Transcriptional Reprogramming of Pea Leaves at Early Reproductive Stages.

Pea (Pisum sativum L.) is an important source of dietary proteins. Nutrient recycling from leaves contributes to the accumulation of seed proteins and is a pivotal determinant of protein yields in this grain legume. The aim of this study was to unveil the transcriptional regulations occurring in pea leaves before the sharp decrease in chlorophyll breakdown. As a prelude to this study, a time-series analysis of 15N translocation at the whole plant level was performed, which indicated that nitrogen recycling among organs was highly dynamic during this period and varied depending on nitrate availability. Leaves collected on vegetative and reproductive nodes were further analyzed by transcriptomics. The data revealed extensive transcriptome changes in leaves of reproductive nodes during early seed development (from flowering to 14 days after flowering), including an up-regulation of genes encoding transporters, and particularly of sulfate that might sustain sulfur metabolism in leaves of the reproductive part. This developmental period was also characterized by a down-regulation of cell wall-associated genes in leaves of both reproductive and vegetative nodes, reflecting a shift in cell wall structure. Later on, 27 days after flowering, genes potentially switching the metabolism of leaves toward senescence were pinpointed, some of which are related to ribosomal RNA processing, autophagy, or transport systems. Transcription factors differentially regulated in leaves between stages were identified and a gene co-expression network pointed out some of them as potential regulators of the above-mentioned biological processes. The same approach was conducted in Medicago truncatula to identify shared regulations with this wild legume species. Altogether the results give a global view of transcriptional events in leaves of legumes at early reproductive stages and provide a valuable resource of candidate genes that could be targeted by reverse genetics to improve nutrient remobilization and/or delay catabolic processes leading to senescence.

High Temperatures During the Seed-Filling Period Decrease Seed Nitrogen Amount in Pea (Pisum sativum L.): Evidence for a Sink Limitation.

Higher temperatures induced by the on-going climate change are a major cause of yield reduction in legumes. Pea (Pisum sativum L.) is an important annual legume crop grown in temperate regions for its high seed nitrogen (N) concentration. In addition to yield, seed N amount at harvest is a crucial characteristic because pea seeds are a source of protein in animal and human nutrition. However, there is little knowledge on the impacts of high temperatures on plant N partitioning determining seed N amount. Therefore, this study investigates the response of seed dry matter and N fluxes at the whole-plant level (plant N uptake, partitioning in vegetative organs, remobilization, and accumulation in seeds) to a range of air temperature (from 18.4 to 33.2°C) during the seed-filling-period. As pea is a legume crop, plants relying on two different N nutrition pathways were grown in glasshouse: N2-fixing plants or NO3 --assimilating plants. Labeled nitrate (15NO3 -) and intra-plant N budgets were used to quantify N fluxes. High temperatures decreased seed-filling duration (by 0.8 day per °C), seed dry-matter and N accumulation rates (respectively by 0.8 and 0.032 mg seed-1 day-1 per °C), and N remobilization from vegetative organs to seeds (by 0.053 mg seed-1 day-1 per °C). Plant N2-fixation decreased with temperatures, while plant NO3 - assimilation increased. However, the additional plant N uptake in NO3 --assimilating plants was never allocated to seeds and a significant quantity of N was still available at maturity in vegetative organs, whatever the plant N nutrition pathway. Thus, we concluded that seed N accumulation under high temperatures is sink limited related to a shorter seed-filling duration and a reduced seed dry-matter accumulation rate. Consequently, sustaining seed sink demand and preserving photosynthetic capacity of stressed plants during the seed-filling period should be promising strategies to promote N allocation to seeds from vegetative parts and thus to maintain crop N production under exacerbated abiotic constraints in field due to the on-going climate change.

Determinism of carbon and nitrogen reserve accumulation in legume seeds.

In legume plants, the determination of individual seed weight is a complex phenomenon that depends on two main factors. The first one corresponds to the number of cotyledon cells, which determines the potential seed weight as the cotyledon cell number is related to seed growth rate during seed filling. Since cell divisions take place between flowering and the beginning of seed filling, any stress occurring before the beginning of seed filling can affect individual seed growth rate (C and N reserve accumulation in seeds), and thus individual seed weights. The second factor concerns carbon and nitrogen supply to the growing seed to support reserve accumulation. Grain legume species produce protein-rich seeds involving high requirement of nitrogen. Since seed growth rate as determined by cotyledon cell number is hardly affected by photoassimilate availability during the filling period, a reduction of photosynthetic activity caused by nitrogen remobilization in leaves (e.g., remobilization of essential proteins involved in photosynthesis) can lead to shorten the duration of the filling period, and by that can provoke a limitation of individual seed weights. Accordingly, any biotic or abiotic stress during seed filling causing a decrease in photosynthetic activity should lead to a reduction of the duration of seed filling.

How does temperature affect C and N allocation to the seeds during the seed-filling period in pea? Effect on seed nitrogen concentration.

The effect of moderate temperature on seed N concentration during the seed-filling period was evaluated in pea (Pisum sativum L.) kept in growth cabinets and the relation between plant assimilate availability and the variation of seed N concentration with temperature was investigated. Seed N concentration of pea was significantly lowered when temperature during the seed-filling period decreased from a day / night temperature of 25 / 20°C to 15 / 10°C. Our results demonstrate that during the seed-filling period mechanisms linked with assimilate availability can modify seed N accumulation rate and / or seed-filling duration between 25 / 20°C and 15 / 10°C. At the lower temperature (15 / 10°C), an increased C availability resulting from an enhanced carbon fixation per degree-day allowed new competing vegetative sinks to grow as pea is an indeterminate plant. Consequently N availability to filling seeds was reduced. Because the rate of seed N accumulation per degree-day mainly depends on N availability to filling seeds, the rate of seed N accumulation was lower at the low temperature of our study (15 / 10°C) than at 25 / 20°C while seed growth rate per degree-day remains unaffected, consequently seed N concentration was reduced. Concomitantly, the increased C availability at the lower temperature prolonged the duration of the seed-filling period.