Transient proliferation of proanthocyanidin-accumulating cells on the epidermal apex contributes to highly aluminum-resistant root elongation in camphor tree.

Aluminum (Al) is a harmful element that rapidly inhibits the elongation of plant roots in acidic soils. The release of organic anions explains Al resistance in annual crops, but the mechanisms that are responsible for superior Al resistance in some woody plants remain unclear. We examined cell properties at the surface layer of the root apex in the camphor tree (Cinnamomum camphora) to understand its high Al resistance mechanism. Exposure to 500 µm Al for 8 d, more than 20-fold higher concentration and longer duration than what soybean (Glycine max) can tolerate, only reduced root elongation in the camphor tree to 64% of the control despite the slight induction of citrate release. In addition, Al content in the root apices was maintained at low levels. Histochemical profiling revealed that proanthocyanidin (PA)-accumulating cells were present at the adjacent outer layer of epidermis cells at the root apex, having distinctive zones for cell division and the early phase of cell expansion. Then the PA cells were gradually detached off the root, leaving thin debris behind, and the root surface was replaced with the elongating epidermis cells at the 3- to 4-mm region behind the tip. Al did not affect the proliferation of PA cells or epidermis cells, except for the delay in the start of expansion and the accelerated detachment of the former. In soybean roots, the innermost lateral root cap cells were absent in both PA accumulation and active cell division and failed to protect the epidermal cell expansion at 25 µm Al. These results suggest that transient proliferation and detachment of PA cells may facilitate the expansion of epidermis cells away from Al during root elongation in camphor tree.

A cell-type-specific defect in border cell formation in the Acacia mangium root cap developing an extraordinary sheath of sloughed-off cells.

BACKGROUND AND AIMS: Root caps release border cells, which play central roles in microbe interaction and root protection against soil stresses. However, the number and connectivity of border cells differ widely among plant species. Better understanding of key border-cell phenotype across species will help define the total function of border cells and associated genes. METHODS: The spatio-temporal detachment of border cells in the leguminous tree Acacia mangium was investigated by using light and fluorescent microscopy with fluorescein diacetate, and their number and structural connectivity compared with that in soybean (Glycine max). KEY RESULTS: Border-like cells with a sheet structure peeled bilaterally from the lateral root cap of A. mangium. Hydroponic root elongation partially facilitated acropetal peeling of border-like cells, which accumulate as a sheath that covers the 0- to 4-mm tip within 1 week. Although root elongation under friction caused basipetal peeling, lateral root caps were minimally trimmed as compared with hydroponic roots. In the meantime, A. mangium columella caps simultaneously released single border cells with a number similar to those in soybean. CONCLUSIONS: These results suggest that cell type-specific inhibitory factors induce a distinct defective phenotype in single border-cell formation in A. mangium lateral root caps.

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.

Primary and secondary aerenchyma oxygen transportation pathways of Syzygium kunstleri (King) Bahadur & R. C. Gaur adventitious roots in hypoxic conditions.

Some plant species develop aerenchyma to avoid anaerobic environments. In Syzygium kunstleri (King) Bahadur & R. C. Gaur, both primary and secondary aerenchyma were observed in adventitious roots under hypoxic conditions. We clarified the function of and relationship between primary and secondary aerenchyma. To understand the function of primary and secondary aerenchyma in adventitious roots, we measured changes in primary and secondary aerenchyma partial pressure of oxygen (pO2) after injecting nitrogen (N2) into the stem 0-3 cm above the water surface using Clark-type oxygen microelectrodes. Following N2 injection, a decrease in pO2 was observed in the primary aerenchyma, secondary aerenchyma, and rhizosphere. Oxygen concentration in the primary aerenchyma, secondary aerenchyma, and rhizosphere also decreased after the secondary aerenchyma was removed from near the root base. The primary and secondary aerenchyma are involved in oxygen transport, and in adventitious roots, they participate in the longitudinal movement of oxygen from the root base to root tip. As cortex collapse occurs from secondary growth, the secondary aerenchyma may support or replace the primary aerenchyma as the main oxygen transport system under hypoxic conditions.

Source of Oxygen Fed to Adventitious Roots of Syzygium kunstleri (King) Bahadur and R.C. Gaur Grown in Hypoxic Conditions.

Syzygium kunstleri, a woody plant species, adapts to hypoxic conditions by developing new adventitious roots. Here, we investigate its morphological adaptation to long-term water level changes and the sources and pathways of O2 supplied to its adventitious roots. Cuttings were cultivated in hydroponic and agar media, and then, the water level was increased by 6 cm following adventitious root emergence; afterward, O2 partial pressure changes were measured using a Clark-type O2 microelectrode. O2 concentrations in the adventitious roots decreased when N2 was injected, regardless of the presence of light, indicating that the O2 source was not photosynthetic when bark was removed. New adventitious roots developed near the surface when the water level increased, and O2 conditions above the raised water level influenced O2 concentrations in adventitious roots. O2 concentrations in adventitious roots that developed before the water level increased were lower than in the newly developed adventitious roots but increased when the O2 concentrations above the original water level increased. Our study highlights morphological changes, such as the development of adventitious roots, as environmental adaptation mechanisms. By revealing O2 sources in S. kunstleri under hypoxic environments, we offer insights into the challenges of long-term adaptation to changing environments in woody plants.

Potassium supply reduces cesium uptake in Konara oak not by an alteration of uptake mechanism, but by the uptake competition between the ions.

Radiocesium contamination of forests has been a severe problem after the Fukushima Daiichi Nuclear Power Plant accident in 2011. Bed logs of Konara oak (Quercus serrata Murray), used for mushroom cultivation, were an economically important product from the forests prior to their contamination. One of the potential countermeasures to reduce radiocesium content in trees is potassium fertilization, but the evidence for the effect of K+ in reducing Cs+ uptake has not been obtained yet in the woody plant. Therefore, we investigated the ability of rhizospheric K+ to suppress uptake and translocation of Cs+ in Konara oak seedlings through hydroponic experiments in order to clarify the effect of K+. Elemental analysis showed that the seedlings cultivated for 4 weeks under low-K (K+ = 50 µM) contained higher amount of Cs comparing to the seedlings cultivated under high-K (K+ = 3 mM). Then, the uptake rate of Cs+ and K+ in the seedlings from the solution having 50 µM K+ and 0.1 µM Cs+ was calculated using radioactive 137Cs+ and 42K+ to evaluate the effect of growth condition on the ion uptake mechanism. The interference between Cs+ and K+ at the site of root uptake was also evaluated based on the Cs+ and K+ uptake rates at K+ concentrations of 50 µM, 200 µM, and 3 mM in the seedlings grown under the medium-K (K+ = 200 µM) condition. As a result, the Cs+ uptake rate at 50 µM K+ was not influenced by the growth condition, whereas Cs+ uptake decreased when the uptake solution itself was supplemented with 3 mM K+. In addition, the Cs/K ratio in the seedlings was found to rise to exceed the Cs/K ratio in the culture solution as the rhizospheric K+ concentration increased, which was in contrast with previous findings in herbaceous plants. Our experiments demonstrated the first direct evidence for woody plants that a high K+ concentration can suppress Cs accumulation in Konara oak and that it was derived from competition for uptake between K+ and Cs+ in the rhizosphere, not from the growth K+ condition.

A comparison of a mini-PEMS and a 1065 compliant PEMS for on-road gaseous and particulate emissions from a light duty diesel truck.

The primary goal of this study was to compare emissions measurements between a 1065 compliant PEMS, and the NTK Compact Emissions Meter (NCEM) capable of measuring NOx, PM, and solid PN. Both units were equipped on a light-duty diesel truck and tested over local, highway, and downtown driving routes. The results indicate that the NOx measurements for the NCEM were within approximately ±10% of those the 1065 compliant PEMS, which suggests that the NCEM could be used as a screening tool for NOx emissions. The NCEM showed larger differences for PM emissions on an absolute level, but this was at PM levels well below the 1 mg/mi level. The NCEM differences ranged from -2% to +26% if the comparisons are based on a percentage of the 1.0 mg/mi standard. Larger differences were also seen for PN emissions, with the NCEM measuring higher PN emissions, which can primarily be attributed to a zero current offset that we observed for the NCEM, which has been subsequently improved in the latest generation of the NCEM system. The comparisons between the 1065 compliant PEMS and the NCEM suggest that there could be applications for the NCEM or other mini-PEMS for applications such as identification of potential issues by regulatory agencies, manufacturer evaluation and validation of emissions under in-use conditions, and potential use in inspection and maintenance (I/M) programs, especially for heavy-duty vehicles.

Seasonal and spatial variations in leaf nitrogen content and resorption in a Quercus serrata canopy.

To elucidate the relationships between spatiotemporal changes in leaf nitrogen (N) content and canopy dynamics, changes in leaf N and distribution in the canopy of a 26-year-old deciduous oak (Quercus serrata Thunb. ex. Murray) stand were monitored throughout the developmental sequence from leaf expansion to senescence, by estimating the leaf mass and N concentrations of all the canopy layers. Seasonal changes were observed in leaf N concentration per unit leaf dry mass (N (m)), which peaked after bud burst, declined for two weeks shortly thereafter, and then remained constant for the rest of the growing season for each canopy layer. Leaf N concentration per unit leaf area (N (a)) was higher in the upper layer than in the lower layer throughout the growing season, and was closely correlated with relative irradiance (RI) in the summer when the air temperature was moderately high. The N concentrations of all leaf layers started to decrease in November, and reached their lowest values in late November, whereas LMA scarcely changed throughout the season. The lowest N concentrations did not differ significantly among the canopy layers. Seasonal changes in the relationship between N (a) and RI were detected, indicating that N (a) is optimized temporally as well as spatially. Nitrogen resorption efficiency was highest in the upper canopy layers where larger amounts of N were invested. Based on the estimates of leaf mass and leaf N concentrations of the canopy layers, total leaf N concentration of the whole canopy was estimated to be 84.1 kg ha(-1) in the summer, and 37.3 kg ha(-1) in late November. Therefore, 46.8 kg ha(-1) of leaf N in the canopy (about 56% of the total N) was resorbed just before leaf abscission.

Leaf turnover and growth responses of shade-grown saplings of four Shorea rain forest species to a sudden increase in light.

We tested the hypothesis that sapling growth following a sudden increase in solar irradiance is related to recovery from photoinhibition and the balance between rate of production of new leaves and rate of abscision of old leaves. Leaf gas exchange, chlorophyll fluorescence and relative growth rate (RGR) of stem basal area were measured following the sudden exposure of shade-grown (7% of full sunlight) saplings of four Shorea species to full sunlight. Sudden exposure to full sunlight resulted in an immediate and substantial reduction in dark-adapted quantum yield of photosystem II (Fv/Fm), followed by a gradual recovery in all species. Near light-saturated net assimilation rate (A max) and area-based leaf chlorophyll concentration ([Chl area]) also declined immediately after exposure. Eleven days after exposure, A max had recovered to pre-exposure values in all species, whereas [Chl area] had not recovered. Across species, RGR of stem basal area increased with increasing RGR of the number of leaves following exposure to full sunlight. The interspecific variations in RGR of stem basal area suggest that new leaf production is crucial for determining the potential growth of saplings following gap formation.

Elevated atmospheric CO2 concentration alters the effect of phosphate supply on growth of Japanese red pine (Pinus densiflora) seedlings.

We demonstrated that the inorganic phosphate (P(i)) requirement for growth of Japanese red pine (Pinus densiflora Sieb. & Zucc.) seedlings is increased by elevated CO(2) concentration ([CO(2)]) and that responses of the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker & Couch to P(i) supply are also altered. To investigate the growth response of non-mycorrhizal seedlings to P(i) supply in elevated [CO(2)], non-mycorrhizal seedlings were grown for 73 days in ambient or elevated [CO(2)] (350 or 700 micromol mol(-1)) with nutrient solutions containing one of seven phosphate concentrations (0, 0.02, 0.04, 0.06, 0.08, 0.10 and 0.20 mM). In ambient [CO(2)], the growth response to P(i) was saturated at about 0.1 mM P(i), whereas in elevated [CO(2)], the growth response to P(i) supply did not saturate, even at the highest P(i) supply (0.2 mM), indicating that the P(i) requirement is higher in elevated [CO(2)] than in ambient [CO(2)]. The increased requirement was due mainly to an altered shoot growth response to P(i) supply. The enhanced P(i) requirement in elevated [CO(2)] was not associated with a change in photosynthetic response to P(i) or a change in leaf phosphorus (P) status. We investigated the effect of P(i) supply (0.04, 0.08 and 0.20 mM) on the ectomycorrhizal fungus P. tinctorius in mycorrhizal seedlings grown in ambient or elevated [CO(2)]. Root ergosterol concentration (an indicator of fungal biomass) decreased with increasing P(i) supply in ambient [CO(2)], but the decrease was far less in elevated [CO(2)]. In ambient [CO(2)] the ratio of extramatrical mycelium to root biomass decreased with increasing P(i) supply but did not change in elevated [CO(2)]. We conclude that, because elevated [CO(2)] increased the P(i) requirement for shoot growth, the significance of the ectomycorrhizal association was also increased in elevated [CO(2)].

Root temperature drives winter acclimation of shoot water relations in Cryptomeria japonica seedlings.

In many temperate evergreen plant species, reductions in turgor loss point of leaves (Psi(tlp)) and leaf osmotic potential at full turgor (pi(sat)) occur from late summer to winter. To test the hypothesis that this seasonal change in leaf water relations is driven by root temperature, we manipulated the temperature of the roots and shoots of Cryptomeria japonica D. Don seedlings separately. Whole-plant warming diminished the seasonal changes in shoot water relations observed in the control plants, whereas shoot warming did not. Compared with the controls, root warming diminished the change in Psi(tlp) but not in pi(sat), whereas cooling accelerated the seasonal changes in shoot water relations. These results indicate that: (1) temperature responses of roots are involved in the seasonal changes in Psi(tlp) from late summer to winter; and (2) root temperature is partly responsible for the simultaneous changes in pi(sat). Whole-plant cooling caused increased root hydraulic resistance, suggesting that seasonal changes in shoot water relations represent adaptive responses to increased root hydraulic resistance at low root temperatures.

Phototropic bending of non-elongating and radially growing woody stems results from asymmetrical xylem formation.

Active phototropic bending of non-elongating and radially growing portion of stems (woody stems) has not been previously documented, whereas negative gravitropic bending is well known. We found phototropic bending in woody stems and searched for the underlying mechanism. We inclined 1-year-old Quercus crispula Blume seedlings and unilaterally illuminated them from a horizontal direction perpendicular to ('normal' illumination) or parallel to ('parallel' illumination) the inclination azimuth. With normal illumination, active phototropic bending and xylem formation could be evaluated separately from the negative gravitropic response and vertical deflection resulting from the weight of the seedlings. One-year-old stems with normal illumination bent significantly, with asymmetrical xylem formation towards the illuminated upper surface and side of the stem, whereas those with parallel illumination showed non-significant lateral bending, with asymmetrical xylem formation only on the upper side. A mechanical model was built on the assumption that a bending moment resulted from the asymmetrical xylem formation during phototropic bending of the woody stems. The model fitted the relationship between the observed spatial distributions of the xylem and the observed lateral bending, and thus supported the hypothesis that phototropic bending of woody stems results from asymmetrical xylem formation, as such occurs during gravitropism.

Stem phototropism of trees: a possible significant factor in determining stem inclination on forest slopes.

BACKGROUND AND AIMS: The main stems of trees on forest slopes incline down the slope to various extents that are characteristic of the species. The inclination has been explained as an active response to a horizontally asymmetrical light environment, but the contributing physiological mechanisms are unknown. The present study tested the hypothesis that stem phototropism, gravitropism, or a combination of the two determines the inclination of tree stems on forest slopes. METHODS: Cryptomeria japonica, Pinus densiflora, Quercus myrsinaefolia and Q. serrata were studied. Measurements were made of stem inclination of mature trees on forest slopes in uniform plantations of each species, and changes in stem inclination of potted seedlings in response to illumination treatments (unilateral or overhead) and inclination treatments (artificially inclined or erect). Indices of phototropic and gravitropic responsiveness were evaluated for each species, calculated from the change in stem inclination in response to artificial inclination with unilateral or overhead illumination. KEY RESULTS: Stem inclination on forest slopes varied significantly among species: Q. serrata inclined most in the down-slope direction, C. japonica inclined the least, and P. densiflora and Q. myrsinaefolia were intermediate. The change in stem inclination of seedlings in each treatment varied significantly among species. One-year-old stems of Q. serrata and 2-year-old stems of Q. myrsinaefolia bent toward the light source. Interspecific variation in the change in stem inclination in response to the unilateral illumination or that in the index of phototropic responsiveness was strongly correlated with the variation in stem inclination on forest slopes. CONCLUSIONS: The orientation of woody stems that have finished elongation can be actively controlled by phototropism. Interspecific variation in phototropic responsiveness of trees is a possible significant determinant of interspecific variation in stem inclination on forest slopes.