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.     

Cloning and Sequence Analysis of a Glucose-6-Phosphate Dehydrogenase Gene PsG6PDH from Freezing-tolerant Populus suaveolens

A 1 207 bp cDNA fragment (PsG6PDH) was amplified by RT-PCR from cold-induced total RNA of the freez- ing-tolerant P. Suaveolens, using primers based on the highly conserved region of published plant glucose-6-phosphate dehydro- genase (G6PDH) genes. The sequence analysis showed that PsG6PDH coding region had 1 101 bp and encoded 367 predicted amino acid residues. Moreover, the nucleotide sequence of PsG6PDH showed 83%, 82%, 79%, 79% and 78% identity, and the derived amino acid sequence shared 44.2%, 44.7%, 42.0%, 40.5% and 43.9% identity with those of the Solanum tuberosum, Nicotiana ta- bacum, Triticum aestivum, Oryza sativa and Arabidopsis thaliana, respectively. The results show that PsG6PDH is a new member of G6PDH gene family and belongs to the cytosolic G6PDH gene. This is the first report on cloning of the G6PDH gene from woody plants.     

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.     

High Level Expression of Glucose-6-phosphate Dehydrogenase Gene PsG6PDH from Populus suaveolens in E. coli

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-a-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·L–1 IPTG treatment for 4 h and that pET-G product was predominately soluble and not extra-cellular secreting.     

Stereochemistry of matairesinol formation by <Emphasis Type="Italic">Daphne</Emphasis> secoisolariciresinol dehydrogenase

Secoisolariciresinol dehydrogenase activity was detected for the first time from Daphne odora and Daphne genkwa (Thymelaeaceae), which are known to produce optically pure (+)-matairesinol. In sharp contrast, (–)-matairesinol was formed selectively over the (+)-antipode by the secoisolariciresinol dehydrogenase preparation from both D. odora callus and D. genkwa shoots.     

The influence of Desert False Indigo, <Emphasis Type="Italic">Amorpha fruticosa</Emphasis>, and three arbuscular mycorrhizae fungi on the growth of <Emphasis Type="Italic">Populus ussuriensis</Emphasis> seedlings

The influence of Desert False Indigo, Amorpha fruticosa, on the growth of Populus ussuriensis seedlings inoculated with three arbuscular mycorrhizae (AM) fungi (Glomus mosseae, Glomus intraradices, Glomus sinuosa) was studied using the nylon net method. The results showed that all three AM fungi infected P. ussuriensis seedlings and G. intraradices and also G. mosseae infected A. fruticosa. The AM fungi promoted growth of P. ussuriensis and Desert False indigo seedlings. Moreover, under co-cultivation with A. fruticosa, the biomass of P. ussuriensis increased significantly. The concentration of nitrogen in P. ussuriensis grown with A. fruticosa and the concentration of soluble nitrogen in the rhizosphere were also higher than when grown alone. Hypha were found on the two plant seedlings inoculated with G. mosseae and G. intraradices, suggesting that AM fungi may transport nutrients from seedlings of A. fruticosa to the rhizosphere of P. ussuriensis seedlings, which may have promoted the growth of P. ussuriensis. The AM fungi played a critical role on the effect of A. fruticosa on growth of P. ussuriensis.     

Establishment of cell suspension line of <Emphasis Type="Italic">Populus tomentosa</Emphasis> Carr.

Leaves of fine Populus tomentosa genotype TC152 were used as explants to establish cell suspension lines. The effects of plant growth regulators on callus induction and establishment of cell suspension lines were studied. The callus induction rate was the highest on a MS solid medium supplemented with 1.0 mg·L^-1 2,4-D. A cell suspension line could be obtained by inoculating calli which were not subcultured into a MS liquid medium supplemented with 1.5 mg·L^-1 2,4-D. The best subculture medium was MS ＋ 0.8 mg＇L-1 2,4-D ＋ 30 g·L^-1 sucrose with a subculture cycle of seven days.     

Impact of addition of soil amendments and microbial inoculants on nursery growth of <Emphasis Type="Italic">Populus deltoides</Emphasis> and <Emphasis Type="Italic">Toona ciliata</Emphasis>

Present study evaluated growth of Populus deltoides G48 and Toona ciliata over a period of 6 months, in nursery soil amended with 10% fly ash (v/v), 5% distillery waste (v/v), 20% farmyard manure (v/v) and microbial consortium of Pseudomonas striata and Azotobacter sp. @ 30 ml/pot in different combinations leading to 12 different treatments with 16 replicates in completely randomized block design. Biometric parameters such as plant height, collar diameter and total dry biomass were analyzed which indicated that the treatment (T8) comprising of fly ash @ 10% (v/v), farmyard manure @ 20% (v/v) and microbial consortium @ 30 ml/pot promoted growth of P. deltoides. The results indicated that combined addition of fly ash, farm yard manure and microbial inoculants can be used as a good potting mixture for improving survival rates and plant growth in forestry nurseries.     

Comparison and early selection of new clones in <Emphasis Type="Italic">Populus tomentosa</Emphasis>

In our study, two experimental plantations, respectively, with 24 and 32 new clones of P tomentosa, were established in Weixian County, Hebei Province and Wuzhi County, Henan Province using a completely randomized block design. A comparative study was conducted on the continuous 5-year-old height and diameter at breast height （DBH） of new clones in the two plantations. As well, based on genetic correlation over the years of testing of these clones, a preliminary study of early selection was carried out. Results indicate that the growth traits of the new clones in Weixian were better than those in Wuzhi. The traits show weak correlation between the two plantations. In some stands, the height, DBH and seedling volume of 5-year-old clones presented statistically significant differences among clones. In both plantations, the new clones showed over 0.6 repeatability of height, DBH and volume, as well as larger coefficients of variation （CV）. The fact that these clones achieved the largest repeatability and CV in the second year suggests that these traits are highly controlled by heredity. Thus, based on the growth traits of the second year, the new clones B305, B307, B303, H75, BT18, BT17 and 21J-1 were considered suitable in Weixian. In Wuzhi, the new clones had variable repeatability and CVs in various years and their correlation of growth traits among different years was not high. We conclude that early selection of new clones was not feasible in Wuzhi.     

Structural characteristics and eco-adaptability of heteromorphic leaves of <Emphasis Type="Italic">Populus euphratica</Emphasis>

The microstructural and ultrastructural traits of three kinds of typical leaves ofPopulus euphratica Olive, including lanceolate, broad-ovate and dentate broad-ovate leaves, were studied by using electron microscope and optical microscope. The resuits showed that with the leaves changing from lanceolate shape to dentate broad-ovate shape, their structure obviously tended to be xeromorph: developed palisade tissue, undeveloped spongy tissue, thick cutin layer and sunken stomas. The amount of mitochondria tended to be increased, and the shape of chloroplasts varied from regular spindle to irregular rotundity or oval. The leaves were covered with wax without cilium, and the stomas on the upper and lower epidermis of the leaves opened unevenly. The stomas on the lower epidermis were deeper than those on the upper epidermis under the scanning electron microscope. The results implied that the structural characteristics of the diversiform-leaves ofP euphratica are related to its eco-adaptability.     

<Emphasis Type="Italic">Bursaphelenchus mucronatus</Emphasis> (Nematoda: Aphelenchoididae) vectored by <Emphasis Type="Italic">Monochamus </Emphasis><Emphasis Type="Italic">urussovi</Emphasis> (Coleoptera: Cerambycidae) in Hokkaido,Japan

Bursaphelenchus mucronatus Mamiya et Enda has been recovered for the first time from adults of the cerambycid beetle, Monochamus urussovi (Fischer), in Hokkaido, Japan. The nematode was also recovered from the inner bark of Picea jezoensis (Siebold et Zuccarini) Carrière and Abies sachalinensis (Fr Schmidt) Masters infested with M. urussovi larvae. PCR–RFLP analysis indicated that B. mucronatus in Hokkaido is the European type.     

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     

<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

Characterization and role of glucose-6-phosphate dehydrogenase of Populus suaveolens in induction of freezing resistance

Glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) was purified from the leaves of 8-week-old Populus suaveolens cuttings. The enzyme activity in the absence and presence of reduced dithiothreitol (DTT_red) was determined. The results show that the G6PDH activity is not inactivated by pre-incubation with DTT_red, indicating that the purified enzyme probably presented in cytosol of P. suaveolens. The catalytic characteristics and kinetic parameters of cytosolic G6PDH purified from P. suaveolens cuttings were also studied. The results show that G6PDH is characterized by K _m value of 360 μmol·L⁻¹ for G6P and 16 μmol·L⁻¹ for NADP, a pH range of 7.3-8.9, and the maximum activity around pH 8.2. The enzyme activity is inhibited by various metabolites such as NADPH, NADH, GTP, UTP, ATP, AMP, ADP, CoA, acetyl CoA, fructose-6-phosphate (F6P), erythrose-4-phosphate (E4P), ribose-5-phosphate (R5P) and 3-phosphoglycerate (3-PG) (all at 1 mmol·L⁻¹ except for NADPH and NADH) to different extents. NADPH is the most effective inhibitor of enzyme activity, with an inhibition of 72.0%. The addition of metal ions such as MgCl₂, CaCl₂ and KCl (all 1.0 mmol·L⁻¹) to the standard reaction mixture has no remarkable influence on the cytosolic G6PDH activity. However, CdCl₂ (1.0 mmol·L⁻¹) causes high inhibitory effect on the enzyme activity. To explore the role of G6PDH on the enhancement of freezing resistance induced by freezing acclimation, the changes in the cytosolic G6PDH activity and freezing resistance (expressed as LT₅₀) of P. suaveolens cuttings during freezing acclimation at −20 °C were investigated. The results reveal that freezing acclimation decreases LT₅₀ of cuttings, and increases the activity of cytosolic G6PDH compared with control ones, while 2 d of de-acclimation at 25 °C result in a decrease in cytosolic G6PDH activity, and caused an increase in LT₅₀. Furthermore, the change in cytosolic G6PDH activity is found to be closely correlated to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that cytosolic G6PDH may be involved in the induction of freezing resistance of cuttings. [Supported by the Foundation of State-designated Base for Biology Researching and Teaching in Beijing Forestry University]     

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     

Model of leaf energy distribution and its experimental validation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> Carr.

Leaf temperature of a plant is the result of heat transfer between the plant and its environment. There are many factors that can affect leaf temperature, such as the solar radiation energy, environmental temperature, wind velocity, evaporation on the leaf surface, photosynthesis, respiration and so on, which have different effects on the temperature of leaves. In first instance, we analyzed the heat transfer on leaves of Populus tomentosa Carr. theoretically and constructed a model of energy distribution. We then validated the model by analyzing seven different kinds of one-year-old P. tomentosa leaves experimentally. The result shows that solar radiation is the main energy input and the dominant ways of thermal diffusion are heat transfer between the upper and lower leaf surfaces and evaporation from the leaf surface.     

Laboratory and field evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for controlling Mulberry whitefly <Emphasis Type="Italic">Pealius mori</Emphasis> Takahashi (Homoptera: Aleyrodidae) in mulberry (<Emphasis Type="Italic">Morus alba</Emphasis> Linn)

In a laboratory screening of 12 isolates of entomopathogenic fungi against nymphs of the mulberry whitefly (Pealius mori Takahashi), Beauveria bassiana (Balsamo-Crivelli) Vuillemin CKB-048 was the most virulent, causing 87 ± 3% mortality at 1 × 10⁶ conidia/ml. Infection was confirmed by growth of the fungus from cadavers and by scanning electron microscopy of treated nymphs. Beauveria bassiana CKB-048 was formulated as a wettable powder (1 × 10⁹ conidia/g) and tested in two mulberry (Morus alba Linn) plantations in central and northeastern Thailand. In both locations, two spray applications of B. bassiana CKB-048 at 3.75 × 10¹² to 6.25 × 10¹² conidia/ha and at 14 day intervals provided good control of whitefly nymphs; control with B. bassiana CKB-048 was comparable to that with the pesticide buprofezin at 250 g of active ingredient/ha. In addition, no mortality of silkworm larvae occurred when the larvae were fed with mulberry leaves sprayed with B. bassiana CKB-048 7, 14, or 21 days earlier.     

Aphidicidal activity of some indigenous plant extracts against bean aphid <Emphasis Type="Italic">Aphis </Emphasis><Emphasis Type="Italic">craccivora</Emphasis> Koch (Homoptera: Aphididae)

Aphidicidal activity of hot and cold water extracts of some indigenous plants, Azadirachta indica A. Juss (neem), Calotropis procera (Aiton) W.T. Aiton (akanda), Polygonum hydropiper L. (biskatali) and Ipomoea sepiaria J. Koenig ex Roxb. (bankalmi), were tested against the bean aphid, Aphis craccivora Koch. Hot water extract of P. hydropiper and A. indica was found to be the most effective (87.6–94.5 and 80.47–89.6% mortality respectively, P < 0.01) among all the extracts. Other hot and cold water extracts also appeared to be useful (59.5–87.5% mortality) as pesticides for A. craccivora. The highest yield (3.25 kg per plant) was obtained using hot water extract of P. hydropiper followed by hot water extract of A. indica (3.15 kg per plant). The lowest yield (0.32 kg per plant) was recorded from the control block. All the phytoproduct treatments had significantly (P < 0.01) better yield than the control block.     

Biological performance of <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> leaf oils from Thailand against the subterranean termite <Emphasis Type="Italic">Coptotermes formosanus</Emphasis> Shiraki

The antitermitic activities of leaf oils and their constituents, taken from three clones of Eucalyptus camaldulensis Dehnh. in Thailand, against Coptotermes formosanus Shiraki were investigated in contact and noncontact tests. The termiticidal mechanism was also examined. Antitermitic tests demonstrated that E. camaldulensis leaf oils were both contact toxicants and fumigants to C. formosanus with LC₅₀ values ranging between 12.68 and 17.50 mg/g by the contact method, and between 12.65 and 17.50 mg/petri dish (100 cm³) by the noncontact method. p-Cymene and γ-terpinene were primarily responsible for the contact toxicity and 1,8-cineole was responsible for fumigation. From the investigation of termiticidal mechanism, E. camaldulensis leaf oils exhibited the inhibition of acetylcholinesterase activity and showed the common symptoms of a neurotoxic mode of action against C. formosanus. Part of this report was presented at the 58th Annual Meeting of the Japan Wood Research Society, Tsukuba, March 2008     

Recessive inheritance of morphological characteristics of utsukushimatsu,<Emphasis Type="Italic">Pinus densiflora</Emphasis> f.<Emphasis Type="Italic">umbraculifera</Emphasis>

Pinus densiflora f.umbraculifera, commonly known as utsukushimatsu, is a distinctively shaped form of Japanese red pine whose growth is restricted to a forest stand in Shiga Prefecture, Japan. The inheritance mode of morphological characteristics of utsukushimatsu was studied to preserve the genetic resource of this pine. As previously reported, F₁ trees grown from open-pollinated seeds harvested from trees inhabiting the native stand showed two phenotypes: one resembling utsukushimatsu, which produces multiple trunks, and the other resembling normalP. densiflora, which produces one or a few trunks. In the present study, controlled pollination was carried out using F₁ and normalP. densiflora trees. Segregation ratios of the two phenotypes observed in the F₂ population showed that the morphological characteristics of utsukushimatsu are inherited recessively. This suggests that the mutation of one gene or a few closely linked genes controls the morphological characteristics of utsukushimatsu. Since multiple trunk formation of utsukushimatsu might be related to a loss of lateral bud inhibition, it follows that a simple gene mutation breaks apical dominance inP. densiflora.