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.     

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.     

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。     

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.     

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     

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     

New piezoelectric moduli of wood: <Emphasis Type="Italic">d</Emphasis><Subscript>31</Subscript> and <Emphasis Type="Italic">d</Emphasis><Subscript>32</Subscript>

This article reports the piezoelectric moduli of wood d ₃₁, d ₃₂, and d ₃₆. The piezoelectric moduli of wood d ₃₁ and d ₃₂ have not been previously reported, although there has been much research on the d ₁₄ and d ₂₅ moduli of wood. The moduli d ₃₁, d ₃₂, and d ₃₆ were measured carefully because their absolute values were considerably smaller than those of d ₁₄ and d ₂₅. For Softwoods, d ₃₆ values were mostly negative, whereas the values for hardwoods had either positive or negative values. The other moduli, d ₃₁ and d ₃₂, were a mixture of positive and negative values in softwoods and hardwoods. The existence of d ₃₁ and d ₃₂ suggests the presence of an electrical polarity of the cellulose crystal in the fiber direction of the wood. The polarities of d ₃₁ and d ₃₂ became clear from wood in the outer part of the trunk, where the crystallinity of cellulose is large and the alignment of the crystals becomes parallel to the fiber direction.     

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.     

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.     

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.     

Replication of wood into biomorphous nanocrystalline Y<Subscript>2</Subscript>O<Subscript>3</Subscript>:Eu<Superscript>3+</Superscript> phosphor materials

Biomorphous Eu³⁺-doped Y₂O₃ was fabricated by replication of wood templates using vacuum-assisted infiltration of a water-based sol–gel mixture and subsequent calcination at 750°C. The precursor sols were prepared from (Y_0.95Eu_0.05)₂O₃ dissolved in 10 vol% nitric acid and adding citric acid as the chelating agent. X-ray powder diffraction analyses and Rietveld refinements confirmed that the calcined samples were solely composed of bixbyite Y₂O₃:Eu³⁺ phase with a mean crystallite size of 16 nm. Scanning electron micrographs and cathodoluminescence imaging showed that the cellular preform anatomy was retained and that the original wood cell walls were completely transformed into phosphor struts with pore sizes ranging from 5 to 20 μm. The optical properties of the biomorphous phosphor materials were analyzed by photoluminescence spectroscopy and assigned to the characteristic Eu³⁺ (4f⁶ → 4f⁶) electric dipole or magnetic dipole transitions. From fluorescence lifetime measurements, the mean lifetime was calculated as 1.62 ms.     

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.     

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.     

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.     

广州碧道水生态设施标识系统优化设计初探

近年来，广州市深入贯彻落实习近平生态文明思想，大力推进碧道高质量规划建设。水生态设施是落实碧道建设“5+1”任务的重要载体，当前，广州碧道建设缺乏完整、规范的水生态设施标识系统，在实地调研碧道标识系统的基础上，综合考虑环境心理学和人体工程学，凝练地域特色等元素符号，对标识系统的结构组成、外观形式、符号元素和界面信息进行优化设计，形成具有系统性、体验性和文化性的水生态设施标识系统，对现有碧道标识系统进行完善，助力广州碧道建设高质量发展。     

Vitality and storage condition of Syringa pollen

The fresh pollen vitality,the effect of different storage conditions on the pollen vitality,and the difference of vitality among the species of seven species of Syringa were determined in Shenyang,China.The results indicated that the pollen vitality (81.5%) of Syringa villosa was the highest among the seven tested species,followed by S.microphylla and S.meyeri,and that of S.oblata var.affinis was the lowest.The low temperature was the best condition for storage of pollen of Syringa,and the most proper temperature for the storage was 0-2℃.The storability of S.microphylla was the best of all,and it could be stored over 60 days at the temperature of 0-2℃,next was S.villosa and S.meyeri.     

Embryo culture and rapid propagation of Syringa

Embryo of lilacs (Syringa L) culture in vitro and the rapid propagation were studied. The orthogonal experiments, including the selection of basal medium, embryo age and other factors such as sugar, benzyladenine (BA), naphthalene acetic acid (NAA) and glutamine (Gln), were carried out. The results indicated that the optimal medium for embryo culture was Monnier medium supplemented with NAA (0.001 mg.L^-1), BA (0.1 mg.L^-1), sugar (50 g.L^-1), and Gin (400 mg.L^-1), with a germination rate of 91.7% at least; the optimal embryo age was 50 d; and Gln had significant effects on the germination rate of embryo.Moreover, the optimal medium for subculture was MS BA (2 mg.L^-1) NAA (0.001 mg.L^-1) Gln (0.5 mg.L^-1), with the propagation coefficient of 3.6 at least.     

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.     

Horse grazing in firebreaks sown with <Emphasis Type="Italic">Trifolium brachycalycinum</Emphasis> (Katznl. &; Morley) and <Emphasis Type="Italic">Cynodon</Emphasis> dactylon (L.) Pers

Firebreaks are useful to reduce the extent of forest fires, however, when they are cleared down the slope, soil erosion is increased and the native tall sward in summer can increase fire risk. The sown mixture Trifolium brachycalycinum (Katznl. & Morley) and Cynodon dactylon L. Pers. has already proved to be useful for both fire prevention (the short sward remains green in summer, creating easy access) and biomass production (clover can give high yields with high palatability and protein levels). This pasture needs to be grazed to be kept short and reduce fuel biomass in summer. Unfortunately, there is not much livestock reared in Central Italy and animals are normally fed indoors. In this article, a trial in a coastal farm of Central Italy is reported to investigate the efficiency of horses in maintaining the sown species botanical composition, soil cover, sward height, biomass (before and after grazing) and fuel biomass in summer in a firebreak over 4 years. The mixture was intensively grazed over short periods during spring and summer. Animals grazed both the sown species and native plants, they trampled on the remaining herbage, breaking the stems, and leaving short and compact dry mulch. Very little combustible vegetation was left in the hot season and horse grazing is recommended for farms that have firebreaks.