Increasing photosynthetic ability as a whole is essential for acquiring higher crop yields. Nonleaf green organs (NLGOs) make important contributions to photosynthate formation, especially under stress conditions. However, there is little information on the pod wall in legume forage related to seed development and yield. This experiment is designed for alfalfa (Medicago sativa) under drought stress to explore the photosynthetic responses of pod walls after 5, 10, 15, and 20 days of pollination (DAP5, DAP10, DAP15, and DAP20) based on ultrastructural, physiological and proteomic analyses. Stomata were evidently observed on the outer epidermis of the pod wall. Chloroplasts had intact structures arranged alongside the cell wall, which on DAP5 were already capable of producing photosynthate. The pod wall at the late stage (DAP20) still had photosynthetic ability under well-watered (WW) treatments, while under water-stress (WS), the structure of the chloroplast membrane was damaged and the grana lamella of thylakoids were blurry. The chlorophyll a and chlorophyll b concentrations both decreased with the development of pod walls, and drought stress impeded the synthesis of photosynthetic pigments. Although the activity of ribulose-1,5-bisphosphate carboxylase (RuBisCo) decreased in the pod wall under drought stress, the activity of phosphoenolpyruvate carboxylase (PEPC) increased higher than that of RuBisCo. The proteomic analysis showed that the absorption of light is limited due to the suppression of the synthesis of chlorophyll a/b binding proteins by drought stress. Moreover, proteins involved in photosystem I and photosystem II were downregulated under WW compared with WS. Although the expression of some proteins participating in the regeneration period of RuBisCo was suppressed in the pod wall subjected to drought stress, the synthesis of PEPC was induced. In addition, some proteins, which were involved in the reduction period of RuBisCo, carbohydrate metabolism, and energy metabolism, and related to resistance, including chitinase, heat shock protein 81-2 (Hsp81-2), and lipoxygenases (LOXs), were highly expressed for the protective response to drought stress. It could be suggested that the pod wall in alfalfa is capable of operating photosynthesis and reducing the photosynthetic loss from drought stress through the promotion of the C4 pathway, ATP synthesis, and resistance ability.

1. The quantification of the allocation of nitrogen (N) and phosphorus (P) among plant organs is essential to improve our understanding of plant growth, lifehistory strategies and ecosystem nutrient and energy cycles. However, allocation strategies of N and P between terrestrial plant leaves and fine roots are unclear.2. Here, we compiled a global dataset comprising 807 terrestrial plant species to analyse the stoichiometric scaling of N and P between leaves and fine roots across species, families, plant groups, biomes and locations.3. We found that N and P, and N:P ratios manifest different scaling exponents (α-values) between leaves and fine roots. Specifically, overall α N and α P were >1 and <1, respectively, and α N:P = 1, indicating a higher proportional increase of N to fine roots and P to leaves. However, there were differences in α N , α P and α N:P among major species, families, plant groups, biomes and locations. In addition, α N and α N:P increased with latitude; there was no clear trend for α P . Mean annual temperature accounted for the greatest proportion of variation in α N , whereas soil total P accounted for the greatest proportion of variation in α P and α N:P . 4. Synthesis. Our results demonstrate a divergent N and P allocation strategy between leaves and fine roots in terrestrial plants. This study improves our understanding of the effects of the environment (including changes in global climate and life-history strategies) on nutrient allocation patterns between these two important plant organs.

The objective of this study is to determine the effect of freeze–thaw condition on the fermentation characteristics, microbial community, and aerobic stability of oat (Avena sativa) silage in the Qinghai-Tibet Plateau. Oat forage was harvested at milk ripening stage, ensiled in vacuum-sealed bags, and then stored at (1) a constant temperature of 20°C, as a control (20 group) or (2) subjected to freeze–thaw condition (alternating 20 and −5°C every 12 h; S group). The quality and microbial community in the silage were measured after 1, 3, 7, 14, and 60 days of ensiling, and the aerobic stability was measured after 60 days of ensiling at room temperature or at the two treatment temperatures. The results showed that the higher the pH, the lower the concentration of lactic acid and the ratio of lactic acid/acetic acid of the samples under freeze–thaw condition, as compared to those stored at 20°C. The dry matter content of 20 groups was significantly higher than S group (p &lt; 0.05). While ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), and water-soluble carbohydrates (WSC) had no significant difference between two groups. Lactobacillus spp., Leuconostoc spp., and Weissella spp. were the most prevalent bacterial genera in all groups. The abundance of Lactobacillus spp. in the 20 group was the highest on day 3 of ensiling (p &lt; 0.05), and it reached the peak on day 14 in the S group, but the abundance in the S group did not exceed 50% during whole fermentation process. The abundance of Enterobacterales and the count of Escherichia coli in the S group was significantly higher than 20 group (p &lt; 0.05). Interestingly, the lactic acid concentration was significant correlated with Lactobacillus spp. in 20 group, while correlated with Leuconostoc spp. in S group. The aerobic stability of the S group was lower than that of the 20 group (p &lt; 0.05). The present study indicates that the freeze–thaw condition led to insufficient fermentation degree of silage by limiting the fermentation of Lactobacillus spp. and severely reduced the aerobic stability of oat silage.

In oat production, the over-application of nitrogen (N) fertilizer in fields due to low N fertilizer use efficiency not only increases production costs but also causes environmental pollution. Currently, mining low N-tolerant oat varieties is an important way to promote sustainable agriculture. In this study, 30 oat varieties were grown in a seedling culture with two treatments of normal N (10 mM NH4NO3) and low N (1.25 mM NH4NO3), and the correlations between agronomic traits and plant N content and low N tolerance coefficients and indices were determined, which can be used as indicators for the evaluation of low N-tolerant oat varieties. Coefficient of variation, correlation analysis, principal component analysis, partial least-squares discrimination analysis, random forest analysis, least absolute shrinkage and selection operator regression and model evaluation, and membership function analysis were used for in-depth analysis of these indicators. Plant N content, root–crown ratio, and dry weight of aboveground plant parts were found to be important indicators of low N tolerance in oats. According to the membership function ranking of the 30 selected oat varieties, Jiayan 2, Qingyongjiu 035, and Qingyin 2 had strong tolerance to low N stress and Qingyongjiu 003, Qingyongjiu 021, and Qingyongjiu 016 had poor tolerance to low N stress. Thus, this study provides a reliable and comprehensive method for evaluating the low N tolerance of oat varieties as well as a reference for screening other low N-tolerant plants.