Impact of two shoot-galling biological control candidates on Russian knapweed, Acroptilon repens

Russian knapweed, Acroptilon repens, is one of the most serious exotic invaders of temperate grasslands in North America. Here we present results from a field experiment in which we quantified the impact of two potential biological control agents, the gall wasp Aulacidea acroptilonica V.Bel. (Hymenoptera, Cynipidae) and the gall midge Jaapiella ivannikovi Fedotova (Diptera, Cecidomyiidae), on A. repens under field conditions in the plant’s native range in Uzbekistan. Attack by A. acroptilonica reduced shoot length by 21%, above-ground biomass by 25% and seed output by 75%, while attack by J. ivannikovi reduced shoot length by 12%, above-ground biomass by 24%, and seed output by 92%. The results of these field experiments are likely to accurately reflect the potential of these two gall formers to reduce above-ground biomass and sexual reproduction of A. repens shoots, since the shoots were part of a clonal network. Despite this, the attacked shoots were not able to compensate for the reallocation of plant resources to gall formation. Moreover, the mean number of galls per shoot obtained in the experiments was within the range of observed gall incidences in the native range. The impact of these two gall-forming insects on Russian knapweed in North America will depend on the population size the species reach and on the timing of attack. The highest impact is likely to occur when the insects attack shoots that have not yet started producing flower-buds.     

Growth of Phragmites australis and Phalaris arundinacea in constructed wetlands for wastewater treatment in the Czech Republic

Common reed (Phragmites australis) and reed canarygrass (Phalaris arundinacea) are two most commonly used plant species in constructed wetlands for wastewater treatment in the Czech Republic. Growth characteristics of both plants (biomass, stem count, and length) have been measured in 13 horizontal sub-surface flow constructed wetlands since 1992. The results revealed that while Phalaris usually reaches its maximum biomass as early as during the second growing season, Phragmites usually reaches its maximum only after three to four growing seasons. The maximum biomass of both species varies widely among systems and the highest measured values (5070 g m⁻² for Phragmites and 1900 g m⁻² for Phalaris) are similar to those found in eutrophic natural stands. The shoot count of Phragmites decreases after the second growing season while length and weight of individual shoots increases over time due to self-thinning process. Number of Phalaris shoots is the highest during the second season and then the shoot count remains about the same. Also the shoot length remains steady over years of constructed wetland operation.     

Leaf scarring by the weevils Neochetina eichhorniae and N. bruchi enhances infection by the fungus Cercospora piaropi on waterhyacinth, Eichhornia crassipes

Additive or synergistic effects among introduced and native insect and plant pathogen agents are necessary to achieve biological control of waterhyacinth (Eichhornia crassipes), a globally damaging aquatic weed. In field plots, plants were infested with waterhyacinth weevils (Neoechetina bruchi and N. eichhorniae) and leaves were scarred by weevil feeding. Subsequent infection by the fungal pathogen Cercospora piaropi caused necrotic lesions to form on leaves. Necrosis development was 7.5- and 10.5-fold greater in plots augmented with both weevils and C. piaropi and weevils alone, respectively, than in plots receiving only C. piaropi. Twenty-four days after weevil infestation, the percentage of laminar area covered by lesions on third-youngest and oldest live leaves was elevated 2.3–2.5-fold in plots augmented with weevils. Scar density and necrosis coverage on young leaf laminae were positively correlated, even though antipathogenic soluble peroxidases were elevated 3-fold in plots augmented with weevils alone or weevils and C. piaropi. Combined weevil and fungal augmentation decreased shoot densities and leaves per plant. In a no-choice bioassay, weevil feeding on oldest but not young leaves was reduced 44 two weeks after C. piaropi inoculation. Protein content and peroxidase activities were elevated 2–6-fold in oldest leaves three weeks after inoculation. Augmentation with both waterhyacinth weevils and C. piaropi led to the development of an additive biological control impact, mediated by one or more direct interactions between these agents, and not plant quality effects.     

Biology of Platycephala planifrons (Diptera: Chloropidae) and its potential effectiveness as biological control agent for invasive Phragmites australis in North America

Phragmites australis is a cosmopolitan clonal grass valued for its support of diversity-rich communities in its native range and feared for its devastating effects on native diversity where the species is introduced. Lack of successful control in North America resulted in the initiation of a biological control program. We used a combination of field surveys and common garden experiments in Europe to study life history and ecology of a chloropid fly, Platycephala planifrons, to assess its potential as a biological control agent. The fly is widely distributed (in non-flooded sites) throughout Eurasia but attack rates are generally low (mean 5–10%; max. 29%). Adults emerge in late June and may live for several months. Females lay eggs at the base of Ph. australis shoots. First instar larvae of this stem-feeding fly overwinter in dormant below-ground shoots of Ph. australis and rapidly complete development in early spring. Larval feeding destroys the growing meristem of the shoot causing premature wilting and 60–70% reductions in shoot biomass production. Early season attack and considerable impact suggest that Pl. planifrons could be a potent biocontrol agent, if it can escape suppression by natural enemies in the introduced range. However, the generally low attack rates in its native range and its dependence on dry sites appear to make the species a “second-choice” candidate for potential release in North America.     

Competitive Interactions between Hydrilla (Hydrilla verticillata) and Vallisneria (Vallisneria americana) as Influenced by Insect Herbivory

Two experiments (winter and summer) were conducted in outdoor tanks using addition-series methods to evaluate the impact of specialized feeding by two biological control agents,Hydrellia pakistanaeDeonier andBagous hydrillaeO'Brien, on competitive interactions between hydrilla [Hydrilla verticillata(L.f.) Royle] and vallisneria (Vallisneria americanaMichx). Competitive abilities of each plant species were determined using the reciprocal-yield model of mean plant weight. In the absence of the biocontrol agents, intraspecific competition from hydrilla on itself was 8.3 times stronger than interspecific competition from vallisneria.Hydrellia pakistanaeinterfered with hydrilla canopy formation by removing as much as 80% of the plant biomass in the top 30 cm of the water column. Damage byH. pakistanaealso caused a 43% reduction in hydrilla tuber production during the winter experiment. Similarly,B. hydrillaecaused up to a 48% reduction in hydrilla plant weight in the summer experiment. Neither insect species damaged vallisneria. As a result, there were significant shifts in the competitive balance between hydrilla and vallisneria due to selective insect feedings. In the presence ofH. pakistanae, hydrilla intraspecific competition was nearly equal to interspecific competition from vallisneria, indicating that hydrilla had lost its competitive edge over vallisneria.Bagous hydrillaealso produced similar, but smaller, shifts in the relative competitive abilities of hydrilla and vallisneria. These results indicate that biological control agents can disrupt the competitive balance between plant species in favor of native species, thus adding another element to the weed biological control strategies.     

Field efficacy and predicted host range of the pickerelweed borer, Bellura densa, a potential biological control agent of water hyacinth

The North American noctuidmoth Bellura densa offers promise as abiological control agent for use in Africa andother countries invaded by water hyacinth. Anaugmentative release at a pond in Florida, USA,eliminated water hyacinth within a few months. Laboratory studies, though, indicated thatoviposition was indiscriminate and thatdevelopment was completed on taro (Colocasia esculenta [Araceae]) as well as onseveral Pontederiaceae. Acceptability of taroas a larval food plant was confirmed in thefield when larvae were found in isolated standsof taro in Florida. Evidence of use of Peltandra virginica (Linnaeus) (Araceae) wasnoted at another site. The distribution oflarval damage was compared at a site containinga mixture of 97% taro and 3% pickerelweed(Pontederia cordata). Larvae damaged87% of the pickerelweed compared to only about5% of the taro, suggesting spillover ontotaro. In another study, 416 larvae wereliberated into a concrete tank containing waterhyacinth (818 plants) surrounded by taro (96plants). Three months later, taro accountedfor only 4% of the damaged plants, less thanthe 11% expected if host selection had beenrandom. In a similar study, larvae wereliberated onto water hyacinth in a large tankdivided into thirds, with pickerelweed or taroat either end and water hyacinth in the middle. The distributions of F₁ egg masses andincidence of damage 3 months later indicatedthat pickerelweed was preferred over taro, but26% of the taro plants were damaged. Weconclude that while B. densa prefersplants in the Pontederiaceae, it is notrestricted to this plant family. Plants in theAraceae would be at risk if this insect werereleased outside of North America, particularlyin cropping situations near water hyacinthinfestations. Bellura densa could beuseful for water hyacinth management in theU.S. if effective augmentation strategies weredeveloped.     

原产地和入侵地紫茎泽兰对泽兰实蝇的选择性、卵巢蛋白质含量与解毒酶活性的影响

[目的] 紫茎泽兰是我国危害严重的恶性入侵杂草。比较专一性天敌泽兰实蝇对该杂草入侵前后植株的适应性,是揭示外来植物入侵后适应性机制的重要科学问题之一。[方法] 比较泽兰实蝇对原产地和入侵地紫茎泽兰植株的寄主选择性,并测定寄生于2类植株的上泽兰实蝇卵巢蛋白质含量及乙酰胆碱酯酶、羧酸酯酶、谷胱甘肽S-转移酶活性。[结果] 泽兰实蝇对原产地和入侵地紫茎泽兰的选择无显著性差异;寄生在紫茎泽兰入侵地植株上的卵巢蛋白质含量较原产地植株上更高。解毒酶活力比较表明,入侵地紫茎泽兰上泽兰实蝇的羧酸酯酶活性低于原产地上的,但谷胱甘肽S-转移酶（雌虫）活性比较则相反,乙酰胆碱酯酶活性比较均无显著性差异。[结论] 紫茎泽兰入侵后,专一性天敌泽兰实蝇的适应性有所下降,丰富了外来植物入侵机制中天敌逃逸假说的内涵。     

Fruit-set and seed weight variation inAnthyllis vulneraria subsp.vulgaris (Fabaceae)

Fruit-set and seed weight variation was studied in a population ofAnthyllis vulneraria subsp.vulgaris (Fabaceae) in northwestern Spain. The plants produce several shoots, each bearing two to four inflorescences that open one at a time from bottom to top. Fruit-set, seed-pod weight and seed weight were found to be significantly higher in proximal inflorescences than in distal inflorescences of the same shoot; mean seed weight was up to three times higher in proximal than in distal inflorescences. By contrast, none of the three variables varied significantly among plants or among shoots of the same plant. Similarly, none of the three variables differed significantly between early- and late-flowering plants, or between plants monitored in 1993 and in 1994. These results are compatible with the view that shoots function as semiautonomous units as regards resource allocation, and that within the shoot resources are preferentially allocated to proximal (= early-opening) inflorescences. In the plants studied, the ratio of seed-pod weight to seed weight was fairly constant, suggesting that the pod is important for seed success.     

Mycobiota associated with larval mines of Thrypticus truncatus and T. sagittatus (Diptera, Dolichopodidae) on waterhyacinth, Eichhornia crassipes, in Argentina

Thrypticus truncatus is a candidate agent for biocontrol of waterhyacinth; the larvae of this diptera mine in the petioles and feed on the phloem in the vascular bundles. The mycobiota associated with T. truncatus and T. sagittatus mines was investigated during two surveys undertaken in the spring and autumn in the Delta of the Paraná River, Argentina. Isolations were made from the mines and larval feeding points, as well as from the larvae, following dissection of the petioles, and plated onto agar. Young and upper parts of the petioles without Thrypticus mines were used as controls. Twenty eight fungal species were isolated from the mines. Pestalotiopsis guepinii, Mucor attenuatus, Phoma tropica, Achlya americana, Fusarium avenaceum, Cladosporium cladosporioides, Clonostachys rosea, Epicoccum purpurascens, Plectosporium tabacinum, Alternaria alternata, and Acremonium sp. were the most common fungi associated with mines and feeding points. Cladosporium cladosporioides, Cytospora sp., Mucor attenuatus, and Phoma tropica were associated with the larval body. The list of fungi in mines was compared with bibliographic information to determine if the species are known pathogens on waterhyacinth or other plant species. This is the first study on mycobiota associated with T. truncatus and T. sagittatus mines in waterhyacinth petioles in Argentina.     

Development of transgenic sorghum for insect resistance against the spotted stem borer (Chilo partellus)

Transgenic sorghum plants expressing a synthetic cry1Ac gene from Bacillus thuringiensis (Bt) under the control of a wound-inducible promoter from the maize protease inhibitor gene (mpiC1) were produced via particle bombardment of shoot apices. Plants were regenerated from the transformed shoot apices via direct somatic embryogenesis with an intermittent three-step selection strategy using the herbicide Basta. Molecular characterisation based on polymerase chain reaction and Southern blot analysis revealed multiple insertions of the cry1Ac gene in five plants from three independent transformation events. Inheritance and expression of the Bt gene was confirmed in T₁ plants. Enzyme-linked immunosorbant assay indicated that Cry1Ac protein accumulated at levels of 1–8 ng per gram of fresh tissue in leaves that were mechanically wounded. Transgenic sorghum plants were evaluated for resistance against the spotted stem borer (Chilo partellus Swinhoe) in insect bioassays, which indicated partial resistance to damage by the neonate larvae of the spotted stem borer. Reduction in leaf damage 5 days after infestation was up to 60%; larval mortality was 40%, with the surviving larvae showing a 36% reduction in weight over those fed on control plants. Despite the low levels of expression of Bt -endotoxin under the control of the wound-inducible promoter, the transgenic plants showed partial tolerance against first instar larvae of the spotted stem borer.     

The effect of two biological control agents, the weevil Neochetina eichhorniae and the mirid Eccritotarsus catarinensis on water hyacinth, Eichhornia crassipes , grown in culture with water lettuce, Pistia stratiotes

We assessed the effect of two biological control agents, the mirid Eccritotarsus catarinensis (Carvalho) and the weevil Neochetina eichhorniae (Warner), singly or in combination, on the competitive ability of their host plant, water hyacinth, Eichhornia crassipes (Mart.) Solms-Laub., grown in a screen house, in competition with another aquatic plant (Pistia stratiotes L.). Water hyacinth plant growth characteristics measured included fresh weight, leaf and petiole lengths, number of inflorescences produced, and new shoots. Without herbivory, water hyacinth was 18 times more competitive than water lettuce (across all experimental combinations of initial plant densities), as estimated from fresh weights. Both insect species, singly or in combination, reduced water hyacinth plant growth characteristics. E. catarinensis alone was less damaging than the weevil and under normal conditions, i.e., floating water hyacinth, is not expected to increase control of water hyacinth beyond that of the weevil. When combined with the weevil, half the inoculum of weevils and half the inoculum of mirids produced the same growth reduction as the full inoculum of the weevil. Under conditions where the weevils are not effective because water hyacinths are seasonally rooted in mud, the mirid, which lives entirely on leaves, should become a useful additional biological control agent. Handling Editor: John Scott.     

The Host Range ofEccritotarsus catarinensis(Heteroptera: Miridae), a Potential Agent for the Biological Control of Waterhyacinth (Eichhornia crassipes)

The host range ofEccritotarsus catarinensiswas determined using 33 plant species to assess the risk of using this insect, a native of South America, for the classical biological control of waterhyacinth (Eichhornia crassipes) in Australia and Papua New Guinea. The results, in conjunction with the results of Hillet al.(1999) who tested 67 species (mostly South African), strongly suggest thatE. catarinensisis restricted to the Pontederiaceae, a family of aquatic plants. All five species of Pontederiaceae in the Australian testing,E. crassipes, Pontederia cordata, Monochoria vaginalis, M. cyanea,andM. australasica,were suitable for insect development. Colonies persisted for at least four generations onE. crassipes, P. cordata,andM. vaginalis.Two-way choice and multiple-choice preference trials were conducted and discussed.E. catarinensisdid not exhibit a clear preference for waterhyacinth over other Pontederiaceae in these trials. Most oviposition occurred into the upper surface of the lamina during laboratory testing despite observations that the underside was preferred in the field. Although not considered suitable for release in Australia, this insect may be useful in other countries where more serious waterhyacinth problems occur and whereM. vaginalisis a serious weed, such as in Southeast Asia.     

The effects of mullein plants (Verbascum thapsus) on the population dynamics of Dicyphus hesperus (Heteroptera: Miridae) in tomato greenhouses

The response of Dicyphus hesperus Knight (Heteroptera: Miridae) to whitefly populations in tomato greenhouses was measured in the presence and absence of mullein (Verbascum thapsus L.) as an alternative host plant. The dynamics of the D. hesperus population on tomato (Lycopersicon esculentum Mill.) and on mullein plants were followed through an entire growing season. In houses with mullein plants, more predators occurred on mullein when whitefly density was low on tomato. A mark-release-recapture experiment where rabbit IgG was used as an external marker showed that D. hesperus adults moved from mullein plants to tomato plants. D. hesperus was always more abundant in houses with mullein than in the houses with tomato plants alone. Movements between tomato and mullein plants are discussed as a strategy to optimize predator foraging. The use of mullein as an alternative host plant may contribute to the establishment of D. hesperus and help to preserve the predator population when prey on tomato crops is scarce.     

Smoke-water improves shoot growth and indigo accumulation in shoots of Isatis indigotica seedlings

Although smoke treatments have successfully been used for promoting the germination of many seeds, there are no reports on the effect of smoke on secondary metabolite production in plants. This study highlights the effects of smoke-water on shoot growth and accumulation of indigo in shoots of Isatis indigotica Fort., a Chinese medicinal plant. Results showed that seedlings treated with smoke-water at a dilution of 1:1000 achieved the highest fresh and dry shoot mass, which was significantly (P < 0.05) different from the control and other smoke-water dilutions tested. A higher concentration of smoke-water (1:500) significantly increased (P < 0.05) the indigo concentration in shoots compared with untreated seedlings. The effect of 1:1000 and 1:2000 dilutions gave non-significant increases in the concentration of indigo. These findings suggest the possible use of smoke-water for promoting the growth and accumulation of secondary metabolites in plants.     

Insecticidal action of Bauhinia monandra leaf lectin (BmoLL) against Anagasta kuehniella (Lepidoptera: Pyralidae), Zabrotes subfasciatus and Callosobruchus maculatus (Coleoptera: Bruchidae)

Bruchid beetle larvae cause major losses in grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil (Callosobruchus maculatus) and the Mexican bean weevil (Zabrotes subfasciatus), are pests that damage stored seeds. The Mediterranean flour moth (Anagasta kuehniella) is of major economic importance as a flour and grain feeder; it is often a severe pest in flour mills. Plant lectins have been implicated as antibiosis factors against insects. Bauhinia monandra leaf lectin (BmoLL) was tested for anti-insect activity against C. maculatus, Z. subfasciatus and A. kuehniella larvae. BmoLL produced ca. 50% mortality to Z. subfaciatus and C. maculatus when incorporated into an artificial diet at a level of 0.5% and 0.3% (w/w), respectively. BmooLL up to 1% did not significantly decrease the survival of A. kuehniella larvae, but produced a decrease of 40% in weight. Affinity chromatography showed that BmoLL bound to midgut proteins of the insect C. maculatus. 33 kDa subunit BmoLL was not digested by midgut preparations of these bruchids. BmoLL-fed C. maculatus larvae increased the digestion of potato starch by 25% compared with the control. The transformation of the genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops.     

Host range of Ceutorhynchus assimilis (Coleoptera: Curculionidae), a candidate for biological control of Lepidium draba (Brassicaceae) in the USA

Ceutorhynchus assimilis has been selected as a potential biological control agent of Lepidium draba, which is a Eurasian invasive weed in North America. Preliminary studies indicated specificity of this weevil collected in southern France on L. draba. This result was in discord with the pest status of C. assimilis found in the literature. Host-specificity tests based both on field and laboratory experiments showed heterogeneity in the host spectrum of the weevils reared from different host-plants as determined by larval development. However, no distinguishable morphological differences could be visually detected between the populations feeding on different host-plants. All sampled populations of weevils were polyphagous as adults. Weevils reared from L. draba were specific to this plant for their complete larval development. Conversely, populations living on other wild and cultivated Brassicaceae species were not able to use L. draba as a host plant. Such differentiation is further highlighted by other biological aspects such as plant infestation rates, sex-ratio, duration of larval development, and differences in the timing of their life cycles. These results demonstrate that C. assimilis, an insect species formerly considered as a pest of Brassicaceae, is characterized by its host-range variability, with one population being potentially useful in the biological control of L. draba. Moreover, this example points to the need to test multiple populations of biological control agents in assessing risk.     

Oviposition and larval survival of Dasineura tetensi on four blackcurrant Ribes cultivars

Monitoring of an unsprayed infested fieldsite using watertraps in S.E. Kent revealed four generations of Dasineura tetensi (Rubs) (Diptera: Cecidomyiidae) occurring between April and September 1996. Ribes nigrum L. cultivars 'Baldwin' (susceptible), 'Ben Alder' (susceptible) and 'Ben Connan' (resistant) were sampled for eggs in the field and assessed for midge damage throughout the season. Oviposition was indiscriminate, but plant damage varied significantly between cultivars. In laboratory choice experiments, mated female midges showed no preference between susceptible shoots of 'Ben Alder' and resistant shoots of 'Ben Connan' for oviposition. Olfactory responses of D. tetensi to leaf volatiles of 'Ben Alder' and 'Ben Connan' were also tested in a 4-way olfactometer. Mated females did not discriminate between volatiles of susceptible and resistant host plants. Larvae reared on cv. 'Ben Connan' shoots were significantly smaller than those reared on shoots of cv. 'Ben Alder'. Larval antibiosis and not female antixenosis appears to be the main mechanism for resistance to D. tetensi in 'Ben Connan'.     

Efficient Plant Regeneration System From Leaf Discs of Zonal (Pelargonium x hortorum) and two scented (P. capitatum and P. graveolens) Geraniums

An efficient and rapid plant regeneration system was established for zonal and scented geraniums using leaf discs as explants. Several explants, medium and culture conditions were studied to optimize shoot induction. Leaf discs taken from 4–5 weeks old in vitro grown plants, whatever the genotype, were more effective for shoot regeneration than those taken from greenhouse grown plants. Darkness proved to be a stimulating factor for shoot regeneration and the combination between NAA and two cytokinins gave the best results. Direct shoot regeneration (100%) was obtained from leaf discs of P. capitatum on half-strength MS medium supplemented with 0.5 mg l⁻¹ NAA in combination with 1 mg l⁻¹ of BAP and zeatin in darkness (11.4 shoots per explant). In the same medium and culture conditions, all P. graveolens leaf discs also exhibited direct shoot regeneration (7.3 shoots per explant). For P. x hortorum, 100% of leaf discs underwent shoot regeneration on a MS medium supplemented with 0.2 mg l⁻¹ NAA in combination with 0.5 mg l⁻¹ of BAP and zeatin in darkness (8.8 shoots per explant) or under low light conditions with 0.2 mg l⁻¹ NAA and 1 mg l⁻¹ of BAP and zeatin (7.5 shoots per explant). For this species, the best results for shoot elongation were obtained on half-strength MS medium gelled with Phytagel 0.3% (v/v). Whatever the genotype, all shoots rooted readily when transferred to diluted MS medium (MS/2) containing 1 mg l⁻¹ IAA. Acclimatized plants grew normally and flowered in greenhouse conditions. Flow cytometry analysis made on leaves of acclimatized plants revealed that all the scented geranium plants are similar to mother plants while 71% of P. x hortorum plants which showed strong growth were tetraploid.     

Thidiazuron promotes high frequency regeneration of peanut (Arachis hypogaea) plants in vitro

Multiple shoots were induced on Valenciatype peanut (Arachis hypogaea L.) explants cultured in vitro on a nutrient medium supplemented with thidiazuron. Zygotic embryos excised from mature seeds were germinated on Murashige-Skoog nutrient medium, and the resulting plantlets (8 days-old) were used as a source of explants. When cultured on a nutrient medium with increasing levels of thidiazuron (0.5 to 30 mg/l), expiants from various parts of the peanut plant (except the root) produced multiple shoot primordia which subsequently developed into individual shoots. Hypocotyl and cotyledon explants produced shoots in higher numbers than other explants (20 shoots per hypocotyl explant at all thidiazuron concentrations and 15 shoots per cotyledon explant at 30 mg/l). Shoots rooted normally on a basal Murashige-Skoog medium containing charcoal and developed into healthy and fertile plants when planted in soil.Abbreviations TDZ thidiazuron - MSO Murashige and Skoog (1962) basal medium - BA 6-benzylaminopurine     

Somatic hybrids between<Emphasis Type="Italic"> Arabidopsis thaliana</Emphasis> and cytoplasmic male-sterile radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis>)

Somatic hybrids were produced by protoplast fusion between Arabidopsis thaliana ecotype Columbia and a male-sterile radish line MS-Gensuke (Raphanus sativus) with the Ogura cytoplasm. Forty-one shoots were differentiated from the regenerated calli and established as shoot cultures in vitro. About 20 of these shoots were judged to be hybrids based on growth characteristics and morphology. Molecular analyses of 11 shoots were performed, confirming the hybrid features. Of these 11 shoots, eight were established as rooted plants in the greenhouse. Polymerase chain reaction and randomly amplified polymorphic DNA analyses of the nuclear genomes of all analyzed shoots and plants confirmed that they contained hybrid DNA patterns. Their chromosome numbers also supported the hybrid nature of the plants. Investigations of the organelles in the hybrids revealed that the chloroplast (cp) genome was exclusively represented by radish cpDNA, while the mitochondrial DNA configuration showed a combination of both parental genomes as well as fragments unique to the hybrids. Hybrid plants that flowered were male-sterile independent of the presence of the Ogura CMS-gene orf138.Abbreviations CMS Cytoplasmic male sterilityCommunicated by M.R. Davey