Flávio Moscardi

Are Chemical Compounds Important for Soybean Resistance to Anticarsia gemmatalis

The identification and quantification of flavonoids (rutin and genistin) present in extracts of soybean genotypes, and their effects on the biology and physiology of Anticarsia gemmatalis Hübner (Lep.: Noctuidae) were studied. Analysis of covariance and bicoordinate utilization plots were used to remove the effect of feeding time from pupal weight and consumption as well as to separate pre- and postingestive effects of treatment on A. gemmatalis growth. Genotypes PI 274454, PI 227687, and “IAC-100” extracts in general, caused higher mortality, negatively influenced initial larval and pupal weight, and elongated larval cycle. Larvae fed on the “IAC-100” extract diet ingested larger amounts of food per unit of time, but were less efficient in its conversion to biomass. Leaf extracts of PI 227687 had the largest concentration of rutin (quercitin 3-O-rhamnosylglucoside), followed by PI 274454, and “IAC-100”; PI 74454 also had the highest genistin (genistein 7-O-glucoside) content. The susceptible cultivar “BR-16” showed only a kaempferol-based flavonoid in its chemical profile, indicating that after successive crosses, secondary compounds responsible for plant defenses were eliminated. Genotypes PI 274454, PI 227687, and “IAC-100” showed accentuated resistance characteristics and were considered inadequate sources for the development of A. gemmatalis. Considering rutin and genistin concentration in these genotypes, it is suggested that flavonoids are important factors conferring resistance to A. gemmatalis.

The impact of fungicides on Nomuraea rileyi (Farlow) Samson epizootics and on populations of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), on soybean

The fungus Nomuraea rileyi (Farlow) Samson is one of the most important natural enemies of soybean caterpillars, mainly under humid weather conditions. Outbreaks of the fungus Microsphaera diffusa Cooke & Peck have demanded fungicide applications on soybeans, which could result in outbreaks of noctuid populations by reduction of the natural inocula of N. rileyi. The recommended fungicides have shown to be detrimental to beneficial fungi, reducing infection, delaying epizootics, and resulting in increased host population densities. In laboratory assays, benomyl, difenoconazole, sulphur and carbendazim affected conidial germination of N. rileyi, being the latter less deleterious. To assess the impact of fungicides used to control M. diffusa, on N. rileyi, two tests were carried on, spraying difenoconazole (75 g a.i./ha) and benomyl (262.5 g a.i./ha) on soybean plots. In the 1997/98 trial, fungicide was sprayed once on soybean plants at R1-R2 developmental stages. In the 1998/99 test, two applications were made, when plants were at V5 and V7 developmental stages, respectively. The number of VBC larvae was significantly higher in the fungicide treated plots than in the control plots. In the 1997/98 test, benomyl treated plots resulted in higher populations of VBC than in the control or in the difenoconazole plots. In the 1998/99 test, VBC population was higher from 7 to 12 days after the first application, and remained high until 19 days after the second application. In general, fungicide treatments delayed the begining of N. rileyi epizootics from 2 to 14 days.

Use of Viruses Against Soybean Caterpillars in Brazil

Among the lepidopterous insects associated with soybeans in Brazil, the velvetbean caterpillar, Anticarsia gemmatalis, is the most important and widespread defoliator, seconded by species of Plusiinae, such as Pseudoplusia includens and Rachiplusia nu, which are of lesser importance but usually occur associated with populations of A. gemmatalis. Research with a nuclear polyhedrosis virus of A. gemmatalis (AgNPV) has led to a wide-scale use of the pathogen at farmer level, estimated in ca. 1,000,000 hectares annually. Most of its production has been carried out by estate organizations and farmer cooperatives, but at present four private companies are commercializing the AgNPV as a biological pesticide. NPVs of the Plusiinae complex are also being developed as a microbial insecticide, through a cooperative effort between Brazilian, Argentinian, and Uruguaian researchers. Research developments with the AgNPV, progress in its use at farmer level, and the work being carried out with baculoviruses of other soybean insects are discussed.

Morphological study of the hindgut in larvae of Anticarsia gemmatalis Hübner(Lepidoptera: Noctuidae)

The velvetbean caterpillar (Anticarsia gemmatalis Hubner) has great economical interest as it affects the soybean crop worldwide. This work describes the morphology of the hindgut in A. gemmatalis larvae emphasizing their histological aspects. Distinct morphological regions, identified as pylorus, ileum, colon and rectum, constitute the hindgut of A. gemmatalis. A thin cuticular intima, a simple epithelium and muscular layer compose the hindgut wall, independent of the region. Microspines project from the cuticular intima in the posterior interstitial ring, between the midgut and the hindgut, and the posterior pyloric region. A single circular layer of large fibers, differing from the other hindgut regions that present two layers of muscular fibers, forms the rectal musculature. The distal ends of Malpighian tubules cross the rectum wall and constitute the cryptonephric excretory system typical in Lepidoptera larvae.

Physical maps and virulence of Anticarsia gemmatalis nucleopolyhedrovirus genomic variants.

Summary. Seventeen plaque purified isolates of two viral preparations of Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), were analyzed in terms of the genomic changes after digestion of their DNAs with HindIII and PstI restriction enzymes. The 1979 AgMNPV wild type preparation (AgMNPV-’79) resulted in six different variants and the 1985 viral commercial preparation (AgMNPV-’85), in eleven. The genomic variation of all the isolates was mapped showing that those from 1985 presented more heterogeneity with changes mapped in additional sites in comparison to the AgMNPV-’79 variants. Their virulence was compared by infecting two Lepidopteran cell lines, Spodoptera frugiperda (IPLB-SF-21AE) and Anticarsia gemmatalis (UFL-AG-286). The results indicated that there was some difference in virulence within the AgMNPV-’85 variants. This commercial preparation had been applied in soybean fields in Brazil over several years to control the velvetbean caterpillar defoliation.

Natural Occurrence of the Entomopathogenic Fungi Metarhizium, Beauveria and Paecilomyces in Soybean Under Till and No-till Cultivation Systems

A ocorrencia de fungos entomopatogenicos Metarhizium, Beauveria e Paecilomyces foi estudada em condicoes de semeadura direta e convencional da soja. Foi determinada a densidade de unidades formadoras de colonia por g de solo e por cm2 de foliolos de soja. Verificou-se que no solo sob semeadura direta ocorreu maior incidencia dos entomopatogenos, mas sobre os foliolos essa diferenca nao ocorreu, proporcionando as mesmas possibilidades de infeccao nos insetos da parte aerea suscetiveis que ocorrem nas duas condicoes de cultivo.

Construction of a recombinant Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV-2D) harbouring the β-galactosidase gene

Summary. We have constructed a transfer vector (pAgGal) containing the β-galactosidase gene under control of the Escherichia coli gpt and AgMNPV polyhedrin (polh) promoters. The transfer vector was cotransfected with wild type Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) DNA into A. gemmatalis (UFL-AG-286) cells and a recombinant baculovirus (vAgGalA2) was isolated. The β-galactosidase gene insertion was checked by polymerase chain reaction (PCR) using DNA from AgMNPV and vAgGalA2 and primers specific for regions upstream and downstream of the polh gene. Insect cells (UFL-AG-286) were infected with the recombinant vAgGalA2 and wild type AgMNPV viruses and the production of the heterologous protein analyzed by SDS-PAGE and Pulse-Chase. β-galactosidase was expressed at high levels late on infection as expected for a gene under the control of the polh promoter. The highly expressed β-galactosidase protein was also shown to be biologically active by a β-galactosidase assay.

Microbial control of insect pests of soybean

Soybean is the major leguminous crop grown worldwide and the most important producers are the United States, Brazil, Argentina, China and India. Insects attacking soybean germinating seeds, roots, stem, leaves, pods and seeds may limit production, depending on their abundance and damage potential in each soybean-growing region (Kogan and Turnipseed, 1987; Moscardi, 1993; Gazzoni et al., 1994; Hoffmann-Campo et al., 2003).

Field efficacy of the nucleopolyhedrovirus of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae): effect of formulations, water pH, volume and time of application, and type of spray nozzle

The efficacy of the nucleopolyhedrovirus of Anticarsia gemmatalis Hubner (AgMNPV) was evaluated in relation to different application parameters of this biological insecticide, to elucidate some problems of its low efficacy in certain regions in Brazil, mainly in the state of Rio Grande do Sul (RS). Each experiment was conducted during four consecutive soybean seasons (1994/95 to 97/98), in Cruz Alta ¾ RS, to evaluate the following parameters on AgMNPV field efficacy: available commercial formulations of the virus; pH of the viral aqueous suspension; spray volume; application time; spray nozzle type; and the mixture of the virus with mineral oil. Each plot consisted of 28 soybean rows of 10 m in length, spaced of 40 cm. The statistical design was the completely randomized block design, with four replicates. Samplings of AgMNPV infected or dead larvae (IL) were periodically made by the shake cloth method. The AgMNPV formulations produced by Nitral, Nova Era, Coodetec and Embrapa were efficient in controlling larval populations of A. gemmatalis when compared to the control. However, these formulations were significantly inferior to the crude extract of AgMNPV (maceration and filtration of dead larvae), indicating that current commercial formulations of this virus need to be improved. The AgMNPV was significantly affected by the water pH in the spray tank, as plots treated with a viral suspension at pH 6 resulted in a significantly higher number of IL than plots treated with suspensions at pH 2 and 10. The spray volume also influenced the control of A. gemmatalis larval populations by the AgMNPV. Volumes of 300, 200 e 100 L/ha were equally efficient in controling the insect. However, at 50 L/ha, the virus efficacy was significantly lower than that provided by the other spray volumes, apparently due to nozzle clogging at the lower application volume. The AgMNPV performance to control the insect was also significantly affected by the time of application, with higher efficiencies when spraying at 2:00 a.m. or 8:00 p.m., in comparison to 8:00 a.m. or 2:00 p.m. On the other hand, no differences were detected on the efficacy of the virus when sprayed with different nozzle types, and mineral oil addition to the viral suspension did not improve the field performance of the formulated AgMNPV.

Production of nuclear polyhedrosis virus of Anticarsia gemmatalis Hübner (Lepidoptera:Noctuidae): effect of vírus dosage, host density and age

The laboratory production of Anticarsia gemmatalis Hubner nuclear polyhedrosis virus (AgNPV) was evaluated for larvae inoculated at 1.0, 1.5, 2.0, and 2.5 cm and with two NPV dosages, 106 and 107 polyhedron inclusion bodies (PIB)/ml, sprayed on the insect diet surface. A fixed population of 25 larvae/300-ml cup was used. Highest virus yield was achieved for larvae inoculated with ca. 2.0 cm at the higher NPV dosage. Losses of NPV-inoculated larvae ranged from 42.4% to 64%, due to high incidence of canibalism, death by other causes, and pupation. In another experiment, larvae were inoculated with two NPV dosages (1.0 x 107 and 4.0 x 107 PIB/ml), using 25,20, and 15 larvae/cup at the beginning of 4th instar (B4L); end of 4th instar (E4L); beginning of 5th instar (B5L); and end of 5th instar (E5L). Highest NPV production was attained for B5L at 25 larvae/cup, for both dosages, with a production efficiency of ca 90%, in spite of a decrease in PIB/larva and an increase in canibalism with the increase in larval density. Better standardization of larvae at inoculation improved efficiency in NPV production.