As a crucial element for plants, calcium (Ca) is involved in photosynthesis and nutrient absorption, and affects the growth of plants. Poplar is an important economic forest and shelter forest species in China. However, the optimum calcium concentration for its growth is still unclear. Herein, we investigated the growth, biomass, photosynthetic pigments, photosynthetic parameters and products, chlorophyll fluorescence parameters, water use efficiency (iWUE), and antioxidant enzyme activity of “Liao Hu NO.1” poplar (P. simonii × P. euphratica) seedlings at 0, 2.5, 5, 10, and 20 mmol·L⁻¹ concentrations of Ca²⁺, and further studied the absorption, distribution, and utilization of nutrient elements (C, N, P, K, and Ca) in plants. We found that with increasing calcium gradient, plant height and diameter; root, stem, leaf, and total biomasses; net photosynthetic rate (Pn); stomatal conductance (Gs); intercellular carbon dioxide (Ci) level; transpiration rate (Tr); Fv/Fm ratio; Fv/F0 ratio; chlorophyll-a; chlorophyll-b; soluble sugar and starch content; superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) levels; and long-term water use efficiency (iWUE) of poplar seedlings first increased and then decreased. These parameters attained maximum values when the calcium concentration was 5 mmol·L⁻¹, which was significantly different from the other treatments (P < 0.05). Moreover, a suitable Ca²⁺ level promoted the absorption of C, N, P, K, and Ca by various organs of poplar seedlings. The absorption of C, N, P, and K increased first and then decreased with the increased calcium concentration, but the optimum calcium concentrations for the absorption of different elements by different organs were different, and the calcium concentration in leaves, stems, and roots increased gradually. Furthermore, the increase in exogenous calcium content led to a decreasing trend in the C/N ratio in different organs of poplar seedlings. C/P and N/P ratios showed different results in different parts, and only the N/P ratio in leaves showed a significant positive correlation with Ca²⁺ concentration. In conclusion, the results of this study indicate that 5 mmol·L⁻¹ concentration of Ca²⁺ is the optimal level, as it increased growth by enhancing photosynthesis, stress resistance, and nutrient absorption.

Phyllosphere microbial communities have an important role in plant growth and resistance to pathogen infection and are partially influenced by leaf characteristics. Pinewood nematode, Bursaphelenchus xylophilus, is one of the greatest threats to pine trees and is spreading all over the world. However, studies on the resistance of plant–microbe interactions to pathogens during the nematode’s pathogenesis and the relationships of leaf chemical characteristics caused by pinewood nematode and phyllosphere microbial communities are limited. In this study, different stages of Pinus tabuliformis that were healthy or infected with B. xylophilus-associated leaf characteristics and phyllosphere bacterial and fungal communities were compared. These results demonstrated that soluble sugar and starch contents decreased based on the extent of infection. Phyllosphere microbial community changes potentially caused by B. xylophilus infection of P. tabuliformis and the fungal community compositions of healthy P. tabuliformis trees (Ya) were clearly different from diseased P. tabuliformis trees at an early stage of B. xylophilus infection (Yb) and P. tabuliformis trees in the last stage of B. xylophilus infection (Yc), particularly along the first coordinate axis. According to a linear discriminant effect size (LEfSe) analysis, the biomarker species in the phyllosphere of Yb were Acidobacteria, Deinococcus-Thermus, and Patescibacteria, while those in the phyllosphere of Ya were Proteobacteria, Aureobasidium, Dictyosporium, Alternariaster, Knufia, Microstroma, and Naganishia. Particularly at the end of PWD (pine wilt disease) infection, the majority of microbial taxa tended to co-exclude rather than co-occur with PWD infection. The result of a canonical correlation analysis (CCA) showed that the chemical properties of leaves, such as carbon and nitrogen, have significant impacts on phyllosphere microbial communities. These results expanded the possible connections between the phyllosphere communities and plant health.

Ca²⁺ is a crucial second messenger in plant cells that plays a vital role in various physiological and biochemical processes, including plant growth and development, photosynthesis, and enzyme regulation. Exogenous calcium concentrations can have different effects on plant growth. The purpose of this study was to determine the appropriate calcium concentration for the growth of four tree seedlings, namely Pinus tabuliformis, Pinus sylvestris var. mongolica, Populus, and Morus alba seedlings, and whether this optimal calcium concentration varies among different tree species. This study utilized five calcium concentration gradients (0, 100, 200, 400, 800 mg·kg⁻¹) for each species with three repeated treatments. To determine each species’ calcium concentration needs, several growth indices, photosynthetic metrics, chlorophyll fluorescence parameters, water usage efficiency, and antioxidant enzyme activities were analyzed. The results showed that exogenous calcium significantly affected the growth and development of each seedling. The growth, biomass, photosynthetic parameters, photosynthetic products, photosynthetic pigments, water use efficiency, and antioxidant enzyme activity all increased initially and then decreased with the increasing calcium concentration. The exogenous calcium supply increased the concentration of calcium in the leaves. Thus, there was an optimal calcium concentration for plant growth, and a high or low calcium concentration was not conducive to plant growth. Furthermore, this study found that the optimum calcium concentration of different tree species was different. The optimal calcium concentration for Pinus tabuliformis, Pinus sylvestris var. mongolica, Populus, and Morus alba seedlings was 100, 100, 200, and 400 mg·kg⁻¹, respectively, with broadleaf seedlings requiring higher calcium concentrations than coniferous seedlings.

In order to investigate the effect of Ca on the biomass allocation strategies of tree species with different growth rates under drought conditions, we treated poplar (Populus canadensis cv) cuttings and mulberry (Morus alba) seedlings with two soil moisture levels (40 ± 5% and 80 ± 5% maximum water holding capacity) and two soil Ca levels (0 and 200 mg·kg⁻¹ Ca²⁺) in a greenhouse experiment, and then measured the Ca uptake, growth, gas exchange parameters, biomass allocation, and leaf traits. Drought induced a reduction in biomass accumulation of poplar cuttings and mulberry seedlings, and the cuttings and seedlings exhibited different biomass allocation patterns in response to drought stress. Under Ca0 treatment, poplar cuttings allocated more biomass to leaves and less biomass to stems under drought conditions, leading to an increased leaf/stem (L/St) ratio and higher SLA than under moist conditions in order to maintain higher Pn, and had enhanced WUE to cope with drought stress. Under the same treatment, mulberry seedlings allocated more biomass to roots and less biomass to stems, leading to an increased root/shoot (R/S) ratio and lower SLA, to improve drought resistance. Ca200 treatment decreased the growth of poplar cuttings and mulberry seedlings, whereas it enhanced the WUE, root growth, and R/S ratio of poplar cuttings and the WUE of mulberry seedlings, and alleviated drought stress in both species.

Increasing soil N not only aggravates calcium (Ca) stress by stimulating Ca leaching from the soil but also impacts the sensitivity of plants to Ca stress. However, how increasing N influences the soil Ca demand of seedlings is largely unknown. We studied the influence of different concentrations of exogenous Ca (i.e., 0, 200, 400, 600, and 800 mg⋅kg⁻¹ Ca²⁺) on the growth, photosynthesis, Ca absorption, and intrinsic water use efficiency (iWUE) of mulberry (Morus alba) seedlings under two N levels (i.e., 200 and 600 mg⋅kg⁻¹ NH₄NO₃). We found that there was an optimal concentration of soil Ca for the growth and net photosynthetic rate (Pn) of mulberry seedlings; the optimal Ca concentration was 200 mg⋅kg⁻¹ under low N conditions and 400 mg⋅kg⁻¹ under high N conditions. Therefore, the application of N fertilizer increased the optimal Ca concentration. Different from the unimodal relationship between biomass and Ca levels, the iWUE of mulberry was significantly and positively correlated with soil Ca levels. At the same time, except under the 800 mg⋅kg⁻¹ Ca treatment, the soil Ca levels were reflected by foliar Ca concentrations. The N deposition, large-scale N fertilizer application, and drought increase Ca demand in plants, thus causing the application of Ca fertilizer to be necessary in low-Ca soil while alleviating Ca stress in high-Ca soil. The balance between the optimal Ca level needed for growth and drought resistance should be considered when determining the amount of Ca fertilizer required.

The concentrations of exogenous nitrogen and calcium can significantly regulate plant growth and photosynthesis and mutually affect their absorption and utilization. However, whether there is an optimal nitrogen–calcium ratio (N:Ca ratio) in poplar seedling growth and physiological adaptation and what the mechanism of changes in the transcriptome is remain unclear. In this study, three different N:Ca ratios were used to grow poplar seedlings, and physiological and transcriptomic methods were used to study the molecular mechanisms of poplar growth under nitrogen–calcium synergy and to determine the optimal N:Ca ratio for poplar seedling growth. The results of this study showed that maximum poplar seedling growth occurred in the treatment with an N:Ca ratio of 2, which resulted in significantly greater induction of growth than the two other treatments (p < 0.05). Under the lowest N:Ca ratio, poplar seedlings can ensure normal development by regulating photosynthesis, while under the highest N:Ca ratio, regulating nitrogen metabolism can achieve the same result. These results contribute to a better understanding of the molecular mechanisms by which poplar seedlings respond to different ratios of N:Ca. This study provides a valuable basis for exploring the synergistic effects of nitrogen and calcium on the growth of poplar shelterbelts.