Effects of genetic and environmental host plant variation on the susceptibility of two noctuids toBacillus thuringiensis

TwoApium graveolens var.rapaceum (L.) cultivars that differ in their suitability for the survival and growth ofSpodoptera exigua (Hübner) andTrichoplusia ni (Hübner) were used to examine the effect of genetic and seasonal environmental variation in host plant suitability on the efficacy ofBacillus thuringiensis subsp.kurstaki (Berliner). The effects of host plant genotype andB. thuringiensis were generally independent, so thatB. thuringiensis efficacy was greatest on the resistant host plant cultivar. Host plant suitability varied within growing season for both insect species but, while host plant suitability decreased with increasing plant age forT. ni, the response ofS. exigua to plant age was not as clear. Within season variation in host plant suitability affectedB. thuringiensis efficacy and the interaction betweenB. thuringiensis and host plant cultivar forS. exigua but not forT. ni. Soluble protein and Folin-Denis phenolic concentrations of host plant tissue were not correlated with changes in host plant suitability to either insect species.     

Host Selection by the Herbivorous Mite Polyphagotarsonemus latus (Acari: Tarsonemidae)

This study examined the host-selection ability of the broad mite Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae). To make long-distance-shifts from one host plant patch to another, broad mites largely depend on phoretic association with whiteflies. However, the host plants of whiteflies and broad mites are not necessarily the same. We determined the host-preference and acceptance of free-moving and phoretic broad mites using two behavioral bioassays. We used a choice test to monitor host selection by free-moving mites. In the case of phoretic mites, we compared their rate of detachment from the phoretic vector Bemisia tabaci placed on leaves taken from various host plants. The suitability of the plant was further determined by monitoring mite’s fecundity and its offspring development. We compared the mites’ responses to young and old cucumber (Cucumis sativus cv. ‘Kfir’) leaves (3rd and 8–9th leaf from the apex, respectively), and two tomato (Solanum lycopersicum cvs. ‘M82’ and ‘Moneymaker’). Free-moving mites of all stages and both sexes preferred young cucumber leaves to old cucumber leaves and preferred young cucumber rather than young tomato leaves, demonstrating for the first time that broad mites are able to choose their host actively. As for phoretic mated females, although eventually most of the mites abandoned the phoretic vector, the rate of detachment from the whitefly vector was host dependent and correlated with the mites’ fitness on the particular host. In general, host preference of phoretic female mites resembled that of the free-moving female. Cues used by mites for host selection remain to be explored.     

Host Preferences of Trichogramma pretiosum and the Influence of Prior Ovipositional Experience on the Parasitism of Plutella xylostella and Pseudoplusia includens Eggs

Trichogramma wasps are generalist egg parasitoids used in biological control efforts. In a multi host situation they may preferentially parasitize a non-target host species to the detriment of the control program. Plutella xylostella Linnaeus (Lepidoptera: Plutellidae) is a very serious pest of cabbage, but is only one in a number of species in the ‘cabbageworm’ complex. We investigated the host preferences of Trichogramma pretiosum Riley (Hymenoptera: Chalcidoidea) when offered the eggs of Plutella xylostella and the eggs of Pseudoplusia includens Hübner (Lepidoptera: Noctuidae), another species in the ‘cabbageworm’ complex. Trichogramma pretiosum reared on the eggs of the factitious host Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) parasitized both Plutella xylostella and Pseudoplusia includens eggs under laboratory conditions. For both choice and no-choice experiments, T. pretiosum parasitized significantly more P. xylostella eggs than P. includens eggs. Prior ovipositional experience with one or other of the two host species had no effect on the subsequent parasitism levels of the two host species. The preference for P. xylostella eggs was also not affected by this prior ovipositional experience.     

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.     

Oviposition site selection on oats: the effect of plant architecture,plant and leaf age,tissue toughness,and hardness on cereal leaf beetle,Oulema melanopus

In sequentially planted oat stands, the cereal leaf beetle (CLB), Oulema melanopus (L.) (Coleoptera: Chrysomelidae: Lemini), is found in greater numbers, and lays more eggs, on later planted (younger) oats (Avena sativa L.) (Poaceae). Plant characteristics that could explain this ovipositional preference were examined in a series of experiments. Cage and open field whole plant preference tests confirmed the attraction of ovipositing females to younger oats. A cage effect illustrated the role of plant architecture (plant height) in CLB host selection. Two multiple‐choice and one no‐choice excised leaf experiments determined that characteristics of individual leaves associated with leaf insertion level (leaf number from base to apex) and age influence ovipositional site selection. Leaves of higher insertion level have higher nitrogen content, but fewer eggs are laid on those leaves. Two experiments examining the interaction between total leaf nitrogen and leaf insertion level showed that only leaf insertion level affected oviposition choice. Published literature suggests variation in secondary plant compounds cannot explain O. melanopus ovipositional preference among leaves. Grass leaves of higher insertion level have more extensively developed cells associated with tissue toughness and hardness. The data and supporting literature suggest tissue toughness and hardness are deterring oviposition on oat leaves of higher insertion level. However, newly eclosed larvae are able to feed on leaves usually avoided as oviposition sites. The explanation for this result may be a lack of correlation between host suitability and ovipositional preference.     

Restoration of vigor and rooting competence in stem tissues of mature citrus by repeated grafting of their shoot apices onto freshly germinated seedlings in vitro

Summary Repeated grafting of 0.2-cm shoot tips from fruiting-age trees ofCitrus reticulata Blanco ‘Ponkan’ mandarin andC. sinensis Osbeck ‘Liu Tseng’ sweet orange onto freshly germinated ‘Troyer’ citrange [Poncirus trifoliata (L.) Raf. X.C. sinensis Osbeck] seedlings in vitro resulted in progressive restoration of rooting competence and vigor of regenerated roots and shoots. The restored traits were retained through the course of the investigation and suggested a phase reversal phenomenon.     

Volatiles from damaged plants as major cues in long-range host-searching by the specialist parasitoidCotesia rubecula

The role of volatile stimuli in the long-range host-searching behaviour of the specialist parasitoidCotesia rubecula Marshall (Hymenoptera: Braconidae) was studied. Components from the plant-host-complex Brussels sprouts (Brassica oleracea L. var.gemmifera (DC.) Schulz. cv. ‘Titurel’)-Pieris rapae L. (Lepidoptera: Pieridae) were compared for their attractiveness in dual choice tests in a windtunnel. Stimuli from cabbage plants that were mechanically damaged or damaged byP. rapae caterpillars were more attractive to this parasitoid species than stimuli emitted by the host larvae or their faeces. Parasitoids preferred leaves from the plant-host-complex over artificially damaged leaves. Undamaged cabbage plants were the least attractive to the foraging females. These results indicate that in-flight searching behaviour ofC. rubecula is guided by plant-derived information and that for this specialist species more reliable and specific host-derived cues play a minor role at longer distances.     

Intertribal hexaploid somatic hybrid plants regeneration from electrofusion between diploids of Citrus sinensis and its sexually incompatible relative, Clausena lansium

 Chinese wampee [Clausena lansium (Lour.) Skeels], a sexually incompatible relative of citrus, is commercially cultivated in South China. In this study, embryogenic protoplasts of ‘Bonanza’ navel orange [Citrus sinensis (L.) Osbeck] were electrically fused with leaf protoplasts isolated from ‘Chicken Heart’ Chinese wampee. After 8 months of culture, fusion products regenerated into shoots. More than 70% of the shoots unexpectedly rooted well. Chromosome counting of several shoot- and root-tips revealed that their chromosome numbers were not 2n=4x=36 as expected, but 2n=6x=54, suggesting that chromosome doubling occurred rather than chromosome elimination in this intertribal fusion combination. RAPD analysis of embryoids and the leaves of unrooted and rooted shoots verified their hybridity. This is the first report of hexaploid somatic hybrid plant regeneration from fusion between diploids in Aurantioideae. Received: 20 June 1998 / Accepted: 18 August 1998     

Oviposition behaviour and larval development of Anastrepha fraterculus from Argentina in citrus

Citrus peel physicochemical attributes are considered the main components conferring partial or even total resistance to fruit fly (Diptera: Tephritidae) infestation. Fruit fly females adapt their ovipositional strategies to overcome such resistance. Here, we explored the effects of citrus species (Rutaceae) on the ovipositional behaviour of the South American fruit fly, Anastrepha fraterculus (Wiedemann), and on its immature development. Particularly, we investigated the effects of (1) citrus species on oviposition behaviour and immature development, (2) citrus species on oviposition preference and on the location of the eggs at different depth in the citrus peel, and (3) harvest season and post‐harvest storage time on oviposition behaviour and immature development in lemon. Citrus species influenced ovipositional behaviour and affected survival of immature stages. Females laid eggs in lemon [Citrus limon (L.) Burm.], orange [Citrus sinensis (L.) Osbeck], and grapefruit (Citrus paradisi Macfadyen). In orange and lemon, larvae were found dead close to the oviposition areas, suggesting chemically mediated resistance mechanisms. Under choice conditions, females preferred grapefruit over lemon and bigger clutches were found in the layers where embryonic development is favoured. Unsuitability of lemon as a medium to complete development was neither affected by harvest season nor by storage time of the fruit after harvest. The physical and chemical characteristics of the peel were distinctive to each citrus species and may have affected the specific levels of resistance of these citrus species to infestation by A. fraterculus.     

Cloning and characterisation of a family of disease resistance gene analogs from wheat and barley

 The most common class of plant disease resistance (R) genes cloned so far belong to the NBS-LRR group which contain nucleotide-binding sites (NBS) and a leucine-rich repeat (LRR). Specific primer sequences derived from a previously isolated NBS-LRR sequence at the Cre3 locus, which confers resistance to cereal cyst nematode (CCN) in wheat (Triticum aestivum L.) were used in isolating a family of resistance gene analogs (RGA) through a polymerase chain reaction (PCR) cloning approach. The cloning, analysis and genetic mapping of a family of RGAs from wheat (cv ‘Chinese Spring’) and barley (Hordeum vulgare L. cvs ‘Chebec’ and ‘Harrington’) are presented. The wheat and barley RGAs contain other conserved motifs present in known R genes from other plants and share between 55–99% amino acid sequence identity to the NBS-LRR sequence at the Cre3 locus. Phylogenetic analysis of the RGAs with other cloned R genes and RGAs from various plant species indicate that they belong to a superfamily of NBS-containing genes. Two of the barley derived RGAs were mapped onto loci on chromosomes 2H (2), 5H (7) and 7H (1) using barley doubled haploid (DH) mapping populations. Some of these loci identified are associated with regions carrying resistance to CCN and corn leaf aphid. Received: 6 January 1998 / Accepted: 1 April 1998     

Nuclear DNA content of Vitis vinifera cultivars and ploidy level analyses of somatic embryo-derived plants obtained from anther culture

Flow cytometry was employed to determine the ploidy level of Vitis vinifera L. somatic embryo-derived plants obtained from anther culture. Only one among the 41 analysed plants (2.4%) presented somaclonal variation (tetraploidy); the other plants were diploid. No significant differences (P≤0.05) were detected between diploid and parental field plants. No haploid or aneuploid plants were observed. The nuclear DNA content of nine V. vinifera cultivars was also estimated using flow cytometry. A non-significant variation was found among the cultivars, with DNA content ranging from 1.17 pg/2C (cv. ‘Tinta Barroca’ and ‘Viosinho’) to 1.26 pg/2C (cv. ‘Cabernet Sauvignon’). These results and previous studies on other Vitis species suggest that Vitis genome is stable with regard to nuclear DNA content.     

Regeneration and analysis of interspecific asymmetric potato –Solanum ssp hybrid plants selected by micromanipulation or fluorescence-activated cell sorting (FACS)

 Recipient protoplasts from three Solanum tuberosum genotypes, cv ‘Folva’ (2n=4x=48), cv ‘Matilda’ (4n) and ‘161 : 14’ (2n), were electrofused with X-ray-irradiated donor protoplasts from two wild species S. spegazzinii (2n) or S. microdontum×S. vernei (2n). Prior to fusion, protoplasts were fluorescence-labelled with either fluorescein diacetate or scopoletin. Fusion products were identified by dual fluorescence and selected by micromanipulation or fluorescence-activated cell sorting (FACS). All putative hybrid plants were analysed by the random amplified polymorphic DNA (RAPD) technique. Our analysis demonstrates that each asymmetric hybrid plant has an individual and stable profile of donor-specific RAPD bands. The irradiation of donor protoplasts hampered the growth of selected heterofusion products in a dose-dependent way. Irradiation resulted in donor chromosome elimination, but not in a dose dependent way, in the tested interval. In asymmetric hybrids with the S. spegazzinii donor 33–68% of the donor-specific RAPD bands were missing, indicating a similar level of chromosome elimination. In asymmetric hybrid plants with the S. microdontum×S. vernei donor 74–95% of the donor RAPD bands were missing. Chromosome countings revealed that these hybrids had chromosome numbers equal to or below the chromosome numbers found in the tetraploid recipients. This is the first time that highly asymmetric hybrid plants between two tetraploid potato recipients and the donor S. microdontum×S. vernei have been obtained. Received : 16 December 1996 / Accepted: 21 February 1997     

Host preference and suitability of grasses for Oebalus pugnax

The rice stink bug, Oebalus pugnax (Fabricius) (Hemiptera: Pentatomidae: Carpocorini), though graminaceous, discriminates among its numerous host grass species. This could represent a feeding preference, it could be related to host suitability for growth and development. To clarify the role of host grass discrimination, two laboratory studies were conducted: (1) free‐choice tests to evaluate preferences of O. pugnax among 11 wild host grass species found in three rice‐producing counties of the central Mississippi Delta (MS, USA), and (2) no‐choice tests to evaluate the impact of rice (Oryza sativa L.), junglerice [Echinochloa colona (L.) Link], and dallisgrass (Paspalum dilatatum Poir.) (all Poaceae), on the development of O. pugnax from second instar to adult. In the free‐choice test, four experiments were conducted, each with four sets of host grass species and observed 1, 2, 4, 8, and 16 h after release in cages. Approximately 4 h was necessary for O. pugnax to settle on preferred host grasses. Oebalus pugnax showed a feeding preference for junglerice over all 10 other grass species. Bahiagrass, Paspalum notatum Flueggé, was the least preferred. The no‐choice tests showed significant effect of host grass species on O. pugnax mean development time of nymphal survival to adults. Survival of nymphs was lower and mean development time was longer on dallisgrass compared to rice and junglerice. Knowledge of O. pugnax rate of growth and development on host grasses could be useful in the future development of rice integrated pest management strategies.     

High-efficiency <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of citrus via sonication and vacuum infiltration

An improved method for the Agrobacterium infiltration of epicotyl segments of ‘Pineapple’ sweet orange [Citrus sinensis (L.) Osbeck] and ‘Swingle’ citrumelo [Citrus paradisi Macf. X Poncirus trifoliata (L.) Raf.] was developed in order to increase transformation frequency. Sonication-assisted Agrobacterium-mediated transformation (SAAT), vacuum infiltration, and a combination of the two procedures were compared with conventional Agrobacterium-mediated inoculation method (‘dipping’ method). It was observed that the morphogenic potential of the epicotyl segments decreased as the duration of SAAT and vacuum treatments increased. Transient GUS expression was not affected by the different SAAT treatments, but it was significantly enhanced by the vacuum infiltration treatments. The highest transformation efficiencies were obtained when the explants were subjected to a combination of SAAT for 2 s followed by 10 min of vacuum infiltration. PCR and Southern blot analysis of the uidA gene were used to confirm the integration of the transgenes. The transformation frequencies achieved in this study (8.4% for ‘Pineapple’ sweet orange and 11.2% for ‘Swingle’ citrumelo) are the highest ones reported for both cultivars.     

Advertising acceptability: is mollusk olfaction important in seedling selection?

Although seedling herbivory is an important selective filter in many plant communities, how and why seedlings are selected is poorly understood. Here, we examined the putative role of herbivore olfaction in dictating seedling selection. Using a Y-tube olfactometer we compared snail (Helix aspersa) preference for pellets derived from 14-day-old macerated seedlings of nine European grassland (‘Test’) species against standard (‘Control’) pellets derived from lettuce. Snail movement towards ‘Test’ pellets was strongly correlated with seedling acceptability (Pearson’s r ² = 0.86, P > 0.01) and where snails exhibited a positive choice for the ‘Test’ species, the choice was made more quickly for highly acceptable species (r ² = 0.86, P > 0.01). In elucidating a link between seedling acceptability and olfactory response to macerated seedlings, our study suggests that even from an early ontogenetic stage plant selection by snails may be governed by olfactory cues. This finding highlights the need for research on the role of plant volatiles in plant–herbivore interactions to consider more fully interactions operating at the seedling stage.     

Oviposition acceptance and larval development of Chilo partellus stemborers in drought‐stressed wild and cultivated grasses of East Africa

Maternal host choices during oviposition by herbivorous insects determine the fitness of their offspring and may be influenced by environmental changes that can alter host‐plant quality. This is of particular relevance to ‘push‐pull’ cropping systems where host preferences are exploited to manage insect pest populations. We tested how drought stress in maize and companion plants that are used in these systems affect oviposition preference, larval feeding, and development of the spotted stemborer, Chilo partellus Swinhoe (Lepidoptera: Crambidae). Five host species were tested (all Poaceae): maize (Zea mays L.), Napier grass (Pennisetum purpureum Schumach), signal grass [Brachiaria brizantha (A. Rich) Stapf], Brachiaria cv. ‘Mulato’, and molasses grass [Melinis minutiflora (Beauv.)]. Under drought stress, maize experienced as much oviposition as control unstressed maize in choice and no‐choice experiments. Similarly, larval leaf damage was not significantly different in drought‐stressed vs. unstressed maize. In contrast, oviposition occurred less on drought‐stressed than on unstressed Napier and signal grass. Oviposition acceptance and leaf damage remained low in both drought‐stressed and unstressed molasses grass and Mulato. Larval survival and development remained high in drought‐stressed maize, but not in Napier, signal, and molasses grass and Mulato, where survival and development were low in both drought‐stressed and unstressed plants. Our results indicate that herbivore responses to drought‐stressed plants depend on the plant species and that drought stress can change host preference and acceptance rankings. In particular, trap‐crops such as Napier grass may not divert oviposition from the main maize crop under drought stress conditions.     

The effect of phosphorus deficiency on nutrient uptake,nitrogen fixation and photosynthetic rate in mashbean,mungbean and soybean

Phosphorous (P) fertilization is the major mineral nutrient yield determinant among legume crops. However, legume crops vary widely in the ability to take up and use P during deficiency. The aim here was to compare P uptake and translocation, biological nitrogen fixing ability and photosynthetic rate among mashbean (Vigna aconitifolia cv. ‘Mash-88’), mungbean (Vigna radiata cv. ‘Moong-6601’) and soybean (Glycine max L. cv. ‘Tamahomare’) during deficiency in hydroponics. Two treatments, the withdrawal of P from the solution (P-deprivation) and continued P at 160 μM (P sufficient) were effected at the pod initiation stage. Plants were grown for 20 days. Short-term labeling with ³²P showed the uptake and distribution of P into plant parts. Withdrawal of P from the solution reduced biomass, photosynthetic activity, and nitrogen fixing ability in mungbean, and mashbean more than in soybean. P deprivation decreased P accumulation more than N accumulation. The decrease was more severe in mungbean and mashbean than soybean. More P was translocated and distributed into leaves in soybean than in mungbean and mashbean. Leaf P amount was more correlated to leaf area than to photosynthetic rate per unit leaf area among all three legume species. The results indicate that selection for increased efficiency of P utilization and leaf area may be used to improve leguminous crops.     

Life history ofAphis gossypii on cucumber: influence of temperature, host plant and parasitism

Life table data forAphis gossypii Glover (Homoptera: Aphididae), an important pest in glasshouse cucumber crops, were studied at 20, 25 and 30°C on two cucumber cultivars (Cucumis sativus L.) in controlled climate cabinets. The development time on the cucumber cv. ‘Sporu’ ranged from 4.8 days at 20°C to 3.2 days at 30°C. Immature mortality was approximately 20% and did not differ between temperatures. Most mortality occurred during the first instar. Reproduction periods did not differ among temperatures, but at 25 and 30°C more nymphs were produced (65.9 and 69.8 nymphs/♀, respectively) than at 20°C (59,9 nymphs/♀) because of a higher daily reproduction. Intrinsic rate of increase was greatest at 25°C (r _m =0.556 day⁻¹). At 20 and 30°C the intrinsic rate of increase was 0.426 and 0.510, respectively. On cv. ‘Aramon’, the development time ofA. gossypii was approximately 20% longer at all temperatures. Immature mortality did not differ between the two cultivars. The intrinsic rate of increase on cv. ‘Aramon’ was 15% smaller than on cv. ‘Sporu’. The use of cucumber cultivars partially resistant to aphids is discussed in relation to biological control of cotton aphid in glasshouses. Development time and immature mortality on leaves of the middle and upper leaf layer of glasshouse grown cucumber plants (cv. ‘Aramon’) were comparable to development in the controlled climate cabinets. On the lower leaves immature mortality was much higher (approximately 82%) than on leaves of the middle (24.0%) and upper leaf layer (24.5%). Reproduction was less on the lower leaf layer (45.9, 70.5 and 70.1 nymphs/♀ on leaves of the lower, middle and upper leaf layer, respectively). Aphids, successfully parasitized byAphidius colemani Viereck (Hymenoptera: Braconidae) only reproduced when they were parasitized after the third instar. Fecundity was 0.1 to 0.9 and 10.5 to 13.3 nymphs/♀ for aphids parasitized in the fourth instar or as adults, respectively. Reproduction of aphids that were stung but survived the attack was lower than for aphids not stung. Average longevity of these aphids was equal to the longevity of aphids not stung byA. colemani.     

Evolutionary relationships of membrers of the generaTaphrina, Protomyces, Schizosaccharomyces, and related taxa within the archiascomycetes: Integrated analysis of genotypic and phenotypic characters

To study the phylogeny and evolution of archiascomycetes, we determined the full sequence of the nuclear 18S rRNA gene from 14Taphrina species and 2Protomyces species, and the partial sequence ofSchizosaccharomyces japonicus var.japonicus. The sequences were phylogenetically analyzed by the neighbor-joining, maximum parsimony, and maximum-likelihood methods. We also looked at their principal phenotypic characters and genotypic character. Relationships within the Ascomycota are concordant with the previously published phylogenies inferred from 18S rDNA sequence divergence and divide the archi-, hemi-and euascomycetes into distinct major lineages. All the trees show that, within the archiascomycete lineage, 11 of the 14Taphrina species and the 2Protomyces species are monophyletic. A core groups ofTaphrina andProtomyces is always monophyletic. The evidence from molecular and phenotypic characters such as cell wall sugar composition, ubiquinone, cell wall ultrastructure, and mode of conidium ontogeny, strongly suggests that ‘T’. californica CBS 374.39, ‘T’. maculans CBS 427.69 and ‘T’. farlowii CBS 376.39 should be excluded from the archiascomycete lineage. ‘Taphrina’ farlowii CBS 376.39 groups withCandida albicans in the Saccharomycetales, whereas ‘T’. californica CBS 374.39 and ‘T’. maculans CBS 427.69 have a basidiomycete affinity and group with Tremellalean members in the hymenomycete lineage.Schizosaccharomyces is monophyletic. The strictly anamorphic yeastSaitoella complicata groups with the apothecial ascomyceteNeolecta vitellina rather than theTaphrina/Protomyces branch.