Natural infection of <Emphasis Type="Italic">Nicandra physaloides</Emphasis> by <Emphasis Type="Italic">Tomato severe rugose virus</Emphasis> in Brazil

Nicandra physaloides, a common weed in South America, was found to be infected by an isolate of Tomato severe rugose virus (ToSRV), a bipartite begomovirus. The plants developed severe yellow rugose mosaic and were collected in São Paulo State, Brazil. This isolate of ToSRV was transmitted by Bemisia tabaci B biotype from infected plants of N. physaloides to healthy plants of N. physaloides and tomato in a glasshouse. This is the first report of natural infection of N. physaloides by ToSRV in Brazil.     

Identification and partial characterization of a <Emphasis Type="Italic">Carica papaya</Emphasis>-infecting isolate of <Emphasis Type="Italic">Alfalfa mosaic virus</Emphasis> in Brazil

A Carica papaya plant with severe yellow leaf mosaic, leaf distortion, and systemic necrosis was found in the municipality of Piracicaba, state of São Paulo, Brazil. Transmission electron microscopy (TEM) analysis revealed the presence of potyvirus-like particles and bacilliform particles similar to those of the Alfamovirus genus. The potyvirus was identified as Papaya ringspot virus-type P (PRSV-P). Biological, serological, and molecular studies confirmed the bacilliform virus as an isolate of Alfalfa mosaic virus (AMV). Partial nucleotide and amino acid sequences of the coat protein gene of this AMV isolate shared 97–98% identity with the AMV isolates in the GenBank database. This report is the first of the natural infection of papaya plants by AMV.     

Occurrence of <Emphasis Type="Italic">Atopozelus opsimus</Emphasis> preying on nymphs and adults of <Emphasis Type="Italic">Glycaspis brimblecombei</Emphasis>

The stink bugs genus Atopozelus (Hemiptera: Reduviidae) is native in the Americas, with five described species of which only three have been recorded in Brazil. In August 2007, Eucalyptus camaldulensis leaves infested by lerp psyllid were collected and taken to the Forest Pest Biological Control Laboratory situated at São Paulo State University. A small green stink bug, identified as Atopozelus opsimus Elkins (Hemiptera: Reduviidae) that fed on nymphs and adult psyllids, was found on the leaves. These stink bugs presented phytophagy, omnivory and ability in opening the lerp (shell) that protect the psyllid. A. opsimus has demonstrated unusual particularities, different from many biological control agents found in nature; this predator thus deserves more attention for its potential to be evaluated.     

Detection of Bemisia tabaci Mediterranean cryptic species on soybean in São Paulo and Paraná States (Brazil) and interaction of cowpea mild mottle virus with whiteflies

Since the detection of Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) species in Brazil in the early 1990s, this whitefly has been the predominant species and the main viral vector in the country. In this work, whiteflies were collected from commercial soybean fields near and far from greenhouses where Mediterranean (MED) species had previously been detected infesting vegetable crops. Results indicated that MEAM1 was the predominant whitefly species in soybean in most sampling sites, while MED was found colonizing soybean plants in open field conditions alone and/or with MEAM1 in several places. Among the tested insects, MED species was mostly detected harbouring the facultative endosymbiont Hamiltonella. We also detected cowpea mild mottle virus (CPMMV) infecting soybean plants. Transmission assays demonstrated that MED species was more efficient transmitting CPMMV from beans to beans and from soybean to soybean plants than MEAM1, while MEAM1 was more efficient in transmitting the virus from soybean to bean than MED. Performance assays indicated that adult emergence of both species on CPMMV-infected soybean plants was higher when compared with the emergence on healthy plants. Moreover, nonviruliferous MED and MEAM1 adults preferred to settle more often on healthy plants, while viruliferous adults settled more often on CPMMV-infected soybean plants. As MED has already been found in soybean open fields in São Paulo and Paraná States, Brazil, and it is a good vector of CPMMV, we anticipate problems related to whitefly management and to increase in the incidence of the virus in soybean.     

A new method for inoculation of fruit with <Emphasis Type="Italic">Guignardia citricarpa</Emphasis>, the causal agent of citrus black spot

Citrus black spot (CBS) is a fungal disease, caused by Guignardia citricarpa, that has a high economic impact on citrus. Although G. citricarpa has been associated with black spot of citrus, an adequate pathogenicity test is still not available. Thus, our objective was to develop and evaluate a simple, safe, and practical pathogenicity test. We used fruits from Pera-Rio and Valencia sweet orange trees from two different orchards, located in the State of São Paulo, Brazil. Inoculation was performed by placing six disks colonized by G. citricarpa, onto the peel of healthy fruits, previously bagged. In the Pera-Rio sweet orange grove, initial symptoms of the false melanose type resulting from the inoculations were observed 55 days after inoculation (dai). In the Valencia grove, initial symptoms also of the false melanose type resulting from the inoculations occurred 73 dai. A total of 92.8% and 86.6% of the Pera Rio and Valencia fruits inoculated, respectively, showed symptoms of CBS. Citrus black spot symptoms were not observed in any of the control fruits.     

Species of Botryosphaeriaceae associated on mango in Brazil

The aim of the present study was to assess diversity in the Botryosphaeriaceae on trees and fruit of mango (Mangifera indica L.) in a semi-arid region in northeastern Brazil in which most exported fruit in the country are produced. Using morphological characteristics and DNA sequence data (ITS-1, ITS-2 and 5.8S rDNA) we confirmed the presence of Lasiodiplodia theobromae in the region, and for the first time report Fusicoccum aesculi and Neofusicoccum parvum. L. theobromae was prevalent in the Assú Valley and F. aesculi and N. parvum were in the São Francisco Valley. In fruit inoculations, L. theobromae and N. parvum were more virulent than F. aesculi.     

Spread of glyphosate‐resistant sourgrass (Digitaria insularis): Independent selections or merely propagule dissemination?

The first reports of glyphosate‐resistant (GR) sourgrass (Digitaria insularis) came from Paraguay (2005) and from South and Southeast Brazil, in Paraná (2008), and São Paulo state (2009). The hypothesis of this research is that the evolution of these resistant populations might have occurred independently. To test this hypothesis, a dose–response experiment was conducted in order to confirm resistance and inter simple sequence repeat markers subsequently were evaluated in five sourgrass populations from Paraguay (PY‐R), Paraná (PR‐S and PR‐R), and São Paulo (SP‐S and SP‐R). Resistant populations were confirmed in PY‐R, PR‐R and SP‐R. The resistant populations presented lower polymorphism than susceptible populations, possibly related to the selection pressure exerted by the herbicide. Both similarity matrix and dendrogram provided evidences that the origin of resistance in the populations from Paraguay and Paraná could be the same, which also is explained by their geographical proximity. In contrast, the evolution of GR populations in São Paulo was suggested to occur independently from other locations because resistance evolved in genetically divergent populations. This study’s data provide evidences that GR sourgrass populations from Paraguay and Paraná share a similar genetic background; therefore, it is possible that resistance from Paraguay spread to Paraná through gene flow. The population from São Paulo probably was selected independently because the high values of its genetic structure and low level of gene flow. Independent selections and seed dispersion of GR sourgrass have contributed to the rapid spread of resistance across South America.     

Phylogenetic analysis of the citrus Huanglongbing (HLB) bacterium based on the sequences of 16S rDNA and 16S/23S rDNA intergenic regions among isolates in China

Phylogenetic analysis of Chinese isolates of the citrus Huanglongbing (HLB) bacterium based on the 16S rDNA and 16S/23S rDNA intergenic regions sequences was carried out. Nine HLB samples collected from different hosts with different symptoms in seven Chinese provinces, were subjected to PCR for amplifying and sequencing the 16S rDNA. The identity level among Chinese isolates was 98.5% to 100% and was the same with the Indian HLB isolate ‘Poona’ (GenBank accession number: L22532). By contrast, identity values were 97.5% to 97.8% with Candidatus Liberibacter africanus strain ‘Nelspruit’ (L22533), 96.3% to 97.3% with Ca. L. africanus subsp. ‘Capensis’ (AF137368), 95.3% to 96.5% with the Ca. Liberibacter sp. ‘LSg2’ (AY919312), and 94.9% to 96.0% with a strain of Ca. L. americanus from Brazil (São Paulo State; AY742824). A phylogenetic tree constructed with 16S rDNA sequences showed that all Chinese isolates belong to Ca. L. asiaticum. Analysis of the 16S/23S rDNA intergenic region was conducted on 18 HLB-diseased citrus samples with different symptoms, collected in seven provinces. These isolates showed no obvious variation and had an identity level >99.0% with one another. Sequence analysis of 16S/23S rDNA intergenic region and the relative phylogenetic tree showed that the Chinese isolates are very close to Ca. L. asiaticus, and distinct from Ca. L. africanus and Ca. L. americanus. These results suggest that the Chinese HLB isolates belong to the species Candidatus Liberibacter asiaticus. This is the first report on the classification of HLB isolates from China based on molecular investigations.     

Bioassay of insecticides on mortality of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> biotype B and transmission of <Emphasis Type="Italic">Tomato severe rugose virus</Emphasis> (ToSRV) on tomatoes

Tomato severe rugose virus (ToSRV) is a serious and prevalent begomovirus that causes severe mosaic and yield loss of tomato plants in Brazil. The virus is transmitted by Bemisia tabaci bitotype B (MEAM1) in a persistent circulative manner. This study evaluated the efficacy of cyantraniliprole foliar spray, cyantraniliprole root drenching, spiromesifen, thiamethoxam, and cartap on the mortality of Bemisia tabaci byotipe B and simulated primary and secondary transmission of ToSRV. None of the insecticides were effective in controlling primary transmission of ToSRV by B. tabaci. Cyantraniliprole root drenching, cartap, and cyantraniliprole foliar spray were effective in controlling secondary transmission of the virus, and infections were reduced by 94.5, 89.6, and 81%, respectively, compared to each of their controls. Thiamethoxam and spiromesifen did not provide effective control of secondary infection. Elimination of external sources of inoculum before starting new plantings and rational use of insecticides to reduce secondary infection may contribute to better disease management of tomato crops.     

Inheritance of resistance to <Emphasis Type="Italic">Meloidogyne enterolobii</Emphasis> and individual selection in segregating populations of <Emphasis Type="Italic">Psidium</Emphasis> spp

Interaction between the phytonematode Meloidogyne enterolobii and the fungus Fusarium solani has caused direct and indirect losses in the entire guava production chain and consequent extermination of guava plantations throughout Brazil. The combined action of these two pathogens is known as “guava decline”. In order to obtain and assess Psidium spp. interspecific hybrids for resistance to the nematode M. enterolobii, interspecific crosses of P. guineense (susceptible araçá) x P. cattleyanum (resistant araçá); P.guineense (susceptible araçá) x P. guajava (susceptible guava) and P. cattleyanum (resistant araçá) x P. guajava (susceptible guava) were conducted. These crosses resulted in hybrid immune, susceptible and resistant to Meloidogyne enterolobii. The chi-square test rejected the hypothesis of monogenic inheritance with incomplete dominance, which corroborates that this trait has polygenic action. Predictions of genetic values ??and parameters were obtained by the REML / BLUP procedure, at individual level. Finally, the 30 selected individuals (immune and resistant) were obtained, which will be backcrossed with guava for the recovery of the agronomic traits desired and subsequent release of a new cultivar.     

Intraspecific Evolution of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG-1 IA Associated with Soybean and Rice in Brazil based on Polymorphisms at the ITS-5.8S rDNA Operon

Rhizoctonia solani AG-1 IA causes leaf blight on soybean and rice. Despite the fact that R. solani AG-1 IA is a major pathogen affecting soybean and rice in Brazil and elsewhere in the world, little information is available on its genetic diversity and evolution. This study was an attempt to reveal the origin, and the patterns of movement and amplification of epidemiologically significant genotypes of R. solani AG-1 IA from soybean and rice in Brazil. For inferring intraspecific evolution of R. solani AG-1 IA sampled from soybean and rice, networks of ITS-5.8S rDNA sequencing haplotypes were built using the statistical parsimony algorithm from Clement et al. (2000) Molecular Ecology 9: 1657–1660. Higher haplotype diversity (Nei M 1987, Molecular Evolutionary Genetics Columbia University Press, New york: 512p.) was observed for the Brazilian soybean sample of R. solani AG-1 IA (0.827) in comparison with the rest of the world sample (0.431). Within the south-central American clade (3-2), four haplotypes of R. solani AG-1 IA from Mato Grosso, one from Tocantins, one from Maranhão, and one from Cuba occupied the tips of the network, indicating recent origin. The putative ancestral haplotypes had probably originated either from Mato Grosso or Maranhão States. While 16 distinct haplotypes were found in a sample of 32 soybean isolates of the pathogen, the entire rice sample (n=20) was represented by a single haplotype (haplotype 5), with a worldwide distribution. The results from nested-cladistic analysis indicated restricted gene flow with isolation by distance (or restricted dispersal by distance in nonsexual species) for the south-central American clade (3-2), mainly composed by soybean haplotypes.     

Special issue “Deepen knowledge in plant pathology for innovative agroecology”

This study aimed to determine the genetic variability of isolates of rotting of pineapple fruitlet core in Brazil on the states of Paraiba, Pernambuco and Rio Grande do Norte, based on phylogenetic analysis of the RPB2 gene, morphocultural markers and aggressiveness of the isolates. The phylogenetic reconstruction of maximum parsimony and bayesian inference of the isolates were performed. Twenty-seven phylogenetic lineages were characterized with morphocultural markers on potato dextrose agar and synthetic nutrient-poor agar. The aggressiveness of these lineages were characterized in leaves and fruits of pineapple ‘Pérola’ cultivar. The Phylogenetic reconstruction showed close relationship between isolates of this study and phylogenetic lineages of F. guttiforme, F. ananatum and F. oxysporum by analysis of RPB2 gene. Phylogenetic lineages of this study shared significant morphocultural markers similar to those described for this species. Overall, the lineages related with Fusarium oxysporum Species Complex were more aggressive to the fruits of the Fusarium lineages related with Fusarium fujikuroi Species Complex. It is possible that F. ananatum and F. guttiforme or other lineages closely related to these species or F. oxysporum are present in the fields causing pineapple rotting fruitlet core in fruits of pineapple ‘Pérola’ in Brazil. The used markers determined high genetic variability in isolates analyzed in leaves and pineapple fruit ‘Pérola’ in the states of Paraíba, Pernambuco and Rio Grande do Norte of Brazil and the pathogenic lineages analyzed were better adapted to the fruits than to the detached leaves in pineapple ‘Pérola’ analyzed.     

Suppressive effect of acetylated glyceride (Bemidetach™EC) on <Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis> transmitted by the whitefly <Emphasis Type="Italic">Bemisia tabaci</Emphasis> on greenhouse tomato plants

Acetylated glyceride (Bemidetach?EC)—a food additive—repels adult sweet potato whiteflies (Bemisia tabaci) and inhibits their mating behavior. We evaluated the effects of acetylated glyceride spraying of greenhouse-grown tomato plants on infestation with B. tabaci and the occurrence of Tomato yellow leaf curl virus (TYLCV) disease under commercial-like conditions. The abundance of adult B. tabaci was significantly reduced by three sprayings of acetylated glyceride, and the TYLCV incidence was significantly suppressed to less than 30% of that in the untreated control. These results suggest that acetylated glyceride sprays suppress the secondary spread of TYLCV in greenhouse-grown tomatoes by lessening B. tabaci adult density.     

Differential reaction of sweet pepper to infection with the crinivirus tomato chlorosis virus probably depends on the viral variant

The tomato chlorosis virus (ToCV), transmitted by whitefly species of the genera Bemisia and Trialeurodes in a semipersistent manner, causes significant losses in solanaceous crops including tomato (Solanum lycopersicum) and sweet pepper (Capsicum annuum). Worldwide reports of natural and experimental infection of sweet pepper plants with ToCV are contradictory, raising the question of whether the critical factor determining infection is related to the susceptibility of sweet pepper cultivars or the genetics of virus isolates. In this work, ToCV isolates obtained from different hosts and geographical origins were biologically and molecularly analysed, transmitted by B. tabaci MEAM1 and MED, and the reaction of different sweet pepper cultivars was evaluated under different environmental conditions. Brazilian ToCV isolates from tomato, potato (S. tuberosum), S. americanum, and Physalis angulata did not infect plants of five sweet pepper cultivars when transmitted by B. tabaci MEAM1. Temperatures did not affect the sweet pepper susceptibility to tomato-ToCV isolates from São Paulo, Brazil, and Florida, USA. However, sweet pepper-ToCV isolates from Spain and São Paulo, Brazil, were transmitted efficiently to sweet pepper plants by B. tabaci MEAM1 and MED. Although the results indicated that ToCV isolates from naturally infected sweet pepper plants seem to be better adapted to plants of C. annuum, phylogenetic analyses based on the complete nucleotide sequences of RNA1 and RNA2 as well as the p22 gene did not reveal significant nucleotide differences among them. Additional studies are needed to identify intrinsic characteristics of ToCV isolates that favour infection of sweet pepper plants.     

Conidial sporulation of <Emphasis Type="Italic">Stemphylium solani</Emphasis> under laboratory conditions and infectivity of the inoculum produced <Emphasis Type="Italic">in vitro</Emphasis>

Potato virus Y (PVY) is the type-species of the genus Potyvirus, family Potyviridae, being reported as a major tomato (Solanum lycopersicum L.) pathogen in several regions of the world. Pepper yellow mosaic virus (PepYMV) was originally described as a resistance-breaking Potato virus Y (PVY) isolate on Capsicum annuum L. cultivars, and afterwards it was also reported infecting tomatoes in Brazil. In the present work, a search for sources of resistance to both PepYMV and PVY was conducted in a collection of 119 accessions belonging to seven Solanum (section Lycopersicon) species. This germplasm was initially evaluated to PepYMV reaction by mechanical inoculation followed by symptom observations and ELISA. Potential PepYMV resistance sources were identified for the first time in S. habrochaites, S. peruvianum, S. corneliomuelleri, S. chilense, S. pimpinellifolium, and one accession derived from an interspecific cross (S. lycopersicum x S. peruvianum). A sub-group of 24 accessions with negative serology for PepYMV was also challenged with a PVY isolate, followed by serological and molecular detection with universal primers. Solanum habrochaites ‘L.03683’ and ‘L.03684’ were the only accessions found with stable resistance to both viruses. These results confirm S. habrochaites as the most important source of multiple resistance factor(s) to distinct Potyvirus species.     

Morphological and molecular characterization of <Emphasis Type="Italic">Diaporthe (</Emphasis>anamorph <Emphasis Type="Italic">Phomopsis)</Emphasis> complex and pathogenicity of <Emphasis Type="Italic">Diaporthe aspalathi</Emphasis> isolates causing stem canker in soybean

The complex of Diaporthe (asexual morph) species occurring on soybean constitutes an important pathogenic group associated with diseases such as pod and stem blight, seed decay and stem canker. Stem canker, caused by Diaporthe aspalathi, has been reported as the most aggressive form of canker and its occurrence has limited soybean crop productivity in the southern United States. The main form of pathogen control is the use of stem canker resistant soybean varieties. In this study, strains of Diaporthe and Phomopsis were isolated from stem and seeds of soybean in different locations in South America during the years 1989–2014. Genomic DNA from 26 isolates were analyzed by PCR-restriction fragment length polymorphism (RFLP) and Amplified Fragment Length Polymorphism (AFLP) techniques, and sequencing of internal transcribed spacer (ITS) regions of ribosomal DNA. The molecular analysis of ITS sequences by alignment with those of ex-type strains deposited in GenBank and morphological characteristics allowed the identification of Phomopsis longicolla, D. phaseolorum var. sojae, D. caulivora and D. aspalathi. An analysis of the pathogenicity of 13 isolates of D. aspalathi inoculated in soybean genotypes carrying different resistance genes to stem canker (Rdm1, Rdm2, Rdm3, Rdm4, Rdm5 and Rdm?) enabled us to identify the occurrence of at least three races of D. aspalathi occurring in Brazil. Among the isolates identified as D. aspalathi, both molecular and phenotypic analyses showed clustering depending on the date of collection and pathogenicity, which revealed the existence of variability of the pathogen.     

Predators of <Emphasis Type="Italic">Marchalina hellenica</Emphasis> (Hemiptera: Marchalinidae) on pine forests in Turkey

The honeydew of Marchalina hellenica is collected by honeybees for pine honey production, which is of great economic importance in Turkey. During 2009–2011 we investigated the predators of M. hellenica, which is distributed mainly in the areas with a Mediterranean climate – in the Aegean, Marmara and Mediterranean regions of Turkey. The findings showed that M. hellenica has many predators in the studied areas and that these play an important role in the forest ecosystem. These predators are: Anystis baccarum (L.) (Acarina: Anystidae); Allothrombium triticium Zhang and Allothrombium pulvinum Ewing (Acarina: Trombidiidae); Neoleucopis kartliana (Tanasijtshuk) (Diptera: Chamaemyiidae); Myrrha octodecimguttata (L.), Rodolia cardinalis Mulstant, Scymnus subvillosus (Goeze) (Coleoptera: Coccinellidae), Dichochrysa genei (Rambur), Dichochrysa prasina (Burmeister) and Chrysoperla lucasina (Lacroix) (Neuroptera: Chrysopidae); Wesmaelius subnebulosus (Stephens) (Neuroptera: Hemerobiidae), Cardiastethus nazarenus Reuter and Elatophilus pachycnemis Horváth (Hemiptera: Anthocoridae). The most efficient and common predator was N. kartliana, here recorded for the first time from Turkey.     

Molecular diversity in <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> isolates from common bean fields in Brazil

The common bean (Phaseolus vulgaris L.) is widely cultivated in Brazil and is known as a very important crop for families in this country. Fusarium wilt severely harms common beans and has become a big issue for this crop. In order to assist the breeding programs that target resistance to this disease, the evaluation of genetic diversity of the pathogen and its molecular characterization are crucial. Thus, the present goal was to identify Fusarium isolates obtained from several places in Brazil using molecular tools; select molecular markers for these isolates; and analyze their diversity. All of isolates were molecularly identified as Fusarium oxysporum f. sp. phaseoli (Fop). By using seven selected SSR markers, the results of diversity obtained by the dendrogram and the Bayesian analysis formed four groups where a large diversity of this fungus was found within each state. However, the groups were more homogenous according to the collection source and the pathogenicity test. More specifically, group 2 was composed of the most virulent strains and originated from Minas Gerais State – UFV, and group 3 was mostly composed by isolates from Goias state. Group I was also more diverse in terms of location and virulence. The overall results indicated a positive correlation between Fusarium diversity and its virulence to common bean. Furthermore, the use of these markers was effective in molecular identification and in detecting polymorphism within F. oxysporum f. sp. phaseoli.