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Kazuyoshi Futai - One of the best experts on this subject based on the ideXlab platform.
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endophytic actinomycetes from Pinus thunbergii and their antifungal activity against cylindrocladium sp
Archives of Phytopathology and Plant Protection, 2011Co-Authors: Ryota Kataoka, Kazuyoshi FutaiAbstract:The inside of Pinus thunbergii could be a reliable screening source for a useful agent in controlling plant disease. Isolation of endophytic actinomycetes from P. thunbergii and their potential as biocontrol agents against the plant pathogen Cylindrocladium sp. were investigated. Two endophytic actinomycetes, Streptomyces sp. and Microbispora sp., were isolated from surface-sterilised root tissues of P. thunbergii seedlings. The recovery test of these two endophytic actinomycetes from pine seedling showed that Streptomyces sp. was isolated from only roots, but Microbispora sp. was isolated from both roots and leaves. Thus, Microbispora sp. is able to move to leaves from roots. Moreover, we evaluated the potential of both strains as biocontrol agents against Cylindrocladium sp. Two weeks after inoculation of Cylindrocladium sp. alone, pine seedlings showed a 50% mortality rate. Co-inoculation of Cylindrocladium sp. and Microbispora sp. did not affect seedling mortality rate. However, inoculation with both ...
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quantification of wautersia ralstonia basilensis in the mycorrhizosphere of Pinus thunbergii parl and its effect on mycorrhizal formation
Soil Biology & Biochemistry, 2009Co-Authors: Ryota Kataoka, Takeshi Taniguchi, Zaki A Siddiqui, Kazuyoshi FutaiAbstract:Abstract The bacterium Wautersia [ Ralstonia ] basilensis has been shown to enhance the mycorrhizal symbiosis between Suillus granulatus and Pinus thunbergii (Japanese black pine). However, no information is available about this bacterium under field conditions. The objectives of this study were to detect W. basilensis in bulk and mycorhizosphere soils in a Japanese pine plantation in the Tottori Sand Dunes, determine the density of W. basilensis in soil, and determine the optimal cell density of W. basilensis for mycorrhizal formation in pine seedlings. We designed and validated 16S rRNA gene-targeted specific primers for detection and quantification of W. basilensis . SYBR Green I real-time PCR assay was used. A standard curve relating cultured W. basilensis cell density (10 3 –10 8 cells ml −1 ) to amplification of DNA showed a strong linear relationship ( R = 0.9968). The specificity of the reaction was confirmed by analyzing DNA melting curves and sequencing of the amplicon. The average cell density of W. basilensis was >4.8 × 10 7 cells g −1 of soil in the mycorrhizosphere and 7.0 × 10 6 cells g −1 in the bulk soil. We evaluated the W. basilensis cell density required for mycorrhizal formation using an in vitro microcosm with various inoculum densities ranging from 10 2 to 10 7 cells g −1 soil (10 4 –10 9 cells ml −1 ). Cell densities of W. basilensis of >10 6 cells g −1 of soil were required to stimulate mycorrhizal formation. In vivo and in vitro experiments showed that W. basilensis was sufficiently abundant to enhance mycorrhizal formation in the mycorrhizosphere of Japanese black pine sampled from the Tottori Sand Dunes.
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diagnosis and quantification of the pine wood nematode bursaphelenchus xylophilus steiner buhner in wood of Pinus thunbergii with real time pcr
Japanese Journal of Nematology, 2009Co-Authors: Yuko Takeuchi, Kazuyoshi FutaiAbstract:A new diagnostic and quantitative real-time PCR assay was developed for the pine wood nematode, Bursaphelenchus xylophilus, which causes a serious epidemic wilt disease of susceptible landscape and forest pine trees. One, 10, 50, 100, 200, and 400 individuals of B. xylophilus in 100 mg of pine tissues were detected and quantified with two sets of primers coding the rDNA-ITS region and β-tubulin (tbb) gene of B. xylophilus, avoiding the need to extract nematodes with Baermann funnels. The assay was efficient, specific for pine wood nematode and sensitive enough to detect a single individual in pine tissues. To test the applicability of the assay for infested hosts, seedlings of the susceptible host species, Pinus thunbergii, were inoculated with B. xylophilus, and the dispersal and multiplication of B. xylophilus inside the seedlings were monitored by both Baermann funnel extraction and realtime PCR. Quantification by real-time PCR gave similar trends in estimates of the nematode population changes inside the host to those obtained by Baermann funnel extraction, although the size of the population estimated was larger for the new method than for the Baermann funnel extraction. The method developed in this study, which was expected to detect and quantify all life stages of the pine wood nematode should be useful for quarantine and field diagnosis.
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fungal selectivity of two mycorrhiza helper bacteria on five mycorrhizal fungi associated with Pinus thunbergii
World Journal of Microbiology & Biotechnology, 2009Co-Authors: Ryota Kataoka, Takeshi Taniguchi, Kazuyoshi FutaiAbstract:The fungal selectivity of helper effect was revealed using four ectomycorrhizal fungi (Rhizopogon sp., Pisolithus tinctorius, Cenococcum geophilum and Suillus granulatus), and one ectoendomycorrhizal fungus (Wilcoxina mikolae). Previously, we isolated a rhizobacteria, Ralstonia basilensis and Bacillus subtilis, which have the ability to enhance the mycorrhizal symbiosis between S. granulatus and Pinus thunbergii. However, the characteristics of each bacterium on mycorrhizal fungi are still unclear. Therefore, we tried to examine the fungal selectivity of helper effect. A confrontation assay revealed that R. basilensis significantly promoted the in vitro hyphal growth of W. mikolae and S. granulatus, but it had no effect on Rhizopogon sp., P. tinctorius and C. geophilum. These results were consistent with the effects shown by R. basilensis on the mycorrhizal formation of these fungi. On the other hand, B. subtilis promoted the hyphal growth of W. mikolae and C. geophilum but suppressed growth of Rhizopogon sp. B. subtilis significantly stimulated the mycorrhizal formation of S. granulatus. Thus the effects of B. subtilis on hyphal growth and on mycorrhizal formation were inconsistent. These results suggest that R. basilensis and B. subtilis have fungal selective and different mechanisms in their roles as mycorrhiza helper bacteria.
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a new mycorrhizal helper bacterium ralstonia species in the ectomycorrhizal symbiosis between Pinus thunbergii and suillus granulatus
Biology and Fertility of Soils, 2009Co-Authors: Ryota Kataoka, Kazuyoshi FutaiAbstract:In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.
Yutaka Gonda - One of the best experts on this subject based on the ideXlab platform.
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regeneration of a coastal pine Pinus thunbergii parl forest 11 years after thinning niigata japan
PLOS ONE, 2012Co-Authors: Yutaka Gonda, Lizhong Yu, Fengqin LiAbstract:To examine the effects of thinning intensity on wind vulnerability and regeneration in a coastal pine (Pinus thunbergii) forest, thinning with intensities of 20%, 30% and 50% was conducted in December 1997; there was an unthinned treatment as the control (total 8 stands). We re-measured the permanent sites to assess the regeneration characteristics 11 years after thinning. In the 50% thinned stand, seedlings aged from 2 to 10 years exhibited the highest pine seedling density and growth. The age composition ranged from 1–3 years with densities of 9.9 and 5.1 seedlings m−2 in 30% and 20% thinned stands; only 1-year-old seedlings with a density of 6.1 seedlings m−2 in the unthinned stand. Similar trends were found for the regeneration of broadleaved species such as Robinia pseudoacacia and Prunus serrulata. We speculate that the canopy openness and moss coverage contributed to the regeneration success in the 50% thinned stand, while the higher litter depth and lack of soil moisture induced the regeneration failure in the unthinned stand. The stands thinned at 20% or 30% were less favourable for pine regeneration than the stands thinned at 50%. Therefore, thinning with less than 30% canopy openness (20% and 30% thinned stands) should be avoided, and thinning at higher than 30% canopy openness (50% thinned stand, approximately 1500 stems ha−1 at ages 40–50 years) is suggested for increasing regeneration in the coastal pine forest. The implications of thinning-based silviculture in the coastal pine forest management are also discussed. The ongoing development of the broadleaved seedlings calls for further observations.
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influence of thinning on regeneration in a coastal Pinus thunbergii forest
Journal of Applied Ecology, 2002Co-Authors: Jiaojun Zhu, Takeshi Matsuzaki, Yutaka GondaAbstract:A coastal forest planted nearby the sea can provide many shelter benefits for the coastal regions. It is ideally if the continuity of the shelter benefits could be preserved through reasonable management. Thinning and regeneration as the most important management techniques for plantations can help the continuity of the shelter benefits of the coastal forest. However, because of the peculiarities of coastal forest, i.e., the coastal plantation nearby the sea is vulnerable to disturbances (thinning as one kind of disturbance), the study on thinning and regeneration within a coastal forest is poorly understood. The purpose of this paper is to give a primary understanding in natural regeneration for the coastal Pinus thunbergii forest with different thinning rates after four growing seasons since thinning. The experiment was carried out at the middle of the shoreline along the Japan Sea, and the investigated sites consisted of four thinning treatments (control, 0% thinned, 20% thinned, 30% thinned and 50% thinned) in a coastal P. thunbergii forest. After thinning, the regenerated seedlings, soil water content, light condition (canopy openness or canopy density), wind regime, and litter depth and quantity were investigated for four growing seasons. The relationships between the regenerated seedlings and light condition, litter, wind profile and soil water content were examined. The results showed that thinning could improve the light condition on the forest floor, increase the exchange of airflow (wind speed) in the coastal forest stand, and ameliorate the water content of the forest soil. These factors accelerated the decomposition of litters, and provided necessary conditions for natural regeneration. The results of regeneration observation indicated that the most intensively thinned treatment (50% thinned with density of about 1500 stems.hm-2) could provide a better condition for regeneration during the four growing seasons. The density and growth of seedling (greater than 1 year) increased significantly with increasing thinning intensities, and the establishment of seedlings was obviously succeeded in the most intensively thinned treatment, but failed in less thinned treatments and understory. The thinned intensity of 50% did not induce wind damage to the coastal forest in the four years after thinning, and did not cause the loss of shelter functions of the coastal forest such as sand blocking, wind breaking and salt preventing etc.. On the contrary, it could provide the suitable conditions for natural regeneration of the pine coastal forest, or for the immigration of other species. Therefore, thinning as the silviculturally created openness is very important for the establishment of seedlings in the coastal forests, which provides a mechanism for the coastal forest from even aged stands dominated by P. thunbergii to stands containing multiple size classes and canopy layers.
Taizo Hogetsu - One of the best experts on this subject based on the ideXlab platform.
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pathway and sink activity for translocation of 14c absorbed as amino acids in the pisolithus extraradical mycelium of ectomycorrhizal Pinus thunbergii seedlings
Mycoscience, 2016Co-Authors: Bingyun Wu, Munemasa Teramoto, Taizo HogetsuAbstract:Abstract To characterize the pathway and sink activity for the translocation of amino acids and their metabolites in the extraradical mycelium (ERM) of ectomycorrhizal (ECM) symbiosis, we locally labeled a part of the ERM of Pinus thunbergii seedlings colonized by a Pisolithus isolate with [ 14 C(U)]- l -alanine ( 14 C-Ala) or [ 14 C(U)]- l -glutamine ( 14 C-Gln) and followed the 14 C distribution in the ERM by autoradiography. After 14 C-Ala or 14 C-Gln labeling, 14 C was mainly distributed in a bundle of rhizomorphs extending from the labeled part towards ECM root tips of the host, but not in thick roots or the aboveground part of the seedling. When 14 C-Ala was added after seedling removal, no 14 C bundle appeared in the remaining ERM. These results indicate that 14 C amino acids and their metabolites are translocated unidirectionally towards the ECM root tips, where strong sink activity for amino acids is localised. 14 C was also distributed diffusely over a broad region in ERM surrounding the labeled part. This result suggests that 14 C amino acids and their metabolites are also translocated multidirectionally in the ERM by sink activity distributed diffusely all over the whole ERM.
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Pathway and sink activity for photosynthate translocation in Pisolithus extraradical mycelium of ectomycorrhizal Pinus thunbergii seedlings
Mycorrhiza, 2016Co-Authors: Munemasa Teramoto, Bingyun Wu, Taizo HogetsuAbstract:The purpose of this study was to identify the pathway and sink activity of photosynthate translocation in the extraradical mycelium (ERM) of a Pisolithus isolate. We labelled ectomycorrhizal (ECM) Pinus thunbergii seedlings with 14CO2 and followed 14C distribution within the ERM by autoradiography. 14C photosynthate translocation in the ERM resulted in 14C distribution in rhizomorphs throughout the ERM, with 14C accumulation at the front. When most radial mycelial connections between ECM root tips and the ERM front were cut, the whole allocation of 14C photosynthates to the ERM was reduced. However, the overall pattern of 14C distribution in the ERM was maintained even in regions immediately above and below the cut, with no local 14C depletion or accumulation. We inferred from this result that every portion in the ERM has a significant sink activity and a definite sink capacity for photosynthates and that photosynthates detour the cut and reach throughout the ERM by translocation in every direction. Next, we prepared paired ECM seedlings, ERMs of which had been connected with each other by hyphal fusion, alongside, labelled the left seedling with 14CO2, and shaded none, one or both of them. 14C photosynthates were acropetally and basipetally translocated from the left ERM to ECM root tips of the right seedling through rhizomorphs in the left and right ERMs, respectively. With the left seedling illuminated, 14C translocation from the left to the right ERM increased by shading the right seedling. This result suggests that reduced photosynthate transfer from the host to its ERM increased sink activity of the ERM.
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The process of epithelial cell death in Pinus thunbergii caused by the pine wood nematode, Bursaphelenchus xylophilus
Nematology, 2014Co-Authors: Joung A. Son, Taizo Hogetsu, Yil-sung MoonAbstract:This study describes a new technique to investigate how the pine wood nematode (PWN), Bursaphelenchus xylophilus, kills pine epithelial cells. After inoculating PWN into 20-cm-long Pinus thunbergii stem cuttings and incubating for 1, 3 or 7 days, the cuttings were split into 2.5 cm segments. The segments were tangentially cut so that the epithelia of several cortical resin canals were exposed, and these were stained with Evans Blue for the detection of dead epithelial cells. While almost no dead epithelial cells were found in the cortical resin canals of non-PWN-inoculated control cuttings up to day 7 of the experiment, dead epithelial cells were distributed sparsely in the epithelium of cortical resin canals throughout pine cuttings inoculated with PWN 1, 3 and 7 days after inoculation. The sparse and sporadic distribution of dead pine cells in the epithelium suggested that individual PWN attacked one epithelial cell at a time with its stylet and migrated between attacks.
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Migration of pine wood nematodes in the tissues of Pinus thunbergii
Journal of Forest Research, 2010Co-Authors: Joung A. Son, Masabumi Komatsu, Norihisa Matsushita, Taizo HogetsuAbstract:The distribution of pine wood nematodes ( Bursaphelenchus xylophilus , PWNs) in Japanese black pine ( Pinus thunbergii ) tissues was investigated by staining with fluorescein isothiocyanate-conjugated wheat germ agglutinin. After PWNs were inoculated to current-year stems of pine seedlings, their distribution at about 5 cm below the inoculation site was confined only to cortical resin canals 1 day after inoculation, and then spread to other tissues, including resin canals of short branches. When PWNs were inoculated onto cross or tangentially cut surfaces of stem segments, maximal PWN migration speed was estimated to be faster through cortical resin canals and xylem axial resin canals vertically (>6.7 and
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migration of pine wood nematodes in the tissues of Pinus thunbergii
Journal of Forest Research, 2010Co-Authors: Joung A. Son, Masabumi Komatsu, Norihisa Matsushita, Taizo HogetsuAbstract:The distribution of pine wood nematodes (Bursaphelenchus xylophilus, PWNs) in Japanese black pine (Pinus thunbergii) tissues was investigated by staining with fluorescein isothiocyanate-conjugated wheat germ agglutinin. After PWNs were inoculated to current-year stems of pine seedlings, their distribution at about 5 cm below the inoculation site was confined only to cortical resin canals 1 day after inoculation, and then spread to other tissues, including resin canals of short branches. When PWNs were inoculated onto cross or tangentially cut surfaces of stem segments, maximal PWN migration speed was estimated to be faster through cortical resin canals and xylem axial resin canals vertically (>6.7 and <2.3 mm/h, respectively) than through cortical tissues both vertically and horizontally (<1.2 and <0.2 mm/h). To examine whether PWNs in cortical resin canals could invade surrounding tissues, segments in which PWNs resided only in cortical resin canals were prepared by removing the top portion 6 h after inoculation. Additional incubation of such segments caused extended PWN distribution to xylem axial resin canals and then to other tissues. A similar experiment with top portions of girdled segments removed 12 h after inoculation also showed extended PWN distribution from xylem axial resin canals and pith to cortical resin canals and then to other tissues. These results provided direct evidence that PWNs have the ability to migrate from cortical resin canals and xylem axial resin canals to other tissues.
Atsushi Watanabe - One of the best experts on this subject based on the ideXlab platform.
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influence of temperature on pine wilt disease progression in Pinus thunbergii seedlings
European Journal of Plant Pathology, 2020Co-Authors: R Yamaguchi, Koji Matsunaga, Atsushi WatanabeAbstract:Pine wilt disease (PWD) affects forests globally and is caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus. It has been proposed that PWD development has a strong association with air temperature. Because symptom development is related to the number of PWNs and they are poikilothermic animals whose development and multiplication are affected by temperature; it appears that temperature conditions affect the PWN multiplication rate, which leads to difference in disease progression. However, limited information is available about the temperatures inside pine trees, and the understanding of how temperature affects PWN multiplication and PWD progression remains incomplete. Therefore, in the present study, we evaluated the temperature, number of PWNs, and external symptoms of PWD in Pinus thunbergii seedlings infected with PWNs at air temperatures of 20 °C, 25 °C, and 30 °C over time. We found that the seedling temperatures were slightly higher than the air temperature under each temperature condition and that the effective accumulated temperature calculated using a base temperature of 10 °C was related to the number of PWNs and symptom development in all temperature test groups. However, at 20 °C, there were very few PWNs in some seedlings and none of the seedlings showed partial or full wilting. These findings suggest that PWD progression is greatly affected by ambient temperature and facilitated by increase in PWN populations with effective accumulated temperature especially in the range of 25 °C–30 °C.
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historical seed use and transfer affects geographic specificity in genetic diversity and structure of old planted Pinus thunbergii populations
Forest Ecology and Management, 2018Co-Authors: Masakazu G Iwaizumi, Tomonori Hirao, Shousuke Miyata, Miho Tamura, Atsushi WatanabeAbstract:Abstract Although most molecular ecology studies examining genetic variation have focused on natural forests, for some major tree species, natural forests are nearly extinct, and the remaining genetic resources are mainly planted forests. In order to manage the genetic variability and develop a conservation strategy for such species, it is important to examine genetic variation resulting from historical processes during repeated artificial population development through plantations. The geographic pattern of genetic diversity and structure of 49 old planted Pinus thunbergii populations (2755 trees) distributed across Japan was examined using seven nuclear microsatellite markers. We found that allelic diversity was generally lower in both northern and eastern populations; however, locally, some populations in other regions also exhibited low allelic diversity. The overall value of the standardized measure of population differentiation (G′ST = 0.206) was higher than that of both other widespread Japanese conifers and continental Pinus species. STRUCTURE software revealed a general gradual cline in genetic structure from southwestern to northeastern populations; however, some populations on the Japan Sea side showed quite a different local proportion of cluster memberships from nearby populations. These observations indicated that most of the preserved, planted populations of P. thunbergii possess regional genetic variation, but some populations were developed from seed pools derived from other regions, possibly through distribution by ship along the Japan Sea. Information on this specific genetic variation as a result of historical seed use and transfer should assist the design of several conservation units and breeding zones, while also taking care of a deep-seated need for conservation of pine forests by local people.
Jiaojun Zhu - One of the best experts on this subject based on the ideXlab platform.
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influence of thinning on regeneration in a coastal Pinus thunbergii forest
Journal of Applied Ecology, 2002Co-Authors: Jiaojun Zhu, Takeshi Matsuzaki, Yutaka GondaAbstract:A coastal forest planted nearby the sea can provide many shelter benefits for the coastal regions. It is ideally if the continuity of the shelter benefits could be preserved through reasonable management. Thinning and regeneration as the most important management techniques for plantations can help the continuity of the shelter benefits of the coastal forest. However, because of the peculiarities of coastal forest, i.e., the coastal plantation nearby the sea is vulnerable to disturbances (thinning as one kind of disturbance), the study on thinning and regeneration within a coastal forest is poorly understood. The purpose of this paper is to give a primary understanding in natural regeneration for the coastal Pinus thunbergii forest with different thinning rates after four growing seasons since thinning. The experiment was carried out at the middle of the shoreline along the Japan Sea, and the investigated sites consisted of four thinning treatments (control, 0% thinned, 20% thinned, 30% thinned and 50% thinned) in a coastal P. thunbergii forest. After thinning, the regenerated seedlings, soil water content, light condition (canopy openness or canopy density), wind regime, and litter depth and quantity were investigated for four growing seasons. The relationships between the regenerated seedlings and light condition, litter, wind profile and soil water content were examined. The results showed that thinning could improve the light condition on the forest floor, increase the exchange of airflow (wind speed) in the coastal forest stand, and ameliorate the water content of the forest soil. These factors accelerated the decomposition of litters, and provided necessary conditions for natural regeneration. The results of regeneration observation indicated that the most intensively thinned treatment (50% thinned with density of about 1500 stems.hm-2) could provide a better condition for regeneration during the four growing seasons. The density and growth of seedling (greater than 1 year) increased significantly with increasing thinning intensities, and the establishment of seedlings was obviously succeeded in the most intensively thinned treatment, but failed in less thinned treatments and understory. The thinned intensity of 50% did not induce wind damage to the coastal forest in the four years after thinning, and did not cause the loss of shelter functions of the coastal forest such as sand blocking, wind breaking and salt preventing etc.. On the contrary, it could provide the suitable conditions for natural regeneration of the pine coastal forest, or for the immigration of other species. Therefore, thinning as the silviculturally created openness is very important for the establishment of seedlings in the coastal forests, which provides a mechanism for the coastal forest from even aged stands dominated by P. thunbergii to stands containing multiple size classes and canopy layers.
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wind speeds within a single crown of japanese black pine Pinus thunbergii parl
Forest Ecology and Management, 2000Co-Authors: Jiaojun Zhu, Takeshi Matsuzaki, Kenji SakiokaAbstract:Abstract Wind speed and crown thickness, which is defined as a horizontal distance from the observed point to outer edge of a crown in windward direction, were measured within a normal crown of a single tree of Japanese black pine (Pinus thunbergii Parl.) at four vertical sections. Simultaneously, wind speed and direction were observed outside the crown. Regression analysis is used to obtain the wind profiles within the single crown. The results suggest that the crown features make a large influence on the wind profiles, therefore, the crown thickness as an indicator of crown feature is employed to develop the horizontal wind profile within the crown. A consistent relationship between relative wind speed and corresponding crown thickness is established for predicting wind speed at a horizontal plane. Additionally, vertical wind profile within the crown is also examined as literature suggested in exponential form. Within a single tree crown, where the distribution of leave and branch is essentially uniform with height, and the tree form is similar to Japanese black pine, the vertical wind profile can be derived from a single wind speed measurement outside the crown with using the parameter derived in this study. Based on the vertical and horizontal profiles, wind speed at any position in a plane within a crown at height z can be reasonably estimated if providing the crown thickness and wind data outside the crown. The wind profiles discussed here may be useful when discussing the ventilation of airflow at any level of a crown, analyzing the transfer processes for questions of disease and solving the problems of the tree damage by salty wind from the sea, etc.