Response of tree growth to nutrient addition is size dependent in a subtropical forest.

Understanding how nutrient addition affects the tree growth is critical for assessing forest ecosystem function and processes, especially in the context of increased nitrogen (N) and phosphorus (P) deposition. Subtropical forests are often considered N-rich and P-poor ecosystems, but few existing studies follow the traditional "P limitation" paradigm, possibly due to differences in nutrient requirements among trees of different size classes. We conducted a three-year fertilization experiment with four treatments (Control, N-treatment, P-treatment, and NP-treatment). We measured soil nutrient availability, leaf stoichiometry, and relative growth rate (RGR) of trees across three size classes (small, medium and large) in 64 plots. We found that N and NP-treatments increased the RGR of large trees. P-treatment increased the RGR of small trees. RGR was mainly affected by N addition, the total effect of P addition was only 10 % of that of N addition. The effect of nutrient addition on RGR was mainly regulated by leaf stoichiometry. This study reveals that nutrient limitation is size dependent, indicating that continuous unbalanced N and P deposition will inhibit the growth of small trees and increase the instability of subtropical forest stand structure, but may improve the carbon sink function of large trees.

Phosphorus addition regulates the growth of Chinese fir by changing needle nitrogen fractions in growing and dormant seasons.

Forest productivity is generally limited by nutrient scarcity. This study aims to reveal seasonal interactions among leaf carbon (C), nitrogen (N) fractions and tree growth driven by nutrient addition in a subtropical forest. Here, a field nutrient addition experiment was conducted with six treatments, namely, +N5 (5 g N m-2 yr-1), +N10 (10 g N m-2 yr-1), +P5 (5 g P m-2 yr-1), +N5 + P5, +N10 + P5, and control (N0 + P0). C fractions (structural and non-structural carbohydrates) and N fractions (soluble N, nucleic N and protein N) in needles as well as tree growth indicated by basal area increment (BAI) were measured in growing and dormant seasons. Total N and protein N in old needles were significantly increased by P addition, while no significant differences of non-structural carbohydrates in young (<1-year old) and old needles (>1-year old) were detected among the treatments in both seasons. N and P addition increased the structural carbohydrates of old needles in dormant season. P addition decreased and increased tree growth in growing and dormant seasons, respectively. The variation of BAI was explained 18.3 % by total N and 17.8 % by protein N in growing season, and was explained 33.9 % by total N and 34.2 % by protein N in dormant season. Our study suggested that the P addition effect on Chinese fir growth mostly depends on needle N fractions. This study highlights tree seasonal growth driven by nutrient alteration might be characterized by leaf N fractions rather than C fractions in subtropical forests.

Isolation and characterization of two phosphate-solubilizing fungi from rhizosphere soil of moso bamboo and their functional capacities when exposed to different phosphorus sources and pH environments.

Phosphate-solubilizing fungi (PSF) generally enhance available phosphorus (P) released from soil, which contributes to plants' P requirement, especially in P-limiting regions. In this study, two PSF, TalA-JX04 and AspN-JX16, were isolated from the rhizosphere soil of moso bamboo (Phyllostachys edulis) widely distributed in P-deficient areas in China and identified as Talaromyces aurantiacus and Aspergillus neoniger, respectively. The two PSF were cultured in potato dextrose liquid medium with six types of initial pH values ranging from 6.5 to 1.5 to assess acid resistance. Both PSF were incubated in Pikovskaya's liquid media with different pH values containing five recalcitrant P sources, including Ca3(PO4)2, FePO4, CaHPO4, AlPO4, and C6H6Ca6O24P6, to estimate their P-solubilizing capacity. No significant differences were found in the biomass of both fungi grown in media with different initial pH, indicating that these fungi could grow well under acid stress. The P-solubilizing capacity of TalA-JX04 was highest in medium containing CaHPO4, followed by Ca3(PO4)2, FePO4, C6H6Ca6O24P6, and AlPO4 in six types of initial pH treatments, while the recalcitrant P-solubilizing capacity of AspN-JX16 varied with initial pH. Meanwhile, the P-solubilizing capacity of AspN-JX16 was much higher than TalA-JX04. The pH of fermentation broth was negatively correlated with P-solubilizing capacity (p<0.01), suggesting that the fungi promote the dissolution of P sources by secreting organic acids. Our results showed that TalA-JX04 and AspN-JX16 could survive in acidic environments and both fungi had a considerable ability to release soluble P by decomposing recalcitrant P-bearing compounds. The two fungi had potential for application as environment-friendly biofertilizers in subtropical bamboo ecosystem.

Soil phosphorus functional fractions and tree tissue nutrient concentrations influenced by stand density in subtropical Chinese fir plantation forests.

Stand density regulation is an important measure of plantation forest management, and phosphorus (P) is often the limiting factor of tree productivity, especially in the subtropics and tropics. However, the stand density influence on ecosystem P cycling is unclear in Chinese fir (Cunninghamia lanceolata) plantations of subtropical China. We collected rhizosphere and bulk soils, leaves and twigs with different ages and roots with different orders to measure P and nitrogen (N) variables in Chinese fir plantations with low density (LDCF) and high density (HDCF) at Fujian and Hunan provinces of subtropical China. Rhizosphere soil labile P, slow P, occluded P and extractable P were higher in LDCF than HDCF at two sites. Meanwhile, P and N concentrations of 1-year-old leaves and twigs were higher in LDCF than HDCF and leaf N/P ratio generally increased with increasing leaf age at two sites. Rhizosphere vs. bulk soil labile P and occluded P were greater in LDCF than HDCF at Fujian. Nitrogen resorption efficiencies (NRE) of leaves and twigs were higher in LDCF than HDCF at Fujian, while their P resorption efficiencies (PRE) were not different between two densities at two sites. The average NRE of leaves (41.7%) and twigs (65.6%) were lower than the corresponding PRE (67.8% and 78.0%, respectively). Our results suggest that reducing stem density in Chinese fir plantations might be helpful to increase soil active P supplies and meet tree nutrient requirements.

Exogenous nutrients and carbon resource change the responses of soil organic matter decomposition and nitrogen immobilization to nitrogen deposition.

It is unclear whether exogenous nutrients and carbon (C) additions alter substrate immobilization to deposited nitrogen (N) during decomposition. In this study, we used laboratory microcosm experiments and (15)N isotope tracer techniques with five different treatments including N addition, N+non-N nutrients addition, N+C addition, N+non-N nutrients+C addition and control, to investigate the coupling effects of non-N nutrients, C addition and N deposition on forest floor decomposition in subtropical China. The results indicated that N deposition inhibited soil organic matter and litter decomposition by 66% and 38%, respectively. Soil immobilized (15)N following N addition was lowest among treatments. Litter (15)N immobilized following N addition was significantly higher and lower than that of combined treatments during the early and late decomposition stage, respectively. Both soil and litter extractable mineral N were lower in combined treatments than in N addition treatment. Since soil N immobilization and litter N release were respectively enhanced and inhibited with elevated non-N nutrient and C resources, it can be speculated that the N leaching due to N deposition decreases with increasing nutrient and C resources. This study should advance our understanding of how forests responds the elevated N deposition.

Nitrogen and phosphorus additions alter nutrient dynamics but not resorption efficiencies of Chinese fir leaves and twigs differing in age.

It is unclear how or even if phosphorus (P) input alters the influence of nitrogen (N) deposition in a forest. In theory, nutrients in leaves and twigs differing in age may show different responses to elevated nutrient input. To test this possibility, we selected Chinese fir (Cunninghamia lanceolata) for a series of N and P addition experiments using treatments of +N1 - P (50 kg N ha(-1) year(-1)), +N2 - P (100 kg N ha(-1) year(-1)), -N + P (50 kg P ha(-1) year(-1)), +N1 + P, +N2 + P and -N - P (without N and P addition). Soil samples were analyzed for mineral N and available P concentrations. Leaves and twigs in summer and their litters in winter were classified as and sorted into young and old components to measure N and P concentrations. Soil mineral N and available P increased with N and P additions, respectively. Nitrogen addition increased leaf and twig N concentrations in the second year, but not in the first year; P addition increased leaf and twig P concentrations in both years and enhanced young but not old leaf and twig N accumulations. Nitrogen and P resorption proficiencies in litters increased in response to N and P additions, but N and P resorption efficiencies were not significantly altered. Nitrogen resorption efficiency was generally higher in leaves than in twigs and in young vs old leaves and twigs. Phosphorus resorption efficiency showed a minimal variation from 26.6 to 47.0%. Therefore, P input intensified leaf and twig N enrichment with N addition, leaf and twig nutrients were both gradually resorbed with aging, and organ and age effects depended on the extent of nutrient limitation.

Topsoil and Deep Soil Organic Carbon Concentration and Stability Vary with Aggregate Size and Vegetation Type in Subtropical China.

The impact of reforestation on soil organic carbon (OC), especially in deep layer, is poorly understood and deep soil OC stabilization in relation with aggregation and vegetation type in afforested area is unknown. Here, we collected topsoil (0-15 cm) and deep soil (30-45 cm) from six paired coniferous forests (CF) and broad-leaved forests (BF) reforested in the early 1990s in subtropical China. Soil aggregates were separated by size by dry sieving and OC stability was measured by closed-jar alkali-absorption in 71 incubation days. Soil OC concentration and mean weight diameter were higher in BF than CF. The cumulative carbon mineralization (Cmin, mg CO2-C kg-1 soil) varied with aggregate size in BF and CF topsoils, and in deep soil, it was higher in larger aggregates than in smaller aggregates in BF, but not CF. The percentage of soil OC mineralized (SOCmin, % SOC) was in general higher in larger aggregates than in smaller aggregates. Meanwhile, SOCmin was greater in CF than in BF at topsoil and deep soil aggregates. In comparison to topsoil, deep soil aggregates generally exhibited a lower Cmin, and higher SOCmin. Total nitrogen (N) and the ratio of carbon to phosphorus (C/P) were generally higher in BF than in CF in topsoil and deep soil aggregates, while the same trend of N/P was only found in deep soil aggregates. Moreover, the SOCmin negatively correlated with OC, total N, C/P and N/P. This work suggests that reforested vegetation type might play an important role in soil OC storage through internal nutrient cycling. Soil depth and aggregate size influenced OC stability, and deep soil OC stability could be altered by vegetation reforested about 20 years.

[Responses of rhizosphere nitrogen and phosphorus transformations to different acid rain intensities in a hilly red soil tea plantation].

Tea (Camellia sinensis) plantation in hilly red soil region has been long impacted by acid deposition, however its effects on nitrogen (N) and phosphorus (P) transformations in rhizosphere soils remain unclear. A 25-year old tea plantation in a typical hilly red soil region was selected for an in situ simulation experiment treated by pH 4.5, pH 3.5, pH 2.5 and control. Rhizosihere and bulk soils were collected in the third year from the simulated acid deposition experiment. Soil mineral N, available P contents and major enzyme activities were analyzed using the chemical extraction and biochemical methods, and N and P mineralization rates were estimated using the indoor aerobic incubation methods. Our results showed that compared to the control, the treatments of pH 4.5, pH 3.5 and pH 2.5, respectively decreased 7.1%, 42.1% and 49.9% NO3(-)-N, 6.4%, 35.9% and 40.3% mineral N, 10.5%, 41.1% and 46.9% available P, 18.7%, 30.1% and 44.7% ammonification rate, 3.6%, 12.7% and 38.8% net N-mineralization rate, and 31.5%, 41.8% and 63.0% P mineralization rate in rhizosphere soils; however, among the 4 treatments, rhizosphere soil nitrification rate was not significantly different, the rhizosphere soil urease and acid phosphatase activities generally increased with the increasing intensity of acid rain (P<0.05). In bulk soil, compared with the control, the treatments of pH 4.5, pH 3.5 and pH 2.5 did not cause significant changes in NO3(-)-N, mineral N, available P as well as in the rates of nitrification, ammonification, net N-mineralization and P mineralization. With increasing the acid intensity, the rhizosphere effects of NH4+-N, NO3(-)-N, mineral N, ammonification and net N-mineralization rates were altered from positive to negative effects, those of urease and acid phosphatease showed the opposite trends, those of available P and P mineralization were negative and that of nitrification was positive. In sum, prolonged elevated acid rain could reduce N and P transformation rates, decrease their availability, alter their rhizosphere effects, and have impact on nutrient cycling in tea plantation.

[Variability of soil water soluble organic carbon content and its response to temperature change in green spaces along urban-to-rural gradient of Nanchang, China].

Topsoil of green space including typical forest, shrub and grassland were collected to measure their water soluble organic carbon ( WSOC) before and after incubation of 30 days at 5, 15, 25, 35 and, 45 °C. The results showed the average values of WSOC were higher in urban than in rural green spaces, but the percentage of WSOC to total organic carbon (TOC) showed an opposite trend. No significant changes were found among the three green space types in WSOC and WSOC/TOC. Response of WSOC in green space to incubation temperature was generally highest in urban sites, followed by suburban sites, and lowest in rural sites at the incubation temperature of 5 °C, but showed an opposite trend at the temperature of 45 °C. Response coefficient of WSOC to temperature change was lower in forest and shrub than in grassland, but increased along the urban-rural gradient. Further analysis showed that WSOC positively correlated with TOC, total nitrogen and available phosphorus, and the response coefficient of WSOC to temperature change negatively correlated with available phosphorus. In summary, exogenous substances input might lead to the accumulation of WSOC in urban green space, however, urban environment was helpful to maintain the stability of WSOC, which might be due to the enrichment of available phosphorus in urban sites.

Variation in carbon storage and its distribution by stand age and forest type in boreal and temperate forests in northeastern China.

The northeastern forest region of China is an important component of total temperate and boreal forests in the northern hemisphere. But how carbon (C) pool size and distribution varies among tree, understory, forest floor and soil components, and across stand ages remains unclear. To address this knowledge gap, we selected three major temperate and two major boreal forest types in northeastern (NE) China. Within both forest zones, we focused on four stand age classes (young, mid-aged, mature and over-mature). Results showed that total C storage was greater in temperate than in boreal forests, and greater in older than in younger stands. Tree biomass C was the main C component, and its contribution to the total forest C storage increased with increasing stand age. It ranged from 27.7% in young to 62.8% in over-mature stands in boreal forests and from 26.5% in young to 72.8% in over-mature stands in temperate forests. Results from both forest zones thus confirm the large biomass C storage capacity of old-growth forests. Tree biomass C was influenced by forest zone, stand age, and forest type. Soil C contribution to total forest C storage ranged from 62.5% in young to 30.1% in over-mature stands in boreal and from 70.1% in young to 26.0% in over-mature in temperate forests. Thus soil C storage is a major C pool in forests of NE China. On the other hand, understory and forest floor C jointly contained less than 13% and <5%, in boreal and temperate forests respectively, and thus play a minor role in total forest C storage in NE China.

[Soil organic carbon density and its influencing factors of major forest types in the forest region of Northeast China].

The five main forest types, i. e. larch forest, birch forest, coniferous mixed forest, broad-leaved mixed forest, and coniferous and broad-leaved mixed forest, with varied age classes (young, mid-aged, pre-mature, and over mature) were selected to investigate the variation of soil organic carbon density in the forest region of Northeast China. Results showed that both soil organic carbon content and density were relatively larger in the top soil layer across the five forest types, decreased gradually with soil depth, and varied insignificantly with either forest type or forest age class. The forest soil carbon density was concentrated in the top 20 cm of soil which accounted for 84.7% - 86.1%, 51.7% - 59.8% and 51.2% - 53.4% of total soil carbon density in the Daxing' an, Xiaoxing' an and Changbai mountains, respectively. The total soil organic carbon density decreased significantly with increasing latitude, which might be attributed mainly to the thickness of soil layers in forest region of Northeast China.

Enhanced expression of cholecystokinin-2 receptor promotes the progression of colon cancer through activation of focal adhesion kinase.

Focal adhesion kinase (FAK) is suggested to be intimately involved in the progression of malignancies. Our previous research has demonstrated that activation of cholecystokinin-2 receptor (CCK2R) by gastrin stimulates a rapid activation of FAK pathway in human colon cancer cells. The purpose of this study is to determine the role of CCK2R and FAK in the progression of colon cancer. In this study, matched tissue samples of primary colon cancer and adjacent normal colon mucosa from the same patient were collected from 45 patients with colon cancer undergoing surgical resection. The gastrin expression was detected using reverse transcription polymerase chain reaction (RT-PCR). The CCK2R expression was examined by in situ hybridization and RT-PCR. The expression of FAK and phosphorylated FAK at tyrosine 397 (phospho-FAK) were detected using immunohistochemistry and immunoblotting. Colo320 and SW787, 2 colon cancer cell lines with or without CCK2R expression, were recruited in this study. Antisense oligonucleotide of FAK was used to block the expression of FAK. Invasiveness and motility of colon cancer cells were detected by Boyden chamber. In this series, enhanced expression of gastrin, CCK2R, FAK and phospho-FAK were observed in colon cancer tissues. CCK2R expression correlated with expression of phospho-FAK. Coexpression of CCK2R and phospho-FAK associated with invasion and lymph node metastasis. Increased invasion and motility was induced by gastrin in Colo320 cells. Overexpression of CCK2R by stable transfection of CCK2R plasmid amplified this increase and incubation with 1 microM L-365,260, a specific CCK2R antagonist, completely inhibited the effect of gastrin. FAK antisense largely blocked the increase of invasion and motility in Colo320 cells. Our data represent the evidence for the CCK2R regulating invasion and motility of colon cancer cells, and support a role of CCK2R in the progression of colon cancer. FAK play a critical role in this CCK2R-mediated effect.