Physiological characteristics of pure cultures of a white-colored truffle Tuber japonicum.

A white-colored truffle Tuber japonicum, indigenous to Japan, is an ascomycetous ectomycorrhizal fungus. To clarify the physiological characteristics of this fungus, we investigated the influence of culture medium, temperature, and sources of nitrogen (N) and carbon (C) on the growth of five strains. Tuber japonicum strains grew better on malt extract and modified Melin-Norkrans medium, and showed peak growth at 20 °C or 25 °C. This fungus utilized inorganic (NH4 + and NO3 -) and organic N sources (casamino acids, glutamine, peptone, urea, and yeast extract). Additionally, this fungus utilized various C sources, such as monosaccharide (arabinose, fructose, galactose, glucose, and mannose), disaccharide (maltose, sucrose, and trehalose), polysaccharide (dextrin and soluble starch), and sugar alcohol (mannitol). However, nutrient sources that promote growth and their effects on growth promotion widely varied among strains. This can result from the strain difference in enzyme activities involved in the assimilation and metabolism of these sources.

Fine Root Growth of Black Spruce Trees and Understory Plants in a Permafrost Forest Along a North-Facing Slope in Interior Alaska.

Permafrost forests play an important role in the global carbon budget due to the huge amounts of carbon stored below ground in these ecosystems. Although fine roots are considered to be a major pathway of belowground carbon flux, separate contributions of overstory trees and understory shrubs to fine root dynamics in these forests have not been specifically characterized in relation to permafrost conditions, such as active layer thickness. In this study, we investigated fine root growth and morphology of trees and understory shrubs using ingrowth cores with two types of moss substrates (feather- and Sphagnum mosses) in permafrost black spruce (Picea mariana) stands along a north-facing slope in Interior Alaska, where active layer thickness varied substantially. Aboveground biomass, litterfall production rate, and fine root mass were also examined. Results showed that aboveground biomass, fine root mass, and fine root growth of black spruce trees tended to decrease downslope, whereas those of understory Ericaceae shrubs increased. Belowground allocation (e.g., ratio of fine root growth/leaf litter production) increased downslope in both of black spruce and understory plants. These results suggested that, at a lower slope, belowground resource availability was lower than at upper slope, but higher light availability under open canopy seemed to benefit the growth of the understory shrubs. On the other hand, understory shrubs were more responsive to the moss substrates than black spruce, in which Sphagnum moss substrates increased fine root growth of the shrubs as compared with feather moss substrates, whereas the effect was unclear for black spruce. This is probably due to higher moisture contents in Sphagnum moss substrates, which benefited the growth of small diameter (high specific root length) fine roots of understory shrubs. Hence, the contribution of understory shrubs to fine root growth was greater at lower slope than at upper slope, or in Sphagnum than in feather-moss substrates in our study site. Taken together, our data show that fine roots of Ericaceae shrubs are a key component in belowground carbon flux at permafrost black spruce forests with shallow active layer and/or with Sphagnum dominated forest floor.

Different Waterlogging Depths Affect Spatial Distribution of Fine Root Growth for Pinus thunbergii Seedlings.

The increase of waterlogged environments at forests and urban greenery is of recent concern with the progress of climate change. Under waterlogging, plant roots are exposed to hypoxic conditions, which strongly affect root growth and function. However, its impact is dependent on various factors, such as waterlogging depth. Therefore, our objective is to elucidate effects of different waterlogging depths on Pinus thunbergii Parl., which is widely used for afforestation, especially at coastal forests. We conducted an experiment to examine growth and morphology of fine roots and transpiration using 2-year-old seedlings under three treatments, (1) control (no waterlogging), (2) partial waterlogging (partial-WL, waterlogging depth = 15 cm from the bottom), and (3) full waterlogging (full-WL, waterlogging depth = from the bottom to the soil surface, 26 cm). As a result, fine root growth and transpiration were both significantly decreased at full-WL. However, for partial-WL, fine root growth was significantly increased compared to control and full-WL at the top soil, where it was not waterlogged. Additionally, transpiration which had decreased after 4 weeks of waterlogging showed no significant difference compared to control after 8 weeks of waterlogging. This recovery is to be attributed to the increase in fine root growth at non-waterlogged top soil, which compensated for the damaged roots at the waterlogged bottom soil. In conclusion, this study suggests that P. thunbergii is sensitive to waterlogging; however, it can adapt to waterlogging by plastically changing the distribution of fine root growth.

Potassium fertilisation reduces radiocesium uptake by Japanese cypress seedlings grown in a stand contaminated by the Fukushima Daiichi nuclear accident.

We analysed suppressive effects of potassium (K) fertilisation on radiocesium (137Cs) uptake by hinoki cypress (Chamaecyparis obtusa) seedlings from soils contaminated after the Fukushima Daiichi Nuclear Power Plant accident. Three-year-old seedlings were planted in a clear-cut forest (ca. 4 ha) during June-July 2014, and potassium chloride fertiliser (83 kg K ha-1) was applied twice (August 2014 and April 2015). 137Cs concentrations in the needles in the fertilised plots were one-eighth of those in the control (unfertilised) plots at the end of the second growing season (October 2015). Our results clearly indicated that K fertilisation reduced radiocesium transfer from soil to planted cypress seedlings. A linear mixed model analysis revealed that 137Cs concentrations in the needles were significantly affected by 137Cs inventory in the soil (Bq m-2) adjacent to the sampled seedlings, exchangeable K concentrations in surface mineral soils (0-5 cm) and fertilisation. The exchangeable K concentrations in surface soils in October 2015 did not differ from those in August 2014 (before fertilisation) in the fertilised plots and in the control plots. These results suggested that the levels of exchangeable K would temporarily increase by fertilisation during the growing season, and radiocesium uptake by tree roots was suppressed.

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.

Biomass and morphology of fine roots of sugi (Cryptomeria japonica) after 3 years of nitrogen fertilization.

Increasing nitrogen (N) deposition may affect carbon and nutrient dynamics in forest ecosystems. To better understand the effects of N deposition, we need to improve our knowledge of N effects on fine roots (roots <2 mm in diameter), as they are a key factor in carbon and nutrient dynamics. In this study, we fertilized 1 × 2 m plots in a sugi (Cryptomeria japonica) stand (336 kg ha(-) (1) y(-) (1)) for 3 years and evaluated the responses of the fine roots to high N load. After fertilization, the concentration of NO3-N in the soil of N-fertilized (NF) plots was five-times as large as that in the control plots and the effect was more remarkable in the subsurface soil than in the surface soil. The biomass of fine roots <2 mm in diameter appeared to be greater in the NF plots (88 ± 19 g m(-) (2)) than in the control plots (56 ± 14 g m(-) (2)), but this difference was not statistically significant. In both plots, 76% of the biomass was accounted for by fine roots that were <1 mm in diameter. In the surface soil, the specific root length of fine roots <1 mm in diameter was significantly greater, and the diameter of those fine roots was marginally smaller, in the NF plots than in the control plots. In addition, the concentration of N in fine roots <1 mm in diameter was marginally greater in the NF plots than in the control plots. There may have been increased production of thinner fine roots or increased root branching in the NF plots. This study suggests that, in general, high N load is likely to have positive effects on sugi in terms of fine root characteristics and the effects on fine-root morphology are more evident than the effects on fine-root biomass.

Arabidopsis boron transporter for xylem loading.

Boron deficiency hampers the productivity of 132 crops in more than 80 countries. Boron is essential in higher plants primarily for maintaining the integrity of cell walls and is also beneficial and might be essential in animals and in yeast. Understanding the molecular mechanism(s) of boron transport is crucial for alleviating boron deficiency. Here we describe the molecular identification of boron transporters in biological systems. The Arabidopsis thaliana mutant bor1-1 is sensitive to boron deficiency. Uptake studies indicated that xylem loading is the key step for boron accumulation in shoots with a low external boron supply and that the bor1-1 mutant is defective in this process. Positional cloning identified BOR1 as a membrane protein with homology to bicarbonate transporters in animals. Moreover, a fusion protein of BOR1 and green fluorescent protein (GFP) localized to the plasma membrane in transformed cells. The promoter of BOR1 drove GFP expression in root pericycle cells. When expressed in yeast, BOR1 decreased boron concentrations in cells. We show here that BOR1 is an efflux-type boron transporter for xylem loading and is essential for protecting shoots from boron deficiency.