Identification and species status of the mango biotype of Colletotrichum gloeosporioides in Ghana

Research work was carried out to identify and ascertain the species status of the mango biotype of Colletotrichum gloeosporioides infecting mangoes in Ghana. Forty five isolates of Colletotrichum species were collected from 12 districts in Ghana while five each were obtained from mango fruits from Florida, Mexico and Puerto Rico. The entire internal transcribed spacer region, partial beta-tubulin gene and partial glyceraldehyde-3-phosphate dehydrogenase gene of isolates were sequenced and used in phylogenetic studies. The results of the sequence analysis of the first ribosomal transcribed spacer (ITS 1) region showed that 35 % of the isolates from Ghana and all the five isolates from Mexico were the mango biotype of C. gloeosporioides, while the others were not. Phylogenetic studies showed that the mango biotype of the pathogen was Colletotrichum asianum but not C. gloeosporioides as previously thought. However, the other isolates that were not the mango biotype were identified as Colletotrichum siamense and Colletotrichum species which had probably cross-infected mango from other fruit crops in the field.     

Detection and partial molecular characterization of Plum pox virus on almond trees in Turkey

Almond (Prunus dulcis) is one of the well known stone fruit species grown for its unripe fruits and delicious seeds in Turkey. In the Trakya region, however, some prevailing virus infections have reduced almond yields and quality. In ten districts of Trakya, 260 leaf samples were collected from affected almond trees in June 2010. DAS–ELISA assays and RT-PCR tests were employed for the identification of viruses. As a result of these detection studies, five of the 260 leaf samples gathered from symptomatic almond trees had Plum pox virus (PPV), 81 of them had Prunus necrotic ringspot virus (PNRSV), and 11 samples contained Prune dwarf virus (PDV). Only four out of 260 samples had a mixture of these viruses. Partial nucleotide sequences of five almond isolates of PPV were determined and compared with 17 other PPV isolates in databases. Computer analysis of obtained and published nucleotide sequences showed identity ranged from 75.72% to 96.87%. Of the five PPV almond isolates obtained, however, there was a close nucleotide identity of 95.82–96.61% to Turkish isolates. Phylogenetic analysis of nucleotides and amino acids showed that five PPV isolates of almond from the Trakya Region of Turkey were clustered in the same subgroup with PPV-T Turkish isolates in GenBank. Therefore we can consider almond isolates of PPV as PPV-T strain, like the two other isolates from apricot trees in Turkey.     

Genetic groups of Clavibacter michiganensis subsp. michiganensis identified by DNA fingerprinting and the effects of inoculation methods on disease development

Clavibacter michiganensis subsp. michiganensis (CMM) strains collected in nine different prefectures in Japan were fingerprinted by inter-simple sequence repeats–polymerase chain reaction (PCR) and repetitive sequence–based PCR. Forty-three CMM strains corresponded to 22 DNA fingerprinting patterns, indicating that the CMM population in Japan has high genetic diversity. Tomato plants were inoculated with CMM by two different methods: defoliation using infected scissors and planting in soil containing infected plant debris. Defoliation resulted in a larger number of diseased plants, disease developed earlier, and plants survived for significantly fewer days. Upon planting in the infected soil, 33.3 % of plants were infected and 6.7 % were diseased, i.e., 20.1 % of infected plants developed disease symptoms. These results indicate that some plants are infected with CMM through plant debris in the soil as the primary inoculum and most infected plants maintain latent infection. Thus, growers may unwittingly carry out disbudding and defoliation of latently infected plants, resulting in secondary inoculum.     

Phenotypic and genotypic structure of Phytophthora infestans populations on tomato and potato in the North of Thailand in 2000–2002

A total of 80 single–lesion isolates of Phytophthora infestans were collected from tomatoes and potatoes in several locations in Chiang Mai and Tak provinces in 2000–2002. These isolates were analyzed for mating type, metalaxyl sensitivity, mitochondrial DNA (mtDNA) haplotype RFLP pattern as determined by probe RG57, and for microsatellite markers. All isolates were A1 mating type. Isolates from tomato were usually sensitive to metalaxyl, but isolates from potato were usually resistant to metalaxyl. With one exception, all tomato isolates were related to the US-1 clonal lineage. With two exceptions, all potato isolates were related to two European lineages. In these two provinces, the populations of P. infestans on tomatoes are clearly different from those on potatoes.     

Alien scale insects (Hemiptera Coccoidea) in European and Mediterranean countries: the fate of new and old introductions

This contribution focuses on recent interceptions and introductions of alien scale insects and their current distribution in European and Mediterranean countries. Data and collections were gathered in markets, nurseries, and botanical gardens, mostly in Italy, either indoors or outdoors. New or recent records of the following alien species are presented: Exallomochlus hispidus (Morrison); Ferrisia virgata (Cockerell) (Pseudococcidae); Coccus viridis (Green); Milviscutulus mangiferae (Green) (Coccidae); Aonidiella orientalis (Newstead); Aspidiotus destructor Signoret; Aulacaspis tubercularis Newstead; Fiorinia fioriniae Targioni Tozzetti; Lepidosaphes pinnaeformis (Bouché); Pseudaulacaspis brimblecombei Williams (Diaspididae). New data and pest status of Phoenicococcus marlatti Cockerell (Phoenicococcidae) and Trabutina mannipara (Hemprich & Ehrenberg) (Pseudococcidae) are also reported. The possible repeated introductions of the latter from North Africa to south Italy by trans-Mediterranean winds, is hypothesized.     

Molecular variability of Apple chlorotic leaf spot virus in Shaanxi,China

Apple chlorotic leaf spot virus (ACLSV) is one of the latent viruses that occur in apple orchards worldwide but usually without visible symptoms. In 2010–2012, a total of 550 apple leaf samples from 12 different major apple-producing areas in Shaanxi, China, were tested by serological assay for ACLSV; the results revealed an infection level of 51.5 %. Because of the known variability in the putative amino acid sequences of the coat protein (CP), and thus the potential for non-detection by serological assay, the molecular variability of isolates of ACLSV collected in Shaanxi was analyzed using PCR and compared with isolates from the rest of the world. Sequences of 504 nt corresponding to 87 % of the CP gene of 12 isolates were acquired by RT-PCR and deposited in GenBank with the accession numbers KF134387–KF134298. Comparisons of the partial CP gene sequences of these 12 isolates as well as isolates previously reported in the world revealed the pairwise identities ranging from 68.9–99.8 % and 73.8–100 % at the nucleotide and amino acid level, respectively. Phylogenetic analysis based on these nucleotide sequences showed that the 72 isolates deposited in GenBank fell into three groups (P205, B6 and Ta Tao 5 Group). Our 12 ACLSV isolates were separated into the P205 and B6 groups, respectively. Multiple alignment analysis of the amino acid sequences of CP revealed that there was a combination of six amino acids at positions 40, 59, 75, 86, 130 and 184 in isolates from each group that could be used to distinguish among the three groups. Two recombination events were identified from all isolates by recombination analysis, and three ACLSV isolates collected in this study participated in these two events. Our results show that molecular variation was present in isolates of ACLSV collected in Shaanxi province and this may reflect introductions of the virus associated with different sources of germplasm.     

Management of wheat blast with synthetic fungicides,partial resistance and silicate and phosphite minerals

Field trials conducted on a yellow-red latossol (pH 6.0), replicated in 2010 and 2011, sought to examine the effect of silicon, phosphite minerals, synthetic fungicides and genetic resistance for wheat blast management (Magnaporthe grisea) in Central Brazil. Disease intensity was measured on cvs. BRS 264 and BR18 subjected to the following Si treatments: pre-plant furrow application of Ca & Mg silicate (300 kg ha^-1); post-plant scattered application of Ca & Mg silicate on top of the soil (1 ton ha^-1); multiple foliar SiO₂ applications (30 g l ^-1); and non-treated control. Blast incidence and severity were scored. Further experiments were conducted on cv. BR-264, for examination of the effect of potassium phosphite and synthetic fungicides on wheat blast intensity, with the following treatments: K₂HPO₃ (1ml l ^-1); epoxinazole + pyraclostrobin (700 ml ha^-1); tebuconazole (600 ml ha^-1); tebuconazole + trifloxystrobin (750 ml ha^-1); and non-treated control. In 2010, disease intensity was lower than in 2011. In the silicate experiments, disease was significantly lower when plants were treated with foliar or furrow silicate. Si applications significantly reduced disease in BRS-264. While BR-18 consistently demonstrated lower disease levels, cv. BRS-264 generally responded more markedly to silicon applications. In the phosphite/fungicide experiment of 2010, all treatments reduced disease when compared with the control, and in 2011 phosphite efficiency was not significantly different from some fungicide treatments. Synthetic fungicides demonstrated an average blast control of 55% by severity values. Yields were increased in the phosphite-treated plots (by 9–80%), in the Si treatments (by 26–92%), and more so, and more consistently, with synthetic fungicides (by 90–121%). Combined results of all field studies, carried out under environmental conditions highly conducive to disease, indicated that control of wheat blast necessitates the joint integration of several alternatives for efficient disease management.     

Analysis of genetic and virulence variability of Stemphylium lycopersici associated with leaf spot of vegetable crops

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46?S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.), pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage.     

The distribution of passionfruit ringspot virus in its main host plants in Ivory Coast

A field survey and testing of leaf samples have given an impression of the distribution of passionfruit ringspot virus in twoAdenia and twoPassiflora spp. in Ivory Coast. The data suggest thatAdenia spp. are the original host plants of the virus. Introduction to Ivory Coast of the virus withP. edulis seems unlikely. Since theAdenia spp. prefer forest and shrub vegetations and aphid flights are shorter in the dryer savannah areas of the North, the cultivatedP. edulis has chances to remain healthy over a prolonged period of growth, which is excluded in the South of the country.     

Synergism of pyrethroid — organophosphorus insecticide mixtures in insects and their toxicity againstspodoptera littoralis larvae

Synergism of mixtures of pyrethroids with organophosphorus (OP) compounds in insects is reviewed, and the toxicity of such combinations againstSpodoptera littoralis (Boisd.) larvae is reported. Mixtures of one of the pyrethroids cypermethrin, fenvalerate or deltamethrin with one of the OP compounds monocrotophos, profenofos, azinphos-methyl or acephate were assayed at different ratios as 24-h-old dipping residues on alfalfa, which was fed toS. littoralis larvae for 48 h. With most of the binary mixtures containing various OP concentrations in excess of those of the pyrethroids, synergism was demonstrated. In the pairs fenvalerate — azinphos-methyl, deltamethrin — azinphos-methyl and deltamethrin — profenofos, however, no synergism was found. In a detailed investigation with pyrethroid concentrations causing 20% mortality and OP concentrations giving a kill of no higher than ;10%, the above findings on synergism were amply confirmed. A cypermethrinmonocrotophos mixture showed synergism also on cotton leaves sprayed in the field. Synergism could not be demonstrated by topical application of pyrethroid — OP mixtures.     

Braconidae (Hymenoptera) associated with forest and ornamental trees and shrubs in Israel

Data are presented on the occurrence of Braconidae (Hymenoptera parasitica) parasitizing insects associated with forest and ornamental trees and shrubs in Israel. Fifty-five genera of plants are listed, the richest in braconid fauna being tamarisk (9 species); acacia, pistachio and poplar (8 species each); carob and oak (7 species each). Of the 95 species of insect hosts mentioned, 53 are Lepidoptera, mostly Gelechiidae (7 species), Pyralidae (6 species), Noctuidae (5 species), Gracillariidae, Tortricidae, Geometridae, Lymantriidae and Lycaenidae (3 species each); 44 are Coleoptera, mostly Cerambycidae (13 species), Scolytidae (12 species), and Bostrichidae (9 species); three are Diptera. Of the 92 species of braconids listed, of which only 65 have been fully named, 56 develop in Lepidoptera, mostly Noctuidae (15 species), Gelechiidae (11 species) and Pyralidae (9 species); 33 species develop in Coleoptera, mostly Cerambycidae (12 species, Bostrichidae (10 species) and Scolytidae (5 species); and three species develop in Diptera. Thirty-eight species are new to the fauna of Israel; at least three of them are new to science,viz., Gnaptodon, Gildoria andDendrosotinus titubatus Papp.     

Detection and estimation of Citrus Tristeza Virus infection rates based on Elisa Assays of packing house fruit samples

Enzyme-linked immunosorbent assay (ELISA) proved to be a sensitive detector for citrus tristeza virus (CTV) in orange fruits (Citrus sinensis (L.) Osbeck). Samples of five fruits were taken from 350-kg packing house containers and tested by ELISA to predict the infection rate of CTV in two infected orange groves. The predicted infection rates, 1% and 11%, were in reasonable agreement with the observed rates of 1% (15/1400) and 16% (324/2053), respectively. The 360 test samples from reputedly uninfected groves all tested negative. These results suggest that the ELISA procedure may provide a general method of detecting viral or other systemic pathogenic infections using the fruit as the test material in place of tree tissue. Fruit samples can be collected routinely at the packing house to reduce test costs.     

Introductions of beneficial insects into Israel by the Institute of Plant Protection Quarantine Laboratory,ARO, during 1971-1978

A list is given of the beneficial insects introduced into Israel during 1971–1978 by the Institute of Plant Protection Quarantine Laboratory for biological control of plant pests. The list includes 15 species of Hymenoptera (14 parasites and one parasite/predator) as well as four coleopterous predators. Included are each shipment’s origin, date and sender, as well as the results of these introductions.     

Arthropod fauna and main insect pests of plane trees in Israel

Fifty-four insect and four mite species are included in a list of the entomofauna of plane trees in Israel. Only two species are monophagous:Phyllonorycter platani (Stgr.) (Lepidoptera: Gracillariidae) andEdwardsiana iranicola Zachv. (Heteroptera: Cicadellidae). Four species are noxious:P. platani, the main insect pest of the plane trees in Israel;Zeuzera pyrina L. (Lepidoptera: Cossidae);Kalotermes flavicollis F. (Isoptera: Kalotermitidae); andE. iranicola. Of much lesser importance areTargionia vitis Sign. (Homoptera: Diaspididae),Heliothrips haemorrhoidalis Bché. andRetithrips syriacus May. (Thysanoptera: Thripidae). About half of the listed species are natural enemies and many are parasites ofP. platani. Details are given on the noxious species, together with recommendations for prevention and control.     

Effect of Cotton leaf curl virus infected plants on the biology of the whitefly,Bemisia tabaci (Hemiptera: Aleyrodidae): Vector–virus mutualism

Cotton leaf curl virus (CLCuV) (Geminiviridae : Begomovirus), the causative agent of leaf curl disease in cotton plants (Gossypium hirsutum), is exclusively transmitted by whitefly species Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). CLCuV transmission occurs in Sriganganagar (Rajasthan), an area endemic with cotton leaf curl disease. The relationships between plant viruses, their herbivore vectors and host plants can be beneficial, neutral, or antagonistic, depending on the species involved. To further understand these relationships, fecundity and life history parameters of an indigenous non- b (Asia II genetic group) biotype whitefly, B. tabaci, were compared on 10, 25, and 40 days post-inoculation (DPI), in CLCuV-infected and healthy cotton plants to determine the effect of virus on its vector. The development time of the immature stages of whiteflies was significantly reduced on CLCuV-infected plants. The development time of the immature stages did not change with severity of symptoms at 25 and 40 DPI (45- and 60-d-old plants). Cotton leaf curl virus infection increased percent egg viability of B. tabaci. Whiteflies deposited significantly fewer eggs on virus-infected plants than on healthy plants. Whiteflies had better egg viability on younger plants than older plants, whereas plant age did not affect the fecundity. Male and female whiteflies had shorter longevity on CLCuV-infected plants than on healthy plants.     

Control of phytophthora leaf blight of taro (Colocasia esculenta) by fungicides and roguing

Six fungicides were evaluated under laboratory and field conditions for control of Phytophthora leaf blight of taro,Coloeasia esculenta, incited byPhytophthora colocasiae. Inin vitro tests Deraosan 65W was the most effective in inhibiting the mycelial growth of the test pathogen followed by Difolatan 80W, Fytolan (copper oxychloride), Apron 35F, Topsin-M 50W and Dithane Z-78 75W. Excellent control was obtained with Demosan 65W and Difolatan 80W, good control with Apron 35F, fair control with Fytolan, and poor control with Topsin-M 50W and Dithane Z-78 75W. Results ofin vivo tests were correlated with those of thein vitro tests. Roguing of infected leaves did not eradicate the pathogen but can only delay epiphytotics.     

Development of conventional and real-time PCR protocols for specific and sensitive detection of Colletotrichum capsici in chilli (Capsicum annuum L.)

Anthracnose, caused by Colletotrichum capsici, is a major disease of chilli (Capsicum annuum L.) affecting both fruit and seed quality. The pathogen is both internally and externally seedborne. However, a rapid and sensitive method for detection of this pathogen in seeds is currently limited. In this study, a polymerase chain reaction (PCR) method based on sequence characterized amplified region (SCAR) marker was developed for specific and sensitive detection of C. capsici in chilli seeds and fruits. The developed SCAR primers were highly specific to C. capsici and resulted in the amplification of an expected 250-bp fragment from genomic DNA of all seven of the C. capsici isolates tested. No amplification occurred when the SCAR primers were tested with genomic DNA from three other fungal isolates and four other Colletotrichum species. The SCAR primers successfully amplified similar sized fragments from DNA derived from C. capsici-infected chilli fruits. The molecular detection sensitivity of C. capsici was 1 pg of purified C. capsici DNA template and 25 ng of DNA from C. capsici-infected chilli fruits. A real-time PCR assay was also developed using SYBR Green chemistry for detection of C. capsici in chilli fruits and seeds. The standard curve obtained showed a linear correlation between copy number of the cloned target DNA sequence of C. capsici and cycle threshold (Ct) values, with R² of 0.98. These PCR-based assays may be highly useful in detection of this important pathogen in chilli seeds and fruits in plant quarantine laboratories.     

The effect of diflubenzuron on Eggs and larvae ofEarias Insulana (Boisd.)

Diflubenzuron was tested for its efficacy against the eggs and larvae ofEarias insulana Boisd. in laboratory experiments. The toxicity obtained on dipping the eggs was only moderate and, moreover, the results were highly inconsistent. The mortality of larvae on an artificial diet into which the wettable powder had been incorporated, was relatively high.     

Potential use of Eucalyptus globulus essential oil against Phytophthora colocasiae the causal agent of taro leaf blight

This study was carried out to determine the antimicrobial activity of essential oil derived from leaves of eucalyptus (Eucalyptus globulus) against Phytophthora colocasiae, the causal agent of taro leaf blight (TLB). Essential oil was obtained by Clevenger-type water distillation. The major compounds in this essential oil were identified using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC/MS). Antimicrobial activity of the essential oil was tested in vitro against mycelial growth, sporangia and zoospores germination of P. colocasiae. Additionally, in situ tests were conducted on detached healthy taro leaves discs and evaluating necrosis symptoms of TLB were assessed. Results of the chemical composition of the essential oil analysis showed that, 1.8-cineole (26.4 %), α-pinene (14.1 %) and p-cymene (10.2 %) are the most abundant compounds. In addition, four components could be identified in noticeable amounts (18.1 % in the total): regulator G1/G2 and its derivatives as well as flavesone, a regulator G3 derivative. G is a generic term for a family of 2,3-dioxabicyclo[4,4,0] decane system growth regulators. Here, G-regulators and derivatives are described for the first time in E. globulus. Antimicrobial activity of the essential oil was recorded with total inhibition of mycelia growth and sporangia germination at 0.625 mg/ml, while the complete inhibition of zoospores germination was recorded at 0.156 mg/ml. In situ results showed that essential oil completely inhibited the appearance of disease symptoms, necrosis development and sporulation at 3.5 mg/ml. These results demonstrated that, the essential oil of Eucalyptus globulus has high ability to inhibit the development of P. colocasiae, and might be used for controlling TLB.