Insect growth-regulating effects of <Emphasis Type="BoldItalic">Withania somnifera</Emphasis> in a polyphagous pest, <Emphasis Type="BoldItalic">Spodoptera litura</Emphasis>

Treatment of sixth instar larvae and pupae of the polyphagous pest Spodoptera litura Fabr. (Lepidoptera: Noctuidae) with an acetone extract of leaves of Ashwagandha, Withania somnifera L. (Solanaceae), caused toxicity, molt disturbances, formation of larval–pupal, pupal–adult intermediates and adultoids. Our results suggest that W. somnifera acts as an insect growth regulator causing disruption of the endocrine mechanism regulating molting and metamorphosis.     

Calcium fertilizers induce soil suppressiveness to <Emphasis Type="BoldItalic">Phytophthora cinnamomi</Emphasis> root rot of <Emphasis Type="BoldItalic">Quercus ilex</Emphasis>

Based on the observation that the root disease caused by P. cinnamomi on Q. ilex has a low incidence and severity in soils with medium-high Ca²⁺ content, we studied the ability of Ca²⁺ fertilizers to induce soil suppressiveness to the pathogen. Studies on cultures of P. cinnamomi exposed to different Ca²⁺ fertilizers in vitro showed significant inhibition of sporangial, chlamydospore and zoospore production at millimolar concentrations while mycelial growth was mainly unaffected. Experiments performed with artificially infested soil showed that some Ca²⁺ fertilizers induce a significant decrease on chlamydospore viability. Additionally, greenhouse experiments using artificially infested soils showed a significant reduction of foliar and root symptom severities in Holm oak seedlings growing in soils amended with Ca²⁺ fertilizers. We suggest that limestone amendments in oak rangelands could enhance the suppressiveness of soils to P. cinnamomi, and it is likely that the inhibition of sporangial production was the main mechanism involved.     

<Emphasis Type="BoldItalic">Didymella glomerata</Emphasis><Emphasis Type="Bold">causing leaf blight on pistachio</Emphasis>

During 2015 and 2016, we detected blighted leaves of pistachio (Pistacia vera) trees in different orchards in Arizona (USA). A Phoma-like species was isolated from pycnidia that appeared embedded in the leaf tissue. The pathogen was identified by means of morphological characteristics and DNA analysis (by sequencing of the ITS, BT, and EF regions) as Didymella glomerata. The optimum temperature for mycelial growth of the pathogen was around 25 °C. Inoculation tests were conducted on healthy and wounded pistachio leaves, fruits, and shoots of the cv. Kerman (female) and the cv. Peters (male). Overall, the pathogen was highly virulent on leaves of both pistachio cultivars and did not need injuries for infection. Conversely, the pathogen did not cause any macroscopic symptoms on the inoculated fruits and shoots but showed a certain endophytic behavior in the shoots. Also, data on the ability of different fungicides to inhibit the in vitro mycelial growth of the pathogen are presented.     

Resistance to <Emphasis Type="Italic">Penicillium</Emphasis><Emphasis Type="Italic">hirsutum</Emphasis> Dierckx in garlic accessions

Among the factors affecting the quality and yield of garlic production, blue mold caused by -- Penicillium spp. -- is responsible for economical losses in many countries. Allicin, present in garlic bulbs, has been suggested as having antifungal activity against some Penicillium species. This study was conducted to evaluate the response of garlic accessions against Penicillium hirsutum infection and to compare this response with bulb allicin content. Twelve garlic accessions were inoculated with P. hirsutum, and assayed in greenhouse and growth chamber experiments. Plant growth parameters and the fungal production of conidia were evaluated. Significant differences were found among the accessions. Accessions Castaño and Morado were most resistant whereas AR-I-125 and Fuego were always severely affected by the disease. A low correlation was found (r = 0.17) between allicin content and tolerance, indicating that allicin is not the main factor involved in the resistance against P. hirsutum.     

Influence of plant stage and organ age on the receptivity of <Emphasis Type="BoldItalic">Pisum sativum</Emphasis> to <Emphasis Type="BoldItalic">Mycosphaerella pinodes</Emphasis>

On spring pea, ascochyta blight (Mycosphaerella pinodes) frequently appears at the plant base on yellowing stipules and disease scores are higher on basal parts of the plants than on the uppermost parts. In order to investigate the relationship between pea plant growth stage and/or organ age, and ascochyta blight on whole plants and detached stipules and pods, two experiments were conducted in 2009 and 2010 under controlled conditions. This study showed that plant stage does not influence receptivity to ascochyta blight before the appearance of the first visual signs of senescence (beginning of yellowing). When stipules were green, regardless of the plant stage, no differences of receptivity was observed (except for the youngest stipule(s) at the top of the plant). Plant stage only had an effect on receptivity by the way of the importance of the visual senescence of the organs and visual senescence has a more predominant effect on plant receptivity than the plant stage considered alone. An effect of the nodal position, linked with the stipule age, was observed on whole plant inoculation, with a decreasing disease severity gradient from the base to the top of the plant which was explained by a greater receptivity for the basal/older stipules which had begun yellowing. On detached stipules, a higher receptivity was observed as soon as visual senescence had been observed. On pods, during their filling, few symptoms appeared and severity was important as soon as they began to yellow. The effect of ascochyta blight on induced senescence of the infected stipules was also observed.     

Comparative biology and life tables of <Emphasis Type="BoldItalic">Trichogramma aurosum</Emphasis> on <Emphasis Type="BoldItalic">Cydia pomonella</Emphasis> at constant temperatures

The influence of constant temperatures on biological parameters of German strains of Trichogramma aurosum Sugonjaev & Sorokina (Hymenoptera: Trichogrammatidae) was evaluated in the laboratory on eggs of Cydia pomonella L. (Lepidoptera: Tortricidae). Development time and longevity of all strains were decreased as temperature increased. Development time of the strains differed significantly only when exposed to 15°, 20°, and 25°C. Cumulative fertility and longevity differed significantly at 15° and 20°C. Realized fertility differed significantly at all constant temperatures. Emergence rates of all strains were less than 65% and were decreased even further as temperature increased. Female-biased sex ratio ranged from 65% to 100% at all constant temperatures. The low temperature threshold for T. aurosum was 10°C and the mean number of degree-days at 15°, 20°, 25° and 30°C was 175, 183, 173 and 185, respectively. The Bavarian strain tolerated high temperatures and had the highest parasitization capability, while the Hessian strain had the lowest parasitization at all temperatures. Fertility life table analysis revealed a major effect of temperature on the population growth parameters. Net reproductive rate was highest at intermediate constant temperatures in all strains, with the highest rate recorded for the Bavarian strains at all constant temperatures. Mean cohort generation time, and population doubling time decreased as temperature increased. The daily intrinsic rate of increase and finite rate of increase were positively correlated with temperature. The relevance of our results is discussed in the context of climatic adaptation, intraspecific variability and biological control.     

Evaluation of the <Emphasis Type="BoldItalic">Sss</Emphasis> AgriStrip rapid diagnostic test for the detection of <Emphasis Type="BoldItalic">Spongospora subterranea</Emphasis> on potato tubers

Spongospora subterranea, f.sp. subterranea (Sss), which causes powdery scab, is mainly spread through infected seed tubers and survives in contaminated soil for many years. The visual assessment of tuber lots by inspectors carries the risk of misidentification due to the difficulty of distinguishing lesions caused by either Sss or Streptomyces spp.. To avoid this, the “Sss AgriStrip”, a rapid and lab-independent test tool based on a lateral flow immunoassay has been developed, and we assessed its accuracy and sensitivity for detecting Sss. The Sss AgriStrip performed as well as other lab-based identification methods. The Sss AgriStrip, microscopy, ELISA, PCR, and real-time PCR techniques identified infection with S. subterranea in all tubers with typical powdery scab lesions. When lots with tubers showing a mixture of typical and atypical (suspicious) symptoms were tested, the presence of S. subterranea was confirmed in all lesions by all methods. The DNA content was generally lower in atypical than in typical lesions. Diverse and suspicious symptoms, which were difficult to assign to either powdery or common scab, tested negative with Sss AgriStrip and the other methods. This was despite microscopic observation of sporosori-like structures in some samples. Isolation and molecular identification confirmed that these lesions were mostly caused by Streptomyces spp. The Sss AgriStrip is as sensitive as DAS-ELISA with a detection limit between 1 and 10 sporosori per ml buffer. It is ideal for rapid and selective detection of Sss on farms and border inspection points to prevent spread of the pathogen.     

Resistance of <Emphasis Type="Italic">Solanum</Emphasis> species to <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> subgroup IA and its vector <Emphasis Type="Italic">Myzus persicae</Emphasis>

Sixty-nine tomato genotypes representing nine Solanum species were evaluated for resistance to Cucumber mosaic virus (CMV) subgroup IA and its aphid vector Myzus persicae. Resistance was assessed by visual scoring of symptoms in the field under natural conditions, and in the greenhouse by artificial inoculations through aphid M. persicae and mechanical transmissions in the year 2007 and 2009. Considerable variation in responses was observed among the evaluation methods used. Field evaluations were found liable to errors as different levels were observed for the same genotypes in the different years, however mechanical inoculation was found to be the most useful in identifying CMV subgroup IA resistance, in contrast aphid transmission was most useful in identifying insect transmission resistance. All genotypes observed as highly resistant to CMV subgroup IA in the field or through vector transmission became systemically infected through mechanical inoculations. Using mechanical inoculation, six genotypes (TMS-1 of S. lycopersicum, LA1963 and L06049 of S. chilense, LA1353, L06145 and L06223 of S. habrochaites) were found resistant and another six (L06188 and L06238 of S. neorickii, L06219 of S. habrochaites, L05763, L05776 and L06240 of S. pennellii) were found tolerant showing mild symptoms with severity index (SI) ranging 1-2 and with delayed disease development after a latent period (LP) of 18–30 days. However, these genotypes were found to be resistant to highly resistant in the field and through inoculation by M. persicae; and they also supported low population levels of M. persicae except TMS-1. Another nine genotypes (LA2184 of S. pimpinellifolium L., LA2727 of S. neorickii, LA0111, L06221, L06127 and L06231 of S. peruvianum L., LA1306, L06057 and L06208 of S. chmielewskii) showing a susceptible response after mechanical inoculation were highly resistant, resistant and tolerant after M. persicae transmission. The resistant genotypes, identified in the present study can be exploited in the breeding programmes aimed at developing tomato varieties resistant to CMV subgroup IA and broadening the genetic base of CMV-resistant germplasm. The differences observed between mechanical and aphid transmission suggests that one should consider both evaluation methods for tomato germplasm screening against CMV subgroup IA.     

Advanced lentil lines screened for resistance to <Emphasis Type="BoldItalic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="BoldItalic">lentis</Emphasis> under greenhouse and field conditions

Fusarium oxysporum f. sp. lentis is the most important pathogen of lentil plants, and most areas under lentil cultivation are reported to have a fusarium wilt disease background. The plants are infected in the seedling stage and later stages of their development. Fusarium wilt disease, which has appeared at high incidence rates during recent years, has caused sharp drops in the yield, especially in Moghan, in the northwest of Iran. Forty-five isolates of the pathogen were collected from different regions of the country with two isolates from ICARDA in the summer of 2008 and identified using Nelson’s key. The pathogenicity of the collected isolates was studied on a sensitive line (ILL 4605) under greenhouse conditions and significant differences in pathogenicity were found among them. The most pathogenic isolates from three provinces, East Azerbaijan (EA 30), Ardebil (Ar 3) and Khorasan (Kh 45), were selected and used in screening of 55 developed lines under greenhouse and field conditions. In the greenhouse, test plants were inoculated by immersing root tips in spore suspension and sowing seeds in pre-infested pot soil. Field tests were carried out in a naturally highly infested farm. At all stages, the plant response to the disease was based on the percentage of dead plants. Cluster analyses of the greenhouse and field data led to the selection of three lines (81S15, FLIP2007-42 L and FLIP2009-18 L) that were resistant under greenhouse and field conditions.     

Resistance to <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> expresses a hypersensitive-like response in <Emphasis Type="Italic">Coffea arabica</Emphasis>

Root-knot nematodes (RKN) are obligate parasite species of the genus Meloidogyne that cause great losses in Arabica coffee (Coffea arabica L.) plantations. Identification of resistant genotypes would facilitate the improvement of coffee varieties aiming at an environmental friendly and costless nematode control. In this work, the C. arabica genotype ‘UFV 408-28’ was found to be resistant to the most destructive RKN species M. incognita. Pathogenicity assays indicated that the highly aggressive populations of M. incognita races 1, 2 and 3 were not able to successfully reproduce on ‘UFV 408-28’ roots and displayed a low gall index (GI = 2). An average reduction of 87% reduction of the M. incognita population was observed on ‘UFV 408-28’ when compared to the susceptible cultivar ‘IAC 15’. By contrast, ‘UFV 408-28’ was susceptible to the related species M. exigua and M. paranaensis (GI = 5 and 4, respectively). Histological observations performed on sections of UFV408-28 roots infected with M. incognita race 1 showed that nematode infection could be blocked right after penetration or during migration and establishment stages, at 6 days, 7 days and 8 days after infection (DAI). Fluorescence and bright field microscopy observations showed that root cells surrounding the nematodes exhibited HR-like features such as accumulation of phenolic compounds and a necrotic cell aspect. In the susceptible ‘IAC 15’ roots, 6 DAI, feeding sites contained giant cells with a dense cytoplasm. Necrotic cells were never observed throughout the entire infection cycle. The HR-like phenotype observed in the ‘UFV 408-28’—M. incognita interaction suggests that the coffee resistance may be mediated by a R-gene based immunity system and may therefore provide new insights for understanding the molecular basis of RKN resistance in perennial crops.     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.     

Characterization of <Emphasis Type="Italic">Inago1</Emphasis> and <Emphasis Type="Italic">Inago2</Emphasis> retrotransposons in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

From the genome of a Japanese field isolate of the rice blast fungus, Magnaporthe oryzae, we newly identified Inago1 and Inago2 LTR retrotransposons. Both elements were found to be Ty3/gypsy-like elements whose copies were dispersed within the genome of Magnaporthe spp. isolates infecting rice and other monocot plants. Southern hybridization patterns of nine re-isolates derived from conidia of the strain Ina168 produced after a methyl viologen treatment were not changed, indicating that the insertion pattern of Inago elements is relatively stable.     

Encapsulation rates of the parasitoid <Emphasis Type="BoldItalic">Anagyrus</Emphasis> sp. nr. <Emphasis Type="BoldItalic">pseudococci</Emphasis>, by three mealybug species (Hemiptera: Pseudococcidae)

Laboratory studies assessing the degree of suitability of the mealybugs Planococcus ficus (Signoret), Planococcus citri (Risso) and Pseudococcus calceolariae (Maskell), towards the Sicilian ecotype of the encyrtid parasitoid Anagyrus sp. nr. pseudococci were carried out. All three species of mealybugs were shown to encapsulate the eggs of the encyrtid; however, significant differences were revealed among them in rates of encapsulation and/or superparasitism. The level of aggregate encapsulation and effective encapsulation of the parasitoid eggs by P. citri (74.95 ± 0.87 and 60.19 ± 1.70, respectively) was significantly higher than that recorded for P. ficus (aggregate encapsulation = 58.43 ± 0.83 and effective encapsulation = 31.31 ± 1.55) and significantly lower than the values found in Ps. calceolariae (93.99 ± 0.97 and 88.61 ± 2.03, respectively). As such, rates of parasitism were significantly lower for P. ficus compared with both P. citri and Ps. calceolariae. The two latter species of mealybugs showed similar rates of parasitism by A. sp. nr. pseudococci. Moreover, a significantly higher rate of superparasitism was found for Ps. calceolariae compared with both P. citri and P. ficus, whereas no significant differences were found between the two species of Planococcus for this parameter.     

Toxicity of butene-fipronil,in comparison with seven other insecticides,in <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> and <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The speed of toxic action of an insecticide is an indicator for control efficacy and has considerable practical importance. For agricultural pest control, fast-acting is an important feature for an insecticide to consistently reduce the amount of feeding damage. Butene-fipronil is a novel compound obtained via the structural modification of fipronil. However, information about the toxicity and speed of toxic action is still limited. In the present paper, we compared the toxic feature of butene-fipronil with seven other insecticides, of which imidacloprid and abamectin are slow-acting insecticides, and acephate, endosulfan, methomyl, α-cypermethrin and spinosad are fast-acting insecticides. We found that the contact and stomach toxicities of butene-fipronil were among the highest ever estimated to Leptinotarsa decemlineata and Drosophila melanogaster. The speed of toxic action of butene-fipronil was determined using median lethal time (LT₅₀) at a dose (concentration) equivalent to LD₈₀ values. For L. decemlineata, the values for butene-fipronil, imidacloprid, abamectin, acephate, endosulfan, methomyl, cypermethrin and spinosad were calculated to be 39.9, 36.5, 37.5, 20.2, 22.4, 23.8, 16.4 and 23.1 h, respectively. Those for D. melanogaster were 29.8, 31.5, 29.4, 14.0, 20.3, 18.1, 13.5, and 20.1 h, respectively. ANOVA analysis showed that butene-fipronil, imidacloprid, abamectin had similar LT₅₀ values, whereas acephate, endosulfan, methomyl, spinosad and cypermethrin had comparable LT₅₀ values. Thus, butene-fipronil belongs to slow-acting insecticides. Our results provide more empirical information for butene-fipronil potential application.     

Characterization of <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis> causing soft-rot disease on <Emphasis Type="Italic">Pinellia ternata</Emphasis> in China

Pinellia ternata is a traditional Chinese herb which has been used in China for over 1,000 years. A soft-rot disease characterized by water-soaked lesions and soft-rot symptoms with a stinking odour was commonly observed in cultivated fields of this plant, and Pectobacterium-like bacteria were consistently isolated from the infected tissues. Two typical strains (SXR1 and ZJR1), isolated from Shanxi and Zhejiang, respectively, were identified. Pathogenicity tests revealed that these strains were virulent to P. ternata and induced the same symptoms as observed in the field. Characterization involving fatty acid profile, metabolic and physiological properties, 16S rDNA sequence and PCR-RFLP identified both isolates as P. carotovorum subsp. carotovorum (Pcc). The 16S rDNA of both isolates shared 97–99% sequence similarity with that of Pcc strains. The phylogenetic trees showed that both isolates were clustered in the group of Pcc and P. carotovorum subsp. odorifera and both PCR-RFLP profiles were consistent with the pattern E produced by the minority of Pcc strains. Thus, isolates SXR1 and ZJR1 were characterized as Pcc in spite of some differences. This is the first report that Pcc has been proven as a causal agent of soft-rot disease on P. ternata.     

Pathogenicity of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> against <Emphasis Type="Italic">Galleria mellonella</Emphasis>

The greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) is occasionally found in beehives and is a major pest of stored wax. Entomopathogenic fungi have recently received attention as possible biocontrol elements for certain insect pests. In this study, 90 isolates of Beauveria bassiana and 15 isolates of Metarhizium anisopliae were screened for proteases and lipases production. The results showed significant variations in the enzymatic action between the isolates. In the bioassay, the selected isolates evinced high virulence against the 4th instar of the G. mellonella larvae. The isolates BbaAUMC3076, BbaAUMC3263 and ManAUMC3085 realized 100% mortality at concentrations of 5.5 × 10⁶ conidia ml⁻¹, 5.86 × 10⁵ conidia ml⁻¹, and 4.8 × 10⁶ conidia ml⁻¹, respectively. Strong enzymatic activities in vitro did not necessarily indicate high virulence against the tested insect pest. The cuticle of the infected larvae became dark and black-spotted, indicating direct attack of fungus on the defense system of the insects. The LC₅₀ values were 1.43 × 10³, 1.04 × 10⁵ and 5.06 × 10⁴ for Bba3263AUMC, Bba3076AUMC and Man3085AUMC, respectively, and their slopes were determined by computerized probit analysis program as 0.738 ± 0.008, 0.635 ± 0.007 and 1.120 ± 0.024, respectively.     

First report of <Emphasis Type="BoldItalic">Chalaropsis punctulata</Emphasis> on date palm in Egypt,comparison with other <Emphasis Type="BoldItalic">Ceratocystis</Emphasis> anamorphs and evaluation of its biological control

Morphological studies of Chalaropsis punctulata and Thielaviopsis paradoxa revealed that the two fungi are similar to each other and produce endoconidia from phialidic conidiophores and thick dark-walled and pigmented aleuroconidia from conidiophores. There are minor morphological characters which separate the fungus Ch. punctulata from T. paradoxa: the latter form aleuroconidia in chains at the tip of specialized hyphae, whereas the former produce aleuroconidia singly. The percent inhibition of mycelial growth and the total count of conidia showed good inhibitory effects in all biocontrol methods used. The antagonist activity of Trichoderma harzianum against Ch. punctulata was due mainly to coil formation and hyphae grew alongside of the pathogen hyphae. Both hyphae and conidia of Ch. punctulata were malformed and swelled abnormally in the case of Bacillus isolates. This is the first report of Ch. punctulata on date palm in Egypt.     

<Emphasis Type="Italic">Eremothecium coryli</Emphasis> and <Emphasis Type="Italic">E.</Emphasis><Emphasis Type="Italic">ashbyi</Emphasis> cause yeast spot of azuki bean

Yeast-like fungi were isolated from lesions on azuki bean (cv. Shin-Kyotodainagon) seeds that had been sucked by bean bugs in Kyoto Prefecture, Japan. On the basis of morphological and physiological characteristics and sequence data of the internal transcribed spacer (ITS) regions including the 5.8S rDNA, these yeasts were identified as Eremothecium coryli and E. ashbyi. Pathogenicity of those yeasts was confirmed by a reinoculation test. To our knowledge, this is the first report of the occurrence of yeast spot in azuki bean in Japan. The nucleotide sequence data reported are available in the GeneBank/EMBL/DDBJ database as accessions AB478291–AB478309 for E. coryli AZC1–19 and AB478310–AB478317 for E. ashbyi AZA1–8.     

Genetic diversity of <Emphasis Type="Italic">Rhizobium vitis</Emphasis> strains in Japan based on multilocus sequence analysis of <Emphasis Type="Italic">pyrG</Emphasis>, <Emphasis Type="Italic">recA</Emphasis> and <Emphasis Type="Italic">rpoD</Emphasis>

Forty-one strains of Rhizobium vitis, either tumorigenic (Ti) or nonpathogenic, were characterized using multilocus sequence analysis (MLSA) of the partial nucleotide sequences of pyrG, recA, and rpoD. The strains separated into seven clades. Rhizobium vitis (Ti) strains isolated from Japan were divided into five genetic groups (A to E), and nonpathogenic R. vitis strains were divided into two genetic groups (F and G). This result suggests that there are new genetic groups of R. vitis in Japan. Among these groups, members of A and B groups are widely distributed throughout Japan.