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.     

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.     

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 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.     

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.     

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.     

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     

Nutrient element contents of cutting seedlings of hybrid species （ Liriodendron chinense x tulipifera）

The cutting seedlings ofLiriodendron chinense xtulipifera were treated with the different concentrations of auxin (treatment₁: IBA of 50 g·kg⁻¹+NAA of 300 g·kg⁻¹; treatment₂; IBA of 100 g·kg⁻¹+NAA of 300 g·kg⁻¹). The biomass nutrient element contents for different organs (root, stem, leaf) of cutting seedling ofLiriodendron chinense xtulipifera were measured by the dry method, Kjeldahl method and Atomic Absorption Spectroscopy method. The result showed that the biomass of root, stem, and leaf of the cutting seedling treated with auxin was all remarkably increased. The contents of element C in root, stem and leaf had no significant difference between the control and auxin treatments, while the contents of N, P, K and Ca in stem were much lower than that in leaf and root. Variance analysis showed that for the same organ with different concentration treatment of auxin, the four nutrient elements (N, P, K, and Ca) had no significant difference in contents, while there existed significant or very significant difference in contents of the four nutrient elements in different organs with the same concentration auxin treatment. The N, P, K and Ca contents were very low in cutting seedlings; as a result, additional fertilizer should be applied to the seedlings when they were planted in the field. Foundation item: This paper was supported by Jiangsu Province Science Foundation (BE96350). Biography: ZHANG Xiao-ping, (1972-), female, Ph. Doctor in Nanjing Forestry University. Nanjing 210037, P. R. China. Responsible editor: Zhu Hong     

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.     

Temperature-dependent development of <Emphasis Type="Italic">Diglyphus isaea</Emphasis> (Hymenoptera: Eulophidae) on <Emphasis Type="Italic">Liriomyza sativae</Emphasis> (Diptera: Agromyzidae) on cucumber

The development of Diglyphus isaea (Walker), a parasitoid of leafminers, was studied under laboratory conditions at seven constant temperatures (10, 15, 20, 25, 30, 35, and 40°C) on Liriomyza sativae Blanchard reared on cucumber (Cucumis sativus L.). The total development period (oviposition to adult emergence) decreased with increasing temperature between 15 and 35°C. In 10 and 40°C no development rate was found in D. isaea, so that it may be claimed that these temperatures fall outside the temperature range for development. Linear regression was used to describe the relationship between development rate and temperature. For egg to adult development, males of D. isaea required 153.8 degree-days (DD) above the theoretical threshold of 9.2°C and females required 161.3 DD above 9.4°C. Data were fitted to four nonlinear temperature-dependent models. Evaluation of the models took place based on the following criteria: fit to data, number and biological value of the fitted coefficient, and accuracy on the estimation of the thresholds. It could be concluded that the Briere-1 and Briere-2 models are suitable for estimating the minimum, maximum and optimal temperature thresholds of D. isaea. Thermal requirements and temperature thresholds can be used to predict the occurrence, number of generations and population dynamics of D. isaea.     

Influence of steam heating on the properties of pine (<Emphasis Type="Italic">Pinus pinaster</Emphasis>) and eucalypt (<Emphasis Type="Italic">Eucalyptus globulus</Emphasis>) wood

Heat treatment of Pinus pinaster and Eucalyptus globulus woods, two important species in Portugal, was performed in the absence of air by steaming, inside an autoclave, for 2–12 h at 190–210°C. Mass losses increased with treatment time and temperature reaching 7.3% for pine and 14.5% for eucalypt wood. The wood behaviour with moisture was improved. The equilibrium moisture content decreased by 46% for pine and 61% for eucalypt, the dimensional stability increased (maximum anti-shrinking efficiency in the radial direction of 57 and 90% for pine and eucalypt, respectively) and the surface wettability was lowered. In relation to mechanical properties, the modulus of elasticity was little affected (maximum decrease of 5% for pine and 15% for eucalypt) but the bending strength was reduced (by 40% at 8% mass loss for pine and 50% at 9% mass loss for eucalypt wood). The variation of properties was related to treatment intensity and mass loss but significant improvements could already be obtained for a 3–4% mass loss without impairing the mechanical resistance. The response of eucalypt was higher than that of pinewood. Heat treatment of eucalypt wood shows an interesting potential to improve the wood quality for solid timber products.     

Spatial distribution of sporocarps and the biomass of ectomycorrhizas of <Emphasis Type="Italic">Suillus pictus</Emphasis> in a Korean pine (<Emphasis Type="Italic">Pinus koraiensis</Emphasis>) stand

 Spatial distribution of sporocarps of Suillus pictus A.H. Smith and Thiers was studied in a plot of 6 × 12 m in size established in a stand of Pinus koraiensis Sieb. et Zucc. in Kyoto, Japan for 4 years, and the biomass of mycorrhizas was examined in the last year. S. pictus was dominant in both sporocarp and ectomycorrhizal community in the study plot. The number of S. pictus sporocarps ranged from 0.94 to 1.26 m⁻² (surface area) in the study plot and did not vary very much during the study period. Sporocarps of S. pictus occurred in clumps and the distributions of clumps were generally random. As the spots of sporocarp occurrence changed gradually from year to year, the distributions of sporocarps that occurred successively in 2-year periods overlapped, especially when analyzed in 9-m² unit size using the m ^* –m method. Mycorrhizas of S. pictus were distributed in more subplots than its sporocarps. Distribution of mycorrhizas and sporocarps of S. pictus generally overlapped well. The biomass of mycorrhizas and mycelia in the mycorrhizas of S. pictus was estimated at 15.5 g DW m⁻² and 6.2 g DW m⁻² (surface area) in this plot, respectively. The biomass of mycorrhizas and mycelia in the mycorrhizas supporting the production of one sporocarp (average dry weight was 0.86 g) of S. pictus was evaluated as about 16.4 and 7.3 g DW, respectively, in this plot. Received: December 20, 2001 / Accepted: August 12, 2002 Acknowledgments We thank Dr. T. Furuno, Mr. N. Kato, and Dr. I. Nakai for their help in preparing the study plot and collecting sporocarps, and Prof. K. Yokoyama for the identification of sporocarps. Thanks are also due to Dr. E. Kuno for his suggestion about analysis. Correspondence to:J. Kikuchi     

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.     

Evaluation of <Emphasis Type="Italic">Serangium parcesetosum</Emphasis> (Col.: Coccinellidae) for biological control of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> under greenhouse conditions

The cotton whitefly, Bemisia tabaci (Genn.) (Hom., Aleyrodidae) is increasingly a very important pest on many vegetables, field crops and ornamental plants. Therefore, controlling of this pest is still needed especially under glasshouse conditions. The specialist whiteflies’ predator, Serangium parcesetosum Sicard (Col., Coccinellidae) appears to have a great potential for the cotton whitefly control. In this study, the dynamic changes in B. tabaci populations in glasshouse cabins in response to S. parcesetosum were monitored. B. tabaci were introduced to cotton plants in three cabins in average of 50 adults per plant. One and two weeks later, adult females and males of S. parcesetosum were introduced at a rate of one female and one male per plant in the first and second cabins, respectively. The third cabin was considered as a control. The results showed that the mean number of whiteflies in the control cabin was found significantly higher than that of either when S. parcesetosum was introduced 1 or 2 weeks after the infestation with the whitefly. Also, the mean number of B. tabaci was significantly higher when the predator was introduced 2 weeks rather than 1 week after B. tabaci infestation. The maximum mean weekly number of whiteflies/plant was 192.3 in the second week, whereas it was 294.6 in the third week and 1136.4 in the fifth week, in first, second and control cabins, respectively. In the last experimental week, the mean weekly numbers were 74.7, 122.9 and 684.7 whiteflies/plant in the three cabins, respectively. S. parcesetosum has been successfully fed, reproduced and established its population on B. tabaci on cotton plants. The mean weekly number of the predatory individuals increased gradually with the progress of the experimental time. The results demonstrated that the maximum reduction percentage in B. tabaci population was 90.7 and 86.5% in the fifth week after B. tabaci infestation, when the predator was introduced 1 and 2 weeks after the infestation with the whiteflies, respectively. Nevertheless, it is speculated that an earlier release of S. parcesetosum would be more effective in the biological control of B. tabaci.     

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.     

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.     

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