Endophytic benefit for a competitive host is neutralized by increasing ratios of infected plants

Leaf endophytes such as Epichloë can affect the competitive ability of host grasses, but the reported responses are inconsistent. We hypothesized that this inconsistency is caused, at least in part, by the following two aspects. One is that a competitive advantage might occur as a result of an increase in storage compounds for both growth and defense. Another is that the effect of the endophyte might be related to both water availability and host density. In a greenhouse experiment, we compared the competitive abilities of endophyte-infected (EI) and endophyte-free (EF) Leymus chinensis, a dominant grass native to the Inner Mongolia Steppe of China, subjected to ten treatments comprised of a factorial combination of two levels of water availability (well-watered and drought) and five proportions of EI to EF plants (12:0, 4:8, 6:6, 8:4, 0:12). The results showed that the competitive ability of EI plants was higher than that of EF under drought. Here, greater belowground biomass and water use efficiency might contribute to better competitiveness of EI plants. When competing under well-watered conditions, endophyte infection did not provide a benefit to the host plant in biomass accumulation, but more carbon was allocated to defense (total phenolics) in EI plants. This scenario could help EI plants suffer less damage than EF when exposed to herbivores in natural habitats. The competitive ability of EI plants was regulated by EI:EF ratios. Competitive ability of EI plants was higher than that of EF plants in mixtures with lower numbers of EI plants, but the beneficial effect of endophyte infection was neutralized in mixtures with higher numbers of EI plants. Overall, endophyte infection improved the competitive ability of the host under either drought or well-watered conditions but in the presence of herbivore, only this benefit was neutralized by increasing ratios of EI plants. We suspect that both the conditional beneficial effects and stabilizing effects of density differences are likely to keep the endophyte infection rate of L. chinensis at an intermediate level.     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L. under drought stress conditions

Endophyte-infected (EI) seeds of Lolium perenne L. were used to attain endophyte-free (EF) population by heating the seeds at 43°C for 15 min and then 57°C for 25 min. Relative water content (RWC), chlorophyll, soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant. Under severe stress, RWC of EI leaf was significantly higher than that of EF leaf, i.e. EI plants took more advantages over EF plants in water-holding ability. Under mild stress, endophyte could enhance soluble sugars in host plants to improve their osmotic ability. With stress intensification, the improvement of endophyte no longer existed, and more photosynthetic products (such as starch) accumulated in EI plants to survive through the undesirable conditions. In the next spring, EI populations will recover more rapidly than EF populations. The biomass of a population is closely related to its photosynthesis. Under severe stress, EI population significantly accumulated more biomass than EF population. As far as photosynthetic pigments were concerned, contents of Chla, Chlb and Car of EI plants were close to those of EF plants, which suggested that endophyte infection didn’t alleviate photosynthetic pigments from being destroyed by drought stress, and endophyte might improve photosynthesis ability of its host plant in other ways.     

Fungal Endophyte (Epichlo? festucae) Alters the Nutrient Content of Festuca rubra Regardless of Water Availability

Festuca rubra plants maintain associations with the vertically transmitted fungal endophyte Epichloë festucae. A high prevalence of infected host plants in semiarid grasslands suggests that this association could be mutualistic. We investigated if the Epichloë-endophyte affects the growth and nutrient content of F. rubra plants subjected to drought. Endophyte-infected (E+) and non-infected (E−) plants of two half-sib lines (PEN and RAB) were subjected to three water availability treatments. Shoot and root biomass, nutrient content, proline, phenolic compounds and fungal alkaloids were measured after the treatments. The effect of the endophyte on shoot and root biomass and dead leaves depended on the plant line. In the PEN line, E+ plants had a greater S:R ratio than E-, but the opposite occurred in RAB. In both plant lines and all water treatments, endophyte-infected plants had greater concentrations of N, P and Zn in shoots and Ca, Mg and Zn in roots than E- plants. On average, E+ plants contained in their shoots more P (62%), Zn (58%) and N (19%) than E- plants. While the proline in shoots increased in response to water stress, the endophyte did not affect this response. A multivariate analysis showed that endophyte status and plant line impose stronger differences in the performance of the plants than the water stress treatments. Furthermore, differences between PEN and RAB lines seemed to be greater in E- than in E+ plants, suggesting that E+ plants of both lines are more similar than those of their non-infected version. This is probably due to the endophyte producing a similar effect in both plant lines, such as the increase in N, P and Zn in shoots. The remarkable effect of the endophyte in the nutrient balance of the plants could help to explain the high prevalence of infected plants in natural grasslands.     

不同浓度和形态磷处理下内生真菌感染对高羊茅的影响

以感染内生真菌(endophyte-infected,EI)和不感染内生真菌(endophyte-free,EF)的高羊茅(Festuca arundinacea Schreb.)为材料,在温室沙培条件下研究内生真菌对高羊茅适应缺磷及利用不同形态磷肥的影响。结果表明,1)缺磷条件下,高羊茅EI和EF植株生长差异不显著;正常供磷条件下,高羊茅EI植株拥有更多分蘖数和绿叶数。说明正常供磷条件下内生真菌改善了宿主高羊茅的生长。2)与水溶性磷相比,高羊茅根有机酸和酸性磷酸酶(acid phosphatase,APase)活性在难溶性磷条件下显著增加,而根总酚含量无显著变化。在水溶性磷条件下,高羊茅EI植株根总酚含量显著高于EF植株,此时EI植株比EF植株拥有更多分蘖数和绿叶数,说明在水溶性磷条件下内生真菌对宿主地上部生长具有一定贡献。在难溶性磷条件下,虽然高羊茅EI植株根总酚含量仍然高于EF植株,但同时EI植株根有机酸含量显著低于EF植株,因此内生真菌感染只是增大了宿主植物的根冠比,而对分蘖数和绿叶数等无显著影响,说明内生真菌对宿主利用难溶性磷贡献不大。可见,内生真菌对宿主植物的生长在水溶性磷条件下更有利。     

Glycine-Glomus-Rhizobium Symbiosis: II. Antagonistic Effects between Mycorrhizal Colonization and Nodulation

Soybean (Glycine max [L.] Merr.) plants grown in pot cultures were inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus mosseae (Nicol. & Gerd.) Gerd. and Trappe and Rhizobium japonicum strain 61A118 at planting (G₁R₁) or at 20 days (G₂₀R₂₀), or with one of the endophytes after the other has colonized the host root (G₁R₂₀, G₂₀R₁). Nodulated (PR₁) and VAM (G₁N) dipartite associations, or nonsymbiotic plants (PN) using nutrient solutions with N, P, or N + P concentrations providing endophyte-equivalent nutrient inputs were used as controls. The delayed tripartite associations received the appropriate N, P, or N + P amendment while one or both endophytes were absent during the first 20 days of growth. Prior inoculation with one endophyte significantly inhibited development of the other. Root hexose sugar concentrations were negatively correlated with VAM colonization (r = −0.89), nodule activity (r = −0.91), and root P content (r = −0.93). Nodule (r = 0.97) and root (r = 0.96) P content correlated positively with VAM colonization. Nodule weight or VAM-fungal biomass were significantly greater in associations grown with only one endophyte. Dry weights of the PN, G₁N, PR₁, and G₂₀R₂₀ plants were significantly greater than those of tripartite plants inoculated at planting with either or both endophytes. Interendophyte inhibition is attributed to competition for root carbohydrates, and this effect apparently also affects overall plant productivity. The objective of the study was to determine if the timing of endophyte introduction and establishment affected the development of the other symbiotic partners.     

内生真菌对羽茅生长及光合特性的影响

通过温室栽培实验,以感染两种内生真菌(Neotyphodium sibiricum和Neotyphodium gansuence)和未感染内生真菌的羽茅(Achnatherum sibiricum)为实验材料,分析感染不同种内生真菌对宿主植物的生长及光合特性的影响。结果表明,感染两种内生真菌羽茅的株高和CO₂补偿点显著低于未染菌的羽茅,而染菌羽茅的蒸腾速率和气孔导度显著高于未染菌羽茅,但对于感染不同种内生真菌的羽茅,无论是分蘖数与生物量的积累还是光合生理值之间均无显著差异。     

Neotyphodium Coenophialum-Infected Tall Fescue and Its Potential Application in the Phytoremediation of Saline Soils

The growth response of endophyte-infected (EI) and endophyte-free (EF) tall fescue to salt stress was investigated under two growing systems (hydroponic and soil in pots). The hydroponic experiment showed that endophyte infection significantly increased tiller and leaf number, which led to an increase in the total biomass of the host grass. Endophyte infection enhanced Na accumulation in the host grass and improved Na transport from the roots to the shoots. With a 15 g l^?1 NaCl treatment, the phytoextraction efficiency of EI tall fescue was 2.34-fold higher than EF plants. When the plants were grown in saline soils, endophyte infection also significantly increased tiller number, shoot height and the total biomass of the host grass. Although EI tall fescue cannot accumulate Na to a level high enough for it to be termed a halophyte, the increased biomass production and stress tolerance suggested that endophyte / plant associations had the potential to be a model for endophyte-assisted phytoextraction in saline soils.     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L. under drought stress conditions

Endophyte-infected (EI) seeds of Lolium perenne L. were used to attain endophyte-free (EF) population by heating the seeds at 43°C for 15 min and then 57°C for 25 min. Relative water content (RWC), chlorophyll, soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant. Under severe stress, RWC of EI leaf was significantly higher than that of EF leaf, i.e. EI plants took more advantages over EF plants in water-holding ability. Under mild stress, endophyte could enhance soluble sugars in host plants to improve their osmotic ability. With stress intensification, the improvement of endophyte no longer existed, and more photosynthetic products (such as starch) accumulated in EI plants to survive through the undesirable conditions. In the next spring, EI populations will recover more rapidly than EF populations. The biomass of a population is closely related to its photosynthesis. Under severe stress, EI population significantly accumulated more biomass than EF population. As far as photosynthetic pigments were concerned, contents of Chla, Chlb and Car of EI plants were close to those of EF plants, which suggested that endophyte infection didn’t alleviate photosynthetic pigments from being destroyed by drought stress, and endophyte might improve photosynthesis ability of its host plant in other ways. __________ Translated from Acta Ecologica Sinica, 2005, 25 (2) [译自: 生态学报, 2005, 25(2)]     

缺磷对黑麦草-内生真菌共生体生长和根中酚含量的影响

培养液中缺磷或内生真菌感染对黑麦草地上部生长的影响不显著,但二者的交互作用却有着显著影响。随着磷浓度的下降,感染内生真菌的黑麦草显示出较强的生长优势,其根长和根中酚类物质的含量显著增加。     

Photosynthetic pigments and photosynthetic products of endophyte-infected and endophyte-free Lolium perenne L.under drought strees conditions

Endophyte-infected fED seeds of Lolium perenne L.were used to attain endophyte-free fED population by heating the seeds at 43℃ for 15 min and then 57℃ for 25 min.Relative water content(RWC),chlorophyll,soluble sugar and starch content of EI and EF populations under normal and drought stress conditions were compared to investigate the effect of endophyte infection on the host plant.Under severe stress.RWC of EI leaf was significantly higher than that Of EF leaf,i.e.EI plants took more advantages over EF plants in water-holding ability.Under mild stress,endophytc could enhance soluble sugars in host plants to improve their osmotic ability.With stress intensification,the improvement of endophyte no longer existed,and more photosynthetic products(such as starch)accumulated in EI plants to survive through the undesirable condidons.In the next spring,EI populations will recover more rapidly than EF populations.The biomass of a population is closely related to its photosynthesis.Under severe stress,EI population significantly accumulated more biomass than EF population.As far as photosynthetic pigments were concerned,contents of Chla,Chlb and Car of EI plants were close to those of EF plants,which suggested that endophyte infection didn't alleviate photosynthetic pigments from being destroyed by drought stress,and endophyte might improve photosynthesis ability of its host plant in other ways.     

内生真菌感染对干旱胁迫下黑麦草生长的影响

 内生真菌是生活在健康植物的茎叶内,形成不明显感染的一类真菌。以黑麦草（Lolium perenne L.）为实验材料,研究在不同强度的干旱胁迫下内生真菌（Neotyphodium lolii）侵染对其叶片延伸生长、分蘖数和生物量的影响。结果表明,与非感染种群相比,内生真菌感染对黑麦草叶片延伸速率无明显促进作用;内生真菌感染种群具有明显较多的分蘖数;在重度胁迫并经过恢复期后,内生真菌感染种群具有较高的根冠比。因而内生真菌可能通过提高植物的分蘖能力和促进有机物向根系的分配来促进宿主植物的营养生长并提高其抗旱性     

Host genotype overrides endophyte infection effects on growth,physiology, and nutrient content of a native grass,Achnatherum sibiricum

The effect of infection by the fungal endophyte Neotyphodium, host genotype, and their interaction on growth and physiology, as well as photosynthesis, was investigated in the native grass Achnatherum sibiricum. We artificially inoculated the endophyte into mature tillers of endophyte-free A. sibiricum. Plants were clipped to 5 cm height after recording growth traits, and analyzed for total nonstructural carbohydrates (TNC %), the percentage of nitrogen (N %), and carbon (C %) in leaves before and after clipping. In our study, the prominent host genotype–endophyte infection interactions detected in A. sibiricum indicates that, for many growth and storage traits, endophyte infection can impact a little change. However, there is no overriding consistently positive effect of the endophyte on growth or storage in A. sibiricum before or after clipping. Our study showed that the interaction between endophyte and host grasses was highly contingent on plant genotypes. We found host genotype overrode fungal endophyte infection in influencing tiller number and photosynthetic properties of A. sibiricum before clipping. After clipping, host genotype accounted for more of the variation in regrowth and above-ground biomass of A. sibiricum than endophyte infection. Our study showed that host genotype affected the total nonstructural carbohydrates of A. sibiricum before and after clipping, whereas endophyte infection increased the carbon content after clipping. Genotype by infection interactions for plant height, leaf mass, total nonstructural carbohydrates, and photosynthetic characteristics indicated genotype-specific effects of endophytes on A. sibiricum physiology and photosynthetic capacity. The host genotype–endophyte infection interactions detected in A. sibiricum suggest that host genotype overrides fungal endophyte infection on growth, physiology, and nutrient content of this native grass. In contrast, endophyte effects did not appear to positively affect growth, physiology, or photosynthetic capacity before or after clipping.     

Effects of simultaneous infections of endophytic fungi and arbuscular mycorrhizal fungi on the growth of their shared host grass Achnatherum sibiricum under varying N and P supply

Grasses can be infected by endophytic fungi and arbuscular mycorrhizal (AM) fungi simultaneously. Here, we investigated the interactions of a native grass, Achnatherum sibiricum, with both endophytic and AM fungi (Glomus mosseae, GM and Glomus etunicatum, GE) at different nitrogen (N) and phosphorus (P) levels. The results showed that endophyte infection significantly suppressed the colonization rates and spore density of GE, but had no effect on those of GM. Endophyte infection increased shoot biomass regardless of the nutrient conditions. The effects of AM fungi on host growth were dependent on mycorrhizal species. There was no significant interaction between endophytic fungi and GE on host growth; however, a significant interaction between endophytic fungi and GE occurred in total phenolic concentrations and P uptake. As for GM, a significant interaction among endophytic fungi, AM fungi and nutrient availability occurred in shoot growth. Under sufficient N and P conditions, endophyte infection alleviated the detrimental effects of GM colonization on host growth.     

Effects of endophytic fungi on some drought tolerance mechanisms of tall fescue in a hydroponics culture

Neotyphodium, a seed-transmissible nonpathogenic fungal endophyte (symbiont) is considered beneficial because endophyte-infected grasses are more drought-tolerant, produce more dry matter, utilize soil nitrogen more efficiently, and deter insects. In this study, the effects of endophytes on physiological mechanisms of drought tolerance in tall fescue (Festuca arundinacea Schreb.) were studied in a greenhouse. Two clonally propagated genotypes of tall fescue (F. arundinacea Schreb.), naturally containing endophyte (EI), and their endophyte-free ramets (EF) were tested at three water stress treatments exerted by PEG 6000 in a hydroponics system. Relative water content (RWC), cell membrane stability (CMS), proline and chlorophyll contents in plant leaves were measured during water stress treatments. After harvest, K⁺, Ca²⁺, and Mg²⁺ contents were measured in plant roots and shoots. After 20 days under stress conditions, plants were transferred to basal hydroponics medium, and their survival after stress relief was evaluated. The results showed that endophyte considerably contributes to host grass water stress tolerance. Both genotypes of EI and EF plants did not differ in RWC, but, regardless of the infection status, genotype 75 had the higher RWC than genotype 83. EI clones of both genotypes maintained slightly higher chlorophyll content and membrane stability than EF clones, although these differences were not significant. The EI plants of genotype 83 concentrated significantly more proline than EF plants, but in the genotype 75, differences between EI and EF clones were not significant. Plant mineral absorption was also influenced by the endophyte presence. EI clones had the higher concentrations of K⁺ in the shoots of both genotypes. The Mg²⁺ and Ca²⁺ contents in EF plants of both genotypes were higher than EI plants in the roots, but in the shoots there were no differences between EI and EF clones. EI clones survived longer after stress removal. These results strongly suggest that Neotyphodium endophytes exert their effects on tall fescue drought tolerance through alteration of various physiological mechanisms involved. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 563–570. This test was submitted by the authors in English.     

Endophyte species influence the biomass production of the native grass Achnatherum sibiricum (L.) Keng under high nitrogen availability

Research on the interaction of endophytes and native grasses normally takes infection status into account, but less often considers the species of endophyte involved in the interaction. Here, we examined the effect of endophyte infection, endophyte species, nitrogen availability, and plant maternal genotype on the performance of a wild grass, Achnatherum sibiricum. Six different Epichloë‐infected maternal lines of A. sibiricum were used in the study; three lines harbored Epichloë gansuensis (Eg), while three lines harbored Epichloë sibirica (Es). These endophytes are vertically transmitted, while Eg also occasionally produces stromata on host tillers. We experimentally removed the endophyte from some ramets of the six lines, with the infected (E+) and uninfected (E?) plants grown under varying levels of nitrogen availability. Eg hosts produced more aboveground biomass than Es hosts only under high nitrogen supply. Endophyte species did not show any influence on the maximum net photosynthetic rate (P_max), photosynthetic nitrogen use efficiency, or total phenolics of A. sibiricum under all nitrogen conditions. However, the plant maternal genotype did influence the P_max and shoot biomass of A. sibiricum. Our results show that endophyte species influenced the shoot biomass of A. sibiricum, and this effect was dependent on nitrogen supply. As with most coevolutionary interactions, A. sibiricum that harbored Eg and Es may show pronounced geographic variation in natural habitats with increased nitrogen deposition. In addition, stroma‐bearing endophyte (Eg) provides positive effects (e.g., higher biomass production) to A. sibiricum plants during the vegetative growth stage.     

Interactive Effects of Nitrogen and Phosphorus on Soil Microbial Communities in a Tropical Forest

Elevated nitrogen (N) deposition in humid tropical regions may exacerbate phosphorus (P) deficiency in forests on highly weathered soils. However, it is not clear how P availability affects soil microbes and soil carbon (C), or how P processes interact with N deposition in tropical forests. We examined the effects of N and P additions on soil microbes and soil C pools in a N-saturated old-growth tropical forest in southern China to test the hypotheses that (1) N and P addition will have opposing effects on soil microbial biomass and activity, (2) N and P addition will alter the composition of the microbial community, (3) the addition of N and P will have interactive effects on soil microbes and (4) addition-mediated changes in microbial communities would feed back on soil C pools. Phospholipid fatty acid (PLFA) analysis was used to quantify the soil microbial community following four treatments: Control, N addition (15 g N m⁻² yr⁻¹), P addition (15 g P m⁻² yr⁻¹), and N&P addition (15 g N m⁻² yr⁻¹ plus 15 g P m⁻² yr⁻¹). These were applied from 2007 to 2011. Whereas additions of P increased soil microbial biomass, additions of N reduced soil microbial biomass. These effects, however, were transient, disappearing over longer periods. Moreover, N additions significantly increased relative abundance of fungal PLFAs and P additions significantly increased relative abundance of arbuscular mycorrhizal (AM) fungi PLFAs. Nitrogen addition had a negative effect on light fraction C, but no effect on heavy fraction C and total soil C. In contrast, P addition significantly decreased both light fraction C and total soil C. However, there were no interactions between N addition and P addition on soil microbes. Our results suggest that these nutrients are not co-limiting, and that P rather than N is limiting in this tropical forest.     

Influence of Nitrogen Fertilizer and Endophyte Infection on Ecophysiological Parameters and Mineral Element Content of Perennial Ryegrass

An expedment was designed to determine the effect of the fungal endophyte Neotyphodium lolii on the growth, physiological parameters and mineral element content of perennial ryegrass (Lolium perennel L.), when growing at two N supply levels. Endophyfe infection had a significant positive effect on both shoot and root growth of ryegrass, but this difference was only significant in the high N supply treatment. At high N supply, endophyte-infected (EI) plants accumulated more soluble sugar in the sheath and the root than endophyte-free (EF) plants. Endophyte infection affected mineral element concentrations In the root more than in the shoot. We found a significant effect of endophyte infection on B, Mn and Mg in the root, but significant effect was only found on B in the shoot. EI plants tended to accumulate less B in the shoot at both N levels, but accumulated more B, Mn and Mg in the root at low N levels. The difference of growth parameters in different periods was significant. The content of soluble sugar and crude protein in the sheath were also dependent on the growth stages of both EI and EF plants.     

Potential antagnosim of cultivated and wild grass–endophyte associations towards Meloidogyne incognita

Potential antagonism towards plant-parasitic nematode Meloidogyne incognita by grass–endophyte associations and their corresponding endophytes was evaluated. The following grass-endophyte associations were studied: (1) the cultivated grass Schedonorus arundinaceus-Neotyphodium coenophialum, (2) the wild grass Leymus chinensis-Neotyphodium sp. and (3) the wild grass Achnatherum sibiricum-Neotyphodium sibiricum. The cultivated grass Schedonorus arundinaceus was used to evaluate the effect of the endophyte fungus on chemotaxis of M. incognita. The results showed a significant negative effect on chemotaxis of the second-stage juveniles (J2) of M. incognita towards S. arundinaceus-N. coenophialum plants (P < 0.05). The associations of the grass S. arundinaceus-N. coenophialum and the wild grass A. sibiricum-N. sibiricum were used for testing their effect on penetration of grasses by J2. The wild grass A. sibiricum-N. sibiricum association appeared to suppress J2 penetration. N. coenophialum, Neotyphodium sp. and N. sibiricum isolated from cultivated and wild grasses respectively were used for measuring their effect on J2 mortality. Undiluted culture filtrates of all endophytic fungi caused significantly high J2 mortality 72.6% for N. coenphialum, 91.7% for Neotyphodium sp. and 66.8% for N. sibiricum. Finally, the penetration of cucumber roots by J2 pretreated by the filtrates of the three endophytic fungi was also tested. N. sibiricum filtrate significantly reduced the number of J2 penetrations by 44.5% and no significant effect in the treatment of the filtrates of Neotyphodium sp. from L. chinensis. Taken together, endophytic fungi of the genus Neotyphodium might be a potential source of biological control agents for plant parasitic nematodes.     

Effects of Nitrogen and Phosphorus Fertilization on Soil Carbon Fractions in Alpine Meadows on the Qinghai-Tibetan Plateau

In grassland ecosystems, N and P fertilization often increase plant productivity, but there is no concensus if fertilization affects soil C fractions. We tested effects of N, P and N+P fertilization at 5, 10, 15 g m⁻² yr⁻¹ (N₅, N₁₀, N₁₅, P₅, P₁₀, P₁₅, N₅P₅, N₁₀P₁₀, and N₁₅P₁₅) compared to unfertilized control on soil C, soil microbial biomass and functional diversity at the 0–20 cm and 20–40 cm depth in an alpine meadow after 5 years of continuous fertilization. Fertilization increased total aboveground biomass of community and grass but decreased legume and forb biomass compared to no fertilization. All fertilization treatments decreased the C:N ratios of legumes and roots compared to control, however fertilization at rates of 5 and 15 g m⁻² yr⁻¹ decreased the C:N ratios of the grasses. Compared to the control, soil microbial biomass C increased in N₅, N₁₀, P₅, and P₁₀ in 0–20 cm, and increased in N₁₀ and P₅ while decreased in other treatments in 20–40 cm. Most of the fertilization treatments decreased the respiratory quotient (qCO₂) in 0–20 cm but increased qCO₂ in 20–40 cm. Fertilization increased soil microbial functional diversity (except N₁₅) but decreased cumulative C mineralization (except in N₁₅ in 0–20 cm and N₅ in 20–40 cm). Soil organic C (SOC) decreased in P₅ and P₁₅ in 0–20 cm and for most of the fertilization treatments (except N₁₅P₁₅) in 20–40 cm. Overall, these results suggested that soils will not be a C sink (except N₁₅P₁₅). Nitrogen and phosphorus fertilization may lower the SOC pool by altering the plant biomass composition, especially the C:N ratios of different plant functional groups, and modifying C substrate utilization patterns of soil microbial communities. The N+P fertilization at 15 g m⁻² yr⁻¹ may be used in increasing plant aboveground biomass and soil C accumulation under these meadows.     

Stoichiometry of Root and Leaf Nitrogen and Phosphorus in a Dry Alpine Steppe on the Northern Tibetan Plateau

Leaf nitrogen (N) and phosphorus (P) have been used widely in the ecological stoichiometry to understand nutrient limitation in plant. However,few studies have focused on the relationship between root nutrients and environmental factors. The main objective of this study was to clarify the pattern of root and leaf N and P concentrations and the relationships between plant nitrogen (N) and phosphorus (P) concentrations with climatic factors under low temperature conditions in the northern Tibetan Plateau of China. We conducted a systematic census of N and P concentrations, and the N∶P ratio in leaf and root for 139 plant samples, from 14 species and 7 families in a dry Stipa purpurea alpine steppe on the northern Tibetan Plateau of China. The results showed that the mean root N and P concentrations and the N∶P ratios across all species were 13.05 mg g⁻¹, 0.60 mg g⁻¹ and 23.40, respectively. The mean leaf N and P concentrations and the N∶P ratio were 23.20 mg g⁻¹, 1.38 mg g⁻¹, and 17.87, respectively. Compared to global plant nutrients concentrations, plants distributing in high altitude area have higher N concentrations and N∶P, but lower P concentrations, which could be used to explain normally-observed low growth rate of plant in the cold region. Plant N concentrations were unrelated to the mean annual temperature (MAT). The root and leaf P concentrations were negatively correlated with the MAT, but the N∶P ratios were positively correlated with the MAT. It is highly possible this region is not N limited, it is P limited, thus the temperature-biogeochemical hypothesis (TBH) can not be used to explain the relationship between plant N concentrations and MAT in alpine steppe. The results were valuable to understand the bio-geographic patterns of root and leaf nutrients traits and modeling ecosystem nutrient cycling in cold and dry environments.