Patricia Valle Pinheiro

The draft genome of whitefly Bemisia tabaci MEAM1, a global crop pest, provides novel insights into virus transmission, host adaptation, and insecticide resistance

BackgroundThe whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is among the 100 worst invasive species in the world. As one of the most important crop pests and virus vectors, B. tabaci causes substantial crop losses and poses a serious threat to global food security.ResultsWe report the 615-Mb high-quality genome sequence of B. tabaci Middle East-Asia Minor 1 (MEAM1), the first genome sequence in the Aleyrodidae family, which contains 15,664 protein-coding genes. The B. tabaci genome is highly divergent from other sequenced hemipteran genomes, sharing no detectable synteny. A number of known detoxification gene families, including cytochrome P450s and UDP-glucuronosyltransferases, are significantly expanded in B. tabaci. Other expanded gene families, including cathepsins, large clusters of tandemly duplicated B. tabaci-specific genes, and phosphatidylethanolamine-binding proteins (PEBPs), were found to be associated with virus acquisition and transmission and/or insecticide resistance, likely contributing to the global invasiveness and efficient virus transmission capacity of B. tabaci. The presence of 142 horizontally transferred genes from bacteria or fungi in the B. tabaci genome, including genes encoding hopanoid/sterol synthesis and xenobiotic detoxification enzymes that are not present in other insects, offers novel insights into the unique biological adaptations of this insect such as polyphagy and insecticide resistance. Interestingly, two adjacent bacterial pantothenate biosynthesis genes, panB and panC, have been co-transferred into B. tabaci and fused into a single gene that has acquired introns during its evolution.ConclusionsThe B. tabaci genome contains numerous genetic novelties, including expansions in gene families associated with insecticide resistance, detoxification and virus transmission, as well as numerous horizontally transferred genes from bacteria and fungi. We believe these novelties likely have shaped B. tabaci as a highly invasive polyphagous crop pest and efficient vector of plant viruses. The genome serves as a reference for resolving the B. tabaci cryptic species complex, understanding fundamental biological novelties, and providing valuable genetic information to assist the development of novel strategies for controlling whiteflies and the viruses they transmit.

Evidence of the Biochemical Basis of Host Virulence in the Greenbug Aphid, Schizaphis graminum (Homoptera: Aphididae)

Biotypes of aphids and many other insect pests are defined based on the phenotypic response of host plants to the insect pest without considering their intrinsic characteristics and genotypes. Plant breeders have spent considerable effort developing aphid-resistant, small-grain varieties to limit insecticide control of the greenbug, Schizaphis graminum. However, new S. graminum biotypes frequently emerge that break resistance. Mechanisms of virulence on the aphid side of the plant-insect interaction are not well understood. S. graminum biotype H is highly virulent on most small grain varieties. This characteristic makes biotype H ideal for comparative proteomics to investigate the basis of biotype virulence in aphids. In this study, we used comparative proteomics to identify protein expression differences associated with virulence. Aphid proteins involved in the tricarboxylic acid cycle, immune system, cell division, and antiapoptosis pathways were found to be up-regulated in biotype H relative to other biotypes. Proteins from the bacterial endosymbiont of aphids were also differentially expressed in biotype H. Guided by the proteome results, we tested whether biotype H had a fitness advantage compared with other S. graminum biotypes and found that biotype H had a higher reproductive fitness as compared with two other biotypes on a range of different wheat germplasms. Finally, we tested whether aphid genetics can be used to further dissect the genetic mechanisms of biotype virulence in aphids. The genetic data showed that sexual reproduction is a source of biotypic variation observed in S. graminum.

Toxicity of neem oil to Bemisia tabaci biotype B nymphs reared on dry bean.

O objetivo deste trabalho foi determinar o estagio ninfal de Bemisia tabaci biotipo B mais suscetivel ao oleo de nim (Azadirachta indica A. Juss.) aplicado em feijoeiro (Phaseolus vulgaris L.), em casa telada. Foram avaliados o tempo letal (TL) e concentracao letal (CL) do oleo comercial de sementes de nim Dalneem. Para CL, concentracoes de 0, 0,1, 0,25, 0,5, 1 e 2% do produto foram pulverizadas diretamente sobre as ninfas em cada instar. Para TL, o produto foi avaliado a 0, 0,5 e 1% de oleo de nim em cada instar. Ninfas vivas e mortas foram contadas cinco dias apos a pulverizacao para CL e diariamente para TL durante seis dias. Para o quarto instar, a CL50 foi de 0,56% de oleo de nim. Considerando todos os instares, CL50 e CL95 foram estimadas em 0,32 e 2,78% de oleo de nim, respectivamente. Os TL50 para 1% de nim foram estimados em 2,46, 4,45, 3,02 e 6,98 dias para o primeiro, segundo, terceiro e quarto instares, respectivamente. Os TL50 estimados para 0,5 e 1% de oleo de nim foram de cinco e quatro dias, respectivamente, considerando todos os instares. No sexto dia, foi observada mortalidade superior a 80% para o primeiro, segundo e terceiro instares a 1% de oleo de nim. Os tres primeiros instares foram mais suscetiveis ao oleo de nim que o quarto instar.

Host plants indirectly influence plant virus transmission by altering gut cysteine protease activity of aphid vectors

The green peach aphid, Myzus persicae, is a vector of the Potato leafroll virus (PLRV, Luteoviridae), transmitted exclusively by aphids in a circulative manner. PLRV transmission efficiency was significantly reduced when a clonal lineage of M. persicae was reared on turnip as compared with the weed physalis, and this was a transient effect caused by a host-switch response. A trend of higher PLRV titer in physalis-reared aphids as compared with turnip-reared aphids was observed at 24 h and 72 h after virus acquisition. The major difference in the proteomes of these aphids was the up-regulation of predicted lysosomal enzymes, in particular the cysteine protease cathepsin B (cathB), in aphids reared on turnip. The aphid midgut is the site of PLRV acquisition, and cathB and PLRV localization were starkly different in midguts of the aphids reared on the two host plants. In viruliferous aphids that were reared on turnip, there was near complete colocalization of cathB and PLRV at the cell membranes, which was not observed in physalis-reared aphids. Chemical inhibition of cathB restored the ability of aphids reared on turnip to transmit PLRV in a dose-dependent manner, showing that the increased activity of cathB and other cysteine proteases at the cell membrane indirectly decreased virus transmission by aphids. Understanding how the host plant influences virus transmission by aphids is critical for growers to manage the spread of virus among field crops.

NEEM OIL ANTIFEEDANT AND INSECTICIDAL EFFECTS ON Oebalus poecilus (HEMIPTERA: PENTATOMIDAE) MALES AND FEMALES 1

The antifeedant and insecticidal effects of two commercial neem ( Azadirachta indica ) oil formulations (Dalneem and Nim-I-Go) to Oebalus poecilus were evaluated on irrigated rice. To evaluate the antifeedant effect, both formulations were tested at 1% and 2% (v/v) concentration levels. The insecticidal effect was evaluated at 0.5%, 1%, 2%, and 4% (v/v) concentrations of Dalneem, by ingestion. Both experiments were conducted with four replications by treatment, containing one panicle and adult insects segregated by gender (two insects per plot for the antifeedant effect and five for the insecticidal effect). Both products were efficient, reducing the damage caused by insects. Insects fed less on the panicles treated with neem oil, causing lower number of feeding sheaths per panicle and lower percentage of damaged grains than the control. Spikelets weight was higher in panicles treated with neem oils. Females caused significantly higher damage than males in controls, for numbers of feeding sheaths and for the feeding deterrence index. Only at the 4% (v/v) concentration level, Dalneem caused adult mortality higher than in the control. Results showed that neem oil formulations, at >= 1% (v/v) concentration, can be used to reduce the quantitative and qualitative damages caused by O. poecilus in lowland rice. KEY-WORDS: Small rice stink bug; Azadirachta indica ; qualitative damages; Oryza sativa ; botanical extract.

Transgene inheritances and genetic similarities of near isogenic lines of genetically modified common beans

(2) Embrapa Recursos Geneticos e Biotecnologia, PqEB W5 Norte (fi nal), CEP 70770-900 Brasilia, DF. E-mail: elsa@cenargen.embrapa.br, aragao@cenargen.embrapa.br Abstract - The objective of the present work was to determine the inheritance and stability of transgenes of a transgenic bean line expressing the genes rep-trap-ren from Bean golden mosaic virus and the bar gene. Crosses were done between the transgenic line and four commercial bean cultivars, followed by four backcrosses to the commercial cultivars. Progenies from each cross were evaluated for the presence of the transgenes by brushing the leaves with glufosinate ammonium and by polymerase chain reaction using speciÞ c oligonucleotides. Advanced generations were rub-inoculated with an isolate of Bean common mosaic necrosis virus (BCMNV). The transgenes were inherited consistently in a Mendelian pattern in the four crosses studied. The analyzed lines recovered close to 80% of the characteristics of the recurrent parent, as determined by the random ampliÞ ed DNA markers used, besides maintaining important traits such as resistance to BCMNV . The presence of the transgene did not cause any detectable undesirable effect in the evaluated progenies.

Populational survey of arthropods on transgenic common bean expressing the rep gene from Bean golden mosaic virus

Genetically modified (GM) crops is considered the fastest adopted crop technology in the history of modern agriculture. However, possible undesirable and unintended effects must be considered during the research steps toward development of a commercial product. In this report we evaluated effects of a common bean virus resistant line on arthropod populations, considered as non-target organisms. This GM bean line (named M1/4) was modified for resistance against Bean golden mosaic virus (BGMV) by expressing a mutated REP protein, which is essential for virus replication. Biosafety studies were performed for a period of three years under field conditions. The abundance of some species was significantly higher in specific treatments in a particular year, but not consistently different in other years. A regular pattern was not observed in the distribution of insects between genetically modified and conventional treatments. Data analyses showed that minor differences observed can be attributed to random variation and were not consistent enough to conclude that the treatments were different. Therefore the present study indicates that the relative abundance of species are similar in transgenic and non-transgenic fields.