From horticulture and biofuel to invasion: the spread of Miscanthus taxa in the USA and Europe

Species of the genus Miscanthus (Poaceae) have recently become widely planted for ornamental and industrial uses. Yet these large perennial grasses have several traits that suggest a high invasion potential. To inform the debate on the benefits and risks of introducing these new biofuel crops, this publication summarises the invasion of Miscanthus species in Europe and the USA. Databases from North America were interrogated, and occurrence data from a variety of European sources were examined. In both continents, M. sinensis and M. sacchariflorus have recently spread rapidly, although invasion started earlier in the USA and both species are more widespread there. Both species mostly colonise grasslands and ruderal habitats. In addition, some escapes of the hybrid M. × giganteus, which has a particularly strong invasion potential because of its vigorous growth and greater capacity to establish in cool climates, have already been recorded. We conclude that the invasion potential of Miscanthus species needs to be thoroughly tested, the incipient invasions in Europe and the USA monitored and, if required, planting restrictions should be adopted.     

Molecular and morphological identification of the mealybug,Phenacoccus solani Ferris (Hemiptera: Pseudococcidae): first report in Egypt

During the summer and autumn of 2016, pumpkins, Cucurbita spp. (Cucurbitaceae), were observed to be infested by mealybug species. Populations were collected on various pumpkin fruits from Rosetta, a port city at the North‐Western Nile River delta, Lower Egypt. This study aimed to identify the principle mealybug species infesting the major pumpkin‐producing regions in Egypt, by molecular and morphological characterization. DNA from 15 specimens and morphological data from 22 specimens were used to confirm the identity of the mealybug species. The species present were identified as Phenacoccus solani Ferris, Phenacoccus solenopsis Tinsley, and Planococcus citri (Risso). This is the first record of Phenacoccus solani Ferris (Hemiptera: Pseudococcidae) in Egypt.     

Miscanthus x giganteus crop fields hide a genotype of the invasive M. sacchariflorus

Despite their putative invasive potential, Miscanthus taxa have been increasingly viewed as promising crops for bioenergy production. Miscanthus x giganteus is cultivated worldwide due to its high productivity, and its non‐spreading growth limits its risk of invasion. However, the genetic uniformity of its unique cultivated clone has recently been questioned. We used morphometry, nuclear microsatellites, AFLP fingerprints and nrDNA ITS barcoding to assess Miscanthus genotypes cultivated in eastern France. As expected, the globally cultivated clone was the main morphotype and genotype of our samplings; however, we also identified a second genotype, assigned to M. sacchariflorus, in the sampled fields. Morphological differentiation partially overlaps genotypic assignation, probably due to disturbance at the crop edges. We also show that Miscanthus taxa are misidentified in commercial nurseries. Our study confirms previous reports of the occurrence of other accessions in fields of M. x giganteus. These taxonomic confusions limit the certainty that a single clone of M. x giganteus is cultivated. In this context, M. x giganteus crop fields may actually promote the dispersal of invasive Miscanthus taxa.     

Identification of economically important Liriomyza species by PCR‐RFLP analysis*

Only adult males of Liriomyza bryoniae, L. huidobrensis, L. sativae and L. trifolii can be identified with certainty on basis of their genitalia. Female adults, pupae and larvae can only be identified on the level of groups of species (L. bryoniae and L. huidobrensis vs. L. sativae and L. trifolii). Species identification in all developmental stages is possible using molecular biological techniques. Our method is a PCR amplification of a 790‐bp fragment of mitochondrial cytochrome oxidase II (COII) DNA followed by RFLP analysis. The method was tested on single larvae, pupae and adults and proved to be applicable to these three life stages. The specificity of the assay was assessed by comparing the results of the PCR‐RFLP analysis with those of morphological analysis using 60 Liriomyza specimens. Molecular and morphological identification agreed for all specimens analysed. PCR‐RFLP is a powerful diagnostic tool for rapid and reliable identification of all life stages of economically important Liriomyza species.     

Ditylenchus gigas n. sp. parasitizing broad bean: a new stem nematode singled out from the Ditylenchus dipsaci species complex using a polyphasic approach with molecular phylogeny

Morphologial, biochemical, molecular and karyological analyses of different populations and races of the stem and bulb nematode Ditylenchus dipsaci have suggested that it represents a species complex, of which only D. dipsaci sensu stricto and its morphologically larger variant, known as the giant race of the stem and bulb nematode, are plant parasites of economic importance. The present study singles out the giant race from this complex, herein described as a new species named Ditylenchus gigas n. sp., on the basis of morphological and molecular data obtained from several populations collected from broad beans in southern Italy, southern Spain and Lebanon. The new species epithet, which refers to the large body size of the nematode with respect to the normal races, must be considered to be conspecific with the D. dipsaci‘giant race’ from Fabaceae in recent literature. Morphologically, the new species is characterized by a body size 1·5–2 times longer than the ‘normal race’, stylet delicate (11·5–13·0 μm long) with knobs distinctly sloping backwards, and long post‐vulval uterine sac (81–150 μm long). Results of molecular analysis of rDNA sequences including the ITS1‐5.8S‐ITS2 region, the D2–D3 fragment of the 28S gene, the small 18S subunit, the partial mitochondrial gene for cytochrome c oxidase I (mtCOI), and hsp90 gene sequences, support the new taxonomic species status for the former D. dipsaci giant race from Vicia faba, and clearly distinguish D. gigas n. sp. from D. dipsaci sensu stricto.     

Detection of <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> and <Emphasis Type="Italic">P. palmivora</Emphasis> in citrus roots using PCR-RFLP in comparison with other methods

Phytophthora nicotianae and P. palmivora are the most important soil-borne pathogens of citrus in Florida. These two species were detected and identified in singly and doubly infected plants using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of internal transcribed spacer (ITS) regions of ribosomal DNA. The sensitivity of the PCR-RFLP was analyzed and the usefulness of the method evaluated as an alternative or supplement to serological methods and recovery on semi-selective medium. In a semi-nested PCR with universal primers ITS4 and ITS6, the detection limit was 1 fg of fungal DNA, which made it 1000× more sensitive than a single-step PCR with primers ITS4 and DC6. The sensitivity of detection for P. nicotianae was shown to be ten-fold lower than for P. palmivora, limiting its detection with restriction profiles in plants infected by both fungal species. Phytophthora nicotianae was detected with species-specific primers in all samples inoculated with this species despite the absence of species-specific patterns in RFLP. In contrast, the incidence of detection of P. palmivora in the presence of P. nicotianae was considerably lower using plating and morphological detection methods. Due to its high sensitivity, PCR amplification of ribosomal ITS regions is a valuable tool for detecting and identifying Phytophthora spp. in citrus roots, provided a thorough knowledge of reaction conditions for the target species is established prior to the interpretation of data.     

Phytophthora oleae sp. nov. causing fruit rot of olive in southern Italy

A homothallic Phytophthora species was found to be consistently associated with a rot of mature fruits of two local cultivars of olive (Olea europaea) in Calabria, southern Italy. The phylogenetic analysis of sequences of the ITS1‐5.8S‐ITS2 region and cox1 gene enabled its identification as a new species of clade 2, with a basal position compared to previously described subclades. The new species is described formally with the epithet Phytophthora oleae, referring to the natural matrix from which it was isolated. A unique combination of molecular and morphological characters clearly separates P. oleae from other already described Phytophthora species. This new species produced semipapillate, occasionally bipapillate, persistent sporangia on simple sympodially branching sporangiophores as well as globose and smooth‐walled oogonia, paragynous antheridia and spherical, plerotic oospores. The pathogenicity of P. oleae was confirmed in inoculation trials on fruits of three olive cultivars, including the two local cultivars from which the pathogen had been isolated.     

Fusarium miscanthi and other Fusarium species as causal agents of Miscanthus × giganteus rhizome rot

In 2010, Miscanthus × giganteus rhizomes were collected from Belgian fields showing emergence reduction and were analyzed for detection and identification of fungal species. All rhizomes were infected by at least one Fusarium species, of which F. culmorum, F. miscanthi, F. crookwellense and F. graminearum were the most abundant. Results of the pathogenicity tests by soil inoculation demonstrate the ability of F. miscanthi, F. culmorum and F. avenaceum to cause rhizome rot disease under greenhouse conditions. This is the first report of F. miscanthi and other Fusarium species isolated from M. × giganteus rhizomes in Belgium.     

Morphological and genetic variability of local Echinochloa accessions and the link with herbicide sensitivity

Echinochloa crus‐galli and Echinochloa muricata are common weeds in Belgian maize fields. Both species are morphologically difficult to distinguish and exhibit high morphological variability. Their response to herbicides varies from field to field. This study investigated whether the considerable morphological polymorphism found among Belgian Echinochloa accessions has a genetic background and whether it is consistently associated with differences in sensitivity to maize herbicides. For this purpose, accessions of E. crus‐galli and E. muricata were compared for morphological and genetic resemblance and tested for herbicide sensitivity. All accessions were planted in the field to examine the morphological traits. A cluster analysis was conducted to assess them for morphological diversity. DNA of leaf material was used for amplified fragment length polymorphism analysis to cluster the accessions genetically. Dose–response pot experiments were conducted in the glasshouse to assess the effectiveness of an acetolactate synthase (nicosulfuron), acetyl‐CoA carboxylase (cycloxydim) and 4‐hydroxyphenyl phosphate dioxygenase (topramezone) inhibiting herbicide. The genetic and morphological clusters were compared with the effective doses obtained from the dose–response bioassays. Morphological variation significantly correlated with genetic variation, but the relation with herbicide sensitivity was weak. Spikelet size and biomass characteristics are reliable discriminating characteristics for (sub)species classification. Intraspecies identification does not seem essential for optimisation of chemical control of E. crus‐galli and E. muricata in the field.     

A new stem nematode associated with peanut pod rot in China: morphological and molecular characterization of Ditylenchus arachis n. sp. (Nematoda: Anguinidae)

Surveys conducted in peanut production areas of China revealed peanut pod rot in several fields in Shandong and Hebei Provinces, China. A large quantity of an unknown stem nematode was isolated from the hulls and seeds of peanuts, herein described as Ditylenchus arachis n. sp. The new species is characterized by a combination of the following features: lateral lip sectors distinctly projected, stylet delicate, 8·4–10 μm in length, six lines in the lateral field, tail elongate–conoid, bursa covering about 68–86% of tail length. Pathogenicity tests showed that D. arachis n. sp. could infect peanut (Arachis hypogaea), but not sweet potato (Ipomoea batatas) or potato (Solanum tuberosum). Morphologically, D. arachis n. sp. appears closest to D. africanus, D. myceliophagus and D. destructor, but can be differentiated based upon a combination of morphological characteristics, host preference and molecular sequence data. The results of the phylogenetic analysis, based on 18S rDNA, the D2–D3 expansion region of 28S rDNA, and the ITS1–5·8S–ITS2 region, confirmed its status as a new species. A sister relationship with D. destructor was appointed, rather than with its ecologically very similar congener D. africanus.     

Lateral transfer of a phytopathogenic symbiont among native and exotic ambrosia beetles

Different ambrosia beetle species can coexist in tree trunks, where their immature stages feed upon symbiotic fungi. Although most ambrosia beetles are not primary pests and their fungal symbionts are not pathogenic to the host tree, exceptional situations exist. Notably, Xyleborus glabratus carries a phytopathogenic symbiont, Raffaelea lauricola, which causes laurel wilt, a lethal disease of some Lauraceae species. Both X. glabratus and R. lauricola are natives of Asia that recently invaded much of the coastal plain of the southeastern USA. This study examined ambrosia beetles that breed in susceptible trees in Florida (USA), including avocado (Persea americana), redbay (P. borbonia) and swampbay (P. palustris). Raffaelea lauricola was recovered from six of eight ambrosia beetle species that emerged from laurel wilt‐affected swampbay trees, in addition to X. glabratus. Controlled infestations with cohorts of the six species other than X. glabratus revealed that each could transmit the pathogen to healthy redbay trees and two could transmit the pathogen to healthy avocado trees; laurel wilt developed in five and one of the respective beetle × host interactions. These results indicate flexibility in the lateral transfer of a non‐native ambrosial fungus to other ambrosia beetles, and for the first time documents the transmission of a laterally transferred phytopathogenic symbiont by new ambrosia beetle species. Additional work is needed to determine whether, or to what extent, the new beetle × R. lauricola combinations play a role in spreading laurel wilt.     

Heritability of Ulmus minor resistance to Dutch elm disease and its relationship to vessel size,but not to xylem vulnerability to drought

Dutch elm disease (DED) is a vascular wilt disease that causes the occlusion and cavitation of xylem vessels. Therefore, it is hypothesized that those elms that are less vulnerable to cavitation by drought might be more resistant to DED. To test this hypothesis, the relationship between xylem vulnerability to cavitation and susceptibility to DED was examined in progenies of crosses between susceptible and resistant individuals of Ulmus minor. Hydraulic conductivity and xylem vulnerability curves were evaluated and anatomical features such as vessel size, length and grouping were measured. Next, elms were inoculated with Ophiostoma novo‐ulmi, the cause of DED, and pre‐dawn and midday water potentials, stomatal conductance and wilting percentages were assessed. Progenies of R × R crosses showed significantly lower mean wilting percentages (30–50%) than the progeny of S × S crosses (75%). Fifty percent conductivity loss was reached at c. ?1 MPa, pointing out a high vulnerability of this species to drought‐induced cavitation. Crown wilting percentage as a result of inoculation and xylem vulnerability to cavitation by water stress did not show any significant correlation. Nevertheless, significant differences in theoretical hydraulic conductivity and vessel size parameters (diameter, length and size distributions) were found among the tested progenies. Susceptible trees had significantly wider and longer vessels. Xylem structure of resistant elms seems to restrict pathogen spread rather than prevent cavitation.     

Phytophthora acerina sp. nov., a new species causing bleeding cankers and dieback of Acer pseudoplatanus trees in planted forests in northern Italy

A severe dieback of Acer pseudoplatanus trees was noticed in planted forest stands in northern Italy in 2010. Affected trees showed collar rot and aerial bleeding cankers along the stems, leading to crown dieback and eventually death. An unknown Phytophthora species was consistently isolated from necrotic bark and xylem tissue and from rhizosphere soil. Based on its unique combination of morphological and physiological characters and phylogenetic analysis, this new taxon is here described as Phytophthora acerina sp. nov. Phylogenetic analysis of ITS, cox1 and β‐tubulin gene regions demonstrated that P. acerina is unique and forms a separate cluster within the ‘P. citricola complex’, closely related to P. plurivora. Phytophthora acerina is homothallic with smooth‐walled oogonia, thick‐walled, mostly aplerotic oospores with a high abortion rate, paragynous antheridia, and persistent, morphologically variable semipapillate sporangia. Four to 5‐week‐old cultures produced globose to subglobose, appressoria‐like and coralloid hyphal swellings and characteristic stromata‐like hyphal aggregations. Optimum and maximum temperatures for growth are 25°C and 32°C, respectively. Genetic uniformity of all 15 studied isolates and the apparent absence of this species in the extensive surveys of nurseries, forests and seminatural ecosystems conducted in the previous two decades across Europe indicate a recent clonal introduction to northern Italy. Under‐bark inoculation tests demonstrated high aggressiveness of P. acerina to A. pseudoplatanus indicating that this pathogen might be a serious risk to maple plantations and forests in Europe.     

Effect of cutting frequency on above‐ and belowground biomass production of Rumex alpinus,R. crispus,R. obtusifolius and the Rumex hybrid (R. patienta × R. tianschanicus) in the seeding year

Rumex species are important weeds in grasslands and on arable land. The Rumex hybrid (R. patienta × R. tianschanicus; cv. OK‐2, Uteusha) has been planted as a forage and energy crop since 2001 in the Czech Republic, but its ecological requirements and its potential to become a new weedy species have never been investigated. In 2010 and 2011, we performed a pot experiment to investigate the effect of none, one and two cuts per year on biomass production of Rumex OK‐2 and common broad‐leaved Rumex species (Rumex obtusifolius, R. crispus and R. alpinus). The higher cutting frequency can reduce the belowground biomass, but no effect on the aboveground biomass was detected. Flowering in the seeding year was recorded in only 50% of R. obtusifolius plants. Non‐flowering R. obtusifolius plants produced significantly more belowground biomass than flowering plants under no cutting or one cut treatments. The growth response of Rumex OK‐2 to different cutting treatments was very similar to R. crispus. These similarities indicate the weed potential of the hybrid to become a troublesome weedy species, similar to R. crispus.     

Morphological variation of Solanum elaeagnifolium in south‐eastern Australia

Solanum elaeagnifolium (silverleaf nightshade) is an invasive perennial weed in Australia, with aerial growth commencing in spring from either the perennial root system or the soil seedbank, with senescence occurring in autumn. A total of 642 S. elaeagnifolium individuals were collected at flowering from 92 locations in south‐eastern Australia to study morphological variation and its implications for management. Large morphological variation was found between individuals from different locations. Leaf length, width and area ranged from 1.44 to 10.6 cm, 0.39 to 4.09 cm and 0.41 to 25.8 cm² respectively. Plants from higher rainfall regions were significantly taller and had larger leaves, suggesting a possible correlation between rainfall and morphology. Scanning electron microscopy comparison of leaf surfaces showed lower trichome and stomatal densities on the adaxial surface (67.0 ± 3.3 trichomes mm^?2 and 603.4 ± 29.2 stomata mm^?2 respectively) than on the abaxial surface (131.9 ± 7.2 trichomes mm^?2 and 813.7 ± 30.5 stomata mm^?2 respectively). The morphological plasticity of S. elaeagnifolium highlighted in this study could probably contribute to its adaptability and partly explain its establishment and continuing expansion in Australia.     

Emergence of four weed species in response to rainfall and temperature

The key to achieving effective weed control is to target them when they are young. Understanding their temperature and moisture requirements for emergence will help growers to develop timely control strategies. This study examined the emergence requirements of four key species: Conyza bonariensis, Sonchus oleraceus, Echinochloa colona and Chloris virgata. Two experiments were conducted in a growth room with temperatures of 25/15 and 30/20°C day/night with a photoperiod of 12 h. The first experiment examined emergence under single rainfall events of 0, 2, 5, 10, 20, 30 and 50 mm. The second experiment examined emergence under accumulated rainfall events of 0 mm, 5 mm × 2 (consecutive) days, 5 mm × 4 days, 5 mm × 6 days, 10 mm × 1 day, 10 mm × 2 days and 10 mm × 3 days. The temperature was not significant on emergence in both experiments. In the first experiment, at least 10 mm of rainfall was required for the emergence of all the species, with significantly higher germination rates at rainfall amounts of >20 mm. In the second experiment, the species had higher emergence rates with accumulated rainfall over consecutive days, rather than single rainfall events of the same amount. The length of time that the soil surface remained above the permanent wilting point, allowing the imbibition of water, was also an important factor. This study has provided information on the drivers for emergence of these species and will assist growers in determining what rainfall events are likely to trigger emergence in the field and make timely weed management decisions.     

Assessment of inoculation methods for screening black alder resistance to Phytophthora ×alni

Identification of resistance to Phytophthora ×alni could provide the basis for a management strategy against alder decline in riparian ecosystems in Europe. This study aimed to test methods to evaluate the resistance of riparian alders to the disease, and to screen alder genotypes for resistance. Phytophthora ×alni isolates were compared for their aggressiveness (lesion length on stem) and sporulation capacity (sporangia). While no difference in lesion lengths was found between isolates, sporangia production was dependent on isolate, highlighting the need for careful selection of isolates used for zoospore inoculation methods. Inoculation tests carried out at different periods of the year revealed a seasonal change in susceptibility to the disease, with the period from June to September being the most efficient for inoculation tests. Stem‐wounded inoculations, carried out on excised shoots, were unreliable for evaluating the level of resistance of alder genotypes to P. ×alni infection, with divergent results between two successive years or between two inoculation periods during the same year. In contrast, a method that mimics the natural conditions of infection, based on flooding of rooted cuttings in artificially infected river water, was found promising. Another method, based on the inoculation of foliated terminal shoots with zoospore suspensions, was found to be repeatable and could be used for high‐throughput analyses. Altogether, the results show a continuous resistance response from highly susceptible to moderately resistant genotypes. This suggests that breeding might be a useful strategy to manage alder decline caused by P. ×alni.     

Mechanism of herbicidal activity of a new cyclohexane-1,3-dione, tepraloxydim to Poa annua L.

Tepraloxydim [(EZ)‐(RS)‐2‐{1‐[(2E)‐3‐chloroallyloxyimino]propyl}‐3‐hydroxy‐5‐perhydropyran‐4‐ylcyclohex‐2‐en‐1‐one] showed high activity against annual bluegrass (Poa annua L.), which is relatively tolerant to sethoxydim [(±)‐2‐(1‐ethoxyiminobutyl)‐5‐[2‐(ethylthio)propyl]‐3‐hydroxycyclohex‐2‐en‐1‐one]. Absorption and translocation rates of tepraloxydim and sethoxydim were higher in P. annua than in Setaria faberi, but the absorption and translocation patterns of tepraloxydim in the two plants were similar to those of sethoxydim. Metabolic rates of tepraloxydim and sethoxydim in P. annua and S. faberi were found to be similar. The concentration for 50% inhibition (I₅₀) of acetyl‐coenzyme A carboxylase (ACCase) with tepraloxydim was approximately 3 × 10^?6 mol L^?1 for P. annua and 7 × 10^?7 mol L^?1 for S. faberi. For sethoxydim, the I₅₀ was found to be 2 × 10^?6 mol L^?1 with the enzyme of S. faberi, while sethoxydim showed a slight effect on ACCase from P. annua activity, even at 10^?4 mol L^?1. The strong inhibition of ACCase with tepraloxydim is considered to be the major factor contributing to the high herbicidal activity against P. annua. Measuring the whole plant growth response, the ratio of the tepraloxydim I₅₀ dose of P. annua to that of S. faberi (P/S) was found to be 2.4, while the P/S ratio of sethoxydim and a tepraloxydim analog with a propyl chain at R₂ were 56.3 and 73.3, respectively. The herbicidal activity against P. annua was remarkably influenced by the length of the R₂ alkyl chain, while the effect on S. faberi was not affected. Acetyl‐coenzyme A carboxylase from P. annua also exhibited a higher resistance to the tepraloxydim analog with a propyl chain than to tepraloxydim. These results suggest that a binding site structure of cyclohexane‐1,3‐diones in the ACCase differs between P. annua and S. faberi.     

Pythium yorkensis sp. nov., a potential soybean pathogen from southeastern Pennsylvania,USA

Biodiversity surveys of natural as well as agricultural ecosystems commonly reveal novel isolates. A new species, Pythium yorkensis sp. nov., is reported here, isolated from field soil during a survey of oomycete diversity in symptomless soybean across southeastern Pennsylvania. Molecular data from both mitochondrial and nuclear loci support a unique phylogenetic position for the isolates collected, and morphological features distinguish this new species from close relatives in Pythium clade J. Pathogenicity assays suggest that this new species may be a potential agricultural pathogen, as isolates were mildly aggressive on soybean. This study highlights the continued need for culture‐based surveys in collaboration with high‐throughput sequence‐based approaches for environmental sampling.     

Effect of methoprene on the progeny production of Tribolium castaneum (Coleoptera: Tenebrionidae)

BACKGROUND: Tribolium castaneum (red flour beetle) is a serious insect pest of stored products around the world. Current control measures for this species have several limitations: loss of registration of insecticides, insecticide resistance and consumer concerns about chemical residues in food. The objective of this study was to determine whether methoprene affects progeny production of T. castaneum. Late‐instar larvae or young adults were exposed to methoprene‐treated wheat, and progeny production was determined. The pairing of male and female adults was performed as untreated × untreated, treated × untreated or treated × treated, to study sex‐based effects. RESULTS: There were three outcomes to late‐instar larvae held on methoprene‐treated wheat kernels (0.001 and 0.0165 ppm): (1) failure to emerge as an adult; (2) emergence as an adult, and almost no offspring produced; (3) emergence as an adult and normal production of offspring. Male larvae were more susceptible to methoprene than female larvae. In contrast, young adults exposed to methoprene (1.67–66.6 ppm) showed no reduction in offspring production. CONCLUSION: Methoprene concentrations will decline with time following its application. However, this research indicates that methoprene can still reduce populations of T. castaneum by reducing their progeny production, even if adults emerge. Copyright © 2011 Society of Chemical Industry