A novel cold-inducible promoter,PThCAP from Tamarix hispida,confers cold tolerance in transgenic Arabidopsis thaliana

Our previous studies have revealed that the Th CAP gene plays a vital role in transgenic Populus(P.davidiana 9 P.bolleana) in response to cold stress.However,the regulatory mechanism of Th CAP gene expression has been unclear.In this study,the 50 flanking region of the Th CAP promoter(PTh CAP) was cloned using a genomewalking method.By analyzing cis-acting regulatory elements of PTh CAP,a DRE motif and MYC and MYB elements were found to be located in the promoter.To identify the regulatory elements that control the expression of the Th CAP gene promoter,a series of deletion derivatives ofPTh CAP,P1–P5,from the translation start code(-1538,-1190,-900,-718 and-375 bp),were fused to the GUS reporter gene,and then each deletion was stably introduced into Arabidopsis thaliana plants.Deletion analysis of the promoter suggested that only the P2 fragment had strong GUS expression in leaves and roots of A.thaliana exposed to low temperature stress.These results suggest that this290-bp region(-1190 to-900 bp),as an important part in PTh CAP,was associated with cold tolerance of A.thaliana.Our results provide evidence for the regulatory mechanism of Th CAP gene involved in the response to cold stress,and that the gene is promising candidate gene for genetic improvement of crops.     

一个新的柽柳类萌芽素基因ThGLP的结构及其表达分析

萌芽素和类萌芽素蛋白在不同植物的各个生长阶段和胁迫相关过程中起到不同的作用。本研究首次从刚毛柽柳cDNA文库中获得类萌芽素蛋白全长基因ThGLP,该基因编码225个氨基酸,含有植物萌芽素和类萌芽素蛋白的功能序列。通过进化树分析发现该基因屑于真正萌芽素亚家族。利用实时定量PCR方法研究了该基因在PEG、NaCl、低温、CdCl2和ABA胁迫下不同时间的表达模式。结果显示PEG、NaCl、低温、CdCl2和ABA处理均能诱导ThGLP基因在柽柳的根和叶中的表达。结果表明ThGLP在柽柳根和叶中表达,参与非生物胁迫应答并由ABA依赖的信号传导途径调控。     

At5g54160 gene encodes <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> 5-hydroxyconiferaldehyde <Emphasis Type="Italic">O</Emphasis>-methyltransferase

The function of an Arabidopsis thaliana gene, At5g54160 annotated as a caffeic acid O-methyltransferase CAOMT gene was characterized. The recombinant enzyme of this gene (AtOMT1) catalyzed the O-methylation of phenylpropanoid and flavonoid substrates. The specificity constants (k _cat/K _m) for 5-hydroxyconiferaldehyde (5-HCAld) and quercetin were both 0.11 μM⁻¹·min⁻¹. On the other hand, lignins of At5g54160-knockout Arabidopsis mutants lacked syringyl units. In addition, we showed that the gene silencing also resulted in significant accumulation of caffeyl alcohol (CaAlc). These results strongly suggested that At5g54160 gene is involved in syringyl lignin synthesis for the methylation of both 5-hydroxyconiferaldehyde and 3,4-dihydroxyphenyl compound(s). Part of this work was presented at the Annual Meeting of Japan Society for Bioscience, Biotechnology, and Agrochemistry, March 24–27, 2007     

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

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

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

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

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

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

Constitutive expression of sense &; antisense <Emphasis Type="Italic">PtAP3</Emphasis>, an <Emphasis Type="Italic">AP3</Emphasis> homologue gene of <Emphasis Type="Italic">Populus tomentosa</Emphasis>, affects growth and flowering time in transgenic tobacco

To analyze the function of PtAP3, an APETALA3 （AP3） homologue gene isolated from Populus tomentosa Carr., the full length sequence （1797 bp） and a fragment （870 bp） of PtAP3 were fused to a CaMV 35S promoter of pBI121 to generate the sense and antisense constructs of PtAP3. These constructs were transformed into tobacco by Agrobacterium infection of leaf disks and selection on kanamycin medium. Some sense and antisense transgenic tobacco plants were obtained by PCR and Southern blot analysis. Great phenotypic differences in transgenic tobacco plants were observed. Almost all of sense PtAP3 to transgenic tobaccos showed a higher growth rate than those of antisense transformants and a few developed pregnancy earlier than wild type seedlings and antisense transformants under the same conditions.     

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

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

Gas exchange responses of <Emphasis Type="Italic">Eucalyptus</Emphasis>, <Emphasis Type="Italic">C. africana</Emphasis> and <Emphasis Type="Italic">G. robusta</Emphasis> to varying soil moisture content in semi-arid (Thika) Kenya

A dramatic decline in forest cover in eastern Africa along with a growing population means that timber and poles for building and fuelwood are in short supply. To overcome this shortage, the region is increasingly turning to eucalyptus. But eucalyptus raises environmental concerns of its own. Fears that it will deplete water supply, affect wildlife and reduce associated crop yields have caused many countries in the region to discourage farmers from planting this exotic. This paper is part of a series of investigations on the growth and water use efficiency of faster growing eucalyptus hybrids, which was introduced from South Africa to Kenya. The hypothesis is that the new hybrids are more efficient in using water and more suitable for the semi-arid tropics than existing eucalyptus and two popular agroforestry species. Gas exchange characteristics of juvenile Eucalyptus grandis (W. Hill ex Maiden), two eucalyptus hybrids (E. grandis × Eucalyptus camaldulensis Dehnh.), Grevillea robusta (A. Cunn) and Cordia africana (Lam) was studied under field and pot conditions using an infrared gas analyzer was used to measure photosynthetic active radiation (PAR), net photosynthetic rate (A), stomatal conductance (g _s) and transpiration rate (E) at CO₂ concentrations of 360 μmol mol⁻¹ and ambient humidity and temperature. A, E and g _s varied between species, being highest in eucalyptus hybrid GC 15 (24.6 μmol m⁻² s⁻¹) compared to eucalyptus hybrid GC 584 (21.0 μmol m⁻² s⁻¹), E. grandis (19.2 μmol m⁻² s⁻¹), C. africana (17.7 μmol m⁻² s⁻¹) and G. robusta (11.1 μmol m⁻² s⁻¹). C. africana exhibited high E values (7.0 mmol m⁻² s⁻¹) at optimal soil moisture contents than G. robusta (3.9 mmol m⁻² s⁻¹) and eucalyptus (5.3 mmol m⁻² s⁻¹) in field experiment and G. robusta (3.2 mmol m⁻² s⁻¹) and eucalyptus (4.2 mmol m⁻² s⁻¹) in pot-grown trees. At very low soil moisture content, extremely small g _s values were recorded in GC 15 and E. grandis (8 mmol m⁻² s⁻¹) and G. robusta (14 mmol m⁻² s⁻¹) compared to GC 584 (46.9 mmol m⁻² s⁻¹) and C. africana (90.0 mmol m⁻² s⁻¹) indicating strong stomatal control by the species. Instantaneous water use efficiency ranged between 3 and 5 μmol mmol⁻¹ and generally decreased with decline in soil moisture in pot-grown trees but increased with declining soil moisture in field-grown trees.     

Comparative efficacy of NeemAzal and local botanicals derived from <Emphasis Type="Italic">Azadirachta indica</Emphasis> and <Emphasis Type="Italic">Plectranthus glandulosus</Emphasis> against <Emphasis Type="Italic">Sitophilus zeamais</Emphasis> on maize

The efficacy of NeemAzal powder, two local neem (Azadirachta indica ) products [neem seed powder (NSP) and neem seed oil (NSO)], as well as a local Lamiaceae, Plectranthus glandulosus leaf powder, applied at four different rates for the control of Sitophilus zeamais was determined. Mortality was recorded 1, 3, 7 and 14 days after S. zeamais infestation, followed by the determination of F₁ progeny production. Grain damage, population increase and grain germination were assessed for treated grains that were stored for 4 months. Grains treated with P. glandulosus powder and NSP had relatively low mortality (5.0–22.5%) after 3 days whereas NeemAzal and NSO had higher mortality (55.0–98.8%). Maximum mortality of 99, 100, 96 and 74% were achieved for NeemAzal (12 g/kg after 14 days), NSO (4 ml/kg after 7 days), NSP (40 g/kg after 14 days) and P. glandulosus powder (40 g/kg, after 14 days), respectively. In the same order, 7-day LC₅₀ values were 0.02 g/kg, 1.46 ml/kg, 12.44 g/kg and 28.9 g/kg. The three neem products greatly reduced progeny emergence, while P. glandulosus powder was less effective. NeemAzal protected the grains against S. zeamais damage better than the local neem products, which in turn provided far better grain protection against the weevil damage than P. glandulosus powder. NeemAzal was superior to the local neem products in grain protection against germination loss, with P. glandulosus being the least effective. NeemAzal and NSO had sufficient efficacy to be a component of an integrated management package for S. zeamais.     

High level expression of glucose-6-phosphate dehydrogenase gene <Emphasis Type="Italic">PsG6PDH</Emphasis> from <Emphasis Type="Italic">Populus suaveolens</Emphasis> in <Emphasis Type="Italic">E. coli</Emphasis>

In order to investigate the functions of the gene PsG6PDH and the mechanisms underlying freezing tolerance of Populus suaveolens, the recombinant expression vector pET-G (pET30a-G6PDH), which contained full encoding region of PsG6PDH gene, was established. The recombinant was identified by lawn-PCR and double enzyme digestion and then transformed into expression host XA90 and induced by isopropyl-â-D-thiogalactoside (IPTG) to express 100 kD polypeptide of G6PDH fusion protein. The results showed that the expressed amount of the fusion protein culminated after 1 mmol&#8226;L^–1 IPTG treatment for 4 h and that pET-G product was predominately soluble and not extra-cellular secreting.     

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

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

Effectiveness of Neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) insecticides against Brassica pod midge (<Emphasis Type="Italic">Dasineura brassicae</Emphasis> Winn<Emphasis Type="Italic">.</Emphasis>)

The efficiency of the botanical insecticide (BI) NeemAzal T/S (containing 1% Azadirachtin A), on the basis of azadirachtin applied in a dose of 20 g a.i. ha⁻¹ against Brassica pod midge (Dasineura brassicae), has been monitored for 4 years. The biological efficiency of BI was compared with the efficiency of some synthetic insecticides. It was ascertained that BI was very efficient in decreasing the number of damaged oilseed rape pods (ranging from 56.5 to 85.9% compared to untreated plants) and its efficiency was comparable with synthetic insecticides based on Chloronicotinyl (Thiacloprid) and Neonicotinoid (Acetamiprid). BI’s efficiency was, in some years, even significantly higher compared to pyrethroid (λ-cyhalothrin). The high biological efficiency of azadirachtin that we ascertained was significantly reflected in increasing potential crop yields. The yield increase of azadirachtin ranged between 9.3 and 19.4% compared to the control sample. Azadirachtin showed the highest yield for the whole time of experimentation, and in some years the yield increase was significantly higher compared to some synthetic agents.     

Electroantennogram responses of <Emphasis Type="Italic">Adosomus</Emphasis> sp. to volatile components from <Emphasis Type="Italic">Artemisia ordosica</Emphasis>

Adosomus sp.eats the leaves and oviposits on the stems of Artemisia ordosica Krasch.Based on an analytical PTI-GC/MS method, 21 volatile organic compounds (VOCs) from the leaves of A.ordosica were identified.The major operating equipment con-sisted of a DB-5 column (30 m×0.32 mm×0.25 μm) and a flame ionization detector (FID).The amounts of the components from VOCs were determined by a peak area normalization method and 17 VOC components were identified by PTI-GC/MS.The elec-troantennogram (EAG) responses of...     

Antifeedant and toxic effects of acetogenins from <Emphasis Type="Italic">Annona montana</Emphasis> on <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis>

Several species of the large family of tropical plants Annonaceae have been intensely investigated over the last 20 years, mainly because of the discovery of annonaceous acetogenins. These compounds are powerful cytotoxics, with potential applications as insecticides, antiparasitics, acaricides, fungicides, and antitumor drugs. Annona montana Macfad. (Annonaceae) grows in Santa Cruz de la Sierra, Bolivia, where an infusion of leaves is used for the treatment of lice, influenza, and insomnia. The major acetogenins from a Bolivian collection of A. montana, annonacin (1), cis-annonacin-10-one (2), densicomacin-1 (3), gigantetronenin (4), murihexocin-B (5), and tucupentol (6), were evaluated for their antifeedant and toxic effects on Spodoptera frugiperda Smith (Lepidoptera: Noctuidae), a serious pest affecting corn crops in Argentina. All the acetogenins produced 100% mortality during the larval or pupal stages at 100 μg of treatment per gram of diet. In addition, compounds 2, 3, and 4 deterred more than 80% feeding at the same dose. Relative toxicity values of LD₅₀ for the strongest larvicidal compounds 1, 2, and 4 were determined, indicating that the three compounds are effective natural insecticides. This is the first report on the antifeedant and toxic effects produced by the particular type of acetogenins, the mono-THF acetogenins, on the lepidopteran S. frugiperda. No correlation was detected between the toxicity of the mentioned compounds to larvae and the known capacity of the acetogenins 1, 2, and 4 to inhibit the NADH oxidase, indicating that the inhibition of the mitochondrial complex I is not the only cause for larval mortality of S. frugiperda.     

<Emphasis Type="Italic">In vitro</Emphasis> morphogenic response of leaf sheath of <Emphasis Type="Italic">Phyllostachys bambusoides</Emphasis>

Nodal segments from secondary branches of saplings of Phyllostachys bambusoides were inoculated in MS medium to assess the in vitro morphogenic response of leaf sheath through the induction to callogenesis by Picloram(4-amino-3,5,6-trichloropicolinic acid) at different concentrations of carbohydrate under the same conditions with presence or absence of luminosity.In our experiment,secondary explants were kept in MS medium containing 8.0 mg·L-1 of Picloram for the callus formation.Calluses were transferred in MS medium supplemented with sucrose,fructose and glucose(control,2%,4% and 6%).Results show that Picloram induced the callogenesis in leaf sheath.The secondary embryogenesis was formed in yellow-globular callus.The sucrose as carbohydrate source in the absence of light was more efficient to induce rhizogenesis.Glucose was more efficiency in the presence of light.Callogenic induction and further embryogenesis evidenced the competence and determination of leaf sheath cells.     

Morphological characteristics of <Emphasis Type="Italic">Quercus</Emphasis><Emphasis Type="Italic">variabilis</Emphasis> charcoal prepared at different temperatures

The morphology of charcoals was investigated by scanning electron microscopy. Charcoal of Quercus variabilis was prepared in an electric furnace under nitrogen gas atmosphere at 400, 600, 800 and 1,000°C. Charring temperature greatly affects the structure of charcoal. In charcoal prepared at 400°C, most of the morphological characteristics remained relatively unchanged with the exception of the cell-wall layering. The cell walls appeared homogenous and glass-like. Above 400°C, there was an increase in cell-wall thinning and volumetric shrinkage with an increase in the charring temperature. These two factors were responsible for most of the observed changes in structure. Fracture surfaces became increasingly rough and disrupted. Vessel elements were increasingly distorted and tyloses disintegrated with increases in temperature. Parenchyma cells exhibited greater shrinkage except at the pits. This resulted in the appearance of distinctive small protuberances over the surface of the parenchyma cells. Rhomboidal calcium crystals were found to be abundant at all the temperatures studied, but at 800 and 1,000°C the crystals had a sponge-like appearance.     

<Emphasis Type="Italic">Cinderella</Emphasis> agroforestry systems

The underlying principles and traditional knowledge concerning indigenous agroforestry systems (AFS) around the world have been used successfully in the design of improved systems; but lately there has been a slow-down in that effort. Recognizing that Cinderella, by analogy, refers to an individual whose attributes were unrecognized or one who unexpectedly achieved success after a period of obscurity and neglect, we propose Cinderella agroforestry systems for such underexploited and “forgotten” AFS. In order to highlight their unrecognized potentials, a few such systems that are location-specific and little-known outside their areas of existence were selected from around the world. Each selected system is unique in terms of its production, environmental, and sociocultural attributes; but none had been described in quantitative terms of ecology and production. A qualitative SWOT (strengths–weaknesses–opportunities–threats) analysis of the systems showed several commonalities among them. While sustainability, multi-functionality, and high sociocultural values are common strengths, low levels of production and lack of research- and technology inputs to improve the system are the major weaknesses. The opportunities emanating from strengths and weaknesses are also common to all the systems; and “threats” to these systems arise mostly from ramifications of government policies. Given the acknowledged role of AFS in climate-smart agriculture and the current global momentum for working with communities to better manage landscapes, learning from success stories of the Cinderella AFS could suggest models for sustainable development, and efforts in that direction should also be incorporated into global agroforestry research agendas.     

Heartwood extractives from the Amazonian trees <Emphasis Type="Italic">Dipteryx odorata,Hymenaea courbaril</Emphasis>, and <Emphasis Type="Italic">Astronium lecointei</Emphasis> and their antioxidant activities

Heartwood extracts from Amazonian trees cumaru-ferro (Dipteryx odorata), jatoba (Hymenaea courbaril), and guarita (Astronium lecointei) exhibit antioxidant activities comparable with that of α-tocopherol, a well-known antioxidant. This article reports the characterization of the antioxidant compounds in the extracts of the three heartwoods. Silica gel column chromatography of the cumaru-ferro EtOAc extract yielded (−)-(3R)-7,2′,3′-trihydroxy-4′-methoxyisoflavan and (+)-(3R)-8,2′,3′-trihydroxy-7,4′-dimethoxyisoflavan. Silica gel column chromatography followed by preparative high-performance liquid chromatography of the jatoba EtOAc extract yielded (−)-fisetinidol and (+)-trans-taxifolin. Chemical structures were assigned using electron-ionization mass spectrometry, ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy including nuclear Overhauser effect spectroscopy (NOESY), as well as optical rotation and circular dichroism. Gas chromatography-mass spectrometry demonstrated that the isolated compounds were predominant in the EtOAc extracts. In the guarita EtOAc extract, catechin and gallic acid were identified by comparing their retention times and mass fragmentation patterns with those of authentic samples. Antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl assay demonstrated that all these compounds had activities comparable with that of α-tocopherol. Part of this report was presented at the 57th Annual Meeting of the Japan Wood Research Society, Hiroshima, August 2007