCOR test provides an integrated framework to analyse adaptive genomic diversity and phenotypic responses to environmental selection pressures that could be used to facilitate the adaptation of plant species to climate change.

Towards intercrop ideotypes: non-random trait assembly can promote overyielding and stability of species proportion in simulated legume-based mixtures.

BACKGROUNDS AND AIMS: A major challenge when supporting the development of intercropping systems remains the design of efficient species mixtures. The ecological processes that sustain overyielding of legume-based mixtures compared to pure crops are well known, but their links to plant traits remain to be unravelled. A common assumption is that enhancing trait divergence among species for resource acquisition when assembling plant mixtures should increase species complementarity and improve community performance. METHODS: The Virtual Grassland model was used to assess how divergence in trait values between species on four physiological functions (namely light and mineral N acquisition, temporal development, and C-N use efficiency) affected overyielding and mixture stability in legume-based binary mixtures. A first step allowed us to identify the model parameters that were most important to interspecies competition. A second step involved testing the impact of convergent and divergent parameter (or trait) values between species on virtual mixture performance. RESULTS: Maximal overyielding was achieved in cases where trait values were divergent for the physiological functions controlling N acquisition and temporal development but convergent for light interception. It was also found that trait divergence should not affect competitive abilities of legume and non-legumes at random. Indeed, random trait combinations frequently led to reduced mixture yields when compared to a perfectly convergent neutral model. Combinations with the highest overyielding also tended to be associated with mixture instability and decreasing legume biomass proportion. Achieving both high overyielding and mixture stability was only found to be possible under low or moderate N levels, using combinations of traits adapted to each environment. CONCLUSIONS: No simple assembly rule based on trait divergence could be confirmed. Plant models able to infer plant-plant interactions can be helpful for the identification of major interaction traits and the definition of ideotypes adapted to a targeted intercropping system.

The History of Domestication and Selection of Lucerne: A New Perspective From the Genetic Diversity for Seed Germination in Response to Temperature and Scarification.

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.

Alternatives to neonicotinoids.

The European Food Safety Authority concluded in February 2018 that "most uses of neonicotinoid insecticides represent a risk to wild bees and honeybees". In 2016, the French government passed a law banning the use of the five neonicotinoids previously authorized: clothianidin, imidacloprid, thiamethoxam, acetamiprid and thiacloprid. In the framework of an expert assessment conducted by the French Agency for Food, Environmental and Occupational Health and Safety to identify possible derogations, we performed a thorough assessment of the available alternatives to the five banned neonicotinoids. For each pest targeted by neonicotinoids use, we identified the main alternative pest management methods, which we then ranked for (i) efficacy for controlling the target pest, (ii) applicability (whether directly useable by farmers or in need of further research and development), (iii) durability (risk of resistance in targeted pests), and (iv) practicability (ease of implementation by farmers). We identified 152 authorized uses of neonicotinoids in France, encompassing 120 crops and 279 pest insect species (or genera). An effective alternative to neonicotinoids use was available in 96% of the 2968 case studies analyzed from the literature (single combinations of one alternative pest control method or product × one target crop plant × one target pest insect). The most common alternative to neonicotinoids (89% of cases) was the use of another chemical insecticide (mostly pyrethroids). However, in 78% of cases, at least one non-chemical alternative method could replace neonicotinoids (e.g. microorganisms, semiochemicals or surface coating). The relevance of non-chemical alternatives to neonicotinoids depends on pest feeding habits. Leaf and flower feeders are easier to control with non-chemical methods, whereas wood and root feeders are more difficult to manage by such methods. We also found that further field studies were required for many promising non-chemical methods before their introduction into routine use by farmers. Our findings, transmitted to policymakers, indicate that non-chemical alternatives to neonicotinoids do exist. Furthermore, they highlight the need to promote these methods through regulation and funding, with a view to reducing pesticide use in agriculture.

A generic individual-based model can predict yield, nitrogen content, and species abundance in experimental grassland communities.

Functional-structural plant models are increasingly being used to analyse relationships between plant functioning and the topological and spatial organisation of their modular structure. In this study, the performance of an individual-based model accounting for the the architecture and population dynamics of forage legumes in multi-species grasslands was assessed. Morphogenetic shoot and root parameters were calibrated for seven widely used species. Other model parameters concerning C and N metabolism were obtained from the literature. The model was evaluated using a series of independent experiments combining the seven species in binary mixtures that were subject to regular defoliation. For all the species, the model could accurately simulate phytomer demography, leaf area dynamics, and root growth under conditions of weak competition. In addition, the plastic changes induced by competition for light and N in terms of plant development, leaf area, N uptake, and total plant biomass were correctly predicted. The different species displayed contrasting sensitivities to defoliation, and the model was able to predict the superior ability of creeping species to sustain regular defoliation. As a result of competition and management, the balance between species changed over time and was strongly dependent on the pair of species used. The model proved able to capture these differences in community dynamics. Overall, the results demonstrate that integrating the individual components of population dynamics in a process-based model can provide good predictive capacity regarding mixtures of cultivated species.

Short-term effects of defoliation intensity on sugar remobilization and N fluxes in ryegrass.

In grassland plant communities, the ability of individual plants to regrow after defoliation is of crucial importance since it allows the restoration of active photosynthesis and plant growth. The aim of this study was to evaluate the effects of increasing defoliation intensity (0, 25, 65, 84, and 100% of removed leaf area) on sugar remobilization and N uptake, remobilization, and allocation in roots, adult leaves, and growing leaves of ryegrass over 2 days, using a 15N tracer technique. Increasing defoliation intensity decreased plant N uptake in a correlative way and increased plant N remobilization, but independently. The relative contribution of N stored before defoliation to leaf growth increased when defoliation intensity was severe. In most conditions, root N reserves also contributed to leaf regrowth, but much less than adult leaves and irrespective of defoliation intensity. A threshold of defoliation intensity (65% leaf area removal) was identified below which C (glucose, fructose, sucrose, fructans), and N (amino acids, soluble proteins) storage compounds were not recruited for regrowth. By contrast, nitrate content increased in elongating leaf bases above this threshold. Wounding associated with defoliation is thus not the predominant signal that triggers storage remobilization and controls the priority of resource allocation to leaf meristems. A framework integrating the sequential events leading to the refoliation of grasses is proposed on the basis of current knowledge and on the findings of the present work.

A Conserved Potential Development Framework Applies to Shoots of Legume Species with Contrasting Morphogenetic Strategies.

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.

Assessing the effects of architectural variations on light partitioning within virtual wheat-pea mixtures.

BACKGROUND AND AIMS: Predicting light partitioning in crop mixtures is a critical step in improving the productivity of such complex systems, and light interception has been shown to be closely linked to plant architecture. The aim of the present work was to analyse the relationships between plant architecture and light partitioning within wheat-pea (Triticum aestivum-Pisum sativum) mixtures. An existing model for wheat was utilized and a new model for pea morphogenesis was developed. Both models were then used to assess the effects of architectural variations in light partitioning. METHODS: First, a deterministic model (L-Pea) was developed in order to obtain dynamic reconstructions of pea architecture. The L-Pea model is based on L-systems formalism and consists of modules for 'vegetative development' and 'organ extension'. A tripartite simulator was then built up from pea and wheat models interfaced with a radiative transfer model. Architectural parameters from both plant models, selected on the basis of their contribution to leaf area index (LAI), height and leaf geometry, were then modified in order to generate contrasting architectures of wheat and pea. KEY RESULTS: By scaling down the analysis to the organ level, it could be shown that the number of branches/tillers and length of internodes significantly determined the partitioning of light within mixtures. Temporal relationships between light partitioning and the LAI and height of the different species showed that light capture was mainly related to the architectural traits involved in plant LAI during the early stages of development, and in plant height during the onset of interspecific competition. CONCLUSIONS: In silico experiments enabled the study of the intrinsic effects of architectural parameters on the partitioning of light in crop mixtures of wheat and pea. The findings show that plant architecture is an important criterion for the identification/breeding of plant ideotypes, particularly with respect to light partitioning.

Comparison of the morphogenesis of three genotypes of pea (Pisum sativum) grown in pure stands and wheat-based intercrops.

Cereal-legume intercrops represent a promising way of combining high productivity and agriculture sustainability. The benefits of cereal-legume mixtures are highly affected by species morphology and functioning, which determine the balance between competition and complementarity for resource acquisition. Studying species morphogenesis, which controls plant architecture, is therefore of major interest. The morphogenesis of cultivated species has been mainly described in mono-specific growing conditions, although morphogenetic plasticity can occur in multi-specific stands. The aim of the present study was therefore to characterize the variability of the morphogenesis of pea plants grown either in pure stands or mixed with wheat. This was achieved through a field experiment that included three pea cultivars with contrasting earliness (hr and HR type) and branching patterns. Results show that most of the assessed parameters of pea morphogenesis (phenology, branching, final number of vegetative organs and their kinetics of appearance) were mainly dependent on the considered genotype, which highlights the importance of the choice of cultivars in intercropping systems. There was however a low variability of pea morphogenesis between sole and mixed stands except for plant height and branching of the long-cycle cultivar. The information provided in the present study at stand and plant scale can be used to build up structural-functional models. These models can contribute to improving the understanding of the functioning of cereal-legume intercrops and also to the definition of plant ideotypes adapted to the growth in intercrops.

Banning of methyl bromide for seed treatment: could Ditylenchus dipsaci again become a major threat to alfalfa production in Europe?

In Europe, the stem and bulb nematode Ditylenchus dipsaci has been listed as a quarantine pest by EPPO: without any control, it may cause complete failure of alfalfa crops. Movement of nematodes associated with seeds is considered to be the highest-risk pathway for the spread of this pest. Since the 2010 official withdrawal of methyl bromide in Europe, and in the absence of any alternative chemical, fumigation of contaminated seed batches is no longer possible, which makes the production of nematode-free alfalfa seeds difficult to achieve and leads to unmarketable seed batches. Thermotherapy is being considered as a realistic alternative strategy, but its efficiency still remains to be validated. The combination of the currently available methods (i.e. use of resistant cultivars, seed production according to a certification scheme, mechanical sieving, seed batch inspection) could significantly reduce the likelihood of seed contamination. However, it does not guarantee a total eradication of the nematode. Although it is already widely distributed all over Europe, reclassification of D. dipsaci as a regulated non-quarantine pest to reduce the possibility of further introductions and the rate of spread of this pest appears to be a risky strategy because of the lack of up-to-date documented data to evaluate damage thresholds and determine acceptable tolerance levels.

How does pea architecture influence light sharing in virtual wheat-pea mixtures? A simulation study based on pea genotypes with contrasting architectures.

BACKGROUND AND AIMS: Light interception is a key factor driving the functioning of wheat-pea intercrops. The sharing of light is related to the canopy structure, which results from the architectural parameters of the mixed species. In the present study, we characterized six contrasting pea genotypes and identified architectural parameters whose range of variability leads to various levels of light sharing within virtual wheat-pea mixtures. METHODOLOGY: Virtual plants were derived from magnetic digitizations performed during the growing cycle in a greenhouse experiment. Plant mock-ups were used as inputs of a radiative transfer model in order to estimate light interception in virtual wheat-pea mixtures. The turbid medium approach, extended to well-mixed canopies, was used as a framework for assessing the effects of leaf area index (LAI) and mean leaf inclination on light sharing. PRINCIPAL RESULTS: THREE GROUPS OF PEA GENOTYPES WERE DISTINGUISHED: (i) early and leafy cultivars, (ii) late semi-leafless cultivars and (iii) low-development semi-leafless cultivars. Within open canopies, light sharing was well described by the turbid medium approach and was therefore determined by the architectural parameters that composed LAI and foliage inclination. When canopy closure started, the turbid medium approach was unable to properly infer light partitioning because of the vertical structure of the canopy. This was related to the architectural parameters that determine the height of pea genotypes. Light capture was therefore affected by the development of leaflets, number of branches and phytomers, as well as internode length. CONCLUSIONS: This study provides information on pea architecture and identifies parameters whose variability can be used to drive light sharing within wheat-pea mixtures. These results could be used to build up the architecture of pea ideotypes adapted to multi-specific stands towards light competition.

Association study between the gibberellic acid insensitive gene and leaf length in a Lolium perenne L. synthetic variety.

BACKGROUND: Association studies are of great interest to identify genes explaining trait variation since they deal with more than just a few alleles like classical QTL analyses. They are usually performed using collections representing a wide range of variability but which could present a genetic substructure. The aim of this paper is to demonstrate that association studies can be performed using synthetic varieties obtained after several panmictic generations. This demonstration is based on an example of association between the gibberellic acid insensitive gene (GAI) polymorphism and leaf length polymorphism in 'Herbie', a synthetic variety of perennial ryegrass. METHODS: Leaf growth parameters, consisted of leaf length, maximum leaf elongation rate (LERmax) and leaf elongation duration (LED), were evaluated in spring and autumn on 216 plants of Herbie with three replicates. For each plant, a sequence of 370 bp in GAI was analysed for polymorphism. RESULTS: Genetic effect was highly significant for all traits. Broad sense heritabilities were higher for leaf length and LERmax with about 0.7 in each period and 0.5 considering both periods than for LED with about 0.4 in each period and 0.3 considering both periods. GAI was highly polymorphic with an average of 12 bp between two consecutive SNPs and 39 haplotypes in which 9 were more frequent. Linkage disequilibrium declined rapidly with distance with r 2 values lower than 0.2 beyond 150 bp. Sequence polymorphism of GAI explained 8-14% of leaf growth parameter variation. A single SNP explained 4% of the phenotypic variance of leaf length in both periods which represents a difference of 33 mm on an average of 300 mm. CONCLUSIONS: Synthetic varieties in which linkage disequilibrium declines rapidly with distance are suitable for association studies using the "candidate gene" approach. GAI polymorphism was found to be associated with leaf length polymorphism which was more correlated to LERmax than to LED in Herbie. It is a good candidate to explain leaf length variation in other plant material.

Using functional­structural plant models to study, understand and integrate plant development and ecophysiology.

Functional­structural plant models (FSPMs) explore and integrate relationships between a plant's structure and processes that underlie its growth and development. In recent years, the range of topics being addressed by scientists interested in functional­structural plant modelling has expanded greatly. FSPM techniques are now being used to dynamically simulate growth and development occurring at the microscopic scale involving cell division in plant meristems to the macroscopic scales of whole plants and plant communities. The plant types studied also cover a broad spectrum from algae to trees. FSPM is highly interdisciplinary and involves scientists with backgrounds in plant physiology, plant anatomy, plant morphology, mathematics, computer science, cellular biology, ecology and agronomy. This special issue of Annals of Botany features selected papers that provide examples of comprehensive functional­structural models, models of key processes such as partitioning of resources, software for modelling plants and plant environments, data acquisition and processing techniques and applications of functional­structural plant models for agronomic purposes.

How good is the turbid medium-based approach for accounting for light partitioning in contrasted grass--legume intercropping systems?

BACKGROUND AND AIMS: Most studies dealing with light partitioning in intercropping systems have used statistical models based on the turbid medium approach, thus assuming homogeneous canopies. However, these models could not be directly validated although spatial heterogeneities could arise in such canopies. The aim of the present study was to assess the ability of the turbid medium approach to accurately estimate light partitioning within grass-legume mixed canopies. METHODS: Three contrasted mixtures of wheat-pea, tall fescue-alfalfa and tall fescue-clover were sown according to various patterns and densities. Three-dimensional plant mock-ups were derived from magnetic digitizations carried out at different stages of development. The benchmarks for light interception efficiency (LIE) estimates were provided by the combination of a light projective model and plant mock-ups, which also provided the inputs of a turbid medium model (SIRASCA), i.e. leaf area index and inclination. SIRASCA was set to gradually account for vertical heterogeneity of the foliage, i.e. the canopy was described as one, two or ten horizontal layers of leaves. KEY RESULTS: Mixtures exhibited various and heterogeneous profiles of foliar distribution, leaf inclination and component species height. Nevertheless, most of the LIE was satisfactorily predicted by SIRASCA. Biased estimations were, however, observed for (1) grass species and (2) tall fescue-alfalfa mixtures grown at high density. Most of the discrepancies were due to vertical heterogeneities and were corrected by increasing the vertical description of canopies although, in practice, this would require time-consuming measurements. CONCLUSIONS: The turbid medium analogy could be successfully used in a wide range of canopies. However, a more detailed description of the canopy is required for mixtures exhibiting vertical stratifications and inter-/intra-species foliage overlapping. Architectural models remain a relevant tool for studying light partitioning in intercropping systems that exhibit strong vertical heterogeneities. Moreover, these models offer the possibility to integrate the effects of microclimate variations on plant growth.