Allometry of the quasi-pipe (qPipe) model for estimating tree leaf area and tree leaf mass applied to plant functional types.

The allometry of the pipe model quantifies the approximate proportionality between the tree leaf amount and the stem cross-sectional area at the crown base (ACB). It is useful for estimating and modeling carbon fixation abilities of trees but requires climbing the tree and is thus unsuitable for large-scale studies. Here, we adopted a previously proposed allometry (hereafter the quasi-pipe (qPipe) model allometry) formulating the relationship between the tree leaf amount and a surrogate of ACB, ACB_Est, calculated from tree dimensions measurable from the ground. Using published/unpublished data for 962 trees of 159 species collected between tropical rainforests and boreal forests, we established pipe and qPipe model allometries for evergreen-conifer, deciduous-conifer, evergreen-broadleaf, and deciduous-broadleaf plant functional types (PFTs). For the leaf area per tree (LA), allometric lines on a log-log plane were almost identical among the four PFTs in both models, with slopes of ~ 1. For the leaf mass per tree (LM), however, the allometric lines separated among the four PFTs in both models and had slopes greater than 1, indicating that the proportionality assumed in the pipe model held for LA but not LM. The applicability of the qPipe model in estimating the stand-scale leaf amount was further examined.

Carbon use strategies in shoot and acorn growth of two evergreen broadleaf trees unraveled by seasonal carbohydrate measurements and carbon isotope analysis.

Woody species have evolved carbon (C) storage processes that meet needs for reserves associated with asynchronies between C supply and demand. However, our understanding of storage dynamics is still elusive in mature trees, especially when reproduction is involved. Integrated analyses of isotope ratios, concentrations, and biomass may enhance understanding of stored C fractions' dynamics and roles. Thus, we monitored starch and soluble sugars (SSs), C isotope ratios, and biomass, in leaves, twigs and reproductive organs of two mature evergreen broadleaf trees, Quercus glauca and Lithocarpus edulis, for two years. During the growing season, no starch was observed in twigs, while constant starch levels were observed in leaves. Increase in SSs for winter hardening was earlier in L. edulis than in Q. glauca, in line with L. edulis acorns' earlier ripening. Decrease in SSs and increase in starch occurred simultaneously in the next spring. In addition, sucrose accounted for less than 10% of total SSs in leaves of both species, whereas mannose accounted for up to 75% in Q. glauca and myo-inositol up to 23% in L. edulis, indicating species specific sugar composition. These results indicate that seasonal variation of SSs fraction was more reflective to climatic change and NSC storage was less influenced by reproduction. No starch was detected in acorn organs of either Q. glauca or L. edulis except in ripening seeds. The biomass of ripe acorns was 1.7- and 6.4-fold greater than that of current-year twigs in Q. glauca and L. edulis, respectively. Bulk twigs and reproductive organs were ca. 1.0‰ 13C enriched relative to bulk leaves, which was lower than in deciduous trees. These results indicate that new photo-assimilate is the predominant C source for reproductive growth. These findings provide new insights into the dynamics of C storage in relation to reproduction in evergreen broadleaf trees.

Nitrate is an important nitrogen source for Arctic tundra plants.

Plant nitrogen (N) use is a key component of the N cycle in terrestrial ecosystems. The supply of N to plants affects community species composition and ecosystem processes such as photosynthesis and carbon (C) accumulation. However, the availabilities and relative importance of different N forms to plants are not well understood. While nitrate (NO3-) is a major N form used by plants worldwide, it is discounted as a N source for Arctic tundra plants because of extremely low NO3- concentrations in Arctic tundra soils, undetectable soil nitrification, and plant-tissue NO3- that is typically below detection limits. Here we reexamine NO3- use by tundra plants using a sensitive denitrifier method to analyze plant-tissue NO3- Soil-derived NO3- was detected in tundra plant tissues, and tundra plants took up soil NO3- at comparable rates to plants from relatively NO3--rich ecosystems in other biomes. Nitrate assimilation determined by 15N enrichments of leaf NO3- relative to soil NO3- accounted for 4 to 52% (as estimated by a Bayesian isotope-mixing model) of species-specific total leaf N of Alaskan tundra plants. Our finding that in situ soil NO3- availability for tundra plants is high has important implications for Arctic ecosystems, not only in determining species compositions, but also in determining the loss of N from soils via leaching and denitrification. Plant N uptake and soil N losses can strongly influence C uptake and accumulation in tundra soils. Accordingly, this evidence of NO3- availability in tundra soils is crucial for predicting C storage in tundra.

Influence of reproduction on nitrogen uptake and allocation to new organs in Fagus crenata.

The contributions of the internal nitrogen (N) cycle and N uptake from soil to growth in mature trees remain poorly understood, especially during reproduction. In order to elucidate how reproduction affects N uptake, allocation and remobilization, we applied pulse 15N labelling to three fruiting (F) and three non-fruiting (NF) Fagus crenata Blume trees after the leaves were fully unfurled. Three-year-old branches were sampled from upper crowns at about 2 week intervals until leaf fall. 15N content per organ dry mass (15Nexcess) and N concentration in all new shoot organs were determined. Fruiting led to greater 15Nexcess uptake from the soil during the first month following application. Cupules absorbed the highest fraction of 15Nexcess initially and nuts contained about half the 15Nexcess at the end of the growing season. Biomass of reproductive organs represented up to 70% of new shoot growth in F trees. This fruit burden led to 34% and 38% reduction in biomass and 15Nexcess, respectively, in mature leaves compared with NF trees. Moreover, the increment of 15Nexcess in new shoots of F relative to NF trees was lower than the increment of biomass between the two. These results indicate that N is a limiting resource during masting in F. crenata. 15Nexcess incorporated into nuts started to increase dramatically once 15Nexcess in leaves, branches and cupules hit seasonal maxima. Similar seasonal biomass growth patterns were also found in these organs, indicating that sink strength drives uptake and allocation of 15Nexcess between new shoot compartments. These results, together with translocation of 15Nexcess from cupules and senescing leaves to nuts (contributing to fruit ripening), suggest that a finely tuned growth phenology alleviated N limitation. Thus, fruiting did not influence the N concentration in leaves or branches. These reproduction-related variations in N uptake and allocation among new shoot compartments have implications for N dynamics in the plant-soil system.

Growth rate reduction causes a decline in the annual incremental trunk growth in masting Fagus crenata trees.

Tree trunk annual increments are markedly reduced in mast years. There are two hypotheses that could explain the mechanism for this phenomenon: (1) a reduction in the duration of growth due to switching the resource allocation from somatic growth to seed production; (2) reduction of growth rate due to resources being shared between somatic growth and reproduction simultaneously. In this study, we aimed to test these hypotheses in Fagus crenata  Blume from the point of view of resource allocation. The radial growth patterns in F. crenata during a year without reproduction (2014) and a masting year (2015) were monitored using a digital dendrometer. At the same time, shoot growth patterns were monitored by sampling branches from the top of the canopy. Data obtained using the digital dendrometer were fitted to a sigmoidal function, and the parameters of the function were evaluated with a hierarchal Bayesian approach; estimated parameters were used to represent the properties of trunk growth phenology. Trunk growth started synchronously just after leaf unfurling in both mass-fruiting (F15) and limited-fruiting (NF15) trees in 2014 and 2015. Reproduction reduced the growth rate in 2015. This was due to the resources being allocated for the development of cupules and for formation of relatively thick branches, both of which occurred simultaneously with trunk growth. There was no clear difference in the duration of radial growth between F15 and NF15 trees in the 2 years, although seed maturation started after trunk growth ceased. As a result, the annual trunk radius increment was reduced in the F15 trees in 2015. These results suggested that reduction of radial growth rate (Hypothesis 2) caused the reduction in annual trunk increment of reproducing trees of this species.

Calculation procedures to estimate fine root production rates in forests using two-dimensional fine root data obtained by the net sheet method.

Several recent studies have used the net sheet method to estimate fine root production rates in forest ecosystems, wherein net sheets are inserted into the soil and fine roots growing through them are observed. Although this method has advantages in terms of its easy handling and low cost, there are uncertainties in the estimates per unit soil volume or unit stand area, because the net sheet is a two-dimensional material. Therefore, this study aimed to establish calculation procedures for estimating fine root production rates from two-dimensional fine root data on net sheets. This study was conducted in a hinoki cypress (Chamaecyparis obtusa (Sieb. & Zucc.) Endl.) stand in western Japan. We estimated fine root production rates in length and volume from the number (RN) and cross-sectional area (RCSA) densities, respectively, for fine roots crossing the net sheets, which were then converted to dry mass values. For these calculations, we used empirical regression equations or theoretical equations between the RN or RCSA densities on the vertical walls of soil pits and fine root densities in length or volume, respectively, in the soil, wherein the theoretical equations assumed random orientation of the growing fine roots. The estimates of mean fine root (diameter <1 mm) production rates were ∼80-100 g m-2 year-1 using the empirically obtained regression equations, whereas those from the theoretical equations were ∼40-50 g m-2 year-1. The difference in the estimates was attributed to larger slope values of the empirical regression equations than those of the theoretical equations, suggesting that fine root orientation was not random in our study site. In light of these results, we concluded that fine root production rates were successfully estimated from two-dimensional fine root data on the net sheets using these calculation procedures, with the empirical regression equations reflecting fine root orientation in the study site.

Reproduction-related variation in carbon allocation to woody tissues in Fagus crenata using a natural 13C approach.

The contribution of new photo-assimilates and stored carbon (C) to plant growth remains poorly understood, especially during reproduction. In order to elucidate how mast seeding affects C allocation to both reproductive and vegetative tissues, we measured biomass increase in each tissue, branch starch concentration and stable C isotope composition (δ13C) in bulk leaves, current-year shoots, 3-year branches and tree rings in fruiting and non-fruiting trees for 2 years, as well as in fruits. We isolated the effect of reproduction on C allocation to vegetative growth by comparing 13C enrichment in woody tissues in fruiting and non-fruiting specimens. Compared with 2‰ 13C enrichment in shoots relative to leaves from non-fruiting trees, fruiting reduced the enrichment to 1‰ and this reduction disappeared in the following year with no fruiting, indicating that new photo-assimilates are preferentially used for woody tissues even with fruiting burden. In contrast, fruits had up to 2.5‰ 13C enrichment at mid-summer, which dropped thereafter, indicating that fruit production relies on C storage early in the growing season then shifts to current photo-assimilates. At this tipping point, growth of shoots and cupules had almost finished and nuts had a second rapid growth period thereafter. Together with shorter shoots but higher biomass increment per length in fruiting trees than non-fruiting trees, these results indicate that the C limitation due to fruit burden is minimized by fine-tuning of allocation of old C stores and new photo-assimilates, along with the growth pattern in various tissues. Furthermore, fruiting had no significant effect on starch concentration in 3-year-old branches, which became fully depleted during leaf and flower flushing but were quickly replenished. These results indicate that reproduction affects C allocation to branches but not its source or storage. These reproduction-related variations in the fate of C have implications for evaluating forest ecosystem C cycles during climate change.

Nitrogen storage dynamics are affected by masting events in Fagus crenata.

It is generally assumed that the production of a large crop of seeds depletes stores of resources and that these take more than 1 year to replenish; this is accepted, theoretically, as the proximate mechanism of mast seeding (resource budget model). However, direct evidence of resource depletion in masting trees is very rare. Here, we trace seasonal and inter-annual variations in nitrogen (N) concentration and estimate the N storage pool of individuals after full masting of Fagus crenata in two stands. In 2005, a full masting year, the amount of N in fruit litter represented half of the N present in mature leaves in an old stand (age 190-260 years), and was about equivalent to the amount of N in mature leaves in a younger stand (age 83-84 years). Due to this additional burden, both tissue N concentration and individual N storage decreased in 2006; this was followed by significant replenishment in 2007, although a substantial N store remained even after full masting. These results indicate that internal storage may be important and that N may be the limiting factor for fruiting. In the 4 years following full masting, the old stand experienced two moderate masting events separated by 2 years, whilst trees in the younger stand did not fruit. This different fruiting behavior may be related to different "costs of reproduction" in the full masting year 2005, thus providing more evidence that N may limit fruiting. Compared to the non-fruiting stand, individuals in the fruiting stand exhibited an additional increase in N concentrations in roots early in the 2007 growing season, suggesting additional N uptake from the soil to supply resource demand. The enhanced uptake may alleviate the N storage depletion observed in the full masting year. This study suggests that masting affects N cycle dynamics in mature Fagus crenata and N may be one factor limiting fruiting.

BioVision: an application for the automated image analysis of histological sections.

We describe a computer application, "BioVision", that can be trained to quickly and effectively classify and quantify user definable histological objects (e.g., senile plaques, neurofibrillary tangles) within single or double-labeled immunocytochemically stained sections. For a given image population, BioVision is interactively trained (in Independent User Mode) by an investigator to perform the desired classifications. This training yields a statistical model of the different types of objects occurring in the target image population. The resulting model can then be used (in Automated User Mode) to classify all objects in any image or images from the target population. BioVision simplifies the quantification of complex visual objects and improves inter-rater reliability. The program accomplishes classification in two major stages: pixel classification and blob classification. In pixel classification, each pixel is assigned to one of some number of substance classes, based on its chromatic properties and local context, reflecting basic histological distinctions of interest. In the blob classification phase, the image's pixels are first partitioned into "blobs": maximal connected sets of pixels assigned to the same substance class. Then, based on its size, shape, textural and contextual properties, each blob is assigned to a histological object class. A Bayesian classifier is used in each of the pixel and blob classification stages. We report several tests of BioVision. First, we applied BioVision to classify senile plaques and neurofibrillary tangles in several test cases of Alzheimer's brain immunostained for beta-amyloid and PHF-tau and compared the results to those produced by experienced investigators. BioVision was trained to classify Plaque-type blobs as either plaques or plaque-type nonentities, and tangle-type blobs as either tangles or tangle-type nonentities. BioVision classified the objects with an accuracy comparable to the trained investigator. Next, we applied BioVision to the task of counting all the tangles in hippocampal images from 22 Alzheimer's disease (AD) cases selected to span a broad range of dementia levels from the tissue repository of UC Irvine's Center for the study of Brain Aging and Dementia. The tangle counts produced by BioVision proved to be significantly better predictors of the cases' adjusted MMSE scores than any of tangle load, age at death, post mortem interval or the interval between the last MMSE score and death.

Progressive and nonprogressive rheumatoid arthritis over a 10-year period in Japan.

We followed 207 patients with rheumatoid arthritis who were registered at our hospital over a 10-year period from between 1989 and 1990. The number of patients who were still treated at our hospital in 2001 was 87. Sixty patients had died, 39 had changed hospitals, 11 had interrupted treatment, and 10 had not had further followup. The patients at our hospital in 2001 were divided into 2 groups: progressive and nonprogressive disease. We compared clinical and laboratory data obtained for the 2 groups from the initial examination against data obtained from the final examination. The progressive group had a greater number of operations, used a greater variety of disease modifying antirheumatic drugs, and received higher dosages of steroids than the nonprogressive group. In the progressive group the levels of C-reactive protein, erythrocyte sedimentation rate, and IgG were significantly higher, while the levels of hemoglobin were lower at final laboratory examination. However, the initial laboratory examination revealed no significant differences between these groups that could be used to predict the likely progression of disease.