Both hypersensitive and non-hypersensitive responses are associated with resistance in Salix viminalis against the gall midge Dasineura marginemtorquens

Hypersensitivity responses (HR) play a major role in plant resistance to pathogens. It is often claimed that HR is also important in plant resistance to insects, although there is little unambiguous documentation. Large genotypic variation in resistance against the gall midge Dasineura marginemtorquens is found in Salix viminalis. Variation in larval performance and induced responses within a full-sib S. viminalis family is reported here; 36 sibling plants were completely resistant (larvae died within 48 h after egg hatch, no gall induction), 11 plants were totally susceptible, 25 plants were variable (living and dead larvae present on the same plant). Resistance was associated with HR, but to different degrees; 21 totally resistant genotypes showed typical HR symptoms (many distinct necrotic spots) whereas the remaining 15 genotypes showed no, or very few, such symptoms. Hydrogen peroxide, used as a marker for HR, was induced in genotypes expressing HR symptoms but not in resistant genotypes without symptoms, or in susceptible genotypes. These data suggest that production of hydrogen peroxide, and accompanying cell death, cannot explain larval mortality in the symptomless reaction. Another, as yet unknown, mechanism of resistance may be present. If so, then it is possible that this unknown mechanism also contributes to resistance in plants displaying HR. The apparent complexity observed in this interaction, with both visible and invisible plant responses associated with resistance against an adapted insect species, may have implications for the study of resistance factors in other plant-insect interactions.     

Rapid gall midge adaptation to a resistant willow genotype

Abstract 1 We conducted two experiments to investigate why a basket willow Salix viminalis L. genotype, known to be highly resistant to the leaf-roller gall midge Dasineura marginemtorquens (Bremi), should support very high gall densities in a field plantation at Tälle, south Sweden.
2 The first experiment was a field test of the hypothesis of fine-scale host adaptation in the gall midge/willow system. Support for the hypothesis would be established if midges originating from resistant willows and those originating from nearby susceptible willows differed in their abilities to initiate galls and complete development on resistant plants.
3 The objective of the second experiment was to explore whether there was a genetic basis to the trait for virulence in the midge population and to investigate any potential trade-offs this trait may entail.
4 Our results indicate that there was a fine-scaled microgeographic genetic structure to the midge population at Tälle. Midges originating from resistant plants had a heritable trait that enabled them to establish galls on resistant plants.
5 Midges able to initiate galls on the resistant genotype had longer developmental time on the susceptible genotype. This suggests that there is a physiological cost associated with being adapted to the resistant willow genotype.
6 We suggest that driving forces behind the observed host adaptation are selection imposed on the midge population by very strong willow resistance and restricted gene flow in the midge populations due to the special life history features of D. marginemtorquens .     

Oviposition choice and larval survival of Dasineura marginemtorquens (Diptera: Cecidomyiidae) on resistant and susceptible Salix viminalis

Abstract.

• 1 Substantial intraspecific variation exists in Salix viminalis resistance to the gall midge Dasineura marginemtorquens. Earlier work has found this variation to have a large genetic component. Willow clones are stable in their resistances between midge generations and different nutrient levels in both field and laboratory culture.
• 2 This study reports the results of laboratory experiments on female oviposition choice and larval survival on potted plants from clones that are very different in resistance as determined in field studies.
• 3 In choice experiments using pairs of plants, the average female midge did not prefer susceptible willow clones over resistant ones for oviposition. In about one third of the replicates, midges actually laid more eggs on the resistant clone. Further work is necessary to examine the nature of variation among midges in discrimination of these plant types.
• 4 Resistance is manifested as great differences in larval survival. Six days after oviposition survival was 92% on susceptible plants but only 6% on resistant ones. Galls developed on all of the susceptible plants, while in 73% of the resistant plants galls were not even initiated.
• 5 The plant traits causing resistance are enigmatic. Larval behaviour suggests that resistant plants interfere with feeding behaviour. On resistant plants, most larvae wander for more than 24 h without initiating any galls before dying. On susceptible plants many first instar larvae begin feeding and initiate galls within this period.

     

Absence of geographical structure of chloroplast DNA variation in sallow, Salix caprea L

In the present study, we have used PCR-RFLP markers to investigate the chloroplast DNA variation in 24 European populations of Salix caprea L. A subset of these populations has also been analysed with chloroplast microsatellites. The main feature of both markers is the absence of a clear geographic structure (G(ST(PCR-RFLP))=0.090, G(ST(microsatellites))=-0.017) and high levels of variation within populations. This lack of phylogeographic structure in S. caprea is suggested to be the consequence of the joint action of several factors: (i) presence of intermediate latitude refugia with large population sizes during the last glacial maximum, (ii) a high speed of recolonisation and dispersal ability, (iii) a high mutation rate and (iv) extensive hybridisation with other willow species. In addition to the S. caprea samples, a limited number of individuals from several other Salix species were also analysed with PCR-RFLP: S. cinerea, S. aurita, S. purpurea, S. atrocinerea, S. appendiculata, S. elaeagnos, S. fragilis and S. alba. Many of the haplotypes found in Salix caprea were also detected in S. cinerea, S. aurita, S. purpurea, S. atrocinerea and/ or S. appendiculata but not in S. alba, S. elaeagnos or S. fragilis. Our data suggest that hybridisation and gene flow have occurred within these two groups but not between them.     

Host–pathogen relationship between Salix and Melampsora sheds light on the parentage of some biomass willows

The association between willow ( Salix ) and rust ( Melampsora ) is highly specific. Willows named Salix burjatica , S. dasyclados ( S. × dasyclados ) and S. × calodendron are important in renewable-energy plantations in the UK and western Europe. There has been much controversy over their origin, species status and nomenclature. It has been suggested that they have originated from hybridization between. S. caprea , S. viminalis and S. cinerea . In the present work, 59 willow clones were investigated through morphological examination and detached leaf inoculation using willow differentials, for their association, in southwest England, with M. capraearum and three pathotypes of Melampsora epitea (Me-A, B and C). M. capraearum was found on all clones of S. caprea and its hybrids with S. aurita ; Me-A on all S. viminalis clones; Me-B on wild S. cinerea , S. × calodendron , S. × dasyclados 'De Biardii 445' and S . 'Spaethii'; Me-C on all S. burjatica clones and most S. × dasyclados clones. Both M. caprearum and Me-A infected all S. × sericans ( S. caprea × viminalis ) clones and S. × dasyclados 'LA041/03'. We suggest that S. × dasyclados 'LA041/03' should be treated as S. × sericans ( S. caprea × S. viminalis ); S. burjatica , S. dasyclados and S. × dasyclados as synonyms; S. × dasyclados 'De Biardii 445' as S. × calodendron 'De Biardii 445'; and S . 'Spaethii' as S. × calodendron 'Spaethii'.     

Life history strategy and taxonomic position of gall midges (Diptera: Cecidomyiidae) inducing leaf galls on Styrax japonicus (Styracaceae)

Four gall midge species (Diptera: Cecidomyiidae) that induce leaf galls on Styrax japonicus (Styracaceae) were identified to generic level based on larval morphology. Three of these gall midges, which induce whitish hemiglobular galls, flattened subglobular galls, and purple globular galls, respectively, were identified as three genetically distinct species of Contarinia, and the remaining species, which induces globular galls with dense whitish hairs, was identified as a species of Dasineura. Field surveys in Fukuoka, Japan, revealed that adults of these gall midges emerged and oviposited in late March to mid‐April at Mount Tachibana (approximately 200 m a.s.l.) and in late April to early May at Mount Sefuri (about 1050 m a.s.l.), coinciding with the leaf‐opening season of S. japonicus. Larvae of these gall midges mostly developed into third instars by June and then left their galls and dropped to the ground. These species therefore have a life history strategy that differs from that of another S. japonicus‐associated gall midge, Oxycephalomyia styraci, which overwinters as the first instar in ovate swellings, matures rapidly in spring, and emerges directly from the galls.     

Nine new species of Dasineura (Diptera: Cecidomyiidae) from flowers of Australian Acacia (Mimosaceae)

Abstract. Thirteen species of Australian acacias are invasive plants in agricultural and native vegetation areas of South Africa. Biological control programmes for Australian acacias in South Africa have been implemented and are aimed at suppressing reproductive vigour and, in some cases, vegetative growth of these weeds. Gall-forming midges are under consideration as potential biological control agents for invasive acacias in South Africa. Entomological surveys in southern Australia found a diverse cecidomyiid fauna associated with the buds, flowers and fruits of Acacia species. Nine new Dasineura species are described and two species, D. acaciaelongifoliae (Skuse) and D. dielsi Rübsaamen, are redescribed. The newly described taxa are D. fistulosa sp.n. , D. furcata sp.n. , D. glauca sp.n. , D. glomerata sp.n. , D. oldfieldii sp.n. , D. oshanesii sp.n. , D. pilifera sp.n. , D. rubiformis sp.n. and D. sulcata sp.n. All eleven species induce galls on ovaries and prevent the formation of fruit. Two general types of gall are caused. Type A comprises woody, tubular galls with larvae living inside ovaries (D. acaciaelongifoliae, D. dielsi, D. fistulosa, D. furcata, D. glauca, D. glomerata, D. oldfieldii). Type B includes soft-tissued, globose galls that belong to four subtypes: inflated, baglike, hairy galls with larvae living between ovaries (D. pilifera); pyriform, pubescent swellings with larvae living inside ovaries (D. rubiformis); globose, hairy, swellings with larvae living superficially on ovaries in ovoid chambers (D. oshanesii); and inconspicuous, glabrous swellings with larvae living superficially on ovaries in shallow groovelike chambers (D. sulcata). The gall types are associated with a particular pupation pattern. In type A galls, larvae pupate within larval chambers in galls, whereas in type B galls pupation takes place between ovaries in galls or in the soil beneath the host tree. Gall midges responsible for the same general gall type are morphologically related and differ from species causing the other gall type. Phylogenetic analysis of a 410 bp fragment of the mitochondrial cytochrome b gene supports the division of the gall midge species into two groups except for D. sulcata, which appears as a subgroup of the group causing type A galls. The interspecific divergence values in group A species were between 0.5 and 3.9% with intraspecific divergence estimates of 0–0.2%. Gall midges causing type B galls had interspecific divergence values of 4.6–7.3% and intraspecific divergence values of 0–3.7%. Closely related biology and morphology together with low cytochrome b divergence estimates suggest a more recent speciation in group A when compared with species of group B. Dasineura rubiformis and D. dielsi are proposed as potential biological control agents for Acacia mearnsii De Wild. and Acacia cyclops A. Cunn. ex G. Don, respectively, in South Africa due to their narrow host range and ability to form high population densities that reduce seed formation. Both species produce galls with low biomass, which makes them compatible with commercial exploitation of their host species in Africa.     

Strategies for the biological control of invasive willows (Salix spp.) in Australia

Abstract  Willows ( Salix spp.) are Weeds of National Significance in Australia where a large number of taxa are naturalised in temperate regions and can cause serious environmental degradation of riparian and wetland habitats. Several species are of economic or ornamental value and conflicts of interest could arise with planning their suppression. Biological control of six willow species ( S. alba L., S. cinerea L . , S. fragilis L., S. nigra Marshall, S. viminalis L., S . x  rubens Schrank) is under consideration in Australia using three approaches based on targeting different morphological structures. A rich phytophagous biota associated with Salix in their regions of origin indicate a range of organisms are available for development as biological control agents. Implementation strategies will depend on the host species targeted for biological control, their reproductive strategies, the level of control required to reduce environmental impacts and the level of conflict of interest associated with a biological control program.     

Interaction between relative humidity and resistance to Dasineura tetensi in blackcurrant

1 Infestation by the blackcurrant leaf midge Dasineura tetensi and galling incidence on a susceptible (cv. ‘Öjebyn’) and a moderately resistant (cv. ‘Storklas’) blackcurrant genotype was studied in the field for two midge generations in the same growing season. On the resistant genotype gall initiation is delayed. 2 The relation between infestation and galling incidence showed considerable variation between the two generations. Galling symptoms produced by the second midge generation were weak on the resistant as compared with the susceptible blackcurrant. 3 Because larval development of that generation coincided with a period of dry and warm weather, it is possible that larvae on the resistant genotype suffered desiccation to a greater extent than larvae on the susceptible genotype where gall development was stronger. 4 The possible interaction of relative humidity and expression of resistance was investigated in controlled environment experiments. The resistant ‘Storklas’ and a susceptible (‘7801–31’) currant genotype were studied at two constant relative humidities, 30 and 70%. 5 Larvae of D. tetensi suffered from higher mortality and reduced growth rate on both genotypes in the low humidity environment. 6 There was also a significant plant genotype by humidity interaction on larval performance; no galls were produced and no larvae completed development on the resistant currant at low humidity. 7 There was a trend for a positive correlation between larval length and larval density on a plant at low humidity but not at high humidity.     

Vole preference for Salix caprea, S. repens, and their F1, F2, and backcross hybrids

In recent years, the influence of plant hybridization on plant–animal interactions has become an important issue; however, these studies have mainly focused on insects or slugs and to a large degree ignored mammalian herbivores despite their importance in many ecosystems. This study aims to determine the preference of voles for pure Salix caprea , S. repens , and their F1, F2, and backcross hybrids to evaluate whether voles select against hybrids in natural willow populations. To address this, we conducted two field studies and a cafeteria experiment in the laboratory with bank voles ( Clethrionomys glareolus ). We found no strong indications of reduced resistance in any of the hybrid taxa. Voles damaged more S. repens than S. caprea individuals. There was a general decreasing trend of utilization from pure S. repens to pure S. caprea , hybrid taxa being intermediate between the parents. Thus, voles seemingly do not have a more detrimental effect on hybrid fitness than on the fitness of pure individuals and probably will not select against hybrids in these willows populations.     

Biotypes of Dasineura tetensi, differing in ability to gall and develop on black currant genotypes

Variation in damage levels on certain black currant, Ribes nigrum L., genotypes, caused by the black currant leaf midge, Dasineura tetensi (Rübs.) (Diptera: Cecidomyiidae), has been observed in northern Sweden. I investigated whether this variation is due to variation in virulence among midges. From a field population of midges, I successfully selected for virulence and avirulence, respectively, on the resistant black currant genotype cultivar `Storklas' (called resistant genotype). The performance of avirulent and virulent midge larvae on two black currant genotypes were studied in experiments where first or second instar larvae were artificially transferred. There were no differences in larval survival and developmental rate between the two midge types when transferred to the susceptible currant genotype `7801–31' (called susceptible genotype). Larvae of the virulent strain established galls and developed on `Storklas' but development was initially slower there than on the susceptible currant genotype. Larvae of the avirulent strain suffered high mortality or remained in first instar on that same currant genotype when transferred alone, but developed readily if transferred together with virulent larvae. Larvae transferred in second instar to host plants susceptible to the larvae resumed feeding and developed further to maturity. Second instar larvae were also able to establish new galls even though these galls were not as well developed as those caused by first instar larvae. Black currant plantations in northern Sweden were surveyed and local midge populations were found to be composed of either avirulent, virulent or a mixture of both midge types. Virulent midges were not restricted to plantations where resistant currant genotypes were grown. I conclude that, at least, two biotypes of the midge exist, and that those two are distinguished by the ability to gall and survive on `Storklas'.     

Preference–performance relationship in the gall midge Rabdophaga rosaria: insights from a common‐garden experiment with nine willow clones

1. Oviposition preferences of herbivorous insects are predicted to match offspring performance on different host taxa or on conspecific host genotypes. In gall‐inducing insects, host‐plant properties such as growth rate and gall size, which are determined by plant genotype and growing conditions, may have a significant impact on offspring performance and, hence, should influence oviposition site selection. 2. The present study investigated host preference of the European rosette willow gall midge Rabdophaga rosaria (Loew) in relation to offspring success on seven clones of Salix myrsinifolia Salisb. and two naturally hybridised S. myrsinifolia × phylicifolia L. clones growing in a replicated design in an experimental field under two fertilisation regimes. For each clone, the average growth rate, number of shoot tips, and leaf and gall size were determined, and their effects on midge preference and larval survival were examined. 3. Main shoot height, number of shoot tips, and gall size were significantly affected by clone. The midges clearly preferred certain clones over the others, but preferences were not related to willow growth traits or to gall size. Survival probability was higher in large than in small galls, but females did not prefer large‐leaved clones that produced the biggest rosette galls. Midge oviposition was also uncorrelated with prior rates of leaf‐rust infection and with feeding preferences of voles and folivorous insects. 4. The weak preference–performance relationship of R. rosaria within S. myrsinifolia is probably explained by evolutionary constraints that prevent generalist insects from achieving an ability to discriminate among conspecific hosts of variable quality.     

An experimental test of hybrid resistance to insects and pathogens using Salix caprea, S. repens and their F1 hybrids

The aim of this study was to assess the responses of herbivores and pathogens to hybrid plants under controlled conditions. F1 hybrids and parental species, produced by hand-pollinating willows in the field, were potted and kept in an experimental field under controlled conditions. In 1997, plant growth and survival were measured along with densities of insects and the degree of pathogen infection on the willows. The survival rate was higher for S. repens than for the hybrids and lowest for S. caprea. Densities of the sawflies Pontania pedunculi and P. brigmanii and the leaf-galling midge Iteomyia capreae were higher on hybrids and on S. caprea than on S. repens. The densities of Crepidodera fulvicornis (Chrysomelidae), chrysomelid larvae and the bud-galling midge Dasineura rosaria did not differ between any of the plant categories. Hybrids were more severely infected by rust (Melampsora sp.) than S. caprea and the totally resistant S. repens. Densities of herbivores on hybrid willows were consistent with the dominance hypothesis (i.e. herbivore densities were similar to densities on one of the parental species) or supported the no-difference hypothesis. Furthermore, herbivore densities on hybrid plants were most similar to densities on the more susceptible parent. The breakdown in rust resistance in hybrid plants suggests that resistance traits are severely disrupted by the genetic re-arrangement in hybrids and that this increased susceptibility could select against hybridisation. Received: 17 February 1998 / Accepted: 15 June 1998     

Distribution and phenology of Dasineura oxycoccana (Diptera: Cecidomyiidae) in Michigan blueberries

The blueberry gall midge, Dasineura oxycoccana Johnson, is a serious pest of rabbiteye blueberries in Florida, Georgia, and Mississippi, and a potential pest of southern and northern highbush blueberries. Its damage has been observed with increasing frequency in highbush blueberry plantings in the Great Lakes region, including in Wisconsin and in Michigan. Unlike in rabbiteye blueberry plantings, where blueberry gall midge primarily damages flowering buds, it is found to damage only the vegetative shoots of northern highbush blueberry. In this study, farms throughout Michigan were surveyed for the presence of blueberry gall midge and it was found in 43 of 46 sampled farms in 11 counties. From 2009-2011, several monitoring techniques, including yellow sticky traps, emergence traps, observational sampling, and vegetative shoot dissections were used to determine the ecology of this species in blueberry fields in southwest Michigan. Emergence traps were most useful in early detection of blueberry gall midge in April, and observational sampling for damage symptoms and vegetative shoot dissections revealed multiple population peaks throughout July and August. Infestation was detected in vegetative shoot tips in all parts of the bushes, with initial infestation greatest at the base of bushes. Degree day accumulations until first midge detection and peak infestation suggest some potential for predicting key events in the pest's phenology. This information about the distribution and timing of infestation will be useful in developing management strategies for blueberry gall midge infestation.     

Description of the soybean pod gall midge,Asphondylia yushimai sp. n. (Diptera: Cecidomyiidae), a major pest of soybean and findings of host alternation

The soybean pod gall midge is an important pest of soybean in Japan and is known to occur also in Indonesia and China. This gall midge is described from Japan as Asphondylia yushimai sp. n. and is clearly distinguished from its congeners by the arrangement of the lower frontal horns of the pupa and the sequence of the mtDNA COI region. It is concluded that Prunus zippeliana Miquel is a winter host of the soybean pod gall midge since haplotypes of the soybean pod gall midge coincide with those of the Prunus fruit gall midge that produces fruit galls on P. zippeliana. In addition, phenological and distributional information on the two gall midges and on their host plants supports the identification of the winter host. In Japan, the soybean pod gall midge overwinters as a first instar in the fruit galls on P. zippeliana and emerges as an adult from the galls in May. In summer and autumn, the soybean pod gall midge has two or more generations in the pods of soybean, Glycine max (L.) Merrill or wild fabaceous and caesalpiniaceous plants. Thus host alternation by A. yushimai is confirmed. This is the second finding of host alternation by a species of Asphondylia, the first instance being that of Asphondylia gennadii (Marchal) in Cyprus.     

Mixed significance of plant vigor: two species of galling Pontania in a hybridizing willow complex

Distributions of leaf galls and offspring performance of two Pontania sawfly species were explored in individual willows of the subarctic Salix caprea – starkeana hybrid complex. The more common sawfly, an undescribed species near the dolichura group (P1), had the highest gall numbers in trees with long shoots both in S. caprea and hybrids. While numbers were high on vigorously growing hybrids, offspring of P1 were aborted significantly more often on hybrids than on pure hosts. Further, non-aborted galls were smaller on hybrids. Fast shoot growth may be important for P1 sawflies, because females oviposit early in summer and larvae develop rapidly compared with the other species, Pontania pedunculi (P2). Distributions of P2 galls were related to tree height and not to shoot length in both parental and hybrid groups of willows. Like P1, also P2 offspring were frequently aborted on hybrids, but not significantly more often than on pure hosts, and P2 galls were equally large among the host groups. Survival of both species was related to abortion rates, while larvae were parasitized equally in all host groups. This study demonstrates that the significance of plant vigor may vary even for closely related galling sawflies exploiting the same hosts, or for the same species on different host plants. Vigorous growth may mislead gallers to oviposit on suboptimal plants.     

Morphological,genetic and symptomatic identification of an invasive jujube pest in Korea,Dasineura jujubifolia Jiao & Bu (Diptera: Cecidomyiidae)

Correct identification of invasive species is an important step for further management of this species. New type of leafcurling gall midge feeding on jujube, Ziziphus jujube Miller (Rhamnaceae), was discovered in Korea in 2011. At that time, this species was not identified as specific level, so it was recorded as Dasineura sp. Since then, the jujube gall midge has become a serious alien insect pest in Korea. In this study, this jujube gall midge was surveyed by collecting damaged leaf samples from different major jujube-producing regions in Korea. Morphological, genetic and symptomatic identification of this invasive jujube pest identified as Dasineura jujubifolia Jiao & Bu. Here we provided its taxonomic status, geographical distribution, morphological characteristics of all stages and infestation symptoms. In addition to classical morphological characters, mitochondrial COI barcoding sequences were generated for several specimens. The possible pathway of invasion and subsequent socioeconomic consequences were discussed.     

Host relationships ofDasineura ingeris: oviposition preferences and juvenile survival on clones ofSalix viminalis, otherSalix species and hybrids

In the present study we assessed the extent to which oviposition preferences and larval survival rates of the gall midge Dasineura ingeris Sylvén & Lövgren (Diptera: Cecidomyiidae) can explain variation in its use of Salix viminalis phenotypes as hosts. Furthermore, based on oviposition preferences and juvenile survival rates we estimated the host potential of various Salix species and species hybrids. Egg distribution patterns explained 67% of the intraspecific variation in damage to S. viminalis shoots in the field. Juvenile survival, on the other hand, was not related to field damage or egg distribution patterns among the S. viminalis clones studied. Clones of complete or partial S. viminalis origin ranked highest in suitability for D. ingeris survival, and four out of these six clones also received the highest egg rankings. Species belonging to the subgenus Salix were not oviposited on at all, whereas others belonging to the same subgenus as S. viminalis, i.e. Vetrix, were intermediate in terms of oviposition rates. Although there was a significant correlation between D. ingeris egg distributions and larval survival rates on the clones in the Salix species/hybrid set, mismatches were evident.     

Nomenclatural remarks on the Salix viminalis group in Norden

Larsson, G. 1995. Nomenclatural remarks on the Salix viminalis group in Norden. —Nord. J. Bot. 15: 343–346. Copenhagen. ISSN 0107–05513.
The taxa in the Salix viminalis group which have been found in Norden are lectotyp-ified: S. viminalis, S. dasyclados, S . x smithiana, S . x stipularis and S . x calodendron . Flora Nordica notes no. 12.