Daily consumption and predation rate of different<Emphasis Type="Italic">Stethorus punctillum</Emphasis> instars feeding on<Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Laboratory experiments were conducted to evaluate the prey stage preference and the daily consumption of each stage of the coccinellid predatorStethorus punctillum Weise (Coleoptera: Coccinellidae) feeding on the two-spotted spider miteTetranychus urticae (Koch) (Acari: Tetranychidae). Groups of different life stages of the prey were offered (eggs, larvae, nymphs and adults). The prey preference varied with the stage ofS. punctillum. First larval instars had no significant preference among theT. urticae stages offered. Second larval instars consumed significantly more spider mite larvae in comparison with nymphs. In contrast, third larval instars indicated a strong preference for mite eggs. Significantly fewerT. urticae larvae were consumed by the fourth larval instars ofS. punctillum, in comparison with the three other mite stages. Finally, adult predators consumed significantly more mite eggs than the other stages offered. This preferential trend was similar for all adults tested, whether during the pre-oviposition or the oviposition period. http://www.phytoparasitica.org posting Feb. 17, 2004.     

Fumigant toxicity of essential oils fromLaurus nobilis andRosmarinus officinalis against all life stages ofTribolium confusum

The essential oils from rosemary (Rosmarinus officinalis L.) and laurel (Laurus nobilis L.) obtained from Mersin Province in Turkey, were tested for their fumigant toxicity against all life stages of confused flour beetle (Tribolium confusum du Val.). GC-MS analysis showed that 1,8-cineole was found to be the major component of both rosemary and laurel essential oils. Vapors of rosemary and laurel essential oils were toxic to all life stages ofT. confusum. Only 65% mortality of the eggs was achieved when exposed to a dose of 172.6 mgl ⁻¹ air of rosemary essential oil at the longest exposure period (144 h); at the same dose, the pupae were the most resistant stage, with LT₉₀ (lethal time) value of 120.2 h. The adults were the most resistant stage to laurel essential oil, with LT₉₀ value of 77.2 h. On the basis of LT₉₀ values, tolerance of the life stages ofT. confusum to rosemary and laurel essential oils was, in descending order: pupa < larva < adult, and larva < adult < egg < pupa, respectively. Based on the concentration × time (Ct) products (g hl ⁻¹), rosemary essential oil was more toxic than laurel to the adults and larvae ofT. confusum. However, laurel essential oil was more toxic than rosemary to the eggs and pupae. Since these essential oils need such high Ct products to obtain complete mortality ofT. confusum compared with the most commonly used commercial fumigants, it would be impossible to use them on their own as a commercial fumigant against stored-product insects.     

Note: Evaluation of antibacterial activity of essential oil of<Emphasis Type="Italic">Rosa damascena</Emphasis> on<Emphasis Type="Italic">Erwinia amylovora</Emphasis>

Significant antibacterial activity was observed in the essential oil (E.O.) ofRosa damascena Mill. and the Minimum Bactericidal Concentration (MBC) of the E.O. was determined as 1386.5 μg ml⁻¹ forErwinia amylovora, the causal agent of fire blight disease. Ooze formation on immature pears and lesion formation in artificially inoculated shoots were completely (100%) prevented by the essential oil ofR. damascena (1500 μg ml⁻¹), essential oil ofThymbra spicata var.spicata (500 μg ml⁻¹) and streptomycin (100 μg ml⁻¹). Copper oxychloride plus maneb significantly reduced ooze formation and lesion formation, but the control was less than that obtained with the essential oils or streptomycin. The essential oil ofR. damascena may be a useful natural bactericide for the control of the fire blight pathogen,E. amylovora. http://www.phytoparasitica.org posting July 14, 2004.     

Insecticidal activity of crude seed extracts of<Emphasis Type="Italic">Annona</Emphasis> spp.,<Emphasis Type="Italic">Lansium domesticum</Emphasis> and<Emphasis Type="Italic">Sandoricum koetjape</Emphasis> against lepidopteran larvae

Crude ethanolic seed extracts ofAnnona muricata, A. squamosa (Annonaceae),Lansium domesticum andSandoricum koetjape (Meliaceae) collected from different locations and years in Maluku, Indonesia, were screened for inhibition of larval growth against the polyphagous lepidopteranSpodoptera litura (Noctuidae). Extracts ofA. squamosa were significantly more active (20-fold) than those ofA. muricata. A. squamosa collected from Namlea yielded the extracts with the greatest inhibitory activity. There were significant differences among locations for bothA. squamosa andA. muricata but not forL. domesticum andS. koetjape. Extracts ofA. squamosa, collected from Namlea, inhibited larval growth in a dose-dependent manner, with a dietary EC₅₀ (effective concentration to inhibit growth by 50% relative to controls) of 191.7 ppm fresh weight. Extracts ofA. squamosa collected from individual trees in Namlea also varied in growth inhibitory effect againstS. litura andTrichoplusia ni larvae. This species is a candidate for development of a botanical insecticide for local use in Indonesia. http://www.phytoparasitica.org posting Dec. 1, 2003.     

Structural conservation of the <Emphasis Type="Italic">hrp</Emphasis> gene cluster in <Emphasis Type="Italic">Xanthomons oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis>

The clustered hrp genes encoding the type III secretion system in the Japanese strains MAFF301237 and MAFF311018 of Xanthomonas oryzae pv. oryzae were sequenced and compared. The strains differ in their pathogenicity, location, and year of isolation. A 30-kbp sequence comprising 29 open reading frames (ORFs) was identical in its structural arrangement in both strains but differed from X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines in certain genes located between the hpaB-hrpF interspace region. The DNA sequence and the putative amino acid sequence in each ORF was also identical in both X. oryzae pv. oryzae strains as were the PIP boxes and the relative sequences. These facts clearly showed that the structure of the hrp gene cluster in X. oryzae pv. oryzae is unique.     

Susceptibility of eggs of <Emphasis Type="Italic">Tribolium confusum</Emphasis>, <Emphasis Type="Italic">Ephestia kuehniella</Emphasis> and <Emphasis Type="Italic">Plodia interpunctella</Emphasis> to four essential oil vapors

Susceptibility of eggs of Tribolium confusum du Val. (Coleoptera: Tenebrionidae), Ephestia kuehniella (Zell.) (Lepidoptera: Phycitidae) and Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) to vapors of essential oil from garlic (Allium sativum L.), birch (Betula lenta L.), cinnamon (Cinnamonum zeylanicum (Blume)) and aniseed (Pimpinella anisum L.) was studied. Preliminary bioassay tests indicated that vapors of the essential oils had a significant effect on the eggs of tested insect species when exposed to a concentration of 20 μl l ⁻¹ air for 24 h. Generally, garlic and birch essential oils were more toxic to the eggs of tested insect species than cinnamon and aniseed essential oils (except for eggs of T. confusum). There was also a significant difference between susceptibility of eggs of T. confusum, E. kuehniella and P. interpunctella to tested essential oils. Toxicity data indicated that eggs of T. confusum were more susceptible to tested essential oils, with LC₉₀ values ranging from 3.11 to 33.49 μl l ⁻¹ air, than those of E. kuehniella and P. interpunctella; eggs of P. interpunctella were the most tolerant to the essential oils, with LC₉₀ values ranging from 22.02 to 72.42 μl l ⁻¹ air. Concentration × time (Ct) products of 0.29, 0.22, 0.13 and 1.37 mg h l ⁻¹ for garlic, birch, cinnamon and aniseed essential oil, respectively, were required to obtain 90% kill of T. confusum eggs. Although cinnamon essential oil had a much closer Ct product value to methyl bromide, garlic and birch essential oils were found to be the most promising ones since they had also high fumigant toxicity on eggs of both E. kuehniella and P. interpunctella.     

Note: Life history of<Emphasis Type="Italic">Lixus bardanae</Emphasis> on curly dock (<Emphasis Type="Italic">Rumex crispus</Emphasis>) in Turkey

The biology ofLixus bardanae (F.) (Coleoptera: Curculionidae) on curly dock (Rumex crispus L.) in northeastern Anatolia (Bayburt, Erzurum and Kars Provinces), Turkey, was studied during the years 2000 and 2001.L. bardanae completes one generation in a year, overwintering as an adult. It feeds on leaves of the host plant. Females lay eggs individually into stems and the young larvae create galleries in stems while feeding. Pupation occurs inside stems in cells fashioned from frass. In late September, adults move into soil and overwinter in an upright position around the roots of the host plant. Infestation levels were found to range between 34% and 84%. Two parasitoids,Exeristes roborator F. andEndromopoda phragmitidis Perve (Hymenoptera: Ichneumonidae), were reared fromL. bardanae. http://www.phytoparasitica.org posting Dec. 21, 2003.     

Reaction of <Emphasis Type="Italic">Leersia</Emphasis> grasses to <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> collected from Japan and Asian countries

The present study was conducted to determine if there is specificity in the host-pathogen relationship between the isolates of Xanthomonas oryzae pv. oryzae, the causal bacterium for rice blight and Leersia grasses, the alternative weed hosts of the disease. Plants of three species of Leersia, namely, L. sayanuka, L. oryzoides and L. japonica, were collected from various parts of Japan and were inoculated with the X. oryzae pv. oryzae isolates obtained from various locations in Japan and from 11 Asian countries. Four L. sayanuka plants were found susceptible to all Race II isolates and some Race I isolates, but were resistant to all Race III isolates. Race III is known to have a wider range pathogenicity to rice cultivar groups compared with Race I and II. Although the reactions of two L. oryzoides plants to Race I and II isolates were similar to that of L. sayanuka, the L. oryzoides plant collected from Niigata Prefecture showed a susceptible reaction to some Race III isolates. On the other hand, L. japonica plants gave reactions different those of L. sayanuka and L. oryzoides, with two plants of L. japonica found to be resistant to all test isolates collected from Japan. The Asian isolates exhibited a wide host range against the international differential rice cultivars, but almost all of them were avirulent to Leersia plants. These results indicate that the relationship between the pathogenicity of the causal bacterium and the resistance of host plants is very complex, and suggest that pathogenic diversity of X. oryzae pv. oryzae might be related to the resistance of Leersia spp.     

Mechanisms of resistance in<Emphasis Type="Italic">Lycopersicon</Emphasis> germplasm to the whitefly<Emphasis Type="Italic">Bemisia argentifolii</Emphasis>

The silverleaf whiteflyBemisia argentifolii Bellows & Perring (Homoptera: Aleyrodidae) [also known as strain B of the sweetpotato whiteflyB. tabaci (Gennadius)] is a major pest of tomatoes due to both feeding damage and transmission of plant viruses. Certain wild species ofLycopersicon have demonstrated high levels of resistance to the pest. Greenhouse studies were undertaken to quantify the effects on whitefly behavior and mortality of individual, resistant plants selected from three accessions ofL. pennellii (Corr.) D’Arcy (LA 1340, LA 1674 and LA 2560), five accessions ofL. hirsutum f.typicum Humb. & Bonpl. (LA 386, LA 1353, LA 1777, PI 127826 and PI 127827) and one accession ofL. hirsutum f.glabratum C.H. Mull. (PI 126449). In no-choice experiments, fewer adults settled on leaflets of the wild species and deposited 75–100% fewer eggs compared to the cultivated tomato,L. esculentum Mill. Adult mortality ranged from 77–100% on wild accessions but was only 1% onL. esculentum. Most dead adults were trapped in glandular trichome exudates. The effects of these resistant accessions onB. argentifolii were mechanically transferable by appressing the trichome exudates onto the leaves of the susceptible tomato, indicating an association between the factors mediating the resistance and the glandular trichomes. Laboratory studies evaluated the repellent, fumigant and residual toxic effects of representative constituents of trichome exudates onB. argentifolii adults by using selected concentrations and probit analyses. RC₅₀ values (estimated concentration to repel 50% of the adults) and LC₅₀ values for fumigant and residual toxicity indicated that 2-tridecanone had low levels of repellent and residual toxicity activity; that 2-undecanone had high levels of repellent and fumigant activity; and that ginger oil (composed, in part, of sesquiterpene hydrocarbons) had high levels of repellent and residual toxicity activity. These studies suggest that multi-factor resistance exists in wild tomato germplasm. By combining genetically the observed chemical constituents of resistance into a single germplasm, the resulting resistance may be more difficult forB. argentifolii to overcome. http://www.phytoparasitica.org     

Note: Tachinid parasitoids of<Emphasis Type="Italic">Ancyrosoma leucogrammes</Emphasis> and notes on parasitization rates of<Emphasis Type="Italic">Clytiomya dupuisi</Emphasis>

The study deals with the tachinid parasitoids ofAncyrosoma leucogrammes (Gmelin) (Heteroptera: Pentatomidae):Clytiomya dupuisi Kugler,Clytiomya sola (Rondani) andGymnosoma clavatum (Rohdendorf) (Diptera: Tachinidae). All species are recorded for the first time as parasitoids ofA. leucogrammes, andC. dupuisi was reared for the first time from a host. The parasitization rates ofC. dupuisi on adults ofA. leucogrammes varied from 7% to 9% between 1994 and 1999.C. sola andG. clavatum were reared in only small numbers. http://www.phytoparasitica.org posting July 10, 2003.     

Hpa1 secretion via type III secretion system in <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis>

In many Gram-negative plant pathogenic bacteria the type III secretion system (TTSS), encoded by hrp genes, is essential for pathogenicity in the host and induction of a hypersensitive reaction (HR) in nonhost plants. The expression of hrp genes has been suggested to be repressed in complex media, whereas it is induced in planta and under certain in vitro conditions. We recently reported that XOM2 medium allows efficient hrp expression by Xanthomonas oryzae pv. oryzae. In this study, we investigated hrp-dependent secretion of proteins by the bacteria in vitro. Using modified XOM2, in which bovine serum albumin was added and the pH was lowered to 6.0, we detected at least 10 secreted proteins and identified one as Hpa1. This is the first evidence of protein secretion via TTSS in X. oryzae pv. oryzae.     

Population structure of <Emphasis Type="Italic">Eleusine</Emphasis> isolates of <Emphasis Type="Italic">Pyricularia oryzae</Emphasis> and its evolutionary implications

Population structure of Eleusine isolates of Pyricularia oryzae (Magnaporthe oryzae) was examined using DNA markers. On the basis of rDNA sequences, Eleusine isolates were divided into two groups. One group clustered with Triticum isolates, while the other clustered with Eragrostis isolates. This grouping was supported by DNA fingerprinting with three repetitive elements: MGR586, MGR583, and grasshopper. These results suggest that the population of Eleusine isolates is composed of at least two groups that evolved independently from the original population of P. oryzae. Most of the isolates that were collected just after an outbreak of finger millet blast in the 1970s had almost identical fingerprint profiles although they were collected in distant prefectures. This result supports the idea that the outbreak was caused by seed transmission of a particular strain of Eleusine isolates.     

Tachinidae parasitoids of<Emphasis Type="Italic">Traumatocampa ispartaensis</Emphasis> from Turkey

Tachinid parasitoids ofTraumatocampa ispartaensis Doğanlar & Avcı (Lepidoptera, Thaumetopoeidae), which was found to be a new species, were collected from the cedar forests around Isparta-Kapıdağ. The species found wereBlondelia nigripes (Fall.),Compsilura concinnata (Meig.),Pales processioneae (Ratz.),Phryxe caudata (Rond.),Exorista segregata (Rond.) andCarcelia iliaca (Ratz.). Within the six species of Tachinidae,B. nigripes was the most common one, parasitizing up to 4.6% ofT. ispartaensis pupae. http://www.phytoparasitica.org     

Generation and characterisation of Tn5-tagged <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> mutants that overcome <Emphasis Type="Italic">Xa23</Emphasis>-mediated resistance to bacterial blight of rice

Xanthomonas oryzae pv. oryzae causes bacterial blight of rice. Xa23, a bacterial blight resistance gene identified originally in wild rice, Oryza rufipogon, is dominant and resistant to all X. oryzae pv. oryzae field isolates tested. The corresponding avirulence gene avrXa23 is unknown. Here we report the generation of a random insertion mutant library of X. oryzae pv. oryzae strain PXO99 using a Tn5-derived transposon tagging system, and identification of mutant strains that are virulent on CBB23, a near-isogenic rice line containing Xa23. A total of 24,192 Tn5 inserted clones was screened on CBB23 by leaf-cutting inoculation and at least eight of them caused lesions on CBB23 comparable to those on JG30, the susceptible recurrent parent of CBB23. Polymerase chain reaction and Southern blot analysis showed that all the eight mutants, designated as P99M1, P99M2, P99M3, P99M4, P99M5, P99M6, P99M7 and P99M8, have a single Tn5-insertion in their genomes. The flanking DNA sequences of the Tn5-insertion sites were isolated by PCR-walking and sequenced. Bioinformatic analysis of the flanking sequences, by aligning them with the whole genome sequences of X. oryzae pv. oryzae strains PXO99, KACC10331 and MAFF311018 through NCBI, revealed that the Tn5-insertions disrupted genes that encode TAL effector AvrBs3/PthA, ISXo1 transposase, Type II secretion system protein-like protein or outer membrane protein, glycogen synthase, cytochrome C5 and conserved hypothetical protein. Further identification of these mutants will facilitate the molecular cloning of avirulence gene avrXa23. The authors C.-L. Wang, A.-B. Xu contributed equally to this work; Y. Gao and Y.-L. Fan contributed equally to this work.     

<Emphasis Type="Italic">In vitro</Emphasis> Selection of Maize Rhizobacteria to Study Potential Biological Control of <Emphasis Type="Italic">Aspergillus</Emphasis> Section <Emphasis Type="Italic">Flavi</Emphasis> and Aflatoxin Production

The aims of this study were to select bacterial isolates from the non-rhizophere of maize soil and to examine their antagonistic activity against Aspergillus section Flavi strains. The first selection was made through ecophysiological responses of bacterial isolates to water activity (a_w) and temperature stress. Subsequently, an Index of Dominance test (I_D), ecological similarity and inhibition of the lag phase prior to growth, growth rate and aflatoxin B₁ accumulation were used as criteria. From the first assay nine bacterial strains were selected. They grew well at 25 and 30 °C, with growth optima between 0.982 and 0.955 a_W using 48 h of incubation. There was ecological similarity between the bacterial strains Bacillus subtilis (RCB 3, RCB 6), Pseudomonas solanacearum RCB 5, Amphibacillus xylanus RCB 27 and aflatoxigenic Aspergillus section Flavi strains at 0.982 at 25 °C. The predominant interaction between all selected bacteria and fungi in dual culture was mutual intermingling at 0.982. Mutual inhibition on contact and mutual inhibition at a distance was observed at 0.955 a_w, between only four bacteria and some Aspergillus strains. Bacillus subtilis RCB 55 showed antifungal activity against Aspergillus section Flavi strains. Amphibacillus xylanus RCB 27, B.␣subtilis RCB 90 and Sporolactobacillus inulinus RCB 196 increased the lag phase prior to growth and decreased the growth rate of Aspergillus section Flavi strains. Bacillus subtilis strains (RCB 6, RCB 55, RCB 90) and P. solanacearum RCB 110 inhibited aflatoxin accumulation. Bacillus subtilis RCB 90 completely inhibited aflatoxin B₁ accumulation at 0.982 a_W. These results show that the bacterial strains selected have potential for controlling Aspergillus section Flavi over a wide range of relevant environmental conditions in the stored maize ecosystem.     

Laboratory evaluation of a botanical natural product (<Emphasis Type="Italic">AkseBio2</Emphasis>) against the pear psylla<Emphasis Type="Italic">Cacopsylla pyri</Emphasis>

A botanical natural product,AkseBio2, was evaluated under laboratory conditions for its oviposition deterrent, ovicidal and larvicidal (nymphicidal) effects against the pear psyllaCacopsylla pyri (L.) (Hemiptera: Psyllidae). The product exhibited a strong oviposition deterrent effect for winterform and summerform females and caused a reduction in the total number of eggs laid in both choice and no-choice assays. Significant mortalities in freshly laid eggs (0–48 h) and various nymphal stages of the pest were recorded in toxicity assays. At a concentration of 0.1% (formulation), the highest biological activity of the product was recorded against the young (1st and 2nd) nymphal stages (up to 87.4% mortality) in comparison with the other biological stages of the pest. It was less active against the older (3rd-5th) nymphs, causing 62.1% mortality at the same concentration. In assays with non-target organisms, a significant negative effect was not observed. There were no significant changes on treated plants up to 7 days after treatment in any trial, nor was there any phytotoxicity on plant tissue as a result ofAkseBio2 treatments. The results suggest that the product can be used in psylla control instead of synthetic insecticides and may serve as an integrated pest management (IPM) component in pear orchards. http://www.phytoparasitica.org posting July 14, 2004.     

Biological control potential of Turkish entomopathogenic nematodes against the Mediterranean fruit fly <Emphasis Type="Italic">Ceratitis capitata</Emphasis>

The entomopathogenic nematodes Steinernema weiseri, S. feltiae, S. carpocapsae and two strains of Heterorhabditis bacteriophora, isolated from Turkish soils, were evaluated against larvae of the Mediterranean fruit fly (medfly) Ceratitis capitata in plastic cups under laboratory conditions with sandy loam soil and 10% moisture level. At a rate of 100 infective juveniles (IJs)/cm², the last instar larvae of C. capitata were susceptible to the entomopathogenic nematodes: the S. feltiae 09-31 strain recovered from Aydin provided 78% mortality, whereas S. weiseri and S. carpocapsae killed 50% and 56% of the larvae, respectively. Both strains of H. bacteriophora species caused less than 50% mortality. Except for S. feltiae, the majority of infected medflies died as prepupae or pupae within the puparia. More than 90% larval mortality was recorded at 200 and 400 IJs/cm² for S. feltiae. None of the nematode isolates infected the medfly pupae within the puparia. In pot experiments containing soil, S. feltiae caused 96% and 97% mortality at 100 and 200 IJs/cm², respectively. In pot experiments with grass present, more than 94% mortality was obtained in the presence of grass roots.     

Elucidating resistance in cotton genotypes to whitefly, <Emphasis Type="Italic">Bemisia tabaci,</Emphasis> by population buildup studies

Plant resistance has become an important component of integrated pest management (IPM) for management of whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae), an important pest of cotton in India. The present studies were undertaken to standardize the plant stage and identify resistant cotton genotypes against whitefly. Nine plant stages of F846, a susceptible cotton genotype, were exposed to whitefly for 25 days under no-choice conditions. The population buildup (eggs, nymphs, pupae and adults) was recorded. The 12-, 14- and 16-leaf stages were suitable for plant resistance studies against whitefly, and the 14-leaf stage was taken for further studies. Ten cotton genotypes of Gossypium hirsutum and two of G. arboreum were covered with split cages in which five pairs of B. tabaci (F₁) were released. The population buildup was recorded to categorize genotypes as resistant, moderately resistant, moderately susceptible or susceptible. The experiment was repeated with F₂ and F₃ generation whiteflies. Based on overall average score of three experiments, LD694 was rated as resistant; LK861, Supriya, RS2013, CNH911 and PA183 as moderately resistant; IS-376/4/1/20/72, NHH44, TxMaroon2-78, Bt 6304 and RS2098 as moderately susceptible; and F846 as susceptible. LD694 was found to be resistant in three consecutive generations of whitefly.