Variation in pitch canker resistance among provenances of <Emphasis Type="Italic">Pinus patula</Emphasis> and <Emphasis Type="Italic">Pinus tecunumanii</Emphasis> from Mexico and Central America

Pinus patula and high-elevation (HE) sources of P. tecunumanii exhibit intermediate levels of resistance to pitch canker (Fusarium circinatum), compared to extremely resistant species such as P. oocarpa, and extremely susceptible species such as P. radiata. Seedlings from 20 P. patula provenances and 15 HE P. tecunumanii provenances were artificially inoculated with the pitch canker fungus at 21 and 12 weeks of age, respectively, and assessed for resistance 12–20 weeks later. There was important provenance variation in pitch canker resistance for both species. The 20-week LiveStem percentage ranged from 70.3% to 43.6% among the P. patula provenances and 59.6% to 11.7% among HE P. tecunumanii provenances. There was a geographic pattern to the provenance variation, and in both species, low altitude sources demonstrated more resistance than those from high elevation. Provenance variation in pitch canker resistance could be useful when making selection and breeding decisions with these species.     

The impact of shade on morphology,growth and biomass allocation in <Emphasis Type="Italic">Picea sitchensis</Emphasis>, <Emphasis Type="Italic">Larix</Emphasis> × <Emphasis Type="Italic">eurolepis</Emphasis> and <Emphasis Type="Italic">Thuja plicata</Emphasis>

To determine the effect of shade on morphology, growth and biomass allocation in Picea sitchensis, Larix × eurolepis and Thuja plicata, seedlings were grown in the open or under shadehouses providing 25%, 50% and 75% reductions of full-light for two growing seasons. For most of the characteristics assessed there was no significant interaction between species and shade indicating that the morphological responses to changing shade treatments were not species-dependent. After two growing seasons the mean height increment for the three species was significantly greater in 25% (76.1 cm) and 50% shade (74.9 cm) than in the open (69.5 cm). Root collar diameter increment, shoot, root and total biomass declined significantly with increasing shade while the opposite was true for the height:diameter ratio. In both western red cedar and hybrid larch the shoot:root ratio was significantly greater in the shade while in Sitka spruce this characteristic was not influenced by shade. While all species had significantly greater specific shoot areas in 75% shade than in 0% shade, this trend was particularly pronounced in hybrid larch. In hybrid larch and western red cedar, the normalised specific projected shoot area increased significantly with increasing shade. The opposite trend was observed for Sitka spruce. We conclude that in the main the species studied demonstrated similar shade acclimation responses despite their reported differences in shade tolerance.     

Agrobacterium-mediated genetic transformation of Camptotheca acuminata

UGPase gene related with wood cellulose synthesis was transferred into C. acuminata using the method of Agrobacte- rium-mediated genetic transformation, and an efficient transformation system was developed for C. acuminata on the basis of evaluations of several factors affecting Agrobacterium-mediated DNA transfer rate. The highest transformation rate was achieved when pre-cultttred leaf explants were infected with an Agrobacterium culture corresponding to OD600 （0.5） for 10 min, and cultured on explant regeneration medium for three days. The results of Southern hybridization showed that genomic DNA of the kanamycin-resistant shoots to an UGPase gene probe substantiated the integration of the transgene. Transformation efficiency （6%） was achieved under the optimized transformation procedure, This system should facilitate the introduction of important useful genes into C, acuminata.     

Problems of using pines in <Emphasis Type="Italic">Tuber melanosporum</Emphasis> culture: soils and truffle harvest associated with <Emphasis Type="Italic">Pinus nigra</Emphasis> and <Emphasis Type="Italic">P. sylvestris</Emphasis>

In Mediterranean pine forests, truffles and mushrooms generate greater profits than any other woodland products. However, there are no studies on Tuber melanosporum Vittad. associated with pines. For this reason, we have carried out a study of this truffle in mountain woods with Pinus sylvestris L. and P. nigra Arnold subsp. salzmannii (Dunal) Franco, in central Spain. Two hundred and eight Tuber melanosporum burns were monitored for 7 years in five different habitats within the same geographical area. An ANOVA test confirmed significant differences in carpophore production. In higher producing habitats, pines were less abundant. We also confirmed that in 433 burns, T. melanosporum was always unequivocally associated with the root base of Quercus or Corylus trees. Similarly, 14 truffle collectors confirmed that they had never found a single burn with carpophore production associated exclusively with pines. Nevertheless, soil analyses indicated that the soil of these pine woods was very favourable to Tuber melanosporum. We therefore conclude that at present Pinus nigra salzmannii and P. sylvestris are of little interest to Tuber melanosporum culture, as they hinder carpophore production. However, this study has also confirmed that Pinus nigra salzmannii and P. sylvestris mycorrhize easily with Tuber melanosporum, both in the laboratory and in natural environments. On this basis, we propose that pines may act as transmitters of T. melanosporum, although they do not induce fruiting. As a result, the commercial cultivation of Pinus nigra salzmannii and P. sylvestris seedlings mycorrhized with Tuber melanosporum is not recommended in truffle culture at the present time.     

Dynamics of seed imbibition and germination of Austrian pine (<Emphasis Type="Italic">Pinus</Emphasis><Emphasis Type="Italic">nigra</Emphasis> Arnold) from extreme habitat conditions within five Balkan provenances

Seed was collected from two habitat types (harsh/rocky and favorable/control) within five Balkan provenances. An experiment was conducted to examine water uptake (grams water per gram of fresh seed) during 48 h after exposure to water. A second experiment investigated the effect of simulated-drought conditions (4% sucrose solution) compared to non-drought conditions (distilled water) on subsequent germination percentage. Seed from rocky habitats had less water uptake than those from control habitats for three of the provenances. However, water uptake was not correlated with subsequent germination. Drought simulation decreased germination but patterns for seed within each provenance were similar to germination under non-drought conditions. With the exception of one provenance, seed from the rocky habitats had lower germination than seed from control habitats. The data suggests some degree of adaptation to environmental conditions although variability was very high within each provenance’s habitat types. This is of particular importance when considering potential impacts of climate change and the anticipated migration of species due to changes in habitat.     

High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli

In order to investigate the enzymatic properties of the 4CL1 of Populus tomentosa, the recombinant expression vector pQE31-4CL1 was constructed. The recombinant was identified by three restriction endonucleases, then the vector pQE31-4CL1 was transformed into expression host M15 (pREP4) and induced by isopropyl-α-D-thiogalactoside (IPTG) to express 60 kD fused protein Pt4CL1. The biologically active Pt4CL1, expressed as soluble protein, was achieved with 0.6 mmol·L-1 IPTG induction as the ex-pression temperature declined from 37 to 28°C. The 6×His tag facilitates affinity binding to Ni2 -nitrolotriacetic acid (NTA) and enables one-step purification to acquire the molecular SDS-PAGE electrophoresis purity of the active 4CL1 protein by agarose cou-pled with Ni2 -NTA affinity chromatography. The optimal substrate for Pt4CL1 was 4-coumarate.     

忍冬和华南忍冬叶表皮扫描电镜观察及其生态适应性研究

利用扫描电镜观察了忍冬（Lonicera jàponica Thunb）和华南忍冬（Lonicera confus）叶表皮形态特征,观察指标包括：气孔器、表皮毛、表皮细胞等.结果表明：气孔仅分布在下表皮,气孔器呈无规则型散生;上表皮细胞的垂周壁呈沟槽状下陷;下表皮密被表皮毛（包括腺状毛和非腺毛）,同时还分布有瘤状的草酸钙簇晶.为进一步证明这种瘤状结构是否为草酸钙簇晶,作者发现利用Fluo-3/AM（钙离子荧光指示剂）处理的活体叶片在激光共聚焦显微镜下能够发出强烈的荧光.此外,通过电子探针对瘤状物进行分析,发现其元素组成为C、O、Ca.上述叶表皮构造特征,有利于减少植物体水分散失,还有助于植物适应富钙的石灰土,从而适应我国西南岩溶区特殊的干旱、富钙的环境.电镜观察结果表明,在忍冬叶上表皮中脉处有呈线状分布的腺状表皮毛,而华南忍冬叶上表皮则无表皮毛分布,这一微形态特征可以作为二者种间特异性差别,从而为二者的合理区分提供一定的依据.     

Evaluation of efficiency of <Emphasis Type="Italic">Trichogramma evanescens</Emphasis> reared on different factitious hosts to control <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

The release of mass propagated egg parasitoids could be one option to control the lepidopteran pests. The efficiency of the parasitoid Trichogramma evanescens (Westwood) reared on eggs of three different factitious hosts; Sitotroga cerealella, Ephestia kuehniella and Galleria mellonella was studied for controlling bollworm Helicoverpa armigera. Efficiency of Trichogramma was studied by measuring parasitism rates, emergence rates, longevity and sex ratio. Wasps reared from each source were tested on the source host and on the target host, H. armigera under laboratory conditions. Rates of parasitism on H. armigera eggs, emergence rates of parasitoids and their longevity were the highest for wasps reared on H. armigera. Wasps reared on S. cerealella gave comparable rates. However, wasps from E. kuehniella gave the lower rates and G. mellonella gave the lowest ones. Parasitized eggs of H. armigera and S. cerealella produced more parasitoid females than eggs of E. kuehniella and G. mellonella. Results are discussed for magnifying efficiency of the parasitoid in controlling H. armigera in the field.     

Stress responses of <Emphasis Type="Italic">Salix gracilistyla</Emphasis> and <Emphasis Type="Italic">Salix subfragilis</Emphasis> cuttings to repeated flooding and drought

To compare the responses to repeated flooding and drought of Salix gracilistyla, which grows on coarse gravel substrates, and Salix subfragilis, which grows on fine silt or clay substrates, we measured pre-dawn leaf water potential (Ψw _pd), osmotic adjustment (Ψw _tlp, Ψo _sat), and biomass production of cuttings under greenhouse conditions. The experimental design involved a control and four treatments that crossed 1 or 3-week flooding (F) with 1 or 2-week droughts (D). Ψw _pd was reduced after 2 weeks of drought when preceded by 1 week of flooding. Neither species increased osmotic adjustment in response to increased duration of drought between repeated 3-week flooding. Moreover, a decrease in the ratio of leaf biomass to total biomass or an increase in the ratio of root biomass to total biomass with longer drought repetitions was not observed for either species. The root ratio of S. gracilistyla was more strongly inhibited by flooding than that of S. subfragilis. The shoot-to-root ratio of S. subfragilis was higher than that of S. gracilistyla in all F combinations. The hypertrophied lenticel ratio of S. gracilistyla after 1 week of flooding was nearly the same as that after 3 weeks of flooding, whereas values for S. subfragilis after 1 week of flooding were lower than those after 3 weeks of flooding. The low allocation to roots and the generation of hypertrophied lenticels by S. gracilistyla in response to flooding, as compared with S. subfragilis, seem to be related to the different habitat substrate conditions of the two species.     

Dynamics of non-structural carbohydrate reserves in pruned <Emphasis Type="Italic">Erythrina poeppigiana</Emphasis> and <Emphasis Type="Italic">Gliricidia sepium</Emphasis> trees

In alley cropping systems, fast growing leguminous trees are pruned to reduce competition with crops for light and to provide organic inputs for crop nutrition. Tree regrowth depends on non-structural carbohydrate reserves in the remaining tree parts. In this study, the dynamics of starch and soluble carbohydrates in roots and stems of completely pruned (all shoots removed), partially pruned (one branch retained on the pruned stump) and unpruned Erythrina poeppigiana (Walp.) O.F. Cook and Gliricidia sepium (Jacq.) Kunth ex Walp. trees were studied under humid tropical conditions in Turrialba, Costa Rica. Measurements on starch and soluble carbohydrates in roots and stems were made at 0, 2, 6 and 12 weeks after pruning during both a “rainy” and a “dry” season. In general, the dynamics of non-structural carbohydrates in roots and stems of pruned E. poeppigiana and G. sepium trees were similar. Starch concentration was highest in unpruned trees and higher in roots than in stems of pruned trees. The effect of pruning intensity was first observed in stems, and starch reserves were more depleted in stems than in roots, an effect more evident during the “dry” season. The critical tree regrowth stage for starch mobilisation was that of vigorous sprout development at six or four weeks after pruning particularly in completely pruned trees. At this time, fine root biomass and length and nodule biomass in pruned trees decreased. Survival of fine roots and nodules was greater in partially pruned than in completely pruned trees. Starch accumulation in roots recommenced at 12 weeks after pruning in G. sepium, and later than 12 weeks after pruning in E. poeppigiana roots. This study showed that E. poeppigiana responded better to pruning regimes than G. sepium. Recovery of trees after pruning is better when trees are partially pruned than when completely pruned.     

Effects of deer browsing on the survival and growth of <Emphasis Type="Italic">Abies homolepis</Emphasis> and <Emphasis Type="Italic">Picea jezoensis</Emphasis> var. <Emphasis Type="Italic">hondoensis</Emphasis> seedlings after fencing on Mt. Ohdaigahara

We evaluated the survival and growth of Abies homolepis and Picea jezoensis var. hondoensis seedlings on Mt. Ohdaigahara, where the population of sika deer (Cervus nippon) is high and an experimental fence has been in place for 13 years. No significant differences were detected in the survival of small seedlings between fenced and unfenced plots. The growth of A. homolepis was significantly higher in the fenced plot, but growth of P. jezoensis var. hondoensis did not show significant differences between fenced and unfenced plots. Seedlings of height ≤5 cm in the forest floor vegetation of the unfenced plot were probably too small for deer to find and browse, so they survived.     

Introduced <Emphasis Type="Italic">Eucalyptus</Emphasis> psyllids in Brazil

In the last 10 years, four species of psyllid were detected in Brazil on eucalypts: Ctenarytaina spatulata in 1994, Blastopsylla occidentalis in 1997, Ctenarytaina eucalypti in 1998 and Glycaspis brimblecombei in 2003. The latter two are serious pests in several countries. In Brazil, G. brimblecombei caused significant damage to the eucalypt plantations in the first years of its introduction. Now this pest is under control due to the programmes of integrated pest management, where the parasitoid is the principal control agent. The four eucalypt psyllid species introduced into Brazil are presented with information on distribution, hosts, biology and control.     

Introduction of mitochondrial DNA from <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> into <Emphasis Type="Italic">Pleurotus pulmonarius</Emphasis> by interspecific protoplast fusion

Interspecific protoplast fusion between two monokaryotic strains (a methionine auxotrophic and chloramphenicol-resistant Pleurotus ostreatus strain and a wild-type strain of Pleurotus pulmonarius) was carried out to introduce mitochondrial DNA (mtDNA) from P. ostreatus into P. pulmonarius. Because mycelial colonies regenerated on minimum medium containing chloramphenicol only after the treatment of protoplast electrofusion, the regenerants were regarded as protoplast fusants. The fusants isolated from regenerated colonies were uninucleate, and their resistance of chloramphenicol seemed to be a stable trait. The fusants mated with P. pulmonarius but not with P. ostreatus, and showed almost identical random amplified polymorphic DNA (RAPD) patterns to that of the parental P. pulmonarius monokaryon. The sizes of mitochondrial genomes were estimated to be 81.5 kb for P. ostreatus monokaryon, 54.9 kb for P. pulmonarius monokaryon, and 84.5 or 86.0 kb for the fusants. BamHI and EcoRI digest patterns of the fusant mtDNAs were almost the same as the parental P. ostreatus monokaryon. From the above results, the fusants seemed to carry nuclei derived from the monokaryon of P. pulmonarius and mtDNA including the chloramphenicol-resistance gene from the P. ostreatus monokaryon, suggesting that the P. ostreatus mtDNA had been introduced into P. pulmonarius cells by interspecific protoplast fusion. Contribution No. 383 of the Tottori Mycological Institute     

思茅松天然群体的遗传多样性及遗传分化

采用凝胶电泳技术,检测了思茅松(Pinuskesiyavar.Langbinaensis)种子胚乳的9种同工酶。通过对9种酶系统编码的16个酶位点的遗传分析,揭示了思茅松三个天然群体的遗传多样性和遗传分化情况。思茅松天然群体的遗传多样性较高,群体的多态位点比例为0.667;平均每个位点的等位基因数为2.13;期望杂合度和观察杂合度分别为0.288和0.197。群体间的遗传分化程度较低,分化系数仅为0.052,群体间的遗传距离为0.015。表5参15。     

Analysis of sawn lumber production from logging residues of branchwood of <Emphasis Type="Italic">Aningeria robusta</Emphasis> and <Emphasis Type="Italic">Terminalia ivorensis</Emphasis>

One way of meeting the supply demands of the wood industry is through the reduction of waste in timber processing. It has been estimated in Ghana that for every tree felled, nearly 50% of the tree volume is left in the forest in the form of branches, crownwood, and stumps. In this study, the potential of utilizing branchwood as raw material from logging residues to help meet the timber demands of the downstream processing sector was analyzed experimentally. The sawing characteristics of the branchwood of Aningeria robusta and Terminalia ivorensis with diameters in the range 10–25 cm using live sawing and cant sawing methods were determined using a randomized complete block design, 2 × 4 factorial with three replications. Experimental results indicate that, the relative lumber value yields for first and second grade boards of A. robusta and T. ivorensis branches were 25 and 20%, respectively, while the green lumber volume yields were 40 and 32%, respectively. A high proportion of sapwood and the occurrence of washboarding in T. ivorensis contributed to that species’ relatively low lumber value and volume yield. The experimental results further indicate that the sawing technique used had no significant influence on lumber yields. However, at 5% level of significance, significant differences in lumber yield existed between some of the diameter classes.     

Crystalline structure of heat-treated Scots pine [<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.] and Uludağ fir [<Emphasis Type="Italic">Abies nordmanniana</Emphasis> (Stev.) subsp. <Emphasis Type="Italic">bornmuelleriana</Emphasis> (Mattf.)] wood

The aim of this study was to determine changes in crystallinity and crystalline unit cell type of heat-treated Scots pine (Pinus sylvestris L.) and Uludağ fir (Abies nordmanniana stev. subsp. bornmuelleriana Mattf.) wood samples by means of FT-IR spectroscopic method. Heat treatment was applied on the test samples in an oven at three different temperatures (120, 150, and 180°C) and for two different periods of time (6 and 10 h) under atmospheric pressure. It was designated that crystallinity of both Scots pine and Uludağ fir wood samples increased during heat treatment depending on the duration. However, monoclinic structure in crystalline unit cells of Scots pine and Uludağ fir wood samples converted to triclinic structure when heat treated. It was estimated that monoclinic structure was dominant in the crystalline unit cell. It was established that the crystalline structure of Scots pine wood samples was more affected by heat treatment than that of Uludağ fir wood samples.     

Effects of drought on isolates of <Emphasis Type="Italic">Bradyrhizobium elkanii</Emphasis> cultured from <Emphasis Type="Italic">Albizia adianthifolia</Emphasis> seedlings of different provenances

Albizia adianthifolia (Schumach) W. F. Wright, a N-fixing legume tree, has a wide distribution in Africa, in Ghana occurring in high rainfall forests and in seasonally droughted forests, and is associated in the Ghanaian forest zone with dry, infertile sites. We hypothesised that A. adianthifolia hosted different rhizobial strains in different forest types, and that these different strains would show different growth responses to moisture stress and different motility and mortality in droughted soil. Three isolates, extracted from seedlings of A. adianthifolia growing in three forest types differing in seasonal drought, were identified as Bradyrhizobium elkanii and exposed to varying levels of osmotic stress. Growth responses varied between the three strains, one of which displayed clear signs of drought tolerance. A novel approach using soil leaching columns was used to test the effects of soil pore water (in terms of neck diameter) on both the survival and movement of wet and dry forest rhizobial isolates through soil columns. The responses of the isolates were significantly different, the pore neck diameter, marginally insignificant and the drought treatment insignificant. Thus the dry forest isolate survived better in all treatments, and showed less response to the treatments, than the isolate from the wet forest. The results offer preliminary evidence that Bradyrhizobium elkanii strains from A. adianthifolia in Ghana have evolved in response to local differences in seasonal water availability. These differences could assist the selection of A. adianthifolia provenances for agroforestry or land rehabilitation.     

Distribution of solanesol in Nicotiana tabacum

Solanesol is an important secondary metabolite in Nicotiana tabacum. Distribution of solanesol in Nicotiana tabacum was investigated by High Performance Liquid Chromatography （HPLC） method. The quantitative distribution of solanesol in various organs and tissues of N. tabacum showed that solanesol content, obviously different in all organs, was 6.8, 18.3, 27.5, 45.8, and 68.0 times higher in leaves than that in the stalks, flowers, seeds, fruits and roots, respectively. The contents of solanesol in various parts of leaf, stalk and flower were determined. The content of solanesol in top leaf, middle leaf and bottom leaf gradually decreased （6.124, 5.813 and 5.687 mg.g^-1, respectively） and the content of solanesol in various leaf-parts （leaf apex, leaf middle and leaf base） also gradually decreased. The content of solanesol in top stalk was 1.19 times and 1.92 times higher than that in the middle stalk and the bottom stalk, respectively. The content of solanesol in various tissues of stalk （epidermis, cortex and stele） dramatically decreased. The sepal contained higher concentration of solanesol （1.192 mg·g^-1） compared to any other parts in flower. The study will provide the base data for the regulation and control of solanesol, moreover, it will provide the scientific evidences for the rational development and utilization of N. tabacum resources.     

Problems of using rockroses in <Emphasis Type="Italic">Tuber melanosporum</Emphasis> culture: soil and truffle harvest associated with <Emphasis Type="Italic">Cistus laurifolius</Emphasis>

The Périgord black truffle (Tuber melanosporum Vittad.) can mycorrhize with rockroses (Cistus L.) in Mediterranean forests. These shrubs may be of interest for truffle cultivation as they produce early carpophores, collaborate in tree mycorrhization and in the survival of Tuber melanosporum mycelium. However, there are very few quantitative studies on truffle production associated with Cistus. For this reason, we have carried out a study on the natural production of Tuber melanosporum associated with Cistus laurifolius L. in central Spain. An Anova test confirmed the existence of significant differences in carpophore production in four habitats, with and without the presence of C. laurifolius. Twenty-one burns associated with C. laurifolius showed less significant production than the rest. We also confirmed that in 122 burns, Tuber melanosporum was always unequivocally associated with the root base of Quercus L. or Corylus L. trees. Similarly, five truffle-collectors confirmed that they had never found a single burn with carpophore production associated exclusively with rockroses. It has also been statistically verified that many Cistus die in Tuber melanosporum burns, whereas in old burns with minimum carpophore production, a new colonization of young Cistus has been observed. This study also confirms that C. laurifolius mycorrhizes easily with Tuber melanosporum. We propose that Cistus laurifolius may act as transmitters of Tuber melanosporum, although they do not induce fruiting. Consequently, the commercial cultivation of Cistus laurifolius mycorrhized with Tuber melanosporum for production is not recommended in truffle culture. The results reported substantiate the negative observations in cultivations of T. melanosporum with other Cistus species.     

Lignin-degrading activity of edible mushroom <Emphasis Type="Italic">Strobilurus ohshimae</Emphasis> that forms fruiting bodies on buried sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) twigs

Strobilurus ohshimae is an edible mushroom, and it specifically forms its fruiting bodies on buried sugi (Cryptomeria japonica) twigs. In this research, we studied lignindegrading activity of S. ohshimae. We isolated 18 strains of S. ohshimae from various regions of Japan, and determined their lignin degradation rates on sugi wood meal medium. All the strains of S. ohshimae degraded approximately 6%–12% of sugi lignin in 30 days, and these lignin degradation rates were 1.5–3 times higher than those of Trametes versicolor, which is a typical lignin-degrading fungus. Among the three main lignin-degrading enzymes, activity of lignin peroxidase and manganese peroxidase was not observed, while 4340U/g of laccase was produced in 30 days. To investigate the effect of wood species on lignin degradation by S. ohshimae, the lignin degradation rate and laccase productivity on sugi wood meal medium were compared with those on beech (Fagus crenata). In T. versicolor, both lignin degradation rate and laccase productivity were higher on beech than on sugi. Conversely, in S. ohshimae, lignin degradation rate and laccase productivity were higher on sugi than on beech. Therefore, it was suggested that coniferous lignin is not always difficult to degrade for the fungi that inhabit softwood. Part of this article presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004