Definition of fine roots on the basis of the root anatomy,diameter,and branch orders of one-year old Fraxinus mandshurica seedlings

Fine roots are important in root absorption of nutrient and water,and in root turnover.Accurate definition of fine roots is a prerequisite to improved estimation of the physiological and ecological functions of forest ecosystems.Root development and physiological functions are reflections of root anatomical structure.In this study,the anatomical structures of different root orders were analyzed by examining paraffin sections of one-year old Fraxinus mandshurica seedlings.One-year-old F.mandshurica seedlings had over five root orders.The root anatomical structures of all orders showed more differences.First and second order roots consisted of four sections: the epidermis,cortex,pericycle,and vascular bundles.Fourth and fifth order roots were mainly composed of the skin and peripheral vascular bundles(including the xylem and phloem).Third order roots had root epidermal and cortical structures,but the quantity and integrity of the cortical cells were inferior to those of the first and second order roots,and superior to those of the fourth and fifth order roots.All the first and second order roots and some third order roots with discontinuous cork layer(0.4 mm in diameter),but not the fourth and fifth order roots,were the fine roots of one-year old F.mandshurica seedlings.Although they had similar diameters,different portions of root systems had different anatomical structures and therefore,vary in capacity to absorb water and nutrients.Fine roots were accurately defined by root diameter,branch orders,and anatomical structural features of one-year old F.mandshurica seedlings.     

Nutrient retranslocation from the fine roots of <Emphasis Type="Italic">Fraxinus mandshurica</Emphasis> and <Emphasis Type="Italic">Larix olgensis</Emphasis> in northeastern China

Nutrient retranslocation in trees is important in nutrient budgets and energy flows in forest ecosystems. We investigated nutrient retranslocation in the fine roots of a Manchurian Ash (Fraxinus mandshurica) and a Larch (Larix olgensis) plantation in northeastern China. Nutrient retranslocation in the fine roots was investigated using three methods, specifically, nutrient concentration, the ratio of Ca to other elements (Ca/other elements ratio) and nutrient content. The method based on nutrient content proved most suitable when investigating nutrient retranslocation from fine roots of the two species. The nutrient-content-based method showed that there were retranslocations of N, P, K and Mg from the fine roots of Manchurian Ash, with retranslocation efficiencies of 13, 25, 65, and 38 %, respectively, whereas there were no Ca retranslocations. There were retranslocations of N, P, K, Ca and Mg from the fine roots of Larch, with retranslocation efficiencies of 31, 40, 52, 23 and 25 %, respectively.     

Responses of fine root mass,length, production and turnover to soil nitrogen fertilization in <Emphasis Type="Italic">Larix gmelinii</Emphasis> and <Emphasis Type="Italic">Fraxinus mandshurica</Emphasis> forests in Northeastern China

The responses of fine root mass, length, production and turnover to the increase in soil N availability are not well understood in forest ecosystems. In this study, sequential soil core and ingrowth core methods were employed to examine the responses of fine root (≤1 mm) standing biomass, root length density (RLD), specific root length (SRL), biomass production and turnover rate to soil N fertilization (10 g N m⁻² year⁻¹) in Larix gmelinii (larch) and Fraxinus mandshurica (ash) plantations. N fertilization significantly reduced fine root standing biomass from 130.7 to 103.4 g m⁻² in ash, but had no significant influence in larch (81.5 g m⁻² in the control and 81.9 g m⁻² in the fertilized plots). Similarly, N fertilization reduced mean RLD from 6,857 to 5,822 m m⁻² in ash, but did not influence RLD in larch (1,875 m m⁻² in the control and 1,858 m m⁻² in the fertilized plots). In both species, N fertilization did not alter SRL. Additionally, N fertilization did not significantly alter root production and turnover rate estimated from sequential soil cores, but did reduce root production and turnover rate estimated from the ingrowth core method. These results suggested that N fertilization had a substantial influence on fine root standing biomass, RLD, biomass production and turnover rate, but the direction and magnitude of the influence depended on species and methods.     

Shoot multiplication of <Emphasis Type="Italic">Syringa reticulata</Emphasis> var. <Emphasis Type="Italic">mandshurica</Emphasis> from in vitro cultured seedlings

We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog (MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine (BA) and indole-3-butyric acid (IBA). The better medium for shoot multiplication and growth was MS + 5 mg L^?1 BA + 0.5 mg L^?1 IBA + 20 g L^?1 sucrose + 7 g L^?1 agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium (macro-elements of MS medium are at half-strength) supplemented with 1 mg L^?1 IBA, and the survival percentage was >80 % at 16 weeks after transplanting.     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

Inoculation with the ectomycorrhizal fungi <Emphasis Type="Italic">Pisolithus arhizus</Emphasis> and <Emphasis Type="Italic">Scleroderma</Emphasis> sp. improves early growth of <Emphasis Type="Italic">Shorea pinanga</Emphasis> nursery seedlings

Trees of the family Dipterocarpaceae are the dominant trees in Southeast Asian tropical forests where they play an important ecological role and are also important commercially. An experiment was conducted to determine the effect of ectomycorrhizal fungi on the growth of dipterocarp species in peat soils. Seedlings of Shorea pinanga were inoculated with spores of two ectomycorrhizal fungi, Pisolithus arhizus and Scleroderma sp. were grown in pots containing sterilized peat soil for 7 months. The percentage of ectomycorrhizal colonization on S. pinanga exceeded 86%. Colonization of S. pinanga roots by ectomycorrhizal fungi resulted in increased shoot height, stem diameter, number of leaves, and shoot fresh and dry weight. Survival rates of S. pinanga were greater for inoculated seedlings than control seedlings. These results suggest that inoculation of ectomycorrhizal fungi can improve the early growth of S. pinanga grown in tropical forests and that this technique will accelerate the rehabilitation of degraded dipterocarp forests.     

Influence of water-stress acclimation and <Emphasis Type="Italic">Tuber melanosporum</Emphasis> mycorrhization on <Emphasis Type="Italic">Quercus ilex</Emphasis> seedlings

Mycorrhizal and non-mycorrhizal holm oak (Quercus ilex L.) seedlings inoculated with black truffle (Tuber melanosporum) were grown under nursery conditions and subjected to drought hardening for 4 months in autumn and winter followed by irrigation for 10 days. Leaf water potential and stomatal conductance were monitored during the 4 months of drought. When the test was completed (March), measurements were made for each treatment (inoculated or non-inoculated), and watering regime (watered and water-stressed). Pressure–volume curves, osmotic potential at full turgor, osmotic potential at zero turgor and the tissue modulus of elasticity near full turgor were calculated. Mycorrhizal colonization and growth, and the content of the main mineral nutrients N, P, K, Ca and Mg were measured. Water stress affected plant growth, caused an elastic adjustment of the plant tissues, and decreased the P and K content, and inoculation improved the nitrogen content. Drought acclimation apparently achieved the goal of improving the drought tolerance of holm oak seedlings, without depressing ectomycorrhizal root colonization by T. melanosporum. José Antonio Rodríguez Barreal—Deceased     

Insecticidal properties of <Emphasis Type="Italic">Thymus persicus</Emphasis> essential oil against <Emphasis Type="Italic">Tribolium castaneum</Emphasis> and <Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Red flour beetle Tribolium castaneum (Herbst) and rice weevil Sitophilus oryzae (L.) are considered to be the major insect pests in storage. Essential oils from aromatic plants are recognized as proper alternatives to fumigants. Thymus persicus (Ronniger ex Rech. f.) is one of these plants that have medicinal properties and is indigenous to Iran. The essential oil was obtained from aerial parts of the plant and analyzed by GC and GC–MS. Carvacrol (44.69%) and thymol (11.05%) were the major constituents of the oil extracted. In this experiment, fumigant toxicity of the essential oil was studied against T. castaneum, S. oryzae at 27 ± 1°C and 60 ± 5% RH in dark condition. The adult insects were exposed to the concentrations of 51.9, 111.1, 207.4 and 370.4 μl/l air to estimate median lethal time (LT₅₀) values. The fumigant toxicity was increased in response to increased essential oil concentrations. The LT₅₀ values at the lowest and the highest concentrations tested were ranged from 28.09 to 13.47 h for T. castaneum, and 3.86 to 2.30 h for S. oryzae. It was found that S. oryzae adults were much more susceptible to the oil than T. castaneum. After 24 h of exposure, the LC₅₀ values (95% fiducial limit) for T. castaneum and S. oryzae were estimated to be 236.9 (186.27–292.81) and 3.34 (2.62–4.28) μl/l air, respectively. These results suggest that T. persicus essential oil merits further study as potential fumigant for the management of these stored-product insects.     

Enzymatic saccharification and ethanol production of <Emphasis Type="Italic">Acacia mangium</Emphasis> and <Emphasis Type="Italic">Paraserianthes falcataria</Emphasis> wood,and <Emphasis Type="Italic">Elaeis guineensis</Emphasis> trunk

We examined the saccharification and fermentation of meals from Acacia mangium wood, Paraserianthes falcataria wood, and Elaeis guineensis trunk. The levels of enzymatic hydrolysis of cellulose and ethanol production were highest for P. falcataria wood and lowest for A. mangium wood. Ultrasonication pretreatment of meal further increased the rates of hydrolysis and ethanol production in meal from P. falcataria wood. Through this pretreatment, hemicelluloses (xylan and xyloglucan) and cellulose were released in the meal from P. falcataria wood. Loosening of hemicellulose associations can be expected to make P. falcataria wood more useful for bioethanol production.     

Photosynthate supply drives soil respiration of <Emphasis Type="Italic">Fraxinus mandshurica</Emphasis> seedlings in northeastern China: evidences from a shading and nitrogen addition experiment

Improved understanding of the link between photosynthesis and below-ground processes is needed to better understand ecosystem carbon (C) cycling and its feedback to climate change. We conducted a short-term shading and nitrogen (N) addition experiment from June to September 2013 to investigate the effect of photosynthate supply by Manchurian Ash (Fraxinus mandshurica) seedlings on soil respiration (SR). Shading significantly reduced SR in early and middle growing season, but not in late growing season, leading to a decrease in mean SR by 24 % in N-unfertilized treatments. N addition increased mean SR by 42 % in un-shaded treatment. The stimulation of SR was largely attributed to accelerated autotrophic respiration by increasing photosynthesis, leaf area index and belowground biomass. Shading reduced mean SR by 32 % in N addition treatment. The strengthened shading effect on SR resulted from N addition was because of more photosynthates supply at low soil temperature. Our findings highlight the predominance of photosynthates supply in regulating the responses of C cycling to global change.     

Fine roots branch orders of <Emphasis Type="Italic">Abies faxoniana</Emphasis> respond differentially to warming in a subalpine coniferous forest ecosystem

Root is an important plant organ and has high heterogeneity. Global warming could change root and affect belowground ecological processes. There is little information on how fine roots branch orders responds to global change. This study examined the growth, morphological and physiological responses of fine roots of a subalpine coniferous species to warming. We investigated biomass, average diameter, specific root length (SRL), triphenyltetrazolium chloride (TTC) reducing capacity, carbon (C), total non-structural carbon (TNC) and fractions of the primal five branch order roots of Abies faxoniana in April, August, October and December. The decrease in total fine roots biomass after a growing season was significantly greater under warming treatment compared to control, suggesting that warming could accelerate the carbon input from root to soil, but the increment depended on tree species. Warming did not affect average diameter and SRL. Responses of biomass, TTC reducing capacity, C, TNC and fractions to warming significantly differed with root order and month. Significant warming effects were only observed in C and starch concentration of the first order and also TNC and soluble sugar concentration of the first three orders. The results indicated that the lower order roots (the first three orders) were more sensitive to warming, probably because they had more frequent, intense interactions with soil and low defense capability. Thus, global warming may dramatically alter root functions such as nutrients and water uptake as well as the cycle of C and nutrients at the whole subalpine coniferous forest ecosystem.     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Fine root biomass and morphology of <Emphasis Type="Italic">Pinus densiflora</Emphasis> under competitive stress by <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>

The fine root (diameter ≤2.0 mm) biomass and morphology of Japanese red pines (Pinus densiflora) grown under different aboveground conditions (i.e., high and low competitive environments) were examined in a pine–cypress mixed forest. All P. densiflora subject trees were about 40 years old, and the aboveground condition (i.e., size) of red pines appeared to be influenced by the surrounding Japanese cypress (Chamaecyparis obtusa). Smaller P. densiflora exhibited lower fine root biomasses, shorter root lengths, and lower root tip densities, but longer specific root lengths and higher specific root tip densities relative to larger pines. These results suggest that P. densiflora may adjust the morphological traits of fine roots to the different conditions in biomass allocation to fine roots of individuals with different aboveground growth.     

Constituents from the roots of <Emphasis Type="Italic">Taxus cuspidata</Emphasis>

The known propelargonidin, afzelechin-(48)-afzelechin (1), the known lignans 7-hydroxynortrachelogenin (2), epinortrachelogenin (3), nortrachelogenin (4), hydroxymatairesinol (5), allohydroxymatairesinol (6), matairesinol (7), oxomatairesinol (8), and isotaxiresinol (9), and the known taxoids taxinine M (10), taxayuntin (11), and 10-deacetyltaxol (12), and 10-deacetylbaccatin III (13) were isolated from the roots of Taxus cuspidata (Japanese yew, Taxaceae). The propelargonidin was isolated from Taxus spp. for the first time, and was detected in the roots, bark, and twigs.     

Response of <Emphasis Type="Italic">Theobroma cacao</Emphasis> and <Emphasis Type="Italic">Inga edulis</Emphasis> seedlings to cross-inoculated populations of arbuscular mycorrhizal fungi

Response of Theobroma cacao L. (cacao) and the shade tree Inga edulis Mart. (inga) seedlings from an organically-grown cacao plantation to inoculation with native arbuscular mycorrhizae forming fungi (AMF) was studied in a cross-inoculation assay under greenhouse conditions. Seedlings of inga and cacao were grown in pots filled with heat-treated soil from the plantation. Control was heat-treated soil without inoculum and roots of cacao and inga from the plantation were applied as AMF inocula. Undisturbed soil blocks were used as a “positive control” of the inoculation potential of untreated soil and roots combined. No AMF structures were observed in the roots of either species in the heat-treated control. All inocula were infective in both hosts and the differences in the total AMF colonization percentage between the hosts were not significant but inga had significantly higher colonization by hyphal coils and arbuscules. Cacao roots but neither inga roots nor soil block inocula stimulated cacao growth. All inocula significantly increased growth of inga, which had higher relative mycorrhizal responsiveness than cacao. Thus, in spite of the strong infectivity of the inocula in both hosts, cacao and inga responded differently to the same AMF populations. The strong conspecific preference of cacao suggests that attention must be paid to the AMF inoculum used for this species. However, the strong response of inga to cacao root inoculum indicates that the two species may share same AMF symbionts, thus enabling positive interactions between them, including formation of common mycelial networks.     

Production of <Emphasis Type="Italic">Pinus halepensis</Emphasis> seedlings inoculated with the edible fungus <Emphasis Type="Italic">Lactarius deliciosus</Emphasis> under nursery conditions

Inoculation with edible fungi bestows an added value on mycorrhized plants since production of mushrooms as a secondary crop can be an economically-valuable resource in forestry management. In order to establish a protocol for controlled mycorrhization of Pinus halepensis with the edible fungus Lactarius deliciosus, several factors such as fertilisation (35, 60 or 120 mg total N and 13.5, 27 or 54 mg total P), potting substrate (sphagnum peat or sphagnum peat/vermiculite) and fungal inoculum (mycelial slurry produced in a bioreactor, alginate beads or peat/vermiculite) were assessed. The most effective inoculum was mycelial slurry at a dose of 10 ml/plant. The two potting substrates assessed were compatible with mycorrhiza formation. The greatest number of mycorrhized seedlings was obtained with a moderate level of N (35 mg/plant) or P fertilisation (27 mg/plant). Inoculation did not produce a consistent growth effect on P. halepensis, but seedlings met the quality requirements, making them suitable for planting out.     

<Emphasis Type="Italic">Bursaphelenchus mucronatus</Emphasis> (Nematoda: Aphelenchoididae) vectored by <Emphasis Type="Italic">Monochamus </Emphasis><Emphasis Type="Italic">urussovi</Emphasis> (Coleoptera: Cerambycidae) in Hokkaido,Japan

Bursaphelenchus mucronatus Mamiya et Enda has been recovered for the first time from adults of the cerambycid beetle, Monochamus urussovi (Fischer), in Hokkaido, Japan. The nematode was also recovered from the inner bark of Picea jezoensis (Siebold et Zuccarini) Carrière and Abies sachalinensis (Fr Schmidt) Masters infested with M. urussovi larvae. PCR–RFLP analysis indicated that B. mucronatus in Hokkaido is the European type.     

Fluorescein-labeled wheat germ agglutinin stains the pine wood nematode, <Emphasis Type="Italic">Bursaphelenchus </Emphasis><Emphasis Type="Italic">xylophilus</Emphasis>

Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), is a major threat to pine forests throughout East Asia. Nonetheless, its mechanism of invasion has not yet been described in detail. To better understand the pathology of this disease, it is important to examine the distribution of PWNs within pine tissue during the course of disease development. We attempted to stain nematodes with fluorescein-conjugated wheat germ agglutinin (F-WGA) as a means to locate and track the spread of PWNs. Although PWNs proliferated on Botrytis cinerea fungus were successfully stained only on their vulvas and spicule holes, PWNs extracted from inoculated Pinus thunbergii seedlings were stained on their surface. Stainability, or the percentage of PWNs stained with F-WGA over more than half of their surface, was about 20% by 1 day after inoculation, but increased to 80% at 10 days. The stainability of PWNs extracted from a 5-cm main stem segment that included the inoculation site was less than that of PWNs extracted from other parts of the main stem farther away (i.e., those that had dispersed). These results suggest that stainability is related to dispersal activity in time. Thus, to raise the stainability of PWNs at shorter timeframes after inoculation, PWNs with more than 80% stainability were re-inoculated into pine seedlings. This resulted in more than 70% stainability from 1 to 6 days after inoculation. In F-WGA stained thin paraffin sections of pine tissue of re-inoculated seedlings, PWNs brightly fluoresced under epifluorescence and were easily detected against the dark background of pine tissue. This staining technique with F-WGA is an excellent tool for detecting PWNs in pine tissue.     

Aboveground biomass equations for individual trees of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>, <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> and <Emphasis Type="Italic">Larix kaempferi</Emphasis> in Japan

Generic equations are proposed for stem, branch and foliage biomass of individual trees in even-aged pure stands of Cryptomeria japonica, Chamaecyparis obtusa and Larix kaempferi. Biomass data was collected from a total of 1,016 individual trees from 247 stands throughout Japan, and five regression models were assessed by root mean square error, mean bias, fit index (FI), and AIC. The results show that a power equation using diameter at breast height (dbh) and height is the most suitable for all species and components. This equation is more accurate than the familiar power equation that uses ‘dbh² height’, and it expresses the greater volume of branch and foliage mass of trees with a lower height/diameter ratio. A power equation using dbh is more reasonable for models with dbh as the only independent variable and more accurate than a power equation using ‘dbh² height’ for estimating branch and foliage mass. Estimating error for branch and foliage mass is larger than that for stem mass, but the entire aboveground biomass can be estimated with an error of less than 19%, except in the case of small trees with dbh less than 10 cm.     

A novel cold-inducible promoter,P<Emphasis Type="Italic">ThCAP</Emphasis> from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, confers cold tolerance in transgenic <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Our previous studies have revealed that the ThCAP gene plays a vital role in transgenic Populus (P. davidiana × P. bolleana) in response to cold stress. However, the regulatory mechanism of ThCAP gene expression has been unclear. In this study, the 5′ flanking region of the ThCAP promoter (PThCAP) was cloned using a genome-walking method. By analyzing cis-acting regulatory elements of PThCAP, a DRE motif and MYC and MYB elements were found to be located in the promoter. To identify the regulatory elements that control the expression of the ThCAP gene promoter, a series of deletion derivatives of PThCAP, P1–P5, from the translation start code (?1538, ?1190, ?900, ?718 and ?375 bp), were fused to the GUS reporter gene, and then each deletion was stably introduced into Arabidopsis thaliana plants. Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A. thaliana exposed to low temperature stress. These results suggest that this 290-bp region (?1190 to ?900 bp), as an important part in PThCAP, was associated with cold tolerance of A. thaliana. Our results provide evidence for the regulatory mechanism of ThCAP gene involved in the response to cold stress, and that the gene is promising candidate gene for genetic improvement of crops.