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.     

Rice–gall midge interactions: Battle for survival

Gall midges are insects specialized in maneuvering plant growth, metabolic and defense pathways for their benefit. The Asian rice gall midge and rice share such an intimate relationship that there is a constant battle for survival by either partner. Diverse responses by the rice host against the midge include necrotic hypersensitive resistance reaction, non-hypersensitive resistance reaction and gall-forming compatible interaction. Genetic studies have revealed that major R (resistance) genes confer resistance to gall midge in rice. Eleven gall midge R genes have been characterized so far in different rice varieties in India. In addition, no single R gene confers resistance against all the seven biotypes of the Asian rice gall midge, and none of the biotypes is virulent against all the resistance genes. Further, the interaction of the plant resistance gene with the insect avirulence gene is on a gene-for-gene basis. Our recent investigations involving suppressive subtraction hybridization cDNA libraries, microarray analyses, gene expression assays and metabolic profiling have revealed several molecular mechanisms, metabolite markers and pathways that are induced, down-regulated or altered in the rice host during incompatible or compatible interactions with the pest. This is also true for some of the pathways studied in the gall midge. Next generation sequencing technology, gene expression studies and conventional screening of gall midge cDNA libraries highlighted molecular approaches adopted by the insect to feed, survive and reproduce. This constant struggle by the midge to overcome the host defenses and the host to resist the pest has provided us with an opportunity to observe this battle for survival at the molecular level.     

Flanking Microsatellite Markers for Breeding Varieties Against Asian Rice Gall Midge

The Asian rice gall midge, Orseolia oryzae Wood-Mason (Cecidomyiidae: Diptera) is a serious pest of wet season rice in South and Southeast Asia. Due to internal feeding habit and presence of biotypes of the pest, the most feasible way to control is breeding varieties resistant against multiple biotypes through marker-assisted breeding (MAB). But very few versatile co-dominant markers linked to the gall midge resistance genes are available. We used a set of F₉ recombinant inbred lines (RILs) of the cross TN1/PTB10 and identified microsatellite markers for the gall midge resistance gene in cv. PTB10 on short arm of rice chromosome 8. Markers RM22550 and RM547 flank the gene at a distance of 0.9 and 1.9 cM, respectively. Amplification of the markers in gall midge resistant and susceptible cultivars showed that these markers can be successfully used in MAB for development of gall midge resistant varieties.     

Genetic,physiological and molecular interactions of rice and its major dipteran pest,gall midge

The gall midge, Orseolia oryzae, is a major dipteran pest of rice affecting most rice growing regions in Asia, Southeast Asia and Africa. Chemical and other cultural methods for control of this pest are neither very effective nor environmentally safe. The gall midge problem is further compounded by the fact that there are many biotypes of this insect and new biotypes are continuously evolving. However, resistance to this pest is found in the rice germ plasm. Resistance is generally governed by single dominant genes and a number of non-allelic resistance genes that confer resistance to different biotypes have been identified. Genetic studies have revealed that there is a gene-for-gene interaction between the different biotypes of gall midge and the various resistance genes found in rice. This review discusses different aspects of the process of infestation by the rice gall midge and its interaction with its host. Identification of the gall midge biotypes by conventional methods is a long and tedious process. The review discusses the PCR-based molecular markers that have been developed recently to speed up the identification process. Similarly, molecular markers have been developed for two gall midge resistance genes in rice – Gm2 and Gm4t – and these markers are now being used for marker-assisted selection. The mapping, tagging and map-based gene cloning of one of these genes – Gm2 – has also been discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

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.     

Marker-trait association analysis for gall midge (Orseolia oryzae) resistance in a diverse rice population

Damage caused by insect herbivores, notably Asian rice gall midge, Orseolia oryzae is more prevalent in the rice-growing belts of India's southern and north-eastern states. As a prelude to resistant cultivar development, the identification of genomic regions for resistance in the source population is crucial. In the present investigation, 202 rice genotypes were phenotyped and assayed with genomic markers reported for gall midge resistance. Positive skewness and platykurtic distribution of response scores suggested the inheritance of gall midge resistance in the study population. The marker gm3del3 contributed the most genetic variation, followed by RM28574 and marker RM22709 explained minimal variation. A marker-trait association analysis with a single marker-trait linear regression approach was performed to discover gall midge resistant genomic region/genes. The marker RM17480 on chromosome 4 reported to be linked with gm3 gene was found significantly associated with the gall midge resistance genomic region with allelic effects in a negative direction favouring resistance reaction. The allelic effects of significantly associated markers were correlated significantly with the phenotypic variation of gall midge damage scores. Genes identified in the vicinity of this marker contribute to stress response reactions in rice plants. The 200 bp allele of the marker was associated with susceptibility, while the 250 bp allele was associated with resistance expression. This allelic association with trait variation suggests the importance of associated marker for utilisation in marker-assisted selection programmes to incorporate resistance alleles into elite rice genotypes.     

垂叶榕果内寄生瘿蚊的发生规律

在云南西双版纳热带地区,一种还未被描述属和种的瘿蚊寄生于垂叶榕果内的雌花子房里,并膨大形成虫瘿,靠取食花胚组织完成发育.通过2010年定时定点的系统观察和研究瘿蚊的发生规律,结果发现:该种瘿蚊在垂叶榕上发生频率较高,抽样的18批果有17批被瘿蚊寄生,其中有13批果的寄生比率超过了60％,部分甚至达到100％.瘿蚊寄生雌花的平均比率不超过6％,平均每个榕果内有瘿蚊1～40只不等.瘿蚊寄生不仅降低了垂叶榕的种子数,还致命性地影响着传粉榕小蜂的繁殖.在对瘿蚊的羽化、交配行为及其成虫的活动规律观察中,结果显示:寄生在一个榕果里瘿蚊需要5～7d才能羽化完,其羽化高峰期出现在第2～3天;一天内羽化的高峰出现在上午8:00-10:00及晚上20:00-0:00;而羽化后瘿蚊活动高峰期出现在上午8:00-11:00.雄蚊会帮助雌蚊离开榕果并优先获得交配权,雌蚊交配后飞离繁殖自身的榕树,去寻找适合产卵的榕果繁殖后代,雄蚊不离开羽化树,而死在繁殖自身的榕树附近.这些结果为深入研究瘿蚊的防治技术提供了科学依据.     

Dissecting the association between a gall midge,Asteromyia carbonifera,and its symbiotic fungus,Botryosphaeria dothidea

The Ambrosia gall midge [Asteromyia carbonifera (Osten Sacken) (Diptera: Cecidomyiidae: Alycaulini)] consists, in part, of a complex of genetically differentiated populations that have diverged in gall morphology on the host plant Solidago altissima L. (Asteraceae). This divergence appears to be an incipient adaptive radiation that may be driven by parasitoid pressure. Understanding the mechanisms driving this genetic and phenotypic diversification requires a close examination of the relationship between the midge and its fungal associate Botryosphaeria dothidea (Moug.) Ces. & De Not. (Ascomycota: Dothideomycetes), whose mycelia actually form the protective gall structure. We used manipulative experiments to test the degree of interdependency of the fungus and the midge, and we employed field and laboratory studies to gain insight into the source of fungal conidia, which our data and observations indicate are collected by females and stored in specialized pockets (mycangia) on the ovipositor. Manipulative experiments demonstrate that fungal proliferation on the host plant is dependent on the midge larvae and larvae exhibit significant growth on the fungus alone. Field observations and experiments were unable to identify the source of mycangial conidia; however, analyses of conidia shape suggest a biotrophic source. We conclude that this association is an obligatory mutualism with respect to successful gall formation. These findings corroborate recent findings that the primary food source of the midge is the gall fungus.     

First known gall midge (Diptera: Cecidomyiidae) from Arecaceae: Normanbyomyia fructivora gen. & sp. n. damaging fruit of the black palm, Normanbya normanbyi, in tropical Australia

Abstract  A new species of gall midge, Normanbyomyia fructivora gen. & sp. n., was found infesting flowers and fruit of the black palm, Normanbya normanbyi (Arecaceae), in the tropical forest of north-eastern Australia. Larvae of the gall midge live between the perianth and the young fruit causing abnormal fruit growth. Infested fruit are substantially smaller than healthy ones, fall off prematurely and decay on the ground without producing seedlings. Normanbyomyia , a new genus erected for the new species is placed within the supertribe Lasiopteridi and is characterised by a conspicuous female abdominal segment 8 that is inflated and strongly sclerotised, and wide male parameres that loosely sheath the aedeagus and bear no papillae. A key to Australian Lasiopteridi is provided. The new species becomes the first described gall midge that feeds on a plant from the palm family, Arecaceae.     

Dasineura ingeris sp.n. (Diptera: Cecidomyiidae) on Salix viminalis in Sweden, including comparisons with some other Dasineura species on Salix

Infestations of an undescribed gall midge species were discovered in Southern Sweden in biomass plantations and nurseries of Salix viminalis L. Terminal leaf buds are damaged and side shoots subsequently develop. This midge, Dasineura ingeris sp.n., closely resembles three other species occurring on Salix: D.terminalis (H. Loew) on S.alba L. and S.fragilis L.; D.iteobia (Kieffer) recorded from S.caprea L. and S.cinerea L.; and D.schreiteri (Stelter) (= comb.n. for Rabdophaga schreiteri Stelter) originally found as an inquiline in the galls of another gall midge, Dasineura rosaria (H. Loew), on S.repens L. No qualitative differences in morphology were observed between these four gall midge species, but results of morphometric analyses show significant differences between all of them. In oviposition preference trials, which included host plants of all four midge species, D.ingeris laid eggs mainly on S. viminalis, D.schreiteri preferably on S.repens , and D.iteobia exclusively on S.caprea. In larval performance trials D.ingeris produced many galls on S. viminalis , one gall also on S.caprea , but no galls on either S.alba or S.repens.     

Gas chromatography mass spectrometry based metabolic profiling reveals biomarkers involved in rice-gall midge interactions

The Asian rice gall midge (Orseolia oryzae Wood-Mason) is a serious pest of rice that causes huge loss in yield. While feeding inside the susceptible host, maggots secrete substances that facilitate th...     

The Switchgrass Gall Midge (Chilophaga virgati Gagné) in the Northern Great Plains

Switchgrass (Panicum virgatum L.) is considered to be a highly promising bioenergy crop. However, little is known about insect pests that impact its utilization for this purpose. The switchgrass gall midge [Chilophaga virgati Gagné (Diptera: Cecidomyiidae)], which was first discovered in 2008 at Brookings, SD, USA, is shown to have a negative impact on biomass and seed yields of switchgrass. Our objectives were to increase knowledge of the biology of the midge by describing its life stages and any parasitoids that have biological control potential. Data collections were made during May to December in 2011 and April to late autumn in 2012. The gall midge adult is active from early June to late July. This insect overwinters as a late instar larva, usually in large aggregations, enclosed in the sheath of the flag leaf of dry tillers. The mean number of larvae was 31, with a range of 6 to 85 per tiller. Infested tillers m^?2 varied among three phenologically distinct cultivars. The late flowering cultivar ‘Cave-In-Rock’ was more heavily infested (>2×) than the early flowering ‘Dacotah’. A newly discovered parasitoid, Platygaster chilophagae Buhl (Hymenoptera: Platygastridae) and a species of Quadrastichus sp. (Hymenoptera: Eulophidae) were reared from gall midge larvae. These results will be valuable to entomologists, switchgrass breeders, and agronomists as a guide to the occurrence and activities of the gall midge.     

Oxycephalomyia, gen. nov., and life history strategy of O. styraci comb. nov. (Diptera: Cecidomyiidae) on Styrax japonicus (Styracaceae)

A new genus Oxycephalomyia is described to contain the gall midge that was previously known as Asteralobia styraci (Shinji). Oxycephalomyia styraci, comb. nov., produces leaf vein galls on Styrax japonicus (Styracaceae). The adult of O. styraci is redescribed, and its full‐grown larva and pupa are described for the first time. The annual life cycle of the gall midge in northern Kyushu was clarified; the first instars overwinter in the galls on the host plant. However, the galls of O. styraci mature much later in the season than those of other gall midges with a similar life history pattern, and the durations of second and third larval instars are remarkably short. Such a life history pattern is considered to have an adaptive significance in avoiding larval parasitism, particularly by early attackers. The number of host axillary buds as oviposition sites decreased in bearing years and increased in off years, but there was no sign of oviposition site shortage even in bearing years, probably due to the low population density of the gall midge. An unidentified lepidopteran that feeds on galled and ungalled host buds and a Torymus sp. that attacks pupae of O. styraci were recognized as mortality factors of the gall midge.     

Analysis of SSH library of rice variety Aganni reveals candidate gall midge resistance genes

The Asian rice gall midge, Orseolia oryzae, is a serious insect pest causing extensive yield loss. Interaction between the gall midge and rice genotypes is known to be on a gene-for-gene basis. Here, we report molecular basis of HR? (hypersensitive reaction—negative) type of resistance in Aganni (an indica rice variety possessing gall midge resistance gene Gm8) through the construction and analysis of a suppressive subtraction hybridization (SSH) cDNA library. In all, 2,800 positive clones were sequenced and analyzed. The high-quality ESTs were assembled into 448 non-redundant gene sequences. Homology search with the NCBI databases, using BlastX and BlastN, revealed that 73% of the clones showed homology to genes with known function and majority of ESTs belonged to the gene ontology category ‘biological process’. Validation of 27 putative candidate gall midge resistance genes through real-time PCR, following gall midge infestation, in contrasting parents and their derived pre-NILs (near isogenic lines) revealed induction of specific genes related to defense and metabolism. Interestingly, four genes, belonging to families of leucine-rich repeat (LRR), heat shock protein (HSP), pathogenesis related protein (PR), and NAC domain-containing protein, implicated in conferring HR+ type of resistance, were found to be up-regulated in Aganni. Two of the reactive oxygen intermediates (ROI)–scavenging-enzyme-coding genes Cytosolic Ascorbate Peroxidase1, 2 (OsAPx1 and OsAPx2) were found up-regulated in Aganni in incompatible interaction possibly suppressing HR. We suggest that Aganni has a deviant form of inducible, salicylic acid (SA)-mediated resistance but without HR.     

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.     

Hypersensitive reaction and induced resistance in rice against the Asian rice gall midge Orseolia oryzae

Rice seedlings of the resistant variety Phalguna showed premature tillering, browning of central leaf, and tissue necrosis at the apical meristem following artificial infestation with avirulent biotype 1 of the Asian rice gall midge, Orseolia oryzae (Wood-Mason) (Diptera: Cecidomyiidae). Tissue necrosis representing a typical hypersensitive reaction (HR), accompanied by maggot mortality, was observed within 4 days after infestation. However, reinfestation of secondary tillers subsequent to HR in primary tiller, did not lead to HR in secondary tillers though maggot mortality was seen. Artificial infestation with the weed gall midge O. fluvialis did not result in HR either in gall midge susceptible TN 1 or resistant Phalguna rice varieties. Resistance in Phalguna against the virulent biotype 4 could be induced by either prior, simultaneous, or subsequent infestation with the avirulent biotype 1. The duration of effectiveness of such induced resistance varied with the sequence and time lag between infestations.     

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 .     

Tripius gyraloura n. sp. (Aphelenchoidea: Sphaerulariidae) parasitic in the gall midge Lasioptera donacis Coutin (Diptera: Cecidomyiidae)

A new nematode, Tripius gyraloura n. sp., is described from the arundo gall midge, Lasioptera donacis Coutin (Diptera: Cecidomyiidae). This gall midge is being considered as a biological control agent for use in North America against the introduced giant reed Arundo donax (L.) (Poaceae: Cyperales). Thus the present study was initiated to investigate a nematode parasite that was unknown at the time studies with L. donacis were initiated. The new species has a rapid development in the fly host and the mature parasitic female nematodes evert their uterine cells in the hosts’ hemolymph. Because large numbers of nematodes sterilise the host, eradication of the parasite from laboratory colonies of the midge may be necessary before populations of the fly are released.     

Polymorphisms flanking the mariner integration sites in the rice gall midge (Orseolia oryzae Wood-Mason) genome are biotype-specific.

In an effort to study genome diversity within and between the Indian biotypes of the Asian rice gall midge, Orseolia oryzae, a major insect pest of rice, we made use of mariner transposable element integration site polymorphisms. Using degenerate primers, the design of which is based on mariner sequences, we amplified a ca. 450 bp mariner sequence from the rice gall midge. The mariner sequence showed homology with that of a mariner element isolated from the Hessian fly, Mayetiola destructor, a major dipteran pest of wheat. Southern hybridization, using this mariner fragment as a probe, revealed that the mariner elements are moderately to highly repetitive in the rice gall midge genome. Based on the sequence information of this 450-bp PCR-amplified fragment, outward-directed primers were designed and used in an inverse PCR (iPCR) to amplify the DNA flanking the conserved regions. To study the regions flanking the mariner integration sites, we employed a novel PCR-based approach: a combination of sequence specific amplification polymorphism (SSAP) and amplified fragment length polymorphism (AFLP). The outward-directed mariner-specific primer was used in combination with adapter-specific primers with 1-3 selective nucleotides at their 3' ends. The amplification products were resolved on an agarose gel, Southern-transferred onto nylon membranes, and probed with the iPCR fragment. Results revealed biotype-specific polymorphisms in the regions flanking the mariner integration sites, suggesting that mariner elements in the rice gall midge may be fixed in a biotype-specific manner. The implications of these results are discussed in the context of biotype differentiation.