Estimation of flavonoid and centelloside accumulation in leaves of Centella asiatica L. Urban by multiparametric fluorescence measurements

Recently, we have shown the relevance of nitrogen (N), phosphorus (P) and potassium (K) supply levels for resource partitioning between primary and secondary metabolism, and the concentration of centellosides in Centella asiatica L. Urban leaves. So far, no efforts have been made to investigate the effects of mineral supply on flavonoid concentrations in this species. Here, we aimed to examine the accumulation of centellosides in C. asiatica leaves in vivo by means of non-destructive fluorescence measurements using products of the secondary metabolism, particularly the epidermal flavonols and anthocyanins, as reference. For this purpose we conducted three discrete experiments in a greenhouse having N, P and K levels as experimental factors. Our results reveal that flavonoid and anthocyanin accumulation is affected by N, P and K fertigation in the same way as the centelloside accumulation. More precisely, limitations in plant growth were accompanied by higher flavonoid and anthocyanin concentrations, confirming the proposed trade-off between the plant's primary and secondary metabolism. The fluorescence-based flavonol (FLAV) and anthocyanin (ANTH_RG) indices correlated fairly with flavonoid and especially with anthocyanin concentrations. Moreover, centellosides were positively correlated with the FLAV and ANTH_RG indices, and with the BFRR_UV index, which is considered as universal ‘stress-indicator’. Thus, here we indicate for the first time, that the fluorescence-based indices FLAV, ANTH_RG as well as BFRR_UV enable the monitoring of flavonoid and centelloside concentrations in leaves of C. asiatica. Our results support and highlight the significant potential for further development and application of fluorescence-based sensors in ecophysiological research as well as in the production of medicinal plants.     

Copper-Induced Modulation of Biomass Growth,Physiological Parameters,Bioactive Centellosides,and Expression of Biosynthetic Pathway Genes in an Important Medicinal Herb, <Emphasis Type="Italic">Centella asiatica</Emphasis>

Hydroponically established Centella asiatica plants were studied under different concentrations of copper (0, 0.32,1.6, and 3.2 µM) for their morphological, physiological, and biochemical characteristics. The plants grown in the medium without any exogenous supply of copper showed improved biomass accumulation with maximum fresh weight (FW?=?24.7 g plant^?1) and dry weight (DW?=?2.35 g plant^?1). The total chlorophyll content, leaf number, and the leaf area were higher (0.27 mg g^?1 DW, 20 and 100.54 cm²) in the plants grown under copper-deficient conditions. The toxic effects of increased levels of copper were evident by significant inhibition in growth and other morphological parameters. Copper treatment showed an increase in malondialdehyde (MDA) content and SOD activity. Bioactive phytochemical profiling using HPLC analysis revealed that higher levels of copper (1.6 and 3.2 µM) inhibit the accumulation of total centelloside content. The differential accumulation of centelloside content was further validated by the consistent pattern of expression of key pathway genes related to centelloside production in copper-treated C. asiatica plants. The study unveiled some of the important facts associated with the mechanism of copper tolerance in this important medicinal herb C. asiatica. The findings of the present study can be further used to provide better production of high value, in demand centellosides.     

Effect of N:K supply ratios on the performance of three grass species from herbaceous wetlands

Shifts from N limitation to P or K limitation in wetlands (e.g. due to mowing and high atmospheric N deposition) are reflected by increased N:P and N:K ratios of plant biomass and changes in species composition. So far, the implications of increased N:K ratios for wetland vegetation have hardly been investigated. We examined how the supply of N and K influences the growth of three wetland grass species (Holcus lanatus, Anthoxanthum odoratum, Deschampsia caespitosa) to determine at what N:K ratios growth is likely to be K limited, how N:K ratios influence biomass allocation and nutrient uptake, and whether the responses to high N:K ratios vary among species. Plants were grown in sand at six N:K supply ratios ranging from 0.17 to 40.5 and combined with two levels of fertility in a factorial design. In 15 weeks of growth, plant biomass increased with increasing N:K supply ratios, indicating that growth was mostly limited by N and not by K across the entire range of N:K ratios. However, there were indications of K deficiency at the highest N:K supply ratio, such as increased leaf mortality and strong reduction of K concentrations during leaf senescence. The response of total plant biomass to nutrient treatments did not differ among the three species. However, other plant traits, such as biomass allocation to roots, leaf senescence and N concentrations, suggested that D. caespitosa is better adapted to extremely low K availability than the other two species.The short-term responses of the three wetland grass species to N and K supply were similar to those found in previous experiments testing different N:P supply ratios. In both cases, growth depended mainly on N supply, despite high N:K or N:P supply ratios and very low K or P concentrations in plant biomass. In those previous experiments, P supply became more important in the second year. There are indications that the same could also be true for K in a longer-term experiment. Hence, the so-called ‘critical’ values for N:K ratios of plant biomass in the field might be lower than indicated by our short-term experiment.     

塔克拉玛干沙漠腹地人工植被及土壤CNP的化学计量特征

生态化学计量学是研究生态过程和生态作用中化学元素平衡的科学。极端环境中进行植物叶片与土壤中营养元素含量及变化研究,对于揭示植物对营养元素的需要和当地土壤的养分供给能力,以及植物对环境的适应与反馈能力具有十分重要的意义。以塔克拉玛干沙漠腹地塔中植物园生长良好的25种人工植被及其生境为研究对象,运用方差分析、相关分析综合研究植物叶片及土壤的化学计量特征及其相互关系。结果显示:塔克拉玛干沙漠腹地25种人工植被叶片C、N、P的平均含量分别为(386.7±46.6)、(24.7±8.1)和(1.8±0.78) mg/g;叶片C:N、C:P及 N:P分别为(17.5±6.7)、(249.2±102.8)、(15.0±5.6)。其中豆科植物N含量极显著高于非豆科植物(P<0.001)。不同生活型植物的C、N、P含量均存在显著差异,C、N、P含量在3种生活型的大小顺序为草本>灌木>乔木。C:N和N:P在不同生活型植物间不存在显著差异(P>0.05),而乔木和灌木的C:P显著高于草本植物(P< 0.05)。相关分析表明植物的叶片C:N、C:P都与相应的N、P含量呈现极显著负相关性(P<0.001),而叶片N含量与P含量的变化并不相关(P> 0.05)。土壤C、N、P养分元素含量远低于全国的平均水平,尤其是N含量(<0.2 mg/g);土壤C与N存在着极显著的正相关关系(P<0.01),而C与P、N与P间的相关性并不显著(P>0.05)。以上研究结果表明,受极端环境的限制,塔克拉玛干沙漠人工植被植物对养分元素的利用效率显著低于全国陆地植物的平均水平,不同科和不同生活型功能群植物对环境的适应能力显著不同,表现出显著的养分适应策略差异性。     

Nitrogen, phosphorus and potassium nutritional status of semiarid steppe grassland in Inner Mongolia

In grazed semiarid steppe ecosystems, much attention has been paid to aspects of growth limitation by water. So far, potential limitation of primary production by plant nutrients was rarely considered. This knowledge is essential for identification of sustainable land-use practices in these large and important ecosystems on the background of over-exploitation and climate change. In the present study plant nutrient concentrations and ratios were investigated with factorial additions of water and N fertilizer at two sites with contrasting soil nutrient availability. Combined analysis of nutrient concentrations, contents, biomass production, and plant N:P ratios consistently confirmed primary growth limitation by water and a strong N limitation when sufficient amounts of water were supplied. P limitation only occurred at the site with low P availability when in addition to the natural supply, water and N fertilizer were given. According to reported thresholds of N:K and K:P ratios, K was not limiting in any plot. The observed nutritional patterns in the plant community were related to the dynamics of species composition and their specific nutrient status. Stipa grandis had the highest N:P ratio whereas Artemisia frigida showed lowest N:P. These nutrient characteristics were related to growth strategies of dominant species. Accordingly, the relative biomass contribution of S. grandis and A. frigida strongly affected the nutrient status of the plant community. Plant N:P ratios indicate the relative limitation by N or P in the semiarid grasslands under sufficient water supply, but other methods of nutritional diagnosis should be used when plant N:P ratios remain below critical values.     

An optimized biotechnological system for the production of centellosides based on elicitation and bioconversion of Centella asiatica cell cultures

Centella asiatica is a herbaceous plant of Asian traditional medicine. Besides wound healing, this plant is recommended for the treatment or care of various skin conditions such as dry skin, leprosy, varicose ulcers, eczema, and/or psoriasis. Triterpene saponins, known as centellosides, are the main metabolites associated with these beneficial effects. Considering the interest in these high value active compounds, there is a need to develop biosustainable and economically viable processes to produce them. Previous work using C. asiatica plant cell culture technology demonstrated the efficient conversion of amyrin derivatives into centellosides, opening a new way to access these biomolecules. The current study was aimed at increasing the production of centellosides in C. asiatica plant cell cultures. Herein, we report the application of a new elicitor, coronatine, combined with the addition of amyrin‐enriched resins as potential sustainable precursors in the centelloside pathway, for a positive synergistic effect on centelloside production. Our results show that coronatine is a powerful elicitor for increasing centelloside production and that treatments with sustainable natural sources of amyrins enhance centelloside yields. This process can be scaled up to an orbitally shaken CellBag, thereby increasing the capacity of the system for producing biomass and centellosides.     

Important foliar traits depend on species-grouping: analysis of a remnant temperate forest at the Keerqin Sandy Lands, China

Foliar traits are often interpreted to reflect strategies for coping with water and nutrient supply limitations. In this study, we measured several important leaf traits for 147 species sampled from a remnant, temperate deciduous broad-leaved forest in Keerqin Sandy Lands, Northeast China to test whether these traits are ‘invariant’ or dependent on water supply limitations. Our data show that average specific leaf area (SLA), nitrogen (N) and phosphorus (P) concentrations, leaf C/N, C/P and N/P were 273 cm²?g^?1, 18.1 mg?g^?1, 1.60 mg?g^?1, 28.2, 343 and 12.4, respectively. However, most of these traits were significantly different (P?<?0.05) for different species groupings based on growth forms, phylogenetic history, photosynthetic pathways, or habitats. SLA was positively correlated with leaf P concentration across the broad spectrum of 118 species and most species functional groupings. However, SLA was not correlated with N concentration across all species or within each species functional group. SLA and N and P concentrations in dry habitats were lower than those in wet habitats, whereas leaf C/N, C/P, and N/P had the opposite trend both across all species and within major species functional groupings (herb, monocots and C3 species). Our data indicate that SLA vs. leaf N and SLA vs. P relationships may be regulated differentially for different species functional groupings and that water limitation may have a greater influence than nutrient limitation for plant growth.     

微量元素硼和锌在作物营养平衡中的作用

许多土壤中B、Zn由于含量或有效性低而供给不足,成为作物营养平衡与大面积产量提高的重要限制因素。采取各种方法直接施用B、Zn,或者配制在尿素中施用,可以调节作物营养平衡,改善作物-环境生态关系,提高产量和品质。在水稻旱育苗地区还可把Zn配在育苗土壤调制剂中施用。在缺Zn褐土上,施入的Zn迅速转化为各种形态,其中碳酸盐结合态是有效Zn的主要仓库。配合施用P肥对Zn的有效性无不良影响,Zn与N、P配合施用有益于作物营养平衡。     

猫儿山三种森林类型林下植物叶片与土壤化学计量特征

为探究猫儿山不同森林类型林下植物叶片与土壤化学计量特征,揭示其林下植物适应策略。该文对猫儿山针阔混交林(ZK)、常绿阔叶次生林(CLC)和常绿阔叶林(CL)林下草本层和灌木层主要植物叶片与土壤的化学元素含量进行测定,分析其化学计量特征及其相互之间的内在联系。结果表明:(1)从总体上看,草本层和灌木层植物叶片的C、N含量差异不显著,草本层植物叶片P、K含量极显著高于灌木层,N:P显著低于灌木层; 草本层植物更易受N限制,灌木层植物更易受P限制且其N和P利用效率更高; 不同森林类型之间的灌木层植物叶片化学计量差异不显著,草本层植物叶片N含量、C:N和C:P差异显著,针阔混交林草本层植物的养分利用效率较高。(2)3种森林类型的土壤C、N含量显示,CL>CLC>ZK且彼此之间差异极显著,针阔混交林土壤的P含量最高而C:P、N:P最低。(3)针阔混交林的土壤显著影响林下植物部分叶片化学计量,另外2种森林类型的土壤影响不显著。综上认为,猫儿山不同森林类型的土壤化学计量存在显著或极显著差异,林下不同层次的植物对营养元素的需求以及环境适应策略不同; 针阔混交林土壤对林下植物叶片化学计量影响较强,由于有机质分解效率较低导致土壤受N限制,因此应加强针阔混交林的N素管理。该研究结果为森林管理提供了数据支持。     

Nutrient resorption of two evergreen shrubs in response to long-term fertilization in a bog

Plant resorption of multiple nutrients during leaf senescence has been established but stoichiometric changes among N, P and K during resorption and after fertilization are poorly understood. We anticipated that increased N supply would lead to further P limitation or co-limitation with N or K [i.e. P-(co)limitation], decrease N resorption and increase P and K resorption, while P and K addition would decrease P and K resorption and increase N resorption. Furthermore, Ca would accumulate while Mg would be resorbed during leaf senescence, irrespective of fertilization. We investigated the effect of N, P and K addition on resorption in two evergreen shrubs (Chamaedaphne calyculata and Rhododendron groenlandicum) in a long-term fertilization experiment at Mer Bleue bog, Ontario, Canada. In general, N addition caused further P-(co)limitation, increased P and K resorption efficiency but did not affect N resorption. P and K addition did not shift the system to N limitation and affect K resorption, but reduced P resorption proficiency. C. calyculata resorbed both Ca and Mg while R. groenlandicum resorbed neither. C. calyculata showed a higher resorption than R. groenlandicum, suggesting it is better adapted to nutrient deficiency than R. groenlandicum. Resorption during leaf senescence decreased N:P, N:K and K:P ratios. The limited response of N and K and the response of P resorption to fertilization reflect the stoichiometric coupling of nutrient cycling, which varies among the two shrub species; changes in species composition may affect nutrient cycling in bogs.     

Effect of shade on the growth and mineral nutrition of a C4 perennial grass under field conditions

The effect of shading by a shrub legume on the growth and nutrient uptake of a C4 tropical grass was studied during four regrowth cycles. Regrowth periods were characterised by contrasting soil water availability. Dichanthium aristatum (Poir.) C. E. Hubbard swards were grown in full sun and under Gliricidia sepium (Jacq.) Walp. and Leucaena leucocephala (Lam.) de Wit with a light transmission level ranging from 80 to 30% of the incoming photosynthetically active radiation (PAR), depending on shrub regrowth. A treatment with high N and water supply was included in one of the cycles to quantify the effect of shade alone on potential growth.Aboveground biomass (DM) and leaf area index (LAI) of swards were not depressed by the reduction of incoming PAR. The reduction in transmitted PAR by shrubs was compensated by an increase in the radiation use efficiency (RUE) of shaded swards. Higher RUE of unfertilised, shaded stands may be explained by higher levels of N availability in the soil. This is supported by the analysis of curves relating sward N accumulation to sward DM accumulation. In fact, for similar measured biomass the accummulated N was higher in shaded stands, a consequence of their higher N concentrations. This allowed shaded leaves to improve their CO2 assimilation rates on a leaf area basis. Higher RUE reported on shaded stands may be the consequence of higher leaf CO2 assimilation rates and also possible changes in the shoot:root ratio. As with N, the amount of K taken up by the sward was higher under shade, whereas P data were higher under shade only during the driest cycle. A positive water balance, alone or in combination with high N fertilisation, eliminated the improvement of the N nutrition of shaded stands. Thus, the positive effects of shade may be only observed when N and water are limiting sward growth in the open.     

Effects of nutrient-limiting supply ratios on toxin content of Karenia brevis grown in continuous culture

Toxins produced as secondary metabolites can play important roles in phytoplankton communities and contribute to the ecological success of harmful algal bloom (HAB) taxa. Toxin composition and content in phytoplankton are affected by a suite of environmental factors, including nutrient availability. Changes in nutrient availability can increase or decrease toxin content and alter toxin composition, depending on toxin stoichiometry and the mechanisms by which nutrient limitation affects toxin production. The studies that have assessed the effects of nutrient availability on brevetoxin content of the HAB species Karenia brevis have reported contradictory results, although there is growing support that nutrient limitation increases brevetoxin content. In this study, we assessed the effects of decreased nitrogen (N) and phosphorus (P) availability on brevetoxin content and composition of K. brevis grown in chemostats at steady state by altering the nutrient supply ratios of incoming media from the Redfield Ratio. Overall, brevetoxin content was greatest in cultures grown at the lowest rate, regardless of the nutrient supply ratio (i.e., under both Redfield and N-limiting supply ratios). Compared to cultures grown at 0.2 d⁻¹, cultures grown at 0.1 d⁻¹ exhibited 5-fold increases in intracellular toxin content. In contrast, at constant growth rates, N-limiting supply ratios decreased intracellular brevetoxin content by approximately one-third, although this result was significant only in cultures growing at the fastest rate of 0.23 d⁻¹. P-limiting supply ratios had no effect on brevetoxin content or composition. In addition, when cultures grown at rates of 0.2 d⁻¹ were supplied with balanced/Redfield N:P supply ratios, but different absolute nutrient concentrations, toxin content was greater under greater nutrient concentrations. These findings suggest that when growth rate is not nutrient limited, there is a positive relationship between nutrient availability and brevetoxin content. This work contributes to previous studies by demonstrating strong growth rates effects on brevetoxin content and that growth rate and nutrient availability can independently or together affect toxin content of K. brevis. Moreover, our work underscores the value of the chemostat as a tool to elucidate the mechanisms by which nutrient availability and growth rate affect toxin production and content of HAB species.     

长期施肥对红壤旱地作物产量和土壤肥力的影响

历时14年的田间定位试验表明,红壤旱地磷素最为缺乏,施用磷肥对提高作物产量效果最好;施用石灰和微量元素对作物产量没有明显增产作用．施肥可以提高花生植株磷、钾的浓度,表明作物的养分含量受土壤养分供应水平的影响．土壤中赢余(亏损)的养分在养分库重建中的作用可以用速效养分库重建效率来表示．研究表明,当红壤旱地N、P2O5、K2O养分赢余1kg·hm^-2时,可分别使土壤中水解氮、有效磷、交换性钾含量提高0．6～6．26、0．20～0．28和1．1～8．5mg·kg^-1．红壤旱地氮和磷通过径流等损失较大,不同处理之间的变化幅度也较大．红壤每年可固定磷43．5kg·hm^-2,通过自身风化作用每年可提供氮48．1kg·hm^-2和钾40．5kg·hm^-2,以满足作物生长所需．     

Clonal integration ameliorates the carbon accumulation capacity of a stoloniferous herb,Glechoma longituba,growing in heterogenous light conditions by facilitating nitrogen assimilation in the rhizosphere

Background and Aims Enhanced availability of photosynthates increases nitrogen (N) mineralization and nitrification in the rhizosphere via rhizodeposition from plant roots. Under heterogeneous light conditions, photosynthates supplied by exposed ramets may promote N assimilation in the rhizosphere of shaded, connected ramets. This study was conducted to test this hypothesis.Methods Clonal fragments of the stoloniferous herb Glechoma longituba with two successive ramets were selected. Mother ramets were subjected to full sunlight and offspring ramets were subjected to 80 % shading, and the stolon between the two successive ramets was either severed or left intact. Measurements were taken of photosynthetic and growth parameters. The turnover of available soil N was determined together with the compostion of the rhizosphere microbial community.Key Results The microbial community composition in the rhizosphere of shaded offspring ramets was significantly altered by clonal integration. Positive effects of clonal integration were observed on NAGase activity, net soil N mineralization rate and net soil N nitrification rate. Increased leaf N and chlorophyll content as well as leaf N allocation to the photosynthetic machinery improved the photosynthetic capability of shaded offspring ramets when the stolon was left intact. Clonal integration improved the growth performance of shaded, connected offspring ramets and whole clonal fragments without any cost to the exposed mother ramets.Conclusions Considerable differences in microbial community composition caused by clonal integration may facilitate N assimilation in the rhizosphere of shaded offspring ramets. Increased N content in the photosynthetic machinery may allow pre-acclimation to high light conditions for shaded offspring ramets, thus promoting opportunistic light capture. In accordance with the theory of the division of labour, it is suggested that clonal integration may ameliorate the carbon assimilation capacity of clonal plants, thus improving their fitness in temporally and spatially heterogeneous habitats.     

Co-limitation of plant primary productivity by nitrogen and phosphorus in a species-rich wooded meadow on calcareous soils

High small-scale species richness of calcareous grasslands is generally thought to result from evening of species competitive potentials by limited N availability, because of relatively low herb N/P ratios in these communities. However, P mobility is low in alkaline soils as well. We studied soil chemistry and productivity of herb and moss layers in a very diverse calcareous meadow (up to 76 vascular plant species per m²) to test the hypotheses of a co-limitation of herb productivity by both soil N and P availabilities and moss productivity primarily by P availability. The effect of nutrient supply on productivity was investigated using both a natural productivity gradient as well as fertilization experiments. We observed strong positive correlations of soil P availability and total soil N with the above-ground productivity of herb layer. A long-term fertilization experiment demonstrated that P alone and N and P together increased productivity of vascular species, and that the productivity continuously declined after cessation of fertilization with the effect of previous fertilization occasionally visible even 14 years after treatment termination. A short-term fertilization experiment further demonstrated that N and P when supplied alone increase productivity of vascular plants, suggesting that both elements were limiting. Furthermore, there was a significant interaction between N and P on productivity, indicating that simultaneous N and P supply increased productivity more than separate nutrient additions. Moss productivity was negatively associated with vascular plant productivity. In particular, N addition decreased moss productivity, but moss productivity did not decline in P addition treatments. P requirements of mosses were larger than those of vascular plants. Our data indicate co-limitation of herb productivity by both soil N and P in this highly diverse grassland, while limitation of moss productivity mainly by P. We suggest that N and P co-limitations are common in calcareous diverse grasslands, and may partly explain the extreme small-scale species diversity in these communities.     

Co-limitation of plant primary productivity by nitrogen and phosphorus in a species-rich wooded meadow on calcareous soils

High small-scale species richness of calcareous grasslands is generally thought to result from evening of species competitive potentials by limited N availability, because of relatively low herb N/P ratios in these communities. However, P mobility is low in alkaline soils as well. We studied soil chemistry and productivity of herb and moss layers in a very diverse calcareous meadow (up to 76 vascular plant species per m²) to test the hypotheses of a co-limitation of herb productivity by both soil N and P availabilities and moss productivity primarily by P availability. The effect of nutrient supply on productivity was investigated using both a natural productivity gradient as well as fertilization experiments. We observed strong positive correlations of soil P availability and total soil N with the above-ground productivity of herb layer. A long-term fertilization experiment demonstrated that P alone and N and P together increased productivity of vascular species, and that the productivity continuously declined after cessation of fertilization with the effect of previous fertilization occasionally visible even 14 years after treatment termination. A short-term fertilization experiment further demonstrated that N and P when supplied alone increase productivity of vascular plants, suggesting that both elements were limiting. Furthermore, there was a significant interaction between N and P on productivity, indicating that simultaneous N and P supply increased productivity more than separate nutrient additions. Moss productivity was negatively associated with vascular plant productivity. In particular, N addition decreased moss productivity, but moss productivity did not decline in P addition treatments. P requirements of mosses were larger than those of vascular plants. Our data indicate co-limitation of herb productivity by both soil N and P in this highly diverse grassland, while limitation of moss productivity mainly by P. We suggest that N and P co-limitations are common in calcareous diverse grasslands, and may partly explain the extreme small-scale species diversity in these communities.     

Responses of wetland graminoids to the relative supply of nitrogen and phosphorus

The biomass production of wetland vegetation can be limited by nitrogen or phosphorus. Some species are most abundant in N-limited vegetation, and others in P-limited vegetation, possibly because growth-related traits of these species respond differently to N versus P supply. Two growth experiments were carried out to examine how various morphological and physiological traits respond to the relative supply of N and P, and whether species from sites with contrasting nutrient availability respond differently. In experiment 1, four Carex species were grown in nutrient solutions at five N:P supply ratios (1.7, 5, 15, 45, 135) combined with two levels of supply (geometric means of N and P supply). In experiment 2, two Carex and two grass species were grown in sand at the same .ve N:P supply ratios combined with three levels of supply and two light intensities (45% or 5% daylight). After 12-13 weeks of growth, plant biomass, allocation, leaf area, tissue nutrient concentrations and rates and nutrient uptake depended signi.cantly on the N:P supply ratio, but the type and strength of the responses differed among these traits. The P concentration and the N:P ratio of shoots and roots as well as the rates of N and P uptake were mainly determined by the N:P supply ratio; they showed little or no dependence on the supply level and relatively small interspeci.c variation. By contrast, the N concentration, root mass ratio, leaf dry matter content and speci.c leaf area were only weakly related to the N:P supply ratio; they mainly depended on plant species and light, and partly on overall nutrient supply. Plant biomass was determined by all factors together. Within a level of light and nutrient supply, biomass was generally maximal (i.e. co-limited by N and P) at a N:P supply ratio of 15 or 45. All species responded in a similar way to the N:P supply ratio. In particular, the grass species Phalaris arundinacea and Molinia caerulea showed no differences in response that could clearly explain why P. arundinacea tends to invade P-rich (N-limited) sites, and M. caerulea P-limited sites. This may be due to the short duration of the experiments, which investigated growth and nutrient acquisition but not nutrient con­servation.     

Leaf nonstructural carbohydrate concentrations of understory woody species regulated by soil phosphorus availability in a tropical forest

Leaf soluble sugars and starch are important components of nonstructural carbohydrates (NSCs), which are crucial for plant growth, development, and reproduction. Although there is a large body of research focusing on the regulation of plant NSC (soluble sugars and starch) concentrations, the response of foliar NSC concentrations to continuous nitrogen (N) and phosphorus (P) addition is still unclear, especially in tropical forests. Here, we used a long‐term manipulative field experiment to investigate the response of leaf NSC concentrations to continuous N and P addition (3‐, 5‐, and 8‐year fertilization) in a tropical forest in southern China. We found significant species‐specific variation in leaf NSC concentrations in this tropical forest. Phosphorus addition dramatically decreased both leaf soluble sugar and starch concentrations, while N addition had no significant effects on leaf soluble sugar and starch concentrations. These results suggest that, in plants growing in P‐limiting tropical soil, leaf NSC concentrations are regulated by soil P availability rather than N availability. Moreover, the negative relationships between NSC concentrations and leaf mass per area (LMA) revealed that NSCs could supply excess carbon (C) for leaf expansion under P addition. This was further supported by the increased structural P fraction after P fertilization in our previous study at the same site. We conclude that soil P availability strongly regulates leaf starch and soluble sugar concentrations in the tropical tree species included in this study. The response of leaf NSC concentrations to long‐term N and P addition can reflect the close relationships between plant C dynamics and soil nutrient availability in tropical forests. Maintaining relatively higher leaf NSC concentrations in tropical plants can be a potential mechanism for adapting to P‐deficient conditions.