DNA polymorphisms and genetic relationship among populations of <Emphasis Type="Italic">Acacia leucophloea</Emphasis> using RAPD markers

RAPD (randomly amplified polymorphic DNA) markers were employed to characterize polymorphisms among 5 provenances of Acacia leucophloea and to detect genetic relatedness of the species with 6 other acacias (A. holosericea, A. auriculiformis, A. mangium, A. dealbata, A. ferruginea, and A. nilotica) widely grown in India. Of 194 markers scored for the provenances, 29.38% exhibited polymorphism. Also, 326 markers were generated among 7 species of Acacia, accounting for 55.82% of the polymorphisms. The fifteen 10-mer primers employed were capable of producing 1–8 polymorphic bands for the provenances, and 6–17 for all seven species of Acacia. The genetic similarity coefficient based on Jaccard’s coefficient revealed that provenances Thirumangalam and Dharmapuri were closely related. The dendrogram based on a sequential agglomerative hierarchical non-overlapping (SAHN) clustering analysis grouped 4 provenances of A. leucophloea (Dharapuram, Thirumangalam, Pudukottai and Dharmapuri) into one cluster and the other provenance, Sendurai, into a separate cluster. The genetic similarity matrix for 7 Acacia species showed that A. nilotica and A. dealbata were distantly related, while A. holosericea and A. ferruginea were very closely related. Cluster analysis grouped the species of Acacias into 3 major groups of which A. dealbata alone formed a separate group. The RAPD markers generated 36 provenance-specific markers and 162 species-specific markers that could have strong applications for species identification and tree breeding programs for A. leucophloea and for other Acacia species included in this study.     

Low within-population genetic diversity and high genetic differentiation among populations of the endangered plant <Emphasis Type="Italic">Tetracentron sinense</Emphasis> Oliver revealed by inter-simple sequence repeat analysis

Key message

Tetracentron sinense Oliver, an endangered species from China, displays a low within-population genetic diversity and high genetic differentiation among populations, and the existing populations could be divided into three conservation and management units.

Context

The endangered tree Tetracentron sinense Oliver has great value; however, little is known regarding the within-population genetic diversity and differentiation among T. sinense populations.

Aims

We examined the genetic diversity and differentiation of T. sinense wild populations, and we tested the effect of small-size population on the level of genetic diversity within these populations.

Methods

Using inter-simple sequence repeat (ISSR), we assessed the genetic variation and structure among 174 individuals from 26 natural populations of T. sinense sampled across its distribution range in China.

Results

The ISSR primers yielded 180 amplified loci (123 were polymorphic). At the species level, the percentage of polymorphic loci (PPL), Nei’s gene diversity (H), and Shannon’s information index (I) were 68.3%, 0.196 and 0.300, respectively. The average population level PPL was 20.0%, and the Na, Ne, H, and I were 1.20, 1.13, 0.076, and 0.112, respectively. AMOVA revealed high genetic differentiation among populations (52.0% of total variance, P?=?0.001), consistent with the gene differentiation coefficient (Gst?=?0.607) and gene flow (Nm?=?0.326). The 174 individuals of the 26 T. sinense populations clustered into three groups, and T. sinense geographic and genetic distance were significantly correlated.

Conclusions

T. sinense exhibited intermediate within-species genetic diversity, indicating preserved evolutionary potential. The low within-population genetic diversity and high genetic differentiation among T. sinense populations may be one of important factors causing endangerment. Three conservation units were determined based on genetic difference and structure. Inter-population introduction of individuals within units via appropriate propagation and seedling management might be an effective strategy for increasing T. sinense within-population genetic diversity and population size.

     

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.     

Genetic effects of historical anthropogenic disturbance on a long-lived endangered tropical tree <Emphasis Type="Italic">Vatica mangachapoi</Emphasis>

The endangered Vatica mangachapoi, a long-lived, tropical tree with economic and ecological importance found in Hainan, China, was used to assess the hypothesis that historical human activities in Hainan’s tropical rain forest could have negative effects on the genetic diversity of V. mangachapoi. Three hundred and twenty individuals from 11 natural populations—which were classified into three groups according to levels of disturbance—were sampled and analyzed with ISSR markers. Although genetic diversity of V. mangachapoi is high at the species level, it is relatively low within populations. A significant genetic differentiation occurs among different disturbance levels. Significant isolation-by-distance indicated relevant historical anthropogenic changes. Our findings showed that historical human disturbances significantly increase the genetic differentiation and slightly decrease the genetic diversity of long-lived tree V. mangachapoi. Relevant targeting conservation actions were recommended.     

The effects of non-host plant extracts on electroantennogram responses,behavior and egg hatching of codling moth, <Emphasis Type="Italic">Cydia pomonella</Emphasis>

The codling moth (Cydia pomonella L.) is a significant pest of pome fruit throughout the world. Behavioral and ovicidal activities of five non-host plant extracts (Arctium lappa, Bifora radians, Humulus lupulus, Verbascum songaricum, Xanthium strumarium), synthetic sex pheromone, (E,E)-8,10-dodecadienol (codlemone), and the plant volatile lure, (2E,4Z)-2,4-decadienoate (pear ester) were evaluated against the codling moth, C. pomonella L. Codlemone elicited the greatest electroantennogram (EAG) response (6.2 ± 1.2 mV) of the compounds tested from male C. pomonella while pear ester elicited 1.7 ± 0.1 mV EAG response in female moths. Codlemone attracted 34.5% of male C. pomonella in olfactometer studies, and it was followed by the X. strumarium extract with 24.8%. There was a significant difference between the behavior of unmated and mated females. V. songaricum extract was the most active extract, attracting 25.4% of unmated females. However, mated C. pomonella females exhibited greatest attraction to pear ester. In a wind tunnel bioassay, combining X. strumarium with codlemone significantly increased the response of male upwind flight and source contact as compared with codlemone alone. All plant extracts, except for V. songaricum, significantly reduced the number of eggs laid. The plant extracts exhibited some toxic effects to eggs, and hatching rate of eggs was reduced as compared with the control. Our results indicate that some of the plant extracts tested are potential candidates for practical use after elucidation and characterization of active compound(s).     

Spittlebugs as vectors of <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> in olive orchards in Italy

The recent introduction of Xylella fastidiosa in Europe and its involvement in the Olive Quick Decline Syndrome (OQDS) in Apulia (Salento, Lecce district, South Italy) led us to investigate the biology and transmission ability of the meadow spittlebug, Philaenus spumarius, which was recently demonstrated to transmit X. fastidiosa to periwinkle plants. Four xylem-sap-feeding insect species were found within and bordering olive orchards across Salento during a survey carried out from October 2013 to December 2014: P. spumarius was the most abundant species on non-olive vegetation in olive orchards as well as on olive foliage and was the only species that consistently tested positive for the presence of X. fastidiosa using real-time PCR. P. spumarius, whose nymphs develop within spittle on weeds during the spring, are likely to move from weeds beneath olive trees to olive canopy during the dry period (May to October 2014). The first X. fastidiosa-infective P. spumarius were collected in May from olive canopy: all the individuals previously collected on weeds tested negative for the bacterium. Experiments demonstrated that P. spumarius transmitted X. fastidiosa from infected to uninfected olive plants. Moreover, P. spumarius acquired X. fastidiosa from several host plant species in the field, with the highest acquisition rate from olive, polygala and acacia. Scanning electron microscopy (SEM) revealed bacterial cells resembling X. fastidiosa in the foreguts of adult P. spumarius. The data presented here are essential to plan an effective IPM strategy and limit further spread of the fastidious bacterium.     

Genetic structure and phylogeography of <Emphasis Type="Italic">Juniperus phoenicea</Emphasis> complex throughout Mediterranean and Macaronesian regions: different stories in one

Key message

The genetic structure of Juniperus phoenicea in the Mediterranean Basin is inferred using amplified fragment length polymorphism markers (AFLP) markers. As other Mediterranean conifers, J. phoenicea populations show moderate levels of genetic diversity and interpopulational differentiation. The pattern of distribution of genetic diversity seems highly influenced by the climatic fluctuations which occurred in the Pleistocene.

Context

It has been stated that the genetic structure of Mediterranean conifers is mediated by the historical climatic changes and the geological rearrangements which occurred in the Mediterranean Basin. J. phoenicea provides an excellent example to test how its genetic structure is influenced by these events.

Aims

In this work, we study the amount and distribution of genetic diversity of J. phoenicea complex, in order to evaluate its taxonomic status and to reveal underlying phylogeographic patterns.

Methods

The molecular diversity was analyzed for 805 individuals from 46 populations throughout its distribution range using AFLP markers. Principal coordinate analysis, analysis of molecular variance (AMOVA), and Bayesian-based analysis were applied to examine the population structure.

Results

AFLP markers revealed moderate levels of intrapopulation genetic diversity, pairwise genetic differentiation, and a clear pattern of isolation by distance. Bayesian analysis of population structure showed five clusters related to the taxonomic status of J. phoenicea and J. turbinata, and a geographic pattern of genetic structure in J. turbinata.

Conclusion

All the analysis separate J. phoenicea from J. turbinata. For J. turbinata, up to four groups can be distinguished from a phylogeographic point of view. The genetic structure of J. turbinata seems highly influenced by climatic and geologic fluctuations occurring since the Oligocene.

     

Genetic relationships among sympatric varieties of <Emphasis Type="Italic">Acer mono</Emphasis> in the Chichibu Mountains and Central Hokkaido,Japan

Acer mono Maxim. is one of the major components of cool temperate forests in Japan. Some of its many varieties are distributed sympatrically. Because of its great variability, the intraspecific taxonomy and nomenclature of the species are controversial. To understand the genetic relationships among these varieties and whether hybridization or introgression occurred among the sympatric varieties, we studied the genetic relationships among sympatric varieties of A. mono in the Chichibu Mountains (A. mono var. ambiguum, A. mono var. connivens, A. mono var. marmoratum) and Central Hokkaido (A. mono var. mayrii and A. mono var. glabrum) in Japan. Our results showed that varieties in Chichibu are genetically close, suggesting that hybridization or introgression might occur between these varieties, which could explain the higher genetic diversity of varieties in Chichibu than in Hokkaido. In contrast to the close relationships between the varieties in Chichibu, varieties in Hokkaido seemed relatively separated from each other; indeed, there may be reproductive isolation between the two varieties. The results provide new insight for the taxonomy of the varieties of A. mono, especially the sympatric varieties, in Japan.     

Effects of <Emphasis Type="Italic">Verticillium nonalfalfae</Emphasis> on <Emphasis Type="Italic">Ailanthus altissima</Emphasis> and associated indigenous and invasive tree species in eastern Austria

The naturally occurring Verticillium nonalfalfae shows promise for biocontrol of the highly invasive Tree of Heaven (Ailanthus altissima), but might also bear a risk for non-target tree species. In this study, we conducted inoculations on potted seedlings of A. altissima as well as on eight indigenous and two invasive tree species associated with Tree of Heaven in Austria. Although vascular discolourations developed in all inoculated tree species, V. nonalfalfae was reisolated from Ailanthus and eight of the ten non-target-species, whereas typical disease symptoms and mortality only occurred on A. altissima. Results confirmed high susceptibility (S) of A. altissima to V. nonalfalfae but indicated tolerance (T) of Acer campestre, Acer pseudoplatanus and Quercus robur, possible resistance (PR) of Fraxinus excelsior, Populus nigra, Tilia cordata, Ulmus laevis and Ulmus minor and resistance (R) of Fraxinus pennsylvanica and Robinia pseudoacacia to this potential biocontrol agent. Results from seedling inoculations were confirmed by cursory field observations in Ailanthus-inoculated forest stands, where admixed A. campestre, A. pseudoplatanus, F. excelsior, Populus alba, R. pseudoacacia and U. laevis canopy trees remained asymptomatic, while mortality was induced in Ailanthus.     

Tree leaves of <Emphasis Type="Italic">Salix babylonica</Emphasis> extract as a natural anthelmintic for small-ruminant farms in a semiarid region in Mexico

The study aimed to test the potential anthelmintic activity of Salix babylonica (SB) extract for the control of gastrointestinal and pulmonary parasites in sheep and goats under field conditions. A representative sample of 20 % of all animals reared in 8 sheep and 7 goat farms was used in the study. Animals from each farm were randomly selected for a total number of 93 sheep and 75 goats. Animals suffered a natural gastrointestinal nematode infection and had never been treated with chemical anthelmintic drugs. The SB extract (20 mL) was orally administered weekly before the morning feeding to each animal for 60 days. Fecal eggs or oocysts were counted at 0, 1, 20, 40, and 60 days after starting the extract administration. Differences (P < 0.01) in the fecal oocyst and egg output of Eimeria, Dictyocaulus, and Moniezia were observed between sheep and goats. In addition, the treatment influenced (P < 0.05) egg outputs of Cooperia, Dictyocaulus, and Trichuris. Fecal egg or oocyst counts of Haemonchus contortus, Eimeria, Cooperia, Chabertia, Dictyocaulus, Moniezia, and Ostertagia were time-dependent (P < 0.05). For sheep, administration of SB decreased (P < 0.05) the fecal eggs count of H. contortus, Cooperia, Chabertia, Dictyocaulus, Moniezia, and Trichuris. After 20 days of treatment, H. contortus, Cooperia, or Moniezia were not detected. For goats, SB reduced (P < 0.05) the fecal egg counts of H. contortus, Cooperia, Chabertia, and Moniezia. Moreover, decreases were observed (P < 0.05) for Chabertia, Trichostrongylus, and Ostertagia. Eggs of H. contortus and Moniezia were not present in the feces after 1 day of administration of the extract. It could be concluded that the weekly administration of SB extract at 20 mL per animal can be used to treat gastrointestinal and lung nematodes of small ruminants in organic and traditional farming systems of tropical regions.     

Analysis of floral scent emitted from <Emphasis Type="Italic">Syringa</Emphasis> plants

Syringa is an important aromatic woody angiosperm that is widely planted in gardens. Its dry flowers are traditionally used to manufacture infusions and spices in China. In this study, the floral volatiles emitted from nine different Syringa species and varieties, viz. S. chinensis, S. p rotolaciniata, S. o blata, S. o blata var. giraldii, S. o blata var. plena, S. v ulgaris ‘Mrs Harry Bickle’, S. v ulgaris ‘Bright Centennial’, S. v ulgaris ‘White Spires’, and S. v ulgaris ‘President Lincoln’ were collected by the dynamic headspace technique, and then identified by automated thermal desorption–gas chromatography/mass spectrometry. There were significant differences in components and corresponding contents of floral scent emitted from the nine Syringa species and varieties. Among the detected components, β-ocimene exhibited the highest content, accounting for more than 70 % of the floral scent of S. p rotolaciniata, S. o blata var. giraldii, and S. v ulgaris ‘Bright Centennial’. A significant daytime variation of floral scent emitted from S. p ekinensis was recorded, especially for benzaldehyde, whose content first increased and then decreased. We studied the scent emitted from the in vitro flowers of S. v ulgaris ‘Pres Lincoln’ during the process of natural drying. Alcohol compounds were released in the greatest content, and the content of 2-ethyl-1-hexanol accounted for about 40 % of the total release amount. The results described the emission pattern and mechanisms of floral scent in Syringa.     

Phylogenetic relationships among five species of <Emphasis Type="Italic">Armeniaca</Emphasis> Scop. (Rosaceae) using microsatellites (SSRs) and capillary electrophoresis

The genus Armeniaca Scop. is well known for its popular cultivated edible fruit trees such as Armeniaca vulgaris Lam. and ornamental flowers such as A. mume Sieb. Another species, A. cathayana Fu et al., one of six important dry fruits (kernel-using apricot), is cultivated for its edible seeds in North China. In the present study, DNA from 70 individuals of Armeniaca, including 38 of A. cathayana, 18 of A. vulgaris, 12 of A. sibirica, 1 of A. dasycarpa and 1 of A. mume, was extracted and analyzed using microsatellites and capillary electrophoresis. For 20 polymorphic loci selected, 339 alleles and 140.7 effective alleles were detected. The number of alleles per locus ranged from 8 to 28, with an average of 16.95 alleles per locus. The observed heterozygosity (Ho) and the expected heterozygosity (He) ranged from 0.427 to 0.971 and from 0.737 to 0.912, respectively. The polymorphism information content varied from 0.708 to 0.905, with an average of 0.827. Based on the genetic similarity among 70 individuals, a UPGMA was used to establish the phylogenetic relationships. The taxonomic positions among five species were clearly revealed, and A. cathayana was more closely related to A. vulgaris than to A. sibirica. The results will provide a scientific basis for research on the taxonomy, germplasm resources and breeding of Armeniaca, especially for A. cathayana.     

Host specificity of Asian parasitoids for potential classical biological control of <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

The Asian spotted wing drosophila, Drosophila suzukii, has recently become a serious pest of soft fruits in Europe. Classical biological control through the introduction of larval parasitoids from its native range in Asia is presently being considered. However, host specificity of potential biological control agents has to be determined to avoid releasing species that may have unintended non-target impacts. Larvae of six different European non-target fly species and the target D. suzukii were exposed either on diet or blueberries to three Asian larval parasitoids, Asobara japonica, Leptopilina japonica, and Ganaspis cf. brasiliensis, and one European species, Leptopilina heterotoma. Asobara japonica showed the lowest specificity, attacking and developing in all Drosophilidae. Leptopilina japonica successfully parasitized two non-target Drosophilidae, D. melanogaster and D. subobscura, with one singly progeny emerging from D. immigrans. Ganaspis cf. brasiliensis had the highest level of specificity but variations occurred between two geographical populations tested. A Japanese population was strictly specific to D. suzukii, whereas another population from China parasitized D. suzukii, D. melanogaster and sporadically D. subobscura. The European L. heterotoma successfully developed in D. melanogaster, D. subobscura and occasionally in D. immigrans, but nearly all eggs and larvae in D. suzukii were encapsulated. These results show that Ganaspis cf. brasiliensis is the species with the highest potential for biological control, but more studies are needed on its taxonomic status and the existence of biotypes or cryptic species varying in their specificity before field releases can be conducted in Europe.     

Symptoms and fungi associated with elm trees decline in Iran

Elm (Ulmus, Ulmaceae) is one of the most popular and important trees in urban and landscape areas in Iran. A severe decline of elm trees has recently been observed in some areas of Kerman and Shiraz cities. However, the identity of the causal agents has not yet been investigated. The purpose of this study, carried out in the years 2012–2014, was to isolate and identify fungal trunk pathogens associated with decline symptoms observed in elm trees. Samples were collected from trunks and branches of trees showing various disease symptoms and internal wood lesions. Fungal isolations were made from discolored or decayed wood tissue. In Kerman some elm trees were attacked by the beetle Aeolesthes sarta; therefore, samples were also collected from larvae and adults of beetles associated with trees showing wood damage. Fungal isolates were identified by morphological, cultural and molecular characteristics. Thirteen fungal species, Phaeoacremonium (P.) minimum, P. parasiticum, P. sicilianum, P. alvesii, P. fraxinopennsylvanicum, Spencermartinsia viticola, Dothiorella (Do.) sarmentorum, Neoscytalidium hyalinum, Diatrype (Di.) whitmanensis, Cosmospora viridescens, Phoma (Pho.) herbarum, Phellinus (Phe.) tuberculosus and Inonotus levis were identified. Phaeoacremonium parasiticum was the only species isolated from both larvae and adult beetles. Pathogenicity trials were performed on detached shoots of elm under greenhouse conditions. Dothiorella sarmentorum was the most virulent species based on the length of wood necrosis. This study is the first report of P. sicilianum, C. viridescens and Phe. tuberculosus in Iran. In addition, most of the isolated species are reported for the first time on elm trees in the world.     

Evolutionary history and phylogeography of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) in Europe based on molecular markers

In this review we summarized recent historical records and molecular studies on evolutionary history and phylogeography of Scots pine with focus on the European highly fragmented distribution area of the species. Fossilized pollen, plant micro- and macrofossil records provided evidences on the large-scale species’ range shifts and demographic changes during the Quaternary. Populations of Scots pine were documented both in the glacial (incl. full glaciation) and interglacial periods. Recolonization of Europe after the glaciation originated from the (Sub) Mediterranean areas like the Balkan Peninsula but also from around the Eastern Alps and the surroundings of the Danube plain. Fennoscandia and northern European Baltic regions were most probably colonized from two main directions, from Western Europe and from the Russian Plain. Modern history of Scots pine was hardly affected by anthropogenic activities that started to strengthen in the Bronze and Iron Age. Along with the fossil records, molecular genetic tools were used to infer the origin and putative history including migration, differentiation and demography of the species. In this paper we compiled the major publications (30) of molecular genetic studies of the past 20 years derived from distinctly inherited organelle genomes (mitochondrial, chloroplast, nuclear) revealed by different marker systems (mtDNA-cox1, -nad1, -nad3, -nad7, ISSR, cpSSR, nSSR, B-SAP, SNP). It is important to consider that different phylogeographic patterns can be drawn by the analysis of different DNA marker types. Accordingly the use of more than one marker simultaneously outlines the most sophisticated phylogeographical pattern on the genetic lineages and can reveal high differentiation of the European distribution. Combined marker systems and markers derived from coding sequences have also been used to detect species’ phylogeographic patterns, but these were rarely applied to Scots pine. Although new molecular techniques can provide higher resolution data for populations, the reviewed results can shape the direction of further studies.     

Genetic diversity in two threatened species in Vietnam: <Emphasis Type="Italic">Taxus chinensis</Emphasis> and <Emphasis Type="Italic">Taxus wallichiana</Emphasis>

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 and 150 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 (F _ST = 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 and 73.12 % 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.     

ISSR and SSR markers reveal sex-specific DNA sequences in three <Emphasis Type="Italic">Calamus</Emphasis> species from India

Calamus species yields the raw materials for the cane industry. However, the extraction of the cane from the forests is being carried out indiscriminately without considering the sustenance of the species. Plants of both sexes should co-exist for reproductive success. The sex of Calamus plants can be identified only after flowering and hence proper planning of managed forestry practices cannot be accomplished. A study was carried out in this background and male specific ISSR markers for C. tenuis and C. flagellum and SSR markers for C. thwaitesii were identified. The diagnostic potential of these markers can be exploited to sex the Calamus species at the seedling stage for proper breeding and agroforestry management.     

Lipophilic extractives of the inner and outer barks from six different <Emphasis Type="Italic">Pinus</Emphasis> species grown in Indonesia

The chemical compositions of the dichloromethane extracts of inner and outer barks from six Pinus species (P. elliotii, P. oocarpa, P. caribeae, P. merkusii, P. montezumae, and P. insularis) grown in Indonesia were investigated by GC and GC–MS. Generally, the amounts of extractive contents were higher in the inner bark than in the outer bark except for P. merksuii. Fatty acids, monoterpenes, sesquiterpenes, resin acids, triterpenoids, and steroids were detected and quantified. Inner and outer barks differed not only in content of these compounds but also in their composition. Fatty acids and alcohols were the major classes of lipophilic compounds in the outer bark of P. caribeae, P. insularis, and P. montezumae. Steroids and triterpenoids were the dominant compounds identified in the inner bark of P. elliotii, P. insularis, and P. merkusii. Resin acids were the most abundant group in the inner bark of P. oocarpa whereas monoterpenes and sesquiterpenes were recorded in minor quantities in both bark layers of all species.     

First record of the invasive pest <Emphasis Type="Italic">Drosophila suzukii</Emphasis> in Ukraine indicates multiple sources of invasion

Drosophila suzukii, commonly known as the spotted-wing Drosophila, is an invasive polyphagous fruit pest, which has emerged as a global threat to agriculture in the Americas and in Europe. Due to the rapid spread, great economic losses and its pest behavior, D. suzukii represents a powerful model for invasion biology and pest management studies. However, its current European distribution, invasion routes and levels of genetic diversity in populations of D. suzukii are poorly understood. We present the first report of D. suzukii from Ukraine, with the invasion likely occurring close to 2014. The pattern of genetic variation at cytochrome oxidase I among D. suzukii populations from Europe, USA and Asia reveals comparatively high genetic diversity in the Ukrainian population of this pest species, suggesting a complex invasion scenario from multiple sources. Further monitoring patterns of genetic variation across space and time, to better understand the invasion routes of this invasive insect pest, will be an essential part for developing successful pest management strategies.     

A novel cold-inducible promoter,P<Emphasis Type="Italic">ThCAP</Emphasis> from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, confers cold tolerance in transgenic <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Our previous studies have revealed that the ThCAP gene plays a vital role in transgenic Populus (P. davidiana × P. bolleana) in response to cold stress. However, the regulatory mechanism of ThCAP gene expression has been unclear. In this study, the 5′ flanking region of the ThCAP promoter (PThCAP) was cloned using a genome-walking method. By analyzing cis-acting regulatory elements of PThCAP, 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 ThCAP gene promoter, a series of deletion derivatives of PThCAP, 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 this 290-bp region (?1190 to ?900 bp), as an important part in PThCAP, was associated with cold tolerance of A. thaliana. Our results provide evidence for the regulatory mechanism of ThCAP gene involved in the response to cold stress, and that the gene is promising candidate gene for genetic improvement of crops.