Efficacy of fiber bands impregnated with Beauveria brongniartii cultures against the Asian longhorned beetle, Anoplophora glabripennis (Coleoptera: Cerambycidae)

Use of 50 × 500 mm non-woven fiber bands impregnated with cultures of entomopathogenic fungi was investigated for control of Anoplophora glabripennis. Fungal bands produced as a commercial pest control product and obtained from industry contained Beauveria brongniartii NBL 851 while lab-produced bands contained B. brongniartii WU 20 or Metarhizium anisopliae VD 1. In the laboratory, adult longevity decreased when adult A. glabripennis were forced to walk for 5 s on bands impregnated with B. brongniartii NBL 851 and for 25 s on B. brongniartii WU 20 and M. anisopliae VD 1 bands. A field experiment was conducted over a 31-day period in Huaiyuan, Anhui, China, by attaching bands impregnated with B. brongniartii NBL 851 or WU 20 around the trunks of willows (Salix sp.) at two sites infested with A. glabripennis. Treatments were compared directly, by monitoring adult longevity, and indirectly, by counting oviposition scars and exit holes per tree to quantify relative oviposition per female. Adults collected from trees treated with B. brongniartii WU 20 died more quickly than adults from the trees treated with B. brongniartii NBL 851 or controls. At the site with higher A. glabripennis population densities, relative oviposition per female was significantly lower for both fungal treatments compared with the control treatment. Between fungal treatments, B. brongniartii NBL 851 had a 5 day earlier negative effect on oviposition per female than B. brongniartii WU 20.     

The Effect of Variability in the Phenology of the Reproductive Stages of Scotch Broom (Cytisus scoparius) on the Synchronization of the Life Stages of Broom Seed Beetle (Bruchidius villosus) in New Zealand

Variability in timing of the reproductive stages of Scotch broom (Cytisus scoparius) may influence synchronization and establishment of the broom seed beetle (Bruchidius villosus), a biological control agent. A sampling scheme was devised to compare the phenologies of Scotch broom at different sites in the same season and in different seasons at the same site. The synchrony of the broom seed beetle's life stages with those of the host plant was also determined. The phenology of Scotch broom varied only slightly from season to season at Lincoln, but could vary considerably between sites in the same season. At both sites where it is established, the broom seed beetle was synchronized with its host; adult beetles were present throughout the flowering period. Eggs were found on suitable green pods. The broom seed beetle appears capable of adapting to the phenology of its host and has the potential to be an effective agent for Scotch broom. Variability in phenology of the reproductive stages of Scotch broom, even at nearby sites, must be taken into account by practitioners of biological control when releasing broom seed beetles and later when sampling beetles to determine establishment.     

Microbial Community Structure at Different Fermentation Stages of Kutajarista, a Herbal Formulation

Kutajarista is an Ayurvedic fermented herbal formulation prescribed for gastrointestinal disorders. This herbal formulation undergoes a gradual fermentative process and takes around 2 months for production. In this study, microbial composition at initial stages of fermentation of Kutajarista was assessed by culture independent 16S rRNA gene clone library approach. Physicochemical changes were also compared at these stages of fermentation. High performance liquid chromatography–mass spectrometry analysis showed that Gallic acid, Ellagic acid, and its derivatives were the major chemical constituents recovered in this process. At 0 day of fermentation, Lactobacillus sp., Acinetobacter sp., Alcaligenes sp., and Methylobacterium sp. were recovered, but were not detected at 8 day of fermentation. Initially, microbial diversity increased after 8 days of fermentation with 11 operational taxonomic units (OTUs), which further decreased to 3 OTUs at 30 day of fermentation. Aeromonas sp., Pseudomonas sp., and Klebsiella sp. dominated till 30 day of fermentation. Predominance of γ- Proteobacteria and presence of gallolyl derivatives at the saturation stage of fermentation implies tannin degrading potential of these microbes. This is the first study to highlight the microbial role in an Ayurvedic herbal product fermentation.     

Glandular trichomes ofSolanum berthaultii alter host preference of the Colorado potato Beetle,Leptinotarsa decemlineata

Choice and no-choice studies were conducted to determine how the glandular trichomes of the wild potato,Solanum berthaultii Hawkes, affect host preference of the Colorado potato beetle,Leptinotarsa decemlineata (Say). Given a feeding choice betweenS. tuberosum andS. berthaultii, larvae and adults preferred the foliage ofS. tuberosum, but adults were more discriminating. When foliage ofS. berthaultii was appressed toS. tuberosum leaflets, fewer adults fed on the appressed leaflets. When given a choice between ‘trichome-intact’ and ‘trichome-removed’S. berthaultii foliage, adults preferred to feed on the latter. The preference for ‘trichome-removed’ foliage and the percent of adults initiating feeding, increased with the degree of trichome removal. These studies provide evidence that the resistance ofS. berthaultii is associated with feeding deterrents localized in the glandular trichomes, thatS. berthaultii possesses more than one mechanism of resistance to the Colorado potato beetle, and that the expression of resistance is dependent on the developmental stage of the insect.     

Microbial Life at 90 C: the Sulfur Bacteria of Boulder Spring

The physiology of the bacteria living in Boulder Spring (Yellowstone National Park) at 90 to 93 C was studied with radioactive isotope techniques under conditions approximating natural ones. Cover slips were immersed in the spring; after a fairly even, dense coating of bacteria had developed, these cover slips were incubated with radioactive isotopes under various conditions and then counted in a gas flow or liquid scintillation counter. Uptake of labeled compounds was virtually completely inhibited by formaldehyde, hydrochloric acid, and mercuric bichloride, and inhibition was also found with streptomycin and sodium azide. The water of Boulder Spring contains about 3 mug of sulfide per ml. Uptake of labeled compounds occurs only if sulfide or another reduced sulfur compound is present during incubation. The pH optimum for uptake of radioactive compounds by Boulder Spring bacteria is 9.2, a value near that of the natural spring water (8.9). Many experiments with a variety of compounds were performed to determine the temperature optimum for uptake of labeled compounds. The results with all the compounds were generally similar, with broad temperature optima between 80 and 90 C, and with significant uptake in boiling (93 C) but not in superheated water (97 C). The results show that the bacteria of Boulder Spring are able to function at the temperature of their environment, although they function better at temperatures somewhat lower. The fine structure of these bacteria has been studied by allowing bacteria in the spring to colonize glass slides or Mylar strips which were immediately fixed, and the bacteria were then embedded and sectioned. The cell envelope structure of these bacteria is quite different from that of other mesophilic or thermophilic bacteria. There is a very distinct plasma membrane, but no morphologically distinct peptidoglycan layer was seen outside of the plasma membrane. Instead, a rather thick diffuse layer was seen, within which a subunit structure was often distinctly visible, and connections frequently occurred between this outer layer and the plasma membrane. The thick outer layer usually consisted of two parts, the outer part of which was sometimes missing. Within the cells, structures resembling ribosomes were seen, and regions lacking electron density which probably contained deoxyribonucleic acid were also visible.     

Succession of Sulfur-Oxidizing Bacteria in the Microbial Community on Corroding Concrete in Sewer Systems

Microbially induced concrete corrosion (MICC) in sewer systems has been a serious problem for a long time. A better understanding of the succession of microbial community members responsible for the production of sulfuric acid is essential for the efficient control of MICC. In this study, the succession of sulfur-oxidizing bacteria (SOB) in the bacterial community on corroding concrete in a sewer system in situ was investigated over 1 year by culture-independent 16S rRNA gene-based molecular techniques. Results revealed that at least six phylotypes of SOB species were involved in the MICC process, and the predominant SOB species shifted in the following order: Thiothrix sp., Thiobacillus plumbophilus, Thiomonas intermedia, Halothiobacillus neapolitanus, Acidiphilium acidophilum, and Acidithiobacillus thiooxidans. A. thiooxidans, a hyperacidophilic SOB, was the most dominant (accounting for 70% of EUB338-mixed probe-hybridized cells) in the heavily corroded concrete after 1 year. This succession of SOB species could be dependent on the pH of the concrete surface as well as on trophic properties (e.g., autotrophic or mixotrophic) and on the ability of the SOB to utilize different sulfur compounds (e.g., H₂S, S⁰, and S₂O₃²⁻). In addition, diverse heterotrophic bacterial species (e.g., halo-tolerant, neutrophilic, and acidophilic bacteria) were associated with these SOB. The microbial succession of these microorganisms was involved in the colonization of the concrete and the production of sulfuric acid. Furthermore, the vertical distribution of microbial community members revealed that A. thiooxidans was the most dominant throughout the heavily corroded concrete (gypsum) layer and that A. thiooxidans was most abundant at the highest surface (1.5-mm) layer and decreased logarithmically with depth because of oxygen and H₂S transport limitations. This suggested that the production of sulfuric acid by A. thiooxidans occurred mainly on the concrete surface and the sulfuric acid produced penetrated through the corroded concrete layer and reacted with the sound concrete below.     

Assessment of Pollution-Induced Microbial Community Tolerance to Heavy Metals in Soil Using Ammonia-Oxidizing Bacteria and Biolog Assay

This study attempted to investigate if the tolerance of soil bacterial communities in general, and autotrophic ammonia-oxidizing bacteria (AOB) in particular, evolved as a result of prolonged exposure to metals, and could be used as an indigenous bioindicator for soil metal pollution. A soil contaminated with copper, chromium, and arsenic (CCA) was mixed with an uncontaminated garden soil (GS3) to make five test soils with different metal concentrations. A modified potential ammonium oxidation assay was used to determine the metal tolerance of the AOB community. Tolerance to Cr, Cu, and As was tested at the beginning and after up to 13 months of incubation. Compared with the reference GS3 soil, the five CCA soils showed significantly higher tolerance to Cr no matter which form of Cr (Cr³⁺, CrO₄ ^2?, or Cr₂O₇ ^2?) was tested, and the Cr tolerance correlated with the total soil Cr concentration. However, the tolerance to Cu²⁺, As³⁺, and As⁵⁺ did not differ significantly between the GS3 soil and the five CCA soils. Community level physiological profiles using Biolog microtiter plates were also used to examine the chromate tolerance of the bacterial communities extracted after six months of exposure. Our results showed that the bacterial community tolerance was altered and increased as the soil Cr concentration was increased, indicating that the culturable microbial community and the AOB community responded in a similar manner.     

Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

Using Age-Based Life History Data to Investigate the Life Cycle and Vulnerability of Octopus cyanea

Octopus cyanea is taken as an unregulated, recreationally fished species from the intertidal reefs of Ningaloo, Western Australia. Yet despite its exploitation and importance in many artisanal fisheries throughout the world, little is known about its life history, ecology and vulnerability. We used stylet increment analysis to age a wild O. cyanea population for the first time and gonad histology to examine their reproductive characteristics. O. cyanea conforms to many cephalopod life history generalisations having rapid, non-asymptotic growth, a short life-span and high levels of mortality. Males were found to mature at much younger ages and sizes than females with reproductive activity concentrated in the spring and summer months. The female dominated sex-ratios in association with female brooding behaviours also suggest that larger conspicuous females may be more prone to capture and suggests that this intertidal octopus population has the potential to be negatively impacted in an unregulated fishery. Size at age and maturity comparisons between our temperate bordering population and lower latitude Tanzanian and Hawaiian populations indicated stark differences in growth rates that correlate with water temperatures. The variability in life history traits between global populations suggests that management of O. cyanea populations should be tailored to each unique set of life history characteristics and that stylet increment analysis may provide the integrity needed to accurately assess this.     

In Vitro Interactions Between Yeasts and Bacteria and the Fungal Symbionts of the Mountain Pine Beetle (Dendroctonus ponderosae)

Multi-trophic interactions between prokaryotes, unicellular eukaryotes, and ecologically intertwined metazoans are presumably common in nature, yet rarely described. The mountain pine beetle, Dendroctonus ponderosae, is associated with two filamentous fungi, Grosmannia clavigera and Ophiostoma montium. Other microbes, including yeasts and bacteria, are also present in the phloem, but it is not known whether they interact with the symbiotic fungi or the host beetle. To test whether such interactions occur, we performed a suite of in vitro assays. Overall, relative yield of O. montium grown with microbes isolated from larval galleries was significantly greater than when the fungus was grown alone. Conversely, the yield of G. clavigera grown with these same microbes was less than or equal to when it was grown alone, suggesting that O. montium, and at least some microbes in larval galleries, have a mutualistic or commensal relationship, while G. clavigera and those same microbes have an antagonistic relationship. A bacterium isolated from phloem not colonized by beetles was found to inhibit growth of both G. clavigera and O. montium and appears to be an antagonist to both fungi. Our results suggest that bacteria and yeasts likely influence the distribution of mycangial fungi in the host tree, which, in turn, may affect the fitness of D. ponderosae.     

Effect of Primers Hybridizing to Different Evolutionarily Conserved Regions of the Small-Subunit rRNA Gene in PCR-Based Microbial Community Analyses and Genetic Profiling

Genetic profiling techniques of microbial communities based on PCR-amplified signature genes, such as denaturing gradient gel electrophoresis or single-strand-conformation polymorphism (SSCP) analysis, are normally done with PCR products of less than 500-bp. The most common target for diversity analysis, the small-subunit rRNA genes, however, are larger, and thus, only partial sequences can be analyzed. Here, we compared the results obtained by PCR targeting different variable (V) regions (V2 and V3, V4 and V5, and V6 to V8) of the bacterial 16S rRNA gene with primers hybridizing to evolutionarily conserved flanking regions. SSCP analysis of single-stranded PCR products generated from 13 different bacterial species showed fewer bands with products containing V4-V5 (average, 1.7 bands per organism) than with V2-V3 (2.2 bands) and V6-V8 (2.3 bands). We found that the additional bands (>1 per organism) were caused by intraspecies operon heterogeneities or by more than one conformation of the same sequence. Community profiles, generated by PCR-SSCP from bacterial-cell consortia extracted from rhizospheres of field-grown maize (Zea mays), were analyzed by cloning and sequencing of the dominant bands. A total of 48 sequences could be attributed to 34 different strains from 10 taxonomical groups. Independent of the primer pairs, we found proteobacteria (α, β, and γ subgroups) and members of the genus Paenibacillus (low G+C gram-positive) to be the dominant organisms. Other groups, however, were only detected with single primer pairs. This study gives an example of how much the selection of different variable regions combined with different specificities of the flanking “universal” primers can affect a PCR-based microbial community analysis.     

Host Range of Mycoleptodiscus terrestris, a Microbial Herbicide Candidate for Eurasian Watermilfoil, Myriophyllum spicatum

A native strain of the fungal plant pathogen Mycoleptodiscus terrestris is capable of causing under experimental conditions necrotic shoot lesions and a generalized decline and disintegration of Eurasian watermilfoil (Myriophyllum spicatum), an exotic submerged aquatic weed in North America. The potential of this fungus to cause disease on nontarget plants was evaluated on 33 species and cultivars in 11 families. The nontarget selection, intended as a Tier I group, was biased toward probable suscepts consisting of submerged, floating, and emergent aquatic species and terrestrial crop plants. The plants were exposed to the fungal mycelium formulated in alginate beads. Pathogenicity rather than phytotoxicity was the primary mode of attack by the fungus. Of the 16 nontarget aquatic species tested, the fungus was pathogenic to Hydrilla verticillata (hydrilla), Myriophyllum aquaticum (parrotfeather), and Ceratophyllum demersum (coontail), but only in hydrilla did it cause plant mortality comparable to that in Eurasian watermilfoil. The remaining 13 submerged, floating, and emergent aquatic species were unaffected. Of the 17 terrestrial species screened, none was significantly affected by the fungus in germination studies, but postemergent disease symptoms developed on seedlings of 10 species. Four of the 10, Medicago sativa (alfalfa), Lotus corniculatus (birdsfoot trefoil), Trifolium hybridum (alyce clover), and T. repens (white clover), developed disease affecting 26-50% of their tissues. Thus, this host range testing scheme, based on the concept of screening plants under the maximum hazard potential, helped to identify susceptible species. Nonetheless, it exaggerated the nontarget risk due to the small number of host species tested. Screening a taxonomically diverse and larger selection of plants as well as testing under conditions of less severe exposure to the fungal inoculum are necessary to obtain a more realistic view of the host range than presently indicated.     

Transmission of acquired adjustments to salinity in Sorghum bicolor

Previous studies showed that exposure of eight-day-old Sorghum bicolor for three weeks to sublethal salinity induces an increase in salinity tolerance, called physiological adaptation (A). During A, plants of a same population differ in reaction and tolerance to salinity. Tolerance levels of the reaction types depend on environmental conditions besides salinity. Reactions observed most frequently in an experiment have generally highest tolerance levels. This phenomenon is defined adaptive determinism (AD). In this study, the relationship between a potential source of the information subjacent to AD and AD itself is analysed in plants first exposed to salt-inducing A. When the reaction types are close variations of one reaction mode, AD is highest. This relationship is inversed in progeny of adapted plants. Results suggest that information relevant to AD is transmitted to the progeny of adapted plants, and that adaptive information is created during A in plants first exposed to adaptation inducing treatment.     

Percent G+C Profiling Accurately Reveals Diet-Related Differences in the Gastrointestinal Microbial Community of Broiler Chickens

Broiler chickens from eight commercial farms in Southern Finland were analyzed for the structure of their gastrointestinal microbial community by a nonselective DNA-based method, percent G+C-based profiling. The bacteriological impact of the feed source and in-farm whole-wheat amendment of the diet was assessed by percent G+C profiling. Also, a phylogenetic 16S rRNA gene (rDNA)-based study was carried out to aid in interpretation of the percent G+C profiles. This survey showed that most of the 16S rDNA sequences found could not be assigned to any previously known bacterial genus or they represented an unknown species of one of the taxonomically heterogeneous genera, such as Ruminococcus or Clostridium. The data from bacterial community profiling were analyzed by t-test, multiple linear regression, and principal-component statistical approaches. The percent G+C profiling method with appropriate statistical analyses detected microbial community differences smaller than 10% within each 5% increment of the percent G+C profiles. Diet turned out to be the strongest determinant of the cecal bacterial community structure. Both the source of feed and local feed amendment changed the bacteriological profile significantly, whereas profiles of individual farms with identical feed regimens hardly differed from each other. This suggests that the management of typical Finnish farms is relatively uniform or that hygiene on the farm, in fact, has little impact on the structure of the cecal bacterial community. Therefore, feed compounders should have a significant role in the modulation of gut microflora and consequently in prevention of gastrointestinal disorders in farm animals.     

Baculovirus-mediated Expression of a Gene for Trehalase of the Mealworm Beetle, Tenebrio molitor, in Insect Cells, SF-9, and Larvae of the Cabbage Armyworm, Mamestra brassicae

It is of interest to understand what kinds of physiological and biochemical changes occur in insects if the homeostasis of trehalose in the hemolymph is disrupted by the infection with a recombinant baculovirus containing a secretory-trehalase gene. For this purpose, two recombinant non-occluded Autographa california multicapsid nucleopolyhedroviruses (AcMNPVs), vTREVL and vERTVL, containing a trehalase cDNA of the mealworm beetle, Tenebrio molitor, were constructed. The trehalase cDNA was inserted in the sense orientation downstream of the polyhedrin promoter for vTREVL, and in the anitsense orientation for vERTVL. The active trehelase of T. molitor was found outside of cells when SF-9 cells or larvae of the cabbage armyworm, Mamestra brassicae, were infected with vTREVL. In the hemolymph of vTREVL-infected larvae, expression of the active trehelase was followed by disappearance of trehalose and appearance of glucose. However, the mortality time of virus-infected 5th instar larvae increased in the following order: AcMNPV C6 (wild-type virus) ≤ vERTVL < vTREVL. The symptoms (the browning and liquefying of the host body) of NPV infection were moderated considerably in vTREVL-infected larvae.     

Phototoxic effect of UVR on wild type, ebony and yellow mutants of Drosophila melanogaster: Life Span, fertility, courtship and biochemical aspects

Melanin plays an important role in protecting organisms from ultraviolet radiation (UVR). Therefore, it is possible that differently colored strains can show different sensitivities to UVR. In the present work, life span, fertility and courtship behavior of wild type (w), ebony (e) and yellow (y) strains of Drosophila melanogaster were studied to evaluate their sensitivity to ultraviolet (UV). Because a range of phototoxic effects of UVR are mediated through generation of free radicals, levels of free radicals, lipid peroxide (malondialdehyde, MDA) and superoxide dismutase (SOD) activity of three strains were examined to indicate their antioxidant defending ability and oxidative status. It was shown that w always had the highest lifespan and fertility not only in the control but also in UV-exposed groups. Moreover, lifespan and fertility of e were significantly higher (P<0.0001) than those of y in the UV-exposed groups, but not for the control. On the other hand, UV exposure had an adverse effect on courtship of flies. Stronger electron paramagnetic resonance (EPR) signals could be detected in w, e and y exposed to 5 min UV. And there were more significant changes of EPR signals in y than in w and e. UVR had no significant (P=0.1782) effect on the SOD activities. After pooling data from the control and UV-exposed groups, we found that w had a significantly (P<0.05) higher level of SOD activity, but e and y were nearly at the same levels (P>0.05). MDA levels were increased in the UV dose-dependent manner (P=0.0495). In conclusion, our results suggested that UVR can decrease life span and fertility of flies and do harm to courtship, which may be due to oxidative damage to flies tissues (e.g. central nervous system) induced by free radicals. w had the highest tolerance to UVR, which may be ascribed to its advantage of survival under the natural condition and at high level of SOD activity. Then differences of pigment between e and y in absorbing UV, shielding against UV and scavenging free radicals produced by UVR should be responsible for their different sensitivity to UVR.     

In Bradyrhizobium japonicum the common nodulation genes, nodABC, are linked to nifA and fixA

Summary Using cloned Rhizobium phaseoli nodulation (nod) genes as hybridization probes homologous restriction fragments were detected in the genome of the slow-growing soybean symbiont, Bradyrhizobium japonicum strain 110. These fragments were isolated from a cosmid library, and were shown to lie 10 kilobasepairs (kb) upstream from the nifA and fixA genes. Specific nod probes from Rhizobium leguminosarum were used to identify nodA-, nodB-, and nodC-like sequences clustered within a 4.5 kb PstI fragment. A mutant was constructed in which the kanamycin resistance gene from Tn5 was inserted into the nodA homologous B. japonicum region. This insertion was precisely located, by DNA sequencing, to near the middle of the nodA gene. B. japonicum mutants carrying this insertion were completely nodulation deficient (Nod^-).     

Sesqui-Ds , the chromosome-breaking insertion at bz-m1 , links double Ds to the original Ds element

The bz-m1 mutation in maize was one of the first to arise by direct transposition of the chromosome-breaking Ds element from its original or `standard' location in chromosome 9S to a known locus in the same chromosome arm. Thus, elucidation of its structure should shed light on the nature of the original Ds element described by McClintock in 1948. The Ds insertion in bz-m1 has been reported to be only 1.2 kb long – much shorter than other chromosome-breaking Ds elements that have been described. We have characterized here the Ds element in our bz-m1 stocks and have confirmed by genetic and molecular tests that, in the presence of Ac, it acts as a chromosome breaker. The Ds insertion at bz-m1 is 1260 bp long. Besides its normal 5′ and 3′ ends, it contains an internal 3′ end at the same junction as the chromosome-breaking double Ds element that has been found in several sh mutations. Thus, it appears to have arisen from the 4.1-kb double Ds by internal deletion of 2.9 kb. Because the element has lost one internal 5′ end, but retains the chromosome-breaking properties of double Ds, we have named it sesqui-Ds (sDs). The origin, structure and properties of sDs vis-à-vis double Ds support the hypothesis that double Ds corresponds to the chromosome-breaking Ds element at the `standard' location in 9S. Received: 10 March 1997 / Accepted: 2 May 1997