Contamination and browning in tissue culture of Platanus occidentalis L.

Twigs of 2–3-year-old Platanus occidentalis L. were used as experimental material to find the causes for the contamination and browning in the initial stages of tissue cultures. To compare the degree of browning of explants picked off from different growing seasons, the experimental material was excised from trees on each of the first ten days in January, March, May and July, 2006. The results indicated that the contamination and browning rates of the material cut off in January (14.2% and 30.6%, respectively) and March were somewhat lower than those in July. The pretreatment of soaking the explants in different anti-oxidants and absorbents at the same time could diminish some side effects. The pretreatment of using 10 g·L–1 vitamin C reduced the contamination and brown-ing rate effectively. An orthogonal experiment showed that the optimal factor and level arrangement is 0.5 mg·L–1 BA, 2.0 g·L–1 ac-tive carbon and 1.5 g·L–1 PVP which resulted in a browning rate of only 16.5%. In general, sampling period, physical properties and pretreatment of explants are the main factors responsible for the contamination and browning of material in the initial stages of P. occidentalis tissue cultures.     

筋骨草内昆虫蜕皮激素类似物对杨干象的杀虫作用

从不同地点、不同时间采集的多花筋骨草植株不同部位中提取出昆虫蜕皮激素类似物,通过向幼虫人工饲料中添加昆虫蜕皮激素类似物的方法,研究了筋骨草中植物源蜕皮激素对杨干象2龄幼虫的杀虫作用。实验结果表明:(1)在人工饲料中加入1-3 ml的筋骨草提取物,在24天内可以杀死 58%~100%的幼虫;(2)从开花前的筋骨草中提取出的物质的杀虫效果明显优于从花期和开花后的植物中提取出的物质的杀虫作用, 其1 mL、2 mL和3 mL剂量全株提取物的杀虫效果分别达到65.22%、85.07%和98.11%(幼虫校正死亡率);(3)植株的根部提取的昆虫蜕皮激素类似物的杀虫效果要比茎部或叶部提取的好,1 mL、2 mL、3 mL剂量提取物的杀虫效果分别达到100%、98.20%和98.32%;(4)筋骨草提取物的杀虫速度较慢,处理杨干象2龄幼虫16天后杀虫率才超过50%;(5)幼虫取食加有筋骨草提取物的饲料时间长短直接影响幼虫死亡率,16天的取食,幼虫死亡率达到80%以上。筋骨草采集地点和填加植物源蜕皮激素的方式对杀虫效果的影响较小。图3表3参21。     

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.     

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.     

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.     

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.     

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.     

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.     

Extraction of <Emphasis Type="Italic">Trigonella foenum-graecum</Emphasis> L. by supercritical fluid CO<Subscript>2</Subscript> and its contact toxicity to <Emphasis Type="Italic">Rhyzopertha dominica</Emphasis> (Fabricius) (Coleoptera: Bostrichidae)

Rhyzopertha dominica (Fabricius) has developed extensive pesticide resistance in the last several decades. We have developed a supercritical fluid extraction method for Trigonella foenum-graecum L. (TFG) and studied the contact toxicities of the extracts to R. dominica. The extraction method was designed with orthogonal experiments to preserve and collect all the possible active components. Contact toxicity and efficiency of extraction were used as standard values to optimize extraction conditions, which were achieved at 55°C under 25 Mpa of pressure. The extraction efficiency for 200 g of dry sample reached 6.21% with 30 ml of 95% alcohol. Extracts loaded on filter paper showed dose and time dependent toxicities to adult R. dominica with a LC₅₀ value of 65.02 μg/cm² after 3 days post treatment. Our extensive in vivo studies indicated the extracts from Trigonella foenum-graecum seeds have high efficacy against pesticide resistant R. dominica. The active ingredient(s) from the extract shows promise as a novel pesticide candidate.     

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。     

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.     

Dynamic changes of telomeric restriction fragment (TRF) lengths in cells during the developmental process from embryos to seedlings and a comparison with the embryonal calli in Ginkgo biloba L.

Telomeres are the structures that locate at the terminals of linear eukaryotic chromosomes. They can play essential roles in many cellular processes. The terminal location of Arabidopsis-type TTTAGGG tandem repeats were thought to be highly conserved. The terminal location of Ginkgo biloba L. consisting of TTTAGGG tandem repeats, were confirmed by Bal31 exonuclease degradation and Southern blotting. By comparing telomeric restriction fragment (TRF) lengths at different developmental stages from embryos to seedlings, a fluctuant tendency towards variation was found in these samples. The TRF length of embryos was also compared with that of embryonal calli and an upward trend was discovered in callus culture. The results suggest that there should be a telomerase mechanism or/and ALT mechanism for the maintenance of telomere length.     

Floral display and breeding system of Jatropha curcas L.

Plant flowering and breeding characteristics are important for us to understand the reproduction of plant populations. In this paper, we studied the reproduction characteristics of Jatropha curcas in Yuanjiang County (23°36′N, 101°00′E), Yunnan Province. The plant produces flowers in dichasial inflorescences. Normally, the flowers are unisexual, and male and female flowers are produced in the same inflorescence. Only a few male flowers are produced in an inflorescence, and fruits are produced only through pollination between different flowers from the same or different plants. By the treatments of emasculation, bagging and artificial pollination in this experiment, there were few but same fruit set ratios when the inflorescences were emasculated, bagged, or bagged with net, except artificial pollination treatments, which showed that Jatropha curcas could produce fruit through apomixis but not wind pollination. When the inflorescences were unbagged, unemasculated and with free pollination treatments, or bagged, emascu- lated and with artificial cross-pollination treatments, or unbagged, emasculated and with free pollination treatments, there were many fruits produced. It showed that Jatropha curcas shows outcrossing, is self-compatible, and demanding for pollinators. Normally, the male flowers open first and a few flowers bloom in one day in a raceme. These flowers last a long time in bloom. However, a large number of female flowers open from the third to the fifth day, with some female flowers opening first in a few raceme. This shows a tendency to promote xenogamy and minimize geitonogamy.     

Allelopathic effects of Lantana camara on germination and growth behavior of some agricultural crops in Bangladesh

An experiment was conducted to understand the growth inhibitory effects of aqueous extracts derived from Lantana camara L. （a globally recognized invasive alien weed） on six popular agricultural crops of Bangladesh. The test was conducted in sterilized petridishes with a photoperiod of 24 hours and an average temperature of 29℃. The effect of different concentrations ofL. camara leaf extracts were recorded and compared with control （i.e., distil water）. Result showed different concentrations of aqueous leaf extracts caused significant inhibitory effect on germination, root and shoot elongation and development of lateral roots of receptor crops. Bioassays also indicated that the inhibitory effect was proportional to the concentrations of the extracts and higher concentration had the stronger inhibitory effect whereas the lower concentration showed stimulatory effect in some cases. The inhibitory effect was much pronounced in root and lateral root development rather than shoot and germination.     

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.     

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.     

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