Genetic diversity and population structure analysis of <Emphasis Type="Italic">Pistacia</Emphasis> species revealed by <Emphasis Type="Italic">phenylalanine ammonia</Emphasis>-<Emphasis Type="Italic">lyase</Emphasis> gene markers and implications for conservation

Information on population genetic structure and crop genetic diversity is important for genetically improving crop species and conserving threatened species. The PAL gene sequence is part of a multigene family that can be utilized to design DNA marker systems for genetic diversity and population structure investigation. In the current study, genetic diversity and population structure of 100 accessions of wild Pistacia species were investigated with 78 PAL markers. A protocol for using PAL sequences as DNA markers was developed. A total of 313 PAL loci were recognized, showing 100% polymorphism for PAL markers. The PAL markers produced relatively more observed and effective alleles in Pistacia falcata and Pistacia atlantica, with a higher Shannon’s information index and expected heterozygosity in P. atlantica, Pistacia vera and Pistacia mutica. Pairwise assessment of Nei’s genetic distance and genetic identity between populations revealed a close association between geographically isolated populations of Pistacia khinjuk and Pistacia chinensis. The accessions of wild Pistacia species had more genetic relationship among studied groups of species. Analysis of molecular variance indicated 19% among-population variation and 81% within-population variation for the PAL gene based DNA marker. Population structure analysis based on PAL revealed four groups with high genetic admixture among populations. The results establish PAL markers as a functional DNA marker system and provide important genetic information about accessions from wild populations of Pistacia species.     

Cloning of <Emphasis Type="Italic">XET</Emphasis> gene from <Emphasis Type="Italic">Anthocephalus chinensis</Emphasis> and its plant expression vector construction

A full-length cDNA sequence of xyloglucan endotransglycosylase gene (XET), abundantly expressed in the cambium of Anthocepha-lus chinensis was cloned by conserved PCR, rapid-amplification of cDNA ends and by chromosome walking. Analytical results of the DNA sequence show that a 912 bp complete open reading frame (ORF) encoded a 303-amino acid protein was in the 1205 bp full cDNA sequence. The deduced amino acid sequence of AcXET, which contained the conserved specific EIDFE catalytic site sequence to XETs was homologous to the other known XET proteins. In order to study the gene function of AcXET and obtain transgenic plants, a plant expression vector pBIAcXET was constructed by recombinating the AcXET fragment from the cloning vector pMD19AcXET and the binary vector pBI121 between the XbaI and SmaI sites. The fragment of AcXET gene was inserted between the CaMV 35S promotor and the coding region of the GUS gene in pBI121. The identification results show that the plant expression binary vector pBIAcXET was constructed successfully. These results lay the foundation for studying the molecular mechanism of AcXET gene during wood formation.     

Genetic diversity in two threatened species in Vietnam:Taxus chinensis and Taxus wallichiana

Taxus chinensis and T. wallichiana in have been threatened in their distribution areas in recent decades because of their over-exploitation and reduction and destruction of native habitats. Determining the genetic diversity in populations of the two species will provide guidelines for their protection and preservation. Two hundred and fifteen trees from six populations of T. chinensis and150 sampled trees of T. wallichiana were sampled. Six microsatellite primer pairs selected from 16 primer pairs were used to investigate genetic variation at the population and species levels. Five yielded polymorphic alleles, and among the 13 putative alleles amplified, 11 were polymorphic(accounting for 76.33 %).Shannon's information index(I) and percentage of polymorphic bands(PPB)(I = 0.202 and PPB = 67.22 % for T. chinensis; I = 0.217 and PPB = 65.03 % for T. wallichiana). Both species had low levels of genetic diversity(mean H_o= 0.107, H_e= 0.121 for T. chinensis; H_o= 0.095, H_e= 0.109 for T. wallichiana). Genetic differentiation among populations was higher(FST= 0.189) for T. chinensis and lower(0.156) for T.wallichiana, indicating limited gene flow(Nm) among populations for T. chinensis(0.68) and T. wallichiana(0.65).Variation among individuals of T. chinensis was 63.59 and73.12 % for T. wallichiana. Thus, the threatened status of the two conifers is related to a lack of genetic diversity. All populations are isolated in small forest remnants. An ex situ conservation site should be established with a new population for these species that comprises all the genetic groups for the best chance to improve their fitness under environmental stresses.     

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.     

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.     

Leaf-level responses to light in two co-occurring <Emphasis Type="Italic">Quercus</Emphasis> (<Emphasis Type="Italic">Quercus ilex</Emphasis> and <Emphasis Type="Italic">Quercus suber</Emphasis>): leaf structure,chemical composition and photosynthesis

We studied morphological, biochemical and physiological leaf acclimation to incident Photon-Photosynthetic-Flux-Density (PPFD) in Quercus ilex (holm oak) and Quercus suber (cork oak) at Mediterranean evergreen oak woodlands of southern Portugal. Specific leaf area (SLA) decreased exponentially with increasing PPFD in both species. Q. ilex had lower SLA values than Q. suber. Leaf nitrogen, cellulose and lignin concentration (leaf area-based) scaled positively with PPFD. Maximum rate of carboxylation (V_cmax), capacity for maximum photosynthetic electron transport (J_max), rate of triose-P utilization (V_TPU) and the rate of nonphotorespiratory light respiration (Rd) were also positively correlated with PPFD in both Quercus species, when expressed in leaf area but not on leaf mass basis. Q suber showed to have higher photosynthetic potential (V_cmax, J_max^m and V_TPU^m) and a higher nitrogen efficient nitrogen use than Q.ilex. Leaf chlorophyll concentration increased with decreasing PPFD, improving apparent quantum use efficiency (Φ) in both Quercus species. We concluded that, in Q.ilex and Q.suber, leaf structural plasticity is a stronger determinant for leaf acclimation to PPFD than biochemical and physiological plasticity.     

<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.     

Constituents from the roots of <Emphasis Type="Italic">Taxus cuspidata</Emphasis>

The known propelargonidin, afzelechin-(48)-afzelechin (1), the known lignans 7-hydroxynortrachelogenin (2), epinortrachelogenin (3), nortrachelogenin (4), hydroxymatairesinol (5), allohydroxymatairesinol (6), matairesinol (7), oxomatairesinol (8), and isotaxiresinol (9), and the known taxoids taxinine M (10), taxayuntin (11), and 10-deacetyltaxol (12), and 10-deacetylbaccatin III (13) were isolated from the roots of Taxus cuspidata (Japanese yew, Taxaceae). The propelargonidin was isolated from Taxus spp. for the first time, and was detected in the roots, bark, and twigs.     

Genetic diversity and mating system of <Emphasis Type="Italic">bracatinga</Emphasis> (<Emphasis Type="Italic">Mimosa scabrella</Emphasis>) in a re-emergent agroforestry system in southern Brazil

Bracatinga (Mimosa scabrella) is a legume tree species common in the early stages of succession in Araucaria angustifolia forests in southern Brazil. Bracatinga can form high-density monospecific stands called bracatingais. Its traditional management for charcoal production involves maintenance of the seed bank. Our objective was to analyze the genetic diversity and structure of bracatingais to understand the mechanisms by which intraspecific diversity of M. scabrella is created and maintained in landscapes managed by family farmers in their agroforestry mosaics. We analyzed 14 bracatingais using 8 allozyme loci. We compared parental and progeny generation indices (7 loci) and described the mating system (9 loci) of two progenies. Overall diversity was high: A = 2.69, H _o = 0.257, H _e = 0.382 and similar between populations. Overall fixation (F = 0.364) was similar to the fixation index (f = 0.329). The genetic divergence among populations was low (Θ_p = 0.052) but significant. The progenies’ genetic diversity values were similar to those of the previous generation (H _{e pop11} = 0.342 vs. 0.420/H _{e pop10} = 0.432 vs. 0.400). Progenies were compatible with half-sib and full-sib crossing expectations (θ _xy  = 0.204 and 0.194). Our data showed that there is a tendency for genetic structuring caused not only by the reproductive system but also by genetic drift. It is very likely that the high genetic diversity is amplified by internal migration within each bracatingal. This study showed that current landscape management can contribute to maintaining high levels of bracatinga genetic diversity, which contributes to its regional conservation.     

Structural diversity of English yew (<Emphasis Type="Italic">Taxus baccata</Emphasis> L.) populations

In Europe, the English yew species (Taxus baccata L.) is endangered. Intensive human land-use, including forest management, has caused a decrease of the yew populations all over Europe. In Austria, gene conservation forests are used for the in situ conservation of populations of this rare tree species by silvicultural treatments. In order to improve the conservation management in these gene conservation forests, this study addresses the relation between competition and viability of yew populations through the use of structural diversity indices. The structural indices, which include mingling, tree–tree distance, diameter, and tree height differentiation, were determined for a structural group of four trees as well as the neighbouring trees of the male and female yews at the monitoring plots on a regular grid in three gene conservation forests. Although the three study sites provided quite different environmental conditions for English yew, the vitality of each individual yew was influenced by the inter-specific competition of the neighbouring tree species at all sites. Low vitality was associated with a small mean distance to neighbours and large tree height differentiation. In conclusion, we suggest that a combination of different structural indicators is needed for an integrative assessment of conservation status in the gene conservation forests. This would help improve the evaluation of the impact management has on yew population viability.     

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     

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.     

Genetic diversity analysis in a seed orchard of <Emphasis Type="Italic">Eucalyptus tereticornis</Emphasis>

India is the largest grower of Eucalyptus with an area of 3.943 million hectares under plantations and E. tereticornis is the predominant species in the plains of southern India with an average productivity of 12–25 m³ ha⁻¹ year⁻¹. With the aim to establish seedling seed orchards of the species, seed lots of fifteen provenances were imported from Australia and a trial was laid. In the present study the genetic diversity existing in the seed orchard was estimated using ISSR–PCR. Seven ISSR primers amplified 663 amplicons in the size ranging from 255 to 2,711 bp. The total number of polymorphic bands varied from 59 to 123 with 100% polymorphic banding profiles. The average gene diversity (Hj) of all the provenances ranged from 0.0589 to 0.1109 and the total gene diversity estimated was low (H _T = 0.130) when compared to the earlier reports from other eucalypts species. Analysis of Molecular Variance partitioned the ISSR variation into inter and intra population components. The inter population component accounted for 55.2% of the variation and the intra population component accounted for 46.3% (P < 0.001). A neighbour-joining analysis was done using the dissimilarity matrix to determine the aggregation of the individuals into clusters. Existence of population structure among the populations was revealed in STRUCTURE analysis but geographical region based clustering was not observed. The assessment of intra and inter genetic variation documented in the present study suggests that, along with the phenotypic traits, knowledge about genetic diversity measured at the DNA level in individuals of seed orchards provide an objective guide for selective culling of trees for maintaining optimal diversity for enhanced genetic gains.     

<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.     

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.     

Chemical alterations induced by <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis>/<Emphasis Type="Italic">Aspergillus flavipes</Emphasis> co-cultures in wood from different tree species

Chemical alterations following inoculation of Acacia mearnsii, Eucalyptus dunnii, E. grandis, and E. macarthurii with a Pycnoporus sanguineus/Aspergillus flavipes co-culture were investigated. Several wood chemical parameters were measured using standard methods from the pulp and paper industry. The data were described and analyzed using univariate as well as multivariate statistical techniques. Boxplots and in particular biplots show clearly how the chemical composition of each tree species was differently affected by the co-culture. Lignin content was significantly decreased in A. mearnsii, while E. dunnii showed a decrease in cellulose content. The results, therefore, indicate that the manner in which wood is degraded by a specific fungal co-culture depends on the tree species involved. This phenomenon should be considered when selecting fungi for bio-pulping.     

The behaviour of the species mingling index<Emphasis Type="Italic"> M</Emphasis><Subscript><Emphasis Type="Italic">sp</Emphasis></Subscript> in relation to species dominance and dispersion

Ecosystem conservation plays an increasingly important role in forest management. The value for such conservation has been directly linked to high diversity of species and structure within systems. In the past, forestry inventory practices have concentrated on aspects that were primarily required for timber management. Now, however, much interest has arisen in measures that will indicate higher or lower diversity. The interspersion of trees of different species reflects diversity at a local level and may serve as an important indicator for the simplification of a forest or woodland system. This article shows that the mingling index M_sp is robust in describing the spatial dispersion of plants; some guidelines for interpreting M_sp are provided.     

Introduced <Emphasis Type="Italic">Tuber aestivum</Emphasis> replacing introduced <Emphasis Type="Italic">Tuber melanosporum</Emphasis>: a case study

A Tuber melanosporum plantation established in 1994/1995 on Kibbutz Bar’am (in the Upper Galilee, Israel) gradually lost its T. melanosporum mycorrhiza. In 1999, T. aestivum inoculated seedlings were inadvertently introduced into the plantation to fill the gaps between trees. A single T. melanosporum fruit body was found in 1999. Although no truffles were found after 1999 and until 2009, in that year and in 2010, truffles were collected. Morphological and molecular analyses proved these to be T. aestivum. Thus, the intentionally introduced T. melanosporum mycorrhiza was replaced by that of another introduced mycorrhizal fungus, T. aestivum. Local oak species produced higher yields compared to introduced host species known to be good T. melanosporum plant symbionts. The yield was comparable to that reported for young commercial orchards, but the fruiting season was earlier than in Europe.     

Effects of <Emphasis Type="Italic">Eucalypt</Emphasis> and <Emphasis Type="Italic">Acacia</Emphasis> plantations on soil water in Sumatra

Indonesia’s pulp and paper industry needs a large area of sustainably grown plantations to support its continued development. Acacia mangium has been the key species underpinning the pulp and paper industries in Sumatra, however increased disease pressure on A. mangium is expected to require large-scale conversion of Acacia plantations to Eucalyptus in the near future. The effect of such a large scale change in plantation species on soil moisture, for both tree production, and catchment hydrology is unknown. In this study we sought to characterize the impacts of plantation species (Acacia or Eucalyptus) and nitrogen management, on soil moisture, soil water depletion and depth to groundwater under stands of Acacia mangium and Eucalyptus pellita over the first 2–3 years after establishment. The study was conducted in experiments at four sites in Sumatra, Indonesia. Soil moisture and soil water depletion were not influenced by plantation species or fertilizer treatment. Soil moisture content and soil water depletion were strongly influenced by shallow groundwater at two of the four sites, however depth to groundwater did not influence stem growth. Results from the field trials cautiously suggest that large scale conversion of Acacia mangium to Eucalypt species in these regions is unlikely to result in increased moisture stress, nor is conversion of plantation species likely to lead to substantial differences in catchment hydrology. This study demonstrated the importance of conducting multi-site studies when investigating biophysical relationships in forest/plantation systems.