Dietary yeast affects preference and performance in <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

Yeasts play an important role in nutrition physiology and host attraction of many Drosophila species, and associations with various yeast species are documented for several drosophilid flies. The pest Drosophila suzukii (Matsumura) has a predominant association with the yeast Hanseniaspora uvarum. However, research has not been conducted on the nutritional physiology of the yeasts associated with D. suzukii (spotted wing drosophila). Therefore, in this study, we determined whether dietary yeast was nutritionally relevant and whether yeast species closely associated with D. suzukii positively affected life-history traits. Our results confirm a crucial role of dietary yeast in the larval development and survival of D. suzukii. Furthermore, we found specific effects of the closely associated yeast species H. uvarum and Candida sp. on larval survival. Observations of the egg-laying behaviour of D. suzukii on cherry fruits artificially colonised with different yeast species revealed that the number of eggs laid increased on fruits colonised with Candida sp. and Saccharomyces cerevisiae.     

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.     

Lure-and-infect and lure-and-kill devices based on <Emphasis Type="Italic">Metarhizium brunneum</Emphasis> for spotted wing Drosophila control

In the current study, we have designed two experimental devices to study the potential of Metarhizium brunneum (Petch) and its crude extract (filtered growth medium) for the control of Drosophila suzukii (Matsumura). An experimental autoinoculation device impregnated with conidia of EAMa 01/58-Su strain was designed for a lure-and-infect strategy. D. suzukii adult mortality resulted in a 62.2% with an average survival time of 3.6 days, with no significant differences among attractiveness to adults of red and black colours used in the devices. Nevertheless, a 48.0% mortality of untreated males was obtained after being coupled with inoculated females, whereas only 24.0% of untreated females was killed after being coupled with inoculated males, thereby revealing the horizontal transmission potential of the strain in these inoculation devices. We observed an 84.7% reduction in fecundity in M. brunneum-challenged D. suzukii females. Finally, we designed a lure-and-kill device to dispense a crude extract of the EAMb 09/01-Su strain that had previously shown acute mortality in D. suzukii, with an exposure time of 6.9 h required to achieve 50.0% mortality. Mixing the extract with feeding attractant caused a 61.6% D. suzukii adult mortality rate. These results show the high potential of using M. brunneum in lure-and-infect and lure-and-kill strategies contributing to an integrated pest management program for D. suzukii control.     

Potential host ranges of three Asian larval parasitoids of Drosophila suzukii

Asobara japonica (Hymenoptera: Braconidae), Ganaspis brasiliensis and Leptopilina japonica (Hymenoptera: Figitidae) are Asian larval parasitoids of spotted wing drosophila, Drosophila suzukii (Diptera: Drosophilidae). This study evaluated these parasitoids’ capacity to attack and develop from 24 non-target drosophilid species. Results showed that all three parasitoids were able to parasitize host larvae of multiple non-target species in artificial diet; A. japonica developed from 19 tested host species, regardless of the phylogenetic position of the host species, L. japonica developed from 11 tested species; and G. brasiliensis developed from only four of the exposed species. Success rate of parasitism (i.e., the probability that an adult wasp successfully emerged from a parasitized host) by the two figitid parasitoids was low in hosts other than the three species in the melanogaster group (D. melanogaster, D. simulans, and D. suzukii). The failure of the figitids to develop in most of the tested host species appears to correspond with more frequent encapsulation of the parasitoids by the hosts. The results indicate that G. brasiliensis is the most host specific to D. suzukii, L. japonica attacks mainly species in the melanogaster group and A. japonica is a generalist, at least physiologically. Overall, the developmental time of the parasitoids increased with the host’s developmental time. The body size of female A. japonica (as a model species) was positively related to host size, and mature egg load of female wasps increased with female body size. We discuss the use of these parasitoids for classical biological control of D. suzukii.

     

Large-scale spatial dynamics of <Emphasis Type="Italic">Drosophila suzukii</Emphasis> in Trentino,Italy

Drosophila suzukii (Diptera: Drosophilidae) is an invasive alien species devastating soft fruit crops in newly invaded territories. Little is known about the importance and potential of long-distance dispersal at a regional scale. The goal of this work is to investigate D. suzukii dispersal ability during different times of the season, and along an elevational gradient in a mountain valley in Trentino Province, Italy. We employed a mark–release–recapture strategy using protein markers. Flies were recaptured using fruit-baited traps. The protein-marked flies were positively identified using ELISA procedure. Additional microsatellite analyses were performed on D. suzukii collected during autumn at different elevations to characterize the population structure. Results suggest that a portion of the local D. suzukii population moves from low to high elevations during spring and summer and travels back to low elevations in autumn. Genetic analysis further revealed that samples collected during autumn at different elevations belong to the same population. These results show that D. suzukii are able to fly up to about 9000 m away from the marking point and that seasonal breezes likely facilitate long-distance movement. We suggest that these migrations have multiple functions for D. suzukii, including conferring the ability to exploit gradual changes of temperature, food, and ovipositional resources in spring and autumn, as well as to assist in the search for suitable overwintering sites in late autumn. Our findings help to unveil the complex ecology of D. suzukii in Italian mountainous regions and provide important clues for improving the efficacy of integrated pest management control techniques to combat this pest.     

<Emphasis Type="Italic">Drosophila suzukii</Emphasis> flight performance reduced by starvation but not affected by humidity

Drosophila suzukii is widely studied because of its status as a global pest of berries and soft fruits. Environmental conditions and access to food resources impact the physiology and fitness of D. suzukii; these factors could also affect dispersal. Flight mills are a convenient tool for measuring and comparing the flight performance of insects. In this study, two experiments examined the effects of diet and humidity on D. suzukii flight performance using custom-built flight mills, and a third experiment compared the energy reserves of D. suzukii flown or not flown on flight mills. Over all flight assays, the median flight distance and duration were 27.16 m and 2.37 min, respectively, and the mean flight velocity was 0.18 m/s. The maximum flight distance and duration by an individual were 1.75 km and 2.35 h, respectively. Drosophila suzukii provisioned with blossoms, fruits, or standard laboratory diets flew farther distances and longer durations than starved flies. While starvation was associated with reduced flight performance, there were no observed differences between diet types. It remains unclear whether D. suzukii consistently use lipids, glycogen, sugar, or another energy source for flight because tethered individuals may not have flown enough to deplete energy reserves. Humidity did not affect flight performance of D. suzukii within a?~?2 h test period. These data indicate that most D. suzukii are likely to remain within limited area (e.g., within a field) but that some individuals can disperse long distances (field to field spread).     

Identification of attractive blend for spotted wing drosophila, <Emphasis Type="Italic">Drosophila suzukii</Emphasis>, from apple juice

Drosophila suzukii, commonly known as the spotted wing drosophila (SWD), is an exotic fruit fly from Southeast Asia that was introduced to the temperate regions of North America and Europe in 2008. It attacks a wide variety of fruits and has become a devastating pest of soft-skinned fruit crops. Due to the rapid spread of SWD across the newly invaded continents, fresh fruit markets have a zero-tolerance policy regarding D. suzukii infestation. Specific and efficient D. suzukii detection tools are urgently needed so that farmers can deliver timely management interventions to meet market demands. Since SWD is known to be attracted to damaged and rotting fruits, headspace volatiles from fresh and fermented apple juices were collected and analyzed by gas chromatography–mass spectrometry. Special attention was given to the compounds produced and/or enriched during the fermentation process. After performing a series of laboataory and field tests, we identified a quinary blend, which is more efficient and selective for D. suzukii than the currently standard apple cider vinegar and commercially available SWD lure under field conditions. Identification of SWD attractant will help growers accurately detect D. suzukii adult infestations in orchards, thereby allowing for timely pest management interventions while reducing conventional insecticidal usage to protect our crops, environment, and ecosystem.     

Sterile insect technique and <Emphasis Type="Italic">Wolbachia</Emphasis> symbiosis as potential tools for the control of the invasive species <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

Drosophila suzukii, a vinegar fly originated from Southeast Asia, has recently invaded western countries, and it has been recognized as an important threat of a wide variety of several commercial soft fruits. This review summarizes the current information about the biology and dispersal of D. suzukii and discusses the current status and prospects of control methods for the management of this pest. We highlight current knowledge and ongoing research on innovative environmental-friendly control methods with emphasis on the sterile insect technique (SIT) and the incompatible insect technique (IIT). SIT has been successfully used for the containment, suppression or even eradication of populations of insect pests. IIT has been proposed as a stand-alone tool or in conjunction with SIT for insect pest control. The principles of SIT and IIT are reviewed, and the potential value of each approach in the management of D. suzukii is analyzed. We thoroughly address the challenges of SIT and IIT, and we propose the use of SIT as a component of an area-wide integrated pest management approach to suppress D. suzukii populations. As a contingency plan, we suggest a promising alternative avenue through the combination of these two techniques, SIT/IIT, which has been developed and is currently being tested in open-field trials against Aedes mosquito populations. All the potential limiting factors that may render these methods ineffective, as well as the requirements that need to be fulfilled before their application, are discussed.     

Seasonal parasitism and host specificity of <Emphasis Type="Italic">Trissolcus japonicus</Emphasis> in northern China

The brown marmorated stink bug, Halyomorpha halys (Stål), native to China, Japan, and Korea, has emerged as a harmful invasive pest of a variety of crops in North America and Europe. The Asian egg parasitoid Trissolcus japonicus has been identified as the most promising agent for classical biological control of invasive H. halys populations. A 4-year study evaluated the fundamental and ecological host ranges of T. japonicus as well as its phenology and impact on H. halys populations in fruit orchards in its native range in northern China. In laboratory no-choice tests, developmental suitability of eight non-target host species for T. japonicus was demonstrated by the successful production of progeny on the majority (>85%) of non-target host species tested. In field-collected, naturally laid egg masses, T. japonicus was the most abundant parasitoid associated with H. halys and Dolycoris baccarum, but was also sporadically found in Plautia crossota. Furthermore, it was regularly reared from sentinel egg masses of Menida violacea, Arma chinensis, and Carbula eoa. The only species that did not support development in the laboratory and field was Cappaea tibialis. Besides the benefit of having a high impact on H. halys populations in Northern China, the risk assessment conducted in the area of origin indicates that native Pentatomidae in North America and Europe could be negatively impacted by T. japonicus. Whether the benefits of T. japonicus outweigh the possible risks will have to be evaluated based on the outcome of additional host range studies in the two invaded regions.     

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.     

Identifying plant DNA in the sponging–feeding insect pest <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

Drosophila suzukii (Diptera: Drosophilidae) is a highly polyphagous invasive pest threatening fruit production in the Americas and Europe. The current knowledge of its host plants is mainly based on oviposition and larval development in fruits, while little is known on the diet of the adult flies. This information is important for developing effective control strategies. Here, we examine DNA-based techniques to determine food plants of D. suzukii. Adult flies were fed with raspberries (Rubus idaeus) and allowed to digest up to 72 h after feeding. Raspberry DNA was detected by diagnostic PCR for up to 48 h post-feeding with a significant negative effect of time on DNA detection success but no significant differences between male and female flies in detection probabilities. As D. suzukii walks on plants, its body surface can get contaminated with DNA. With a bleaching experiment, we succeeded to remove contaminating external plant DNA, while the DNA in the gut content stayed unaffected. Finally, field-collected flies were subjected to a next-generation sequencing approach, demonstrating that plant DNA from different host plants can be efficiently detected in both bleached and non-bleached specimens. In order to safeguard against erroneous host plant detections, we recommend bleaching flies before they are subjected to DNA extraction. The current findings encourage the use of DNA-based gut content analysis in D. suzukii to obtain a better understanding of its feeding ecology which is a prerequisite for developing successful control strategies.     

The populations and distribution of <Emphasis Type="Italic">Pieris japonica</Emphasis>, a poisonous tree protected from herbivore browsing pressure,increase slowly but steadily

Pieris japonica is a poisonous tree species that is rarely eaten by herbivorous animals, a fact that could enable the expansion of its distribution range and influence ecosystems into which it encroaches. In a regional-scale study, 300 P. japonica trees from 13 populations were sampled at the University of Tokyo Chichibu Forest, Japan, and were analyzed using 11 microsatellite markers. Genetic differentiation among the populations was low (F _ST = 0.022 and G′ _ST = 0.024). A plot (30 × 30 m) was established for a fine-scale study, in which all P. japonica trees and saplings were measured and genetically analyzed using the microsatellite markers. Using this approach, we detected 84 genotypes among the 121 P. japonica trees in the plot. A few genotypes had expanded by more than 5 m, indicating that the ability to reproduce asexually could facilitate P. japonica trees to remain in a given location. Autocorrelation analysis showed that the extent of nonrandom spatial genetic structure was approximately 7.0 m, suggesting that seed dispersal was limited. KINGROUP analyses showed that 44 genotype pairs were full siblings, 23 were half-siblings, and 40 were parent-offspring. Only 32 seedlings were observed, of which 15 had reproduced asexually. The number of P. japonica trees has been increasing gradually for more than half a century in our study areas.     

Incorporating anthropogenic variables into ecological niche modeling to predict areas of invasion of <Emphasis Type="Italic">Popillia japonica</Emphasis>

Niche conservatism providing support for using ecological niche modeling in biological invasions has been widely noticed; nonetheless, the impacts of human disturbance on niche changes and model predictions are not fully recognized. In this study, climate niche changes during the invasion of Japanese beetle (Popillia japonica) were studied under the framework of niche unfilling, stability, and expansion situations. The beetle might undergo niche expansion because of the small native range (Japan) relative to the broad invade range (North America). Relative roles of climatic and anthropogenic factors and their interactions in determining the spread of P. japonica were investigated. Results suggest that the climate niche was conserved during P. japonica invasion, climate niche unfilling rather than niche expansion was observed, suggesting there are suitable climate space remain unoccupied. Inclusion of anthropogenic factors into climate niche model has improved niche model prediction values in areas of high human activity. During P. japonica invasion, anthropogenic factors influenced the climate niche expression and altered the effect of climate suitability. In areas of poor or unsuitable climate, the human activities facilitated the establishment of P. japonica and functioned synergistically, whereas in highly suitable areas, including anthropogenic variables decreased climate suitability. These findings are pivotal to our understanding of climate niche conservatism and niche model transferability when predicting areas of invasion of P. japonica and can be applied elsewhere for predicting other pest’s invasion potential.     

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.     

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.     

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.     

Assessment of a commercial spider venom peptide against spotted-wing Drosophila and interaction with adjuvants

Chemical control of insect pests in food crops is dominated by broad-spectrum insecticides from a few classes, and there is an urgent need for alternative modes of action. We examined the efficacy of a spider venom peptide, GS-omega/kappa-Hxtx-Hv1a (hereafter, Hv1a) for control of spotted-wing Drosophila and evaluated the importance of phagostimulants and adjuvants for its efficacy. Topical and residual activity of Hv1a was low, with only 17.5% of exposed adult D. suzukii dying after 72 h. In contrast, 100% adult mortality was observed after 24 h when three adjuvants were added to Hv1a. Survival of eggs of D. suzukii oviposited into blueberries was also reduced by exposure to Hv1a combined with the same adjuvants, indicating that Hv1a activity against D. suzukii in the laboratory, but requires penetration of the insect cuticle for efficacy. In a field trial in blueberries, Hv1a gave comparable control to phosmet, and significantly reduced infestation in fruit. This biopesticide adds a new mode of action to the options available for integrated pest management of this and other insect’s pests.     

Rapid harvest schedules and fruit removal as non-chemical approaches for managing spotted wing Drosophila

Spotted wing Drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), has caused significant economic losses to small fruit and berry growers throughout the USA and Europe since its invasion. This pest can lay many eggs over its lifetime within ripening and ripe berries, causing yield loss and the risk of fruit contamination. Zero tolerance for this pest has led to increased use of broad-spectrum insecticides to control it, which are costly and pose many other sustainability and pest management concerns. There is an urgent need to evaluate management strategies that can decrease reliance on chemical controls and mitigate economic losses. Over two growing seasons, we compared harvest schedules for their effect on infestation by D. suzukii, revealing that fruit harvested every 1 or 2 days had significantly fewer D. suzukii larvae than a 3-day harvest schedule. Furthermore, we found that yield per unit effort was highest on a 2-day schedule. Sanitation of the crop is another important component of a successful integrated pest management program, and we found that bagging infested waste berries killed 99% of larvae after 32 h, with higher fruit temperatures in clear bags than white or black bags. In combination, these methods can reduce the effects of this invasive pest on raspberry production. This study will provide guidance to growers on culturally based IPM tactics to decrease reliance on chemical management.     

Flammability of some companion species in cork oak (<Emphasis Type="Italic">Quercus suber</Emphasis> L.) forests

Key message

The high flammability of some companion species in Quercus suber forests, estimated in laboratory tests, could potentially generate an increase in fire vulnerability and in fire risk.

Context

Recurrent wildfire is one of the main causes of forest degradation, especially in the Mediterranean region. Increased fire frequency and severity due to global change could reduce the natural resilience of cork oak to wildfire in the future. Hence, it is important to evaluate the flammability of companion species in cork oak forests in the particularly dry bioclimatic conditions of North Africa.

Aims

This study aimed to assess and compare flammability parameters at laboratory scale among ten companion frequent species in cork oak forests.

Methods

Fuel samples were collected in a cork oak (Quercus suber L) forest in the southern part of the mountains of Tlemcen (Western Algeria). A series of flammability tests were carried out using a Mass Loss Calorimeter device (FTT ®). A cluster analysis to classify flammability of the selected species was conducted using the K-means algorithm.

Results

The results revealed differences in the four flammability parameters (ignitability, sustainability, combustibility and consumability), in both fresh and dried fine fuel samples from Quercus suber, Pinus halepensis, Quercus ilex, Quercus faginea, Erica arborea, Arbutus unedo, Pistacia lentiscus, Calicotome spinosa, Juniperus oxycedrus and Tetraclinis articulata. Application of the K-means clustering algorithm showed that C. spinosa, T. articulata, J. oxycedrus and P. halepensis are highly flammable because of their high combustibility and sustainability.

Conclusion

The findings identify species that could potentially increase the vulnerability of cork oak forests to forest fires.

     

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.