Celso Omoto

Herança da resistência de Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) a lambda-cialotrina

Inheritance of lambda-cyhalothrin resistance in Spodoptera frugiperda (J.E. Smith) was investigated in a strain of S. frugiperda showing an intensity of resistance of approximately 13-fold. Results from reciprocal crosses between susceptible (S) and resistant (R) strains of S. frugiperda to lambda-cyhalothrin revealed that resistance was autossomal and conferred by an incompletely recessive gene. Then, backcrosses of F1 progenies to the R strain were performed to determine the number of genes involved in the resistance. From interpretation of chi-square analysis of responses of progenies from backcrosses suggested that the resistance was conferred by a single major gene under a influence of some minor genes.

Assessment of the high-dose concept and level of control provided by MON 87701 × MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil.

BACKGROUND Genetically modified MON 87701 × MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP-synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high-dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS The purified Cry1Ac protein was more active against A. gemmatalis [LC(50) (FL 95%) = 0.23 (0.15-0.34) µg Cry1Ac mL(-1) diet] than P. includens [LC(50) (FL 95%) = 3.72 (2.65-4.86) µg Cry1Ac mL(-1) diet]. In bioassays with freeze-dried MON 87701 × MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf-disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701 × MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS The MON 87701 × MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high-dose event only to A. gemmatalis.

Molecular variability of Spodoptera frugiperda (Lepidoptera: Noctuidae) populations associated to maize and cotton crops in Brazil.

The molecular variability among 10 populations of Spodoptera frugiperda (J.E. Smith), collected from maize, Zea mays L., or cotton Gossypium hirsutum L. crops located at distinctive geographical regions in Brazil, was assessed through random amplification of polymorphic DNA (RAPD)-polymerase chain reaction (PCR). In total, 208 RAPD markers were evaluated, and 98% of them were polymorphic. The mean genetic similarity was 0.6621 and 0.2499 by the Simple Matching and Jaccard matrices, respectively. In general, the unweighted pair-group method with arithmetic average dendrograms separated the populations into clusters related to the geographical origin of the samples. No branch of the dendrograms underpinning a molecular association of S. frugiperda has been identified to either of the two host plants. The molecular variance analysis showed that 18 and 82% of the genetic variation was distributed among and within the groups of populations, respectively. The principal coordinate analysis reinforced the pattern of population clustering found with the unweighted pair-group method with arithmetic average method. These results suggest the occurrence of considerable gene flow between S. frugiperda populations from maize and cotton fields located in the same region in Brazil. Therefore, for an effective management of this pest, there is an urgent need for a better understanding of the gene flow of S. frugiperda populations associated to different host plants along the distribution range of this pest over time in a specific cropping system.

Field-evolved resistance to Cry1Ab maize by Spodoptera frugiperda in Brazil.

BACKGROUND The first Bt maize in Brazil was launched in 2008 and contained the MON 810 event, which expresses Cry1Ab protein. Although the Cry1Ab dose in MON 810 is not high against fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), MON 810 provided commercial levels of control. To support insect resistance management in Brazil, the baseline and ongoing susceptibility of FAW was examined using protein bioassays, and the level of control and life history parameters of FAW were evaluated on MON 810 maize. RESULTS Baseline diet overlay assays with Cry1Ab (16 µg cm(-2) ) caused 76.3% mortality to field FAW populations sampled in 2009. Moderate mortality (48.8%) and significant growth inhibition (88.4%) were verified in leaf-disc bioassays. In greenhouse trials, MON 810 had significantly less damage than non-Bt maize. The surviving FAW larvae on MON 810 (22.4%) had a 5.5 day increase in life cycle time and a 24% reduction in population growth rate. Resistance monitoring (2010-2015) showed a significant reduction in Cry1Ab susceptibility of FAW over time. Additionally, a significant reduction in the field efficacy of MON 810 maize against FAW was observed in different regions from crop season 2009 to 2013. CONCLUSIONS The decrease in susceptibility to Cry1Ab was expected, but the specific contributions to this resistance by MON 810 maize cannot be distinguished from cross-resistance to Cry1Ab caused by exposure to Cry1F maize. Technologies combining multiple novel insecticidal traits with no cross-resistance to the current Cry1 proteins and high activity against the same target pests should be pursued in Brazil and similar environments.

Demographics and Genetic Variability of the New World Bollworm (Helicoverpa zea) and the Old World Bollworm (Helicoverpa armigera) in Brazil

Helicoverpa armigera is one of the primary agricultural pests in the Old World, whereas H. zea is predominant in the New World. However, H. armigera was first documented in Brazil in 2013. Therefore, the geographical distribution, range of hosts, invasion source, and dispersal routes for H. armigera are poorly understood or unknown in Brazil. In this study, we used a phylogeographic analysis of natural H. armigera and H. zea populations to (1) assess the occurrence of both species on different hosts; (2) infer the demographic parameters and genetic structure; (3) determine the potential invasion and dispersal routes for H. armigera within the Brazilian territory; and (4) infer the geographical origin of H. armigera. We analyzed partial sequence data from the cytochrome c oxidase subunit I (COI) gene. We determined that H. armigera individuals were most prevalent on dicotyledonous hosts and that H. zea were most prevalent on maize crops, based on the samples collected between May 2012 and April 2013. The populations of both species showed signs of demographic expansion, and no genetic structure. The high genetic diversity and wide distribution of H. armigera in mid-2012 are consistent with an invasion period prior to the first reports of this species in the literature and/or multiple invasion events within the Brazilian territory. It was not possible to infer the invasion and dispersal routes of H. armigera with this dataset. However, joint analyses using sequences from the Old World indicated the presence of Chinese, Indian, and European lineages within the Brazilian populations of H. armigera. These results suggest that sustainable management plans for the control of H. armigera will be challenging considering the high genetic diversity, polyphagous feeding habits, and great potential mobility of this pest on numerous hosts, which favor the adaptation of this insect to diverse environments and control strategies.

Monitoring the susceptibility to insecticides in Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations from Brazil

Monitoring the susceptibility of a pest population to pesticides is essential for resistance management programs. The objective of this research was to evaluate the genetic variability in pesticide susceptibility in populations of Bemisia tabaci (Gennadius) collected from different Brazilian agricultural regions through the use of two different tests. Four whitefly populations, two from Goiás state (GO-1 and GO-2) e two from Bahia state (BA-1 and BA-2), were tested against a susceptible reference one (SusIAC). A residual contact bioassay was used to evaluate the pesticide susceptibility of each population by using diagnostic concentration bioassays and by estimating the baseline susceptibility data to each one of the tested insecticides, acetamiprid, imidacloprid, thiamethoxam, chlorpyrifos and endosulfan. Adult insects of unknown age and sex were tested. Evaluations were performed after 24h for endosulfan and 48 h for the other chemicals. Both procedures showed significant differences in the susceptibility to the pesticides among B. tabaci populations. However, the discrimination among B. tabaci populations was more evident with the use of diagnosis tests. The population GO-2 was significantly less susceptible to the tested pesticides than SusIAC, mainly to neonicotinoids. The most critical resistance situation of B. tabaci was detected to thiamethoxam, followed by imidacloprid.

Dominance of Cry1F resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) on TC1507 Bt maize in Brazil

BACKGROUND Dominance of resistance has been one of the major parameters affecting the rate of evolution of resistance to Bt crops. High dose is the capacity of Bt crops to kill heterozygous insects and has been an essential component of the most successful strategy to manage resistance to these crops. Experiments were conducted to evaluate directly and indirectly whether the TC1507 event is high dose to Spodoptera frugiperda (JE Smith). RESULTS About 8% of heterozygote neonate larvae were able to survive, complete larval development and emerge as normal adults on TC1507 leaves, while susceptible larvae could not survive for 5 days. The estimated dominance of resistance was 0.15 ± 0.09 and significantly higher than zero; therefore, the resistance to Cry1F expressed in TC1507 was not completely recessive. A 25-fold dilution of TC1507 maize leaf tissue in an artificial diet was able to cause a maximum mortality of only 37%, with growth inhibition of 82% at 7 days after larval infestation. CONCLUSION Resistance to Cry1F in TC1507 maize is incompletely recessive in S. frugiperda. TC1507 maize is not high dose for S. frugiperda. Additional or alternative resistance management strategies, such as the replacement of single-trait Bt maize with pyramided Bt maize, which produces multiple proteins targeting the same insect pests, should be implemented wherever this technology is in use and S. frugiperda is the major pest.

Resistance to hexythiazox in Brevipalpus phoenicis (Acari: Tenuipalpidae) from Brazilian citrus.

The objective of this study was to collect baseline information for implementing an acaricide resistance management program of Brevipalpus phoenicis (Geijskes) to hexythiazox in Brazilian citrus groves. The egg susceptibility of B. phoenicis to hexythiazox was measured by a direct contact bioassay. The estimated LC50 for the S strain was 0.89 mg hexythiazox L−1 of water (95%FL 0.75–1.03). After hexythiazox selection of a field-collected population associated with intense hexythiazox use, a resistance ratio greater than10,000-fold was detected. Results from a survey revealed a great variability in the frequency of resistance in populations of B. phoeniciscollected from citrus groves located in the State of São Paulo. No relationship was observed between the intensity of hexythiazox use and the frequency of resistance. Studies on dynamics of resistance showed that the resistance of B. phoenicis to hexythiazox is stable under laboratory conditions. Therefore, there is an urgent need to implement resistance management of B. phoenicis to hexythiazox in order to prolong its effective use in Brazilian citrus groves.

Cross-Resistance between Cry1 Proteins in Fall Armyworm (Spodoptera frugiperda) May Affect the Durability of Current Pyramided Bt Maize Hybrids in Brazil.

Genetically modified plants expressing insecticidal proteins from Bacillus thuringiensis (Bt) offer valuable options for managing insect pests with considerable environmental and economic benefits. Despite the benefits provided by Bt crops, the continuous expression of these insecticidal proteins imposes strong selection for resistance in target pest populations. Bt maize (Zea mays) hybrids have been successful in controlling fall armyworm (Spodoptera frugiperda), the main maize pest in Brazil since 2008; however, field-evolved resistance to the protein Cry1F has recently been reported. Therefore it is important to assess the possibility of cross-resistance between Cry1F and other Cry proteins expressed in Bt maize hybrids. In this study, an F2 screen followed by subsequent selection on MON 89034 maize was used to select an S. frugiperda strain (RR) able to survive on the Bt maize event MON 89034, which expresses the Cry1A.105 and Cry2Ab2 proteins. Field-collected insects from maize expressing the Cry1F protein (event TC1507) represented most of the positive (resistance allele-containing) (iso)families found. The RR strain showed high levels of resistance to Cry1F, which apparently also conferred high levels of cross resistance to Cry1A.105 and Cry1Ab, but had only low-level (10-fold) resistance to Cry2Ab2. Life history studies to investigate fitness costs associated with the resistance in RR strain revealed only small reductions in reproductive rate when compared to susceptible and heterozygous strains, but the RR strain produced 32.2% and 28.4% fewer females from each female relative to the SS and RS (pooled) strains, respectively. Consistent with the lack of significant resistance to Cry2Ab2, MON 89034 maize in combination with appropriate management practices continues to provide effective control of S. frugiperda in Brazil. Nevertheless, the occurrence of Cry1F resistance in S. frugiperda across Brazil, and the cross-resistance to Cry1Ab and Cry1A.105, indicates that current Cry1-based maize hybrids face a challenge in managing S. frugiperda in Brazil and highlights the importance of effective insect resistance management for these technologies.

Mecanismos de dispersão de Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae) em pomares de citros

The knowledge of pest dispersal is crucial for understanding the dynamics of resistance to pesticides. Therefore, the main objective of this research was to study the dispersal of Brevipalpus phoenicis (Geijskes) under laboratory, greenhouse and citrus groves conditions. Dispersal studies of B. phoenicis on sandy surface conducted under greenhouse conditions showed that dispersal capacity by crawling is relatively limited. After the release of 6,000 mites in one point, approximately 3% reached distances of 40 to 50 cm from the release point. This mite moved less than 1 cm.day-1. Wind speed of 23 km.h-1 was not enough to trigger mite dispersal from the surface of citrus fruits under laboratory conditions. Wind speeds of 30 and 40 km.h-1 were capable to trigger the dispersal of less than 1% of mites on fruits. Mite dispersal studies with the use of sticky traps under field conditions also showed that B. phoenicis dispersal is limited when compared to other mite species from citrus groves. In this experiment, from a total of 2,420 and 661 mites collected from each of two citrus fields in Descalvado, region, State of Sao Paulo, Brazil, 0.4 and 11.8% of the mites belonged to the family Tenuipalpidae. The limited dispersal of B. phoenicis requires considerable attention in the establishment of acaricide resistance management strategies.