Timing of forest fine root production advances with reduced snow cover in northern Japan: implications for climate-induced change in understory and overstory competition.

To investigate the effect of reduced snow cover on fine root dynamics in a cool-temperate forest in northern Japan because of decreases in snowfall at high latitudes due to global warming, we monitored root length, production, and mortality before and after snow removal with an in-ground root scanner. We measured root dynamics of both overstory deciduous oak (Quercus crispula) and understory evergreen dwarf bamboo (Sasa nipponica), the two major species in the forest. Snow removal advanced the timing of peak root production by a month both in total and in Sasa, but not in oak. There was a significant interaction between snow removal and plant form on root production; this indicates that enhanced Sasa root production following snow removal might increase its ability to compete with oak. In contrast, snow removal did not enhance root mortality, suggesting that the roots of these species tolerate soil freezing. The earlier snow disappearance in the snow removal plot expanded the growing season in Sasa. We speculate that this change in the understory environment would advance the timing of root production by Sasa by extending the photosynthetic period in spring. We propose that different responses of root production to reduced snow cover between the two species would change the competitive interactions of overstory and understory vegetation, influencing net primary production and biogeochemistry (e.g., carbon and nitrogen cycles) in the forest ecosystem.

The Impacts of Soil Fertility and Salinity on Soil Nitrogen Dynamics Mediated by the Soil Microbial Community Beneath the Halophytic Shrub Tamarisk.

Nitrogen (N) is one of the most common limiting nutrients for primary production in terrestrial ecosystems. Soil microbes transform organic N into inorganic N, which is available to plants, but soil microbe activity in drylands is sometimes critically suppressed by environmental factors, such as low soil substrate availability or high salinity. Tamarisk (Tamarix spp.) is a halophytic shrub species that is widely distributed in the drylands of China; it produces litter enriched in nutrients and salts that are thought to increase soil fertility and salinity under its crown. To elucidate the effects of tamarisks on the soil microbial community, and thus N dynamics, by creating "islands of fertility" and "islands of salinity," we collected soil samples from under tamarisk crowns and adjacent barren areas at three habitats in the summer and fall. We analyzed soil physicochemical properties, inorganic N dynamics, and prokaryotic community abundance and composition. In soils sampled beneath tamarisks, the N mineralization rate was significantly higher, and the prokaryotic community structure was significantly different, from soils sampled in barren areas, irrespective of site and season. Tamarisks provided suitable nutrient conditions for one of the important decomposers in the area, Verrucomicrobia, by creating "islands of fertility," but provided unsuitable salinity conditions for other important decomposers, Flavobacteria, Gammaproteobacteria, and Deltaproteobacteria, by mitigating salt accumulation. However, the quantity of these decomposers tended to be higher beneath tamarisks, because they were relatively unaffected by the small salinity gradient created by the tamarisks, which may explain the higher N mineralization rate beneath tamarisks.

Root endophytic bacterial and fungal communities in a natural hot desert are differentially regulated in dry and wet seasons by stochastic processes and functional traits.

Dryland ecosystems experience seasonal cycles of severe drought and moderate precipitation. Desert plants may develop symbiotic relationships with root endophytic microbes to survive under the repeated wet and extremely dry conditions. Although community coalescence has been found in many systems, the colonization by functional microbes and its relationship to seasonal transitions in arid regions are not well understood. Here we examined root endophytic microbial taxa, and their traits in relation to their root colonization, during the dry and wet seasons in a hot desert of the southwestern United States. We used high-throughput DNA sequencing of 16S rRNA and internal transcribed spacer gene profiling of five desert shrubs, and analyzed the seasonal change in endophytic microbial lineages. Goodness of fit to the neutral community model in relationship to microbial traits was evaluated. In summer, Actinobacteria and Bacteroidia increased, although this was not genus-specific. For fungi, Glomeraceae selectively increased in summer. In winter, Gram-negative bacterial genera, including those capable of nitrogen fixation and plant growth promotion, increased. Neutral model analysis revealed a strong stochastic influence on endophytic bacteria but a weak effect for fungi, especially in summer. The taxa with higher frequency than that predicted by neutral model shared environmental adaptability and symbiotic traits, whereas the frequency of pathogenic fungi was at or under the predicted value. These results suggest that community assembly of bacteria and fungi is regulated differently. The bacterial community was affected by stochastic and deterministic processes via bacterial response to drought (response trait), beneficial effect on plants (effect trait), and likely stable mutualistic interactions with plants suggested by the frequency of nodule bacteria. For fungi, mycorrhizal fungi were selected by plants in summer. The regulation of beneficial microbes by plants in both dry and wet seasons suggests the presence of plant-soil positive feedback in this natural desert ecosystem.

Seasonal accumulation of photoassimilated carbon relates to growth rate and use for new aboveground organs of young apple trees in following spring.

Deciduous trees accumulate carbon (C) in woody parts during the growth season which is subsequently used for the initial development and growth of newly formed organs in the following season; however, it is unclear which period during the growth season contributes to C accumulation. Three-year-old potted Malus domestica (apple) trees were grown in controlled growth chambers during the growth season and exposed to 13CO2 in an exposure chamber at seven different periods of the growth season, including vegetative and reproductive growth periods. Approximately half of the trees were harvested in late autumn, and the remaining trees were grown in a field in the following year. The 13C accumulation in the different organs in late autumn, and its concentration in the new aboveground growth during the following growth season, was determined. The concentration of the photoassimilated 13C in woody parts (shoots, trunk, rootstock and coarse roots) in the late autumn was higher in the trees labeled during the period of vigorous vegetative growth than in those labeled during other periods of growth. Furthermore, 13C concentration in the leaves, annual shoots, flower buds and flowers in the following early spring was also high in the trees labeled during this period. The concentration of 13C in the flower buds and flowers was positively correlated with that in the woody parts in the late autumn and old shoots in the following spring. Hence, the seasonal accumulation of photoassimilated C in woody parts in late autumn is related to growth rates during the growth season and its use for the initial development of newly formed organs in the following spring. These results suggest that under non-stressed conditions, C accumulated during the period of vigorous vegetative growth largely contributes to the C reserves that are used for the development of new organs in the following year.

DIRECT ASSIMILATION OF ATMOSPHERIC CARBON BY IMMATURE APPLE FRUITS.

Although fruit development primarily depends on photoassimilation by leaves, immature green fruits can also directly assimilate atmospheric CO2. To elucidate the process of C accumulation due to direct assimilation by fruit, we conducted a 13CO2 exposure experiment in an orchard in late June with immature 'Fuji' apples (Malus domestica). Four fruits from three trees were enclosed in transparent plastic bags and exposed to 13CO2 using an in-situ exposure system. Fruits were collected prior to and immediately following exposure in early July, late September and mid-November, and 13C concentrations in the peduncle, skin, flesh and core (including seeds) were measured. The higher assimilated 13C concentrations measured following exposure indicated that the fruits directly assimilated atmospheric 13C. The 13C concentration in fruit skin was higher immediately after exposure and in early July compared with that prior to exposure. In late September and mid-November, 13C concentrations were close to natural levels.

EXPERIMENTAL EVALUATION OF DISTRIBUTION OF 14C PHOTOASSIMILATED INTO CARBOHYDRATES IN DIFFERENT GROWTH STAGES OF FRUIT-BEARING APPLE SHOOTS USING A 13CO2 IN-SITU EXPOSURE SYSTEM.

In this study, we aimed to investigate the photoassimilation process of 14CO2 into agricultural plants through determining the photoassimilated carbohydrate-13C in each part (leaves, current branch and fruit) of the fruit-bearing apple shoots exposed to 13CO2 in different growth stages (early and late fruit development stages). The carbohydrate content was assessed as soluble (ethanol-extracted fraction) and other (HCl-extracted and residual fractions) components. The total (i.e. sum of the three fractions) bulk carbohydrate concentrations in all parts of the shoots were statistically similar between different growth stages. The changes in the concentration of 13C-labeled soluble carbohydrate (i.e. ethanol-extracted fraction) to the total content between different growth stages were statistically unclear among all parts of the shoot. These results suggest that the distribution ratio of photoassimilated 13C in soluble and other components in the apple shoot was thereabout constantly independent of the growth stages.

Investigation of ratio of carbon to hydrogen (C/H ratio) in agricultural plants for further estimation of their productivity of organically bound tritium.

The carbon to hydrogen ratio (C/H ratio, w/w) in plants is a key factor in estimating the amount of hydrogen in the photosynthetic product. The amount of hydrogen calculated from photosynthetic model estimation associated with the C/H ratio is an essential parameter of the estimation model of productivity of organically bound tritium (OBT) by plants. To propose a sophisticated estimation model of OBT by agricultural plants, temporal changes in the C/H ratio of six plant species (Japanese radish, cabbage, orchard grass, paddy field rice, apple, and radish) during their cultivation were investigated for each plant part. The C/H ratio in the plants cultivated in the field and growth chamber generally exceeded 6, which is the value for the primary photosynthetic monosaccharides, such as glucose and fructose (both chemical formulae, C6H12O6). In the vegetative parts (e.g. Japanese radish leaves, cabbage leaves and roots, rice leaves and roots, and radish leaves and fine roots) the C/H ratio fluctuated irregularly or remained constant within an approximate range of 6.6-7.3 during cultivation. The C/H ratio in enlarged organs (e.g. Japanese radish root, rice ear, apple fruit, and radish main root) decreased continuously, approaching 6. These results suggest that the C/H ratio can be generally set as approximately 6.9 except for enlarged organs, in which the ratio may change over time during cultivation, within an approximate range of 6-7.

DEVELOPMENT OF IN-SITU SYSTEMS FOR 13CO2 EXPOSURE AND DETERMINATION OF 13C NET ASSIMILATION RATE OF FRUIT-BEARING SHOOTS AND WHOLE TREE AND CHAMBER FOR PRECISION 13CO2 EXPOSURE OF YOUNG POTTED TREES.

14C is the most important radionuclide for assessing exposure dose around the Rokkasho nuclear fuel reprocessing plant, Aomori Prefecture, Japan. A simplified model with ample margins has been used so far for the assessment of the dose derived from 14C. Realistic dose estimate of 14C using more realistic model is necessary for safety. Apple production is an important core industry in Aomori Prefecture. To construct the dynamic model for apple, using 13C as a tracer to substitute for 14C, we collected data on 13C abundance in organs including fruit after 13CO2 exposure at various fruit growth stages. We developed 13CO2 exposure systems for three intact fruit-bearing shoots (1), whole mature tree (2) and young potted trees (3). Systems (1) and (2) also can determine net amounts of carbon and 13C photoassimilated. System (3) is capable of precise feedback control of 13CO2 and 12CO2 concentrations based on on-time determination.

In situ experimental exposure of fruit-bearing shoots of apple trees to 13CO2 and construction of a dynamic transfer model of carbon.

Evaluating the transfer and metabolism of carbon (C) in apple fruit is key to estimating the potential accumulation of atmospheric 14C in fruit near and around nuclear facilities. We developed a dynamic compartment model for apple fruit-bearing shoots, assuming that the shoots are a simple unit of source and sink for photoassimilates. Fruit-bearing shoots of Malus domestica "Fuji" at different fruit growth stages were exposed to 13CO2in situ, followed by sampling at 72 h after exposure or at harvest. The 13C/(13C+12C) mole ratio in fruits, leaves, and current branch were measured to construct a five-compartment model of 13C (fruit, each fast and slow component of leaves, and current branch). The C inventories in the compartments were presented in accordance with the measured growth curves of C in the organs. The model simulated the 13C dynamics in plant tissues well. Simulation results of photoassimilate distribution using the model indicated that the retention of photoassimilated C at the harvest depended on the growth rate of C in the organs at the exposure.

Effect of soil salinity and nutrient levels on the community structure of the root-associated bacteria of the facultative halophyte, Tamarix ramosissima, in southwestern United States.

Tamarix ramosissima is a tree species that is highly resistant to salt and drought. The Tamarix species survives in a broad range of environmental salt levels, and invades major river systems in southwestern United States. It may affect root-associated bacteria (RB) by increasing soil salts and nutrients. The effects of RB on host plants may vary even under saline conditions, and the relationship may be important for T. ramosissima. However, to the best of our knowledge, there have been no reports relating to T. ramosissima RB and its association with salinity and nutrient levels. In this study, we have examined this association and the effect of arbuscular mycorrhizal colonization of T. ramosissima on RB because a previous study has reported that colonization of arbuscular mycorrhizal fungi affected the rhizobacterial community (Marschner et al., 2001). T. ramosissima roots were collected from five locations with varying soil salinity and nutrient levels. RB community structures were examined by terminal restriction fragment (T-RF) length polymorphism, cloning, and sequencing analyses. The results suggest that RB richness, or the diversity of T. ramosissima, have significant negative relationships with electrical conductivity (EC), sodium concentration (Na), and the colonization of arbuscular mycorrhizal fungi, but have a significant positive relationship with phosphorus in the soil. However, at each T-RF level, positive correlations between the emergence of some T-RFs and EC or Na were observed. These results indicate that high salinity decreased the total number of RB species, but some saline-tolerant RB species multiplied with increasing salinity levels. The ordination scores of nonmetric multidimensional scale analysis of RB community composition show significant relationships with water content, calcium concentration, available phosphorus, and total nitrogen. These results indicate that the RB diversity and community composition of T. ramosissima are affected by soil salinity and nutrient levels. Sequence analysis detected one Bacteroidetes and eight Proteobacteria species. Most 16S rRNA gene sequences had high similarities with the bacteria isolated from saline conditions, indicating that at least a portion of the RB species observed in T. ramosissima was halotolerant.