Design of a Near Optimum Regulator for a Power Plant

For systems which can be decomposed into slow and fast subsystems, a near optimum linear state regulator consisting of two subsystem regulators can be developed. Depending upon the desired criteria, either a short term (fast controller) or a long term controller (slow controller) can be easily designed with minimum computational costs. Using this approach an example of a power system supplying a cyclic load is studied and the performance of the different controllers are compared.

Refining the process of agent selection through understanding plant demography and plant response to herbivory

Understanding plant demography and plant response to herbivory is critical to the selection of effective weed biological control agents. We adopt the metaphor of 'filters' to suggest how agent prioritisation may be improved to narrow our choices down to those likely to be most effective in achieving the desired weed management outcome. Models can serve to capture our level of knowledge (or ignorance) about our study system and we illustrate how one type of modelling approach (matrix models) may be useful in identifying the weak link in a plant life cycle by using a hypothetical and an actual weed example (Parkinsonia aculeata). Once the vulnerable stage has been identified we propose that studying plant response to herbivory (simulated and/or actual) can help identify the guilds of herbivores to which a plant is most likely to succumb. Taking only potentially effective agents through the filter of host specificity may improve the chances of releasing safe and effective agents. The methods we outline may not always lead us definitively to the successful agent(s), but such an empirical, data-driven approach will make the basis for agent selection explicit and serve as testable hypotheses once agents are released.

Relative host plant species use by the lantana biological control agent Aconophora compressa (Membracidae) across its native and introduced ranges

Aconophora compressa Walker (Hemiptera: Membracidae) was released in 1995 against the weed lantana in Australia, and is now found on multiple host plant species. The intensity and regularity at which A. compressa uses different host species was quantified in its introduced Australian range and also its native Mexican range. In Australia, host plants fell into three statistically defined categories, as indicated by the relative rates and intensities at which they were used in the field. Fiddlewood (Citharexylum spinosum L.: Verbenaceae) was used much more regularly and at higher densities than any other host sampled, and alone made up the first group. The second group, lantana (Lantana camara L.: Verbenaceae; pink variety) and geisha girl (Duranta erecta L.: Verbenaceae), were used less regularly and at much lower densities than fiddlewood. The third group, Sheena’s gold (another variety of D. erecta), jacaranda (Jacaranda mimosifolia D. Don: Bignoniaceae) and myoporum (Myoporum acuminatum R. Br.: Myoporaceae), were used infrequently and at even lower densities. In Mexico, the insect was found at relatively low densities on all hosts relative to those in Australia. Densities were highest on L. urticifolia, D. erecta and Tecoma stans (L.) Juss. ex Kunth (Bignoniaceae), which were used at similar rates to one another. It was found also on a few other verbenaceous and non-verbenaceous host species but at even lower densities. The relative rate at which Citharexylum spp. and L. urticifolia were used could not be assessed in Mexico because A. compressa was found on only one plant of each species in areas where these host species co-occurred. The low rate at which A. compressa occurred on fiddlewood in Mexico is likely to be an artefact of the short-term nature of the surveys or differences in the suites of Citharexylum and Lantana species available there. These results provide further incentive to insist on structured and quantified surveys of non-target host use in the native range of potential biological control agents prior to host testing studies in quarantine.

Modulation of arginine decarboxylase activity in cucumber (Cucumis sativus) cotyledons in short-term organ culture

Among the various amines administered to excisedCucumis sativus cotyledons in short-term organ culture, agmatine (AGM) inhibited arginine decarboxylase (ADC) activity to around 50%, and putrescine was the most potent entity in this regard. Homoarginine (HARG) dramatically stimulated (3- to 4-fold) the enzyme activity. Both AGM inhibition and HARG stimulation of ADC were transient, the maximum response being elicited at 12 h of culture. Mixing experiments ruled out involvement of a macromolecular effector in the observed modulation of ADC. HARG-stimulated ADC activity was completely abolished by cycloheximide, whereas AGM-mediated inhibition was unaffected. Half-life of the enzyme did not alter on treatment with either HARG or AGM. The observed alterations in ADC activity are accompanied by change in Km of the enzyme. HARG-stimulated ADC activity is additive to that induced by benzyladenine (BA) whereas in presence of KCl, HARG failed to enhance ADC activity, thus demonstrating the overriding influence of K+ on amine metabolism.

Improved understanding of the damage, ecology, and management of mirids and stinkbugs in Bollgard II

In recent years mirids and stinkbugs have emerged as important sucking pests in cotton. While stinkbugs are causing damage to bolls, mirids are causing damage to seedlings, squares and bolls. With the increasing adoption of Bollgard II and IPM approaches the use of broad-spectrum chemicals to kill Helicoverpa has been reduced and as a result mirids and stinkbugs are building to levels causing damage to bolls later in crop growth stages. Studies on stinkbugs by Dr Moazzem Khan revealed that green vegetable bug (GVB) caused significant boll damage and yield loss. A preliminary study by Dr Khan on mirids revealed that high mirid numbers at later growth stages also caused significant boll damage and that damage caused by mirids and GVB were similar. Mirids and stinkbugs therefore demand greater attention in order to minimise losses caused by these pests and to develop IPM strategies against these pests to enhance gains in IPM that have been made with Bt-transgenic cotton. Progress in this area of research will maintain sustainability and profitability of the Australian cotton industry. Mirid damage at early growth stages of cotton (up to squaring stage) has been studied in detail by Dr Khan. He found that all ages of mirids cause damage to young plants and damage by mirid nymphs is cumulative. Maximum damage occurs when the insect reaches the 4th and 5th nymphal stages. He also found that mirid feeding causes shedding of small and medium squares, and damaged large squares develop as ‘parrot beak’ bolls. Detailed studies at the boll stage, such as which stage of mirids is most damaging or which age boll is most vulnerable to feeding, is lacking. This information is a prerequisite to developing an IPM strategy for the pest in later crop growth stages. Understanding population change of the pest over time in relation to crop development is an important aspect for developing management strategies for the pest which is lacking for mirids in BollgardII. Predators and parasitoids are integral components of any IPM system and play an important part in regulating pest populations. Some generalist predators such as ants, spiders, damsel bugs and assassin bugs are known to predate on mirids. Nothing is known about parasitoids of mirids. Since green mirid (GM), Creontiades dilutus, is indigenous to Australia it is likely that we have one or more parasitoids of this mirid in Australia, but that possibility has not been investigated yet. The impact of the GVB adult parasitoid, Trichopoda giacomelli, has been studied by Dr Khan who found that the fly is established in the released areas and continues to spread. However, to get wider and greater impact, the fly should be released in new locations across the valleys. The insecticides registered for mirids and stinkbugs are mostly non-selective and are extremely disruptive to a wide range of beneficial insects. Use of these insecticides at stage I and II will minimise the impact of existing IPM programs. Therefore less disruptive control tactics including soft chemicals for mirids and stinkbugs are necessary. As with soft chemicals, salt mixtures, biopesticides based on fungal pathogens and attractants based on plant volatiles may be useful tools in managing mirids and stinkbugs with less or no disruption. Dr Khan has investigated salt mixture against mirids and GVB. While salt mixtures are quite effective and less disruptive, they are quite chemical specific. Not all chemicals mixed with salt will give the desired benefit. Therefore further investigation is needed to identify those chemicals that are effective with salt mixture against mirids and 3 of 37 GVB. Dr Caroline Hauxwell of DPI&F is working on fungal pathogen-based biopesticides against mirids and GVB and Drs Peter Gregg and Alice Del Socorro of Australian Cotton CRC are working on plant volatile-based attractants against mirids. Depending on their findings, inclusion of fungal-based biopestcides and plant volatile-based attractants in developing a management system against mirids and stinkbugs in cotton could be an important component of an IPM approach.

CRC50089 Grain Insect Ecology

Resistance to phosphine in target pests threatens market access for Australian grain. While the grains industry is now attempting to develop an effective and sustainable strategy to manage this resistance, action is severely limited by significant gaps in our knowledge of the key ecological factors that influence the development of resistance. There is a need to research this information as a foundation for a rational approach to managing phosphine resistance in the Australian grains industry. Research outcomes: The project has provided critical research methodologies and preliminary data to fill the large gaps in our knowledge of the ecology of two key pests, Rhyzopertha dominica and Tribolium castaneum, and how this may drive the development of phosphine resistance. This information will contribute to the groundwork for future research needed to provide a scientific basis for a rational resistance management strategy.

Fen ecosystem carbon gas dynamics in changing hydrological conditions

Northern peatlands are thought to store one third of all soil carbon (C). Besides the C sink function, peatlands are one of the largest natural sources of methane (CH4) to the atmosphere. Climate change may affect the C gas dynamics as well as the labile C pool. Because the peatland C sequestration and CH4 emissions are governed by high water levels, changes in hydrology are seen as the driving factor in peatland ecosystem change. This study aimed to quantify the carbon dioxide (CO2) and CH4 dynamics of a fen ecosystem at different spatial scales: plant community components scale, plant community scale and ecosystem scale, under hydrologically normal and water level drawdown conditions. C gas exchange was measured in two fens in southern Finland applying static chamber and eddy covariance techniques. During hydrologically normal conditions, the ecosystem was a CO2 sink and CH4 source to the atmosphere. Sphagnum mosses and sedges were the most important contributors to the community photosynthesis. The presence of sedges had a major positive impact on CH4 emissions while dwarf shrubs had a slightly attenuating impact. C fluxes varied considerably between the plant communities. Therefore, their proportions determined the ecosystem scale fluxes. An experimental water level drawdown markedly reduced the photosynthesis and respiration of sedges and Sphagnum mosses and benefited shrubs. Consequently, changes were smaller at the ecosystem scale than at the plant group scale. The decrease in photosynthesis and the increase in respiration, mostly peat respiration, made the fen a smaller CO2 sink. CH4 fluxes were significantly lowered, close to zero. The impact of natural droughts was similar to, although more modest than, the impact of the experimental water level drawdown. The results are applicable to the short term impacts of the water level drawdown and to climatic conditions in which droughts become more frequent.

Short-range resource location by Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) demonstrates a strong preference for fungi associated with cotton seed

Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is a common stored grain pest for which a wide range of suitable resources has been recorded. These beetles are facultatively fungivorous and their resource range may extend to fungi associated with non-grain resources (e.g. cotton seed) and other decaying plant matter. Little is known with respect to fungi in terms of resource location by these beetles in the field. We, therefore, conducted a series of experiments in laboratory arenas, glasshouse cages and the field to determine how beetles respond to grain resources in relation to cotton seed (together with its lint stubble and associated fungal flora). Results from the tests conducted in relatively small arenas and cages in the laboratory and glasshouse reveal that the responses of T. castaneum adults to food resources were twice as strong when walking as when flying (as measured by the proportion of the released beetles that were trapped). Also, a clear preference for linted cotton seeds was evident in walking T. castaneum, especially in small-scale arenas in the laboratory, where at least 60% of beetles released preferred linted cotton seeds over wheat and sorghum. Similarly, in cages (1 m3) they responded five times more strongly to linted cotton seed than to conventional grain resources. However, this pattern was not consistent with those obtained from field trapping over 20 m and the beetles did not show any particular preference to any of the resources tested above. Our results suggest a focus on walking beetles in trapping studies for population estimations and, for developing effective food-based trapping lures, the potential use of active volatiles from the fungi associated with linted cotton seed. © 2012 Elsevier Ltd.

The Significance of Wild Plants in the Evolutionary Ecology of Three Major Viruses Infecting Cultivated Sweetpotato in Uganda

The studies presented in this thesis contribute to the understanding of evolutionary ecology of three major viruses threatening cultivated sweetpotato (Ipomoea batatas Lam) in East Africa: Sweet potato feathery mottle virus (SPFMV; genus Potyvirus; Potyviridae), Sweet potato chlorotic stunt virus (SPCSV; genus Crinivirus; Closteroviridae) and Sweet potato mild mottle virus (SPMMV; genus Ipomovirus; Potyviridae). The viruses were serologically detected and the positive results confirmed by RT-PCR and sequencing. SPFMV was detected in 24 wild plant species of family Convolvulacea (genera Ipomoea, Lepistemon and Hewittia), of which 19 species were new natural hosts for SPFMV. SPMMV and SPCSV were detected in wild plants belonging to 21 and 12 species (genera Ipomoea, Lepistemon and Hewittia), respectively, all of which were previously unknown to be natural hosts of these viruses. SPFMV was the most abundant virus being detected in 17% of the plants, while SPMMV and SPCSV were detected in 9.8% and 5.4% of the assessed plants, respectively. Wild plants in Uganda were infected with the East African (EA), common (C), and the ordinary (O) strains, or co-infected with the EA and the C strain of SPFMV. The viruses and virus-like diseases were more frequent in the eastern agro-ecological zone than the western and central zones, which contrasted with known incidences of these viruses in sweetpotato crops, except for northern zone where incidences were lowest in wild plants as in sweetpotato. The NIb/CP junction in SPMMV was determined experimentally which facilitated CP-based phylogenetic and evolutionary analyses of SPMMV. Isolates of all the three viruses from wild plants were genetically similar to those found in cultivated sweetpotatoes in East Africa. There was no evidence of host-driven population genetic structures suggesting frequent transmission of these viruses between their wild and cultivated hosts. The p22 RNA silencing suppressor-encoding sequence was absent in a few SPCSV isolates, but regardless of this, SPCSV isolates incited sweet potato virus disease (SPVD) in sweetpotato plants co-infected with SPFMV, indicating that p22 is redundant for synergism between SCSV and SPFMV. Molecular evolutionary analysis revealed that isolates of strain EA of SPFMV that is largely restricted geographically in East Africa experience frequent recombination in comparison to isolates of strain C that is globally distributed. Moreover, non-homologous recombination events between strains EA and C were rare, despite frequent co-infections of these strains in wild plants, suggesting purifying selection against non-homologous recombinants between these strains or that such recombinants are mostly not infectious. Recombination was detected also in the 5 - and 3 -proximal regions of the SPMMV genome providing the first evidence of recombination in genus Ipomovirus, but no recombination events were detected in the characterized genomic regions of SPCSV. Strong purifying selection was implicated on evolution of majority of amino acids of the proteins encoded by the analyzed genomic regions of SPFMV, SPMMV and SPCSV. However, positive selection was predicted on 17 amino acids distributed over the whole the coat protein (CP) in the globally distributed strain C, as compared to only 4 amino acids in the multifunctional CP N-terminus (CP-NT) of strain EA largely restricted geographically to East Africa. A few amino acid sites in the N-terminus of SPMMV P1, the p7 protein and RNA silencing suppressor proteins p22 and RNase3 of SPCSV were also submitted to positive selection. Positively selected amino acids may constitute ligand-binding domains that determine interactions with plant host and/or insect vector factors. The P1 proteinase of SPMMV (genus Ipomovirus) seems to respond to needs of adaptation, which was not observed with the helper component proteinase (HC-Pro) of SPMMV, although the HC-Pro is responsible for many important molecular interactions in genus Potyvirus. Because the centre of origin of cultivated sweetpotato is in the Americas from where the crop was dispersed to other continents in recent history (except for the Australasia and South Pacific region), it would be expected that identical viruses and their strains occur worldwide, presuming virus dispersal with the host. Apparently, this seems not to be the case with SPMMV, the strain EA of SPFMV and the strain EA of SPCSV that are largely geographically confined in East Africa where they are predominant and occur both in natural and agro-ecosystems. The geographical distribution of plant viruses is constrained more by virus-vector relations than by virus-host interactions, which in accordance of the wide range of natural host species and the geographical confinement to East Africa suggest that these viruses existed in East African wild plants before the introduction of sweetpotato. Subsequently, these studies provide compelling evidence that East Africa constitutes a cradle of SPFMV strain EA, SPCSV strain EA, and SPMMV. Therefore, sweet potato virus disease (SPVD) in East Africa may be one of the examples of damaging virus diseases resulting from exchange of viruses between introduced crops and indigenous wild plant species. Keywords: Convolvulaceae, East Africa, epidemiology, evolution, genetic variability, Ipomoea, recombination, SPCSV, SPFMV, SPMMV, selection pressure, sweetpotato, wild plant species Author s Address: Arthur K. Tugume, Department of Agricultural Sciences, Faculty of Agriculture and Forestry, University of Helsinki, Latokartanonkaari 7, P.O Box 27, FIN-00014, Helsinki, Finland. Email: tugume.arthur@helsinki.fi Author s Present Address: Arthur K. Tugume, Department of Botany, Faculty of Science, Makerere University, P.O. Box 7062, Kampala, Uganda. Email: aktugume@botany.mak.ac.ug, tugumeka@yahoo.com

An overview on the role of Hexanchiformes in marine ecosystems: biology, ecology and conservation status of a primitive order of modern sharks

The large size, high trophic level and wide distribution of Hexanchiformes (cow and frilled sharks) should position this order as important apex predators in coastal and deep-water ecosystems. This review synthesizes available information on Hexanchiformes, including information not yet published, with the purpose of evaluating their conservation status and assessing their ecological roles in the dynamics of marine ecosystems. Comprising six species, this group has a wide global distribution, with members occurring from shallow coastal areas to depths of c. 2500 m. The limited information available on their reproductive biology suggests that they could be vulnerable to overexploitation (e.g. small litter sizes for most species and suspected long gestation periods). Most of the fishing pressure exerted on Hexanchiformes is in the form of commercial by-catch or recreational fishing. Comprehensive stock and impact assessments are unavailable for most species in most regions due to limited information on life history and catch and abundance time series. When hexanchiform species have been commercially harvested, however, they have been unable to sustain targeted fisheries for long periods. The potentially high vulnerability to intense fishing pressure warrants a conservative exploitation of this order until thorough quantitative assessments are conducted. At least some species have been shown to be significant apex predators in the systems they inhabit. Should Hexanchiformes be removed from coastal and deep-water systems, the lack of sympatric shark species that share the same resources suggests no other species would be capable of fulfilling their apex predator role in the short term. This has potential ecosystem consequences such as meso-predator release or trophic cascades. This review proposes some hypotheses on the ecology of Hexanchiformes and their role in ecosystem dynamics, highlighting the areas where critical information is required to stimulate research directions.

Origin of leaf rust adult plant resistance gene Rph20 in barley

Rph20 is the only reported, simply inherited gene conferring moderate to high levels of adult plant resistance (APR) to leaf rust (Puccinia hordei Otth) in barley (Hordeum vulgare L.). Key parental genotypes were examined to determine the origin of Rph20 in two-rowed barley. The Dutch cultivar 'Vada' (released in the 1950s) and parents, 'Hordeum laevigatum' and 'Gull' ('Gold'), along with the related cultivar 'Emir' (a derivative of 'Delta'), were assessed for APR to P. hordei in a disease screening nursery. The marker bPb-0837-PCR, co-located with Rph20 on the short arm of chromosome 5H (5HS), was used to screen genotypes for the resistance allele, Rph20.ai. Results from phenotypic assessment and DNA analysis confirmed that Rph20 originated from the landrace 'H. laevigatum' (i.e., Hordeum vulgare subsp. vulgare). Tracing back this gene through the pedigrees of two-rowed barley cultivars, indicated that Rph20 has contributed APR to P. hordei for more than 60 years. Although there have been no reports of an Rph20-virulent pathotype, the search for alternative sources of APR should continue to avoid widespread reliance upon a single resistance factor.

Mapping Quantitative Trait Loci for Partial Resistance to Powdery Mildew in an Australian Barley Population

Genomic regions influencing resistance to powdery mildew [Blumeria graminis (DC.) E.O. Speer f. sp. hordei Em. Marchal] were detected in a doubled haploid (DH) barley (Hordeum vulgare L.) population derived from a cross between the breeding line ND24260 and cultivar Flagship when evaluated across four field environments in Australia and Uruguay. Significant quantitative trait loci (OIL) for resistance to B. graminis were detected on six of the seven chromosomes (1H, 2H, 3H, 4H, 5H, and 7H). A QTL with large effect donated by ND24260 mapped to the short arm of chromosome 1H (1 HS) conferring near immunity to B. graminis in Australia but was ineffective in Uruguay. Three OIL donated by Flagship contributed partial resistance to B. graminis and were detected in at least two environments. These OIL were mapped to chromosomes 3H, 4H, and 5H (5HS) accounting for up to 18.6, 3.4, and 8.8% phenotypic variation, respectively. The 5HS QTL contributed partial resistance to B. graminis in all field environments in both Australia and Uruguay and aligned with the genomic region of Rph20, a gene conferring adult plant resistance (APR) to leaf rust (Puccinia hordei Otth), which is found in some cultivars having Vada' or 'Emir' in their parentage. Selection for favorable marker haplotypes within the 3H, 4H, and 5H QTL regions can be performed even in the presence of single (major) gene resistance. Pyramiding such QTL may provide an effective and potentially durable form of resistance to B. graminis.

Response of Tribolium castaneum and Rhyzopertha dominica to various resources, near and far from grain storage

Tribolium castaneum (Herbst) and Rhyzopertha dominica (F.) are common cosmopolitan pests of stored grain and grain products. We evaluated the relative attraction of T.castaneum and R.dominica to wheat, sorghum and cotton seeds in the field, near grain storage facilities and well away from storages in southern and central Queensland using multiple trapping techniques. The results show that T.castaneum is more strongly attracted to linted cotton seed relative to wheat, whereas R.dominica did not respond to cotton seed at all and was attracted only to wheat. Significantly more adults of T.castaneum (10-15 times) were attracted to traps placed on the ground, near grain storage, than to equivalent traps that were suspended (1.5m above the ground) nearby. These results suggest that Tribolium beetles detect and respond to resources towards the end of their dispersal flight, after which they localize resources while walking. By contrast R.dominica was captured only in suspended traps, which suggests they fly directly onto resources as they localize them. The ability of both species to colonize and reproduce in isolated resource patches within the relatively short time of 1month is illustrated by the returns from the traps deployed in the field (at least 1km from the nearest stored grain) even though they caught only a few beetles. The results presented here provide novel insights about the resource location behaviours of both T.castaneum and R.dominica. In particular, the relationship of T.castaneum with non-cereal resources that are not conventionally associated with this species suggests an emphasis on these other resources in investigating the resource location behaviour of these beetles. This new perspective on the ecology of T. castaneum highlights the potential role of non-cereal resources (such as the lint on cotton seed) in the spread of grain pest infestations.

Plot size matters: interference from intergenotypic competition in plant phenotyping studies

Genetic and physiological studies often comprise genotypes diverse in vigour, size and flowering time. This can make the phenotyping of complex traits challenging, particularly those associated with canopy development, biomass and yield, as the environment of one genotype can be influenced by a neighbouring genotype. Limited seed and space may encourage field assessment in single, spaced rows or in small, unbordered plots, whereas the convenience of a controlled environment or greenhouse makes pot studies tempting. However, the relevance of such growing conditions to commercial field-grown crops is unclear and often doubtful. Competition for water, light and nutrients necessary for canopy growth will be variable where immediate neighbours are genetically different, particularly under stress conditions, where competition for resources and influence on productivity is greatest. Small hills and rod-rows maximise the potential for intergenotypic competition that is not relevant to a crop’s performance in monocultures. Response to resource availability will typically vary among diverse genotypes to alter genotype ranking and reduce heritability for all growth-related traits, with the possible exception of harvest index. Validation of pot experiments to performance in canopies in the field is essential, whereas the planting of multirow plots and the simple exclusion of plot borders at harvest will increase experimental precision and confidence in genotype performance in target environments.

An inordinate fondness for Fusarium: Phylogenetic diversity of fusaria cultivated by ambrosia beetles in the genus Euwallacea on avocado and other plant hosts

Ambrosia beetle fungiculture represents one of the most ecologically and evolutionarily successful symbioses, as evidenced by the 11 independent origins and 3500 species of ambrosia beetles. Here we document the evolution of a clade within Fusarium associated with ambrosia beetles in the genus Euwallacea (Coleoptera: Scolytinae). Ambrosia Fusarium Clade (AFC) symbionts are unusual in that some are plant pathogens that cause significant damage in naive natural and cultivated ecosystems, and currently threaten avocado production in the United States, Israel and Australia. Most AFC fusaria produce unusual clavate macroconidia that serve as a putative food source for their insect mutualists. AFC symbionts were abundant in the heads of four Euwallacea spp., which suggests that they are transported within and from the natal gallery in mandibular mycangia. In a four-locus phylogenetic analysis, the AFC was resolved in a strongly supported monophyletic group within the previously described Cade 3 of the Fusarium solani species complex (FSSC). Divergence-time estimates place the origin of the AFC in the early Miocene similar to 21.2 Mya, which coincides with the hypothesized adaptive radiation of the Xyleborini. Two strongly supported clades within the AFC (Clades A and B) were identified that include nine species lineages associated with ambrosia beetles, eight with Euwallacea spp. and one reportedly with Xyleborus ferrugineus, and two lineages with no known beetle association. More derived lineages within the AFC showed fixation of the clavate (club-shaped) macroconidial trait, while basal lineages showed a mix of clavate and more typical fusiform macroconidia. AFC lineages consisted mostly of genetically identical individuals associated with specific insect hosts in defined geographic locations, with at least three interspecific hybridization events inferred based on discordant placement in individual gene genealogies and detection of recombinant loci. Overall, these data are consistent with a strong evolutionary trend toward obligate symbiosis coupled with secondary contact and interspecific hybridization. (C) 2013 Elsevier Inc. All rights reserved.