Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Biological characteristics of Telenomus alecto and Trichogramma galloi reared on eggs of the sugarcane borer Diatraea flavipennella

Diatraea flavipennella (Box) (Lepidoptera, Crambidae) is one of the most destructive pests in sugarcane plantations in the Northeast Region of Brazil. Developmental characteristics and parasitism potential of the egg parasitoids Telenomus alecto Crawford (Hymenoptera, Scelionidae) and Trichogramma galloi Zucchi (Hymenoptera, Trichogrammatidae) were compared with the aim of selecting a suitable species for biological control of D. flavipennella. Both T. alecto and T. galloi developed well and were readily adapted to D. flavipennella eggs as host. Although, T. galloi presented higher viability, with more adults emerging per host egg and higher sex ratio, the developmental period (egg-adult) was shorter in T. alecto and female longevity was extended. In addition, T. alecto exhibited significant higher levels of parasitism during the first three days after emergence than T. galloi. Thus, both parasitoids studied here offer considerable potential for the control of D. flavipennella in sugarcane.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Ultrastructural and quantitative studies of hemocytes in the sugarcane borer, Diatraea saccharalis (Lepidoptera: Pyralidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Morphological aspects of cluster formation in the germarium of the sugarcane borer Diatraea saccharalis Fabricius (Lepidoptera : Pyralidae)

Diatraea saccharalis F. is one of the greatest pests of the sugar cane culture. This report aimed to characterize the germanium region of the sugarcane borer by light and transmission electron microscopy, emphasizing the morphological steps of the ovarian cluster formation. In the germanium of this insect, four zones could be morphologically identified during the cluster formation. In the most apical end of each ovariole - Zone I - the germ line stem cells undergo complete mitotic division, originating the cystoblasts. In the Zone II, each cystoblast produces a group of eight cells, the cystocytes, which are interconnected by the ring canals. Clusters containing all the cystocytes in the meiosis, characterizes the Zone III. Germ cells with ultrastructural features of apoptosis are also detected in this Zone. In the Zone IV the cystocytes differentiate, morphologically, into one oocyte and seven nurse cells. Interstitial somatic cells and pre-follicle cells exhibit, in their cytoplasm, heterogeneous vacuoles containing degenerated cellular fragments, characterized as apoptotic bodies. Our results pointed out to the morphological evidences related with important control mechanisms for new clusters/follicles production and for the cellular arrangement into the germanium, resulting from the programmed cell death. We believe that the morphological characterization of ovarian cluster formation in D. saccharalis provided valuable information for the understanding of the initial steps of oogenesis and contributed for the knowledge of the cellular mechanisms related with the oocyte production and with reproduction in insects.

Mitochondria in the midgut epithelial cells of sugarcane borer parasitized by Cotesia flavipes (Cameron, 1891)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spherites in the midgut epithelial cells of the sugarcane borer parasitized by Cotesia flavipes

Diatraea saccharalis, the main pest of sugarcane, has been controlled by Cotesia flavipes. Very little is known about the effect of parasitism on the host organs, including the midgut. The Lepidoptera midgut epithelium is composed of columnar, goblet, regenerative, and endocrine cells. Spherites have been described in columnar and regenerative cells of several Lepidoptera species, and presented a lot of functional meaning. We identified spherites in the midgut epithelial cells of non-parasitized D. saccharalis larvae analyzed the effect of parasitism on spherite morphology and distribution along the length of the midgut. Midgut fragments of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. All the midgut epithelial cells showed spherites, but they were not preferentially located in a particular part of the cells. Parasitized larvae had more spherites, mainly in the columnar cells, than non-parasitized larvae. This observation was associated with an ionic imbalance within the insect host. Spherites were more abundant in the anterior midgut region than in other regions, which suggests that this region is involved in ion transport by intracellular and/or paracellular route. The morphological variability of spherites in the cells of parasitized larvae was related to the developmental stages of these structures.

Ant diversity (Hymenoptera: Formicidae) and predation by ants on the different stages of the sugarcane borer life cycle Diatraea saccharalis (Lepidoptera: Crambidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The sugarcane borer Diatraea saccharalis (Fabr.) (Lep., Crambidae) and its parasitoids: a synchrony approach to spatial and temporal dynamics

A data set on Diatraea saccharalis and its parasitoids, Cotesia flavipes and tachinid flies, was analysed at five spatial scales-sugarcane mill, region, intermediary, farm and zone-to determine the role of spatial scale in synchrony patterns, and on temporal population variability. To analyse synchrony patterns, only the three highest spatial scales were considered, but for temporal population variability, all spatial scales were adopted. The synchrony-distance relationship revealed complex spatial structures depending on both species and spatial scale. Temporal population variability [SD log(x+1)] levels were highest at the smallest spatial scales although, in the majority of the cases, temporal variability was inversely dependent on sample size. All the species studied, with a few exceptions, presented spatial synchrony independent of spatial scale. The tachinid flies exhibited stronger synchrony dynamics than D. saccharalis and C. flavipes in all spatial scales with the latter displaying the weakest synchrony levels, except when mill spatial scales were compared. In some cases spatial synchrony may at first decay and then increase with distance, but the presence of such patterns can change depending on the spatial scale adopted.

Ultrastructure of the excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis (Lepidoptera : Pyralidae)

The excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis consists of two morphologically distinct regions, recognized by scanning and transmission electron microscopy. The thin posterior region, adjacent to the glandular region, presents a regular surface. Secretory vesicles containing either electron-dense or fibrillar cuticular-like materials are observed in their apical cytoplasm; the same cuticular materials were detected as extracellular deposits among the microvilli. The short anterior region, near the common duct, exhibits surface protrusions; there are no secretory vesicles in their apical cytoplasm. These results show that only the duct cells at the posterior region are involved in the secretion of the cuticular intima elements. Desmosome-like structures were visualized linking together adjacent microvillar membranes only in the cells of anterior duct region, with unknown function. The transition between the duct and the glandular region is abrupt; the cells of the glandular and posterior duct regions present large amounts of microtubules. Nerve fibers can be observed between the duct cells in their two regions, suggesting that control of silk secretion may occur in the excretory duct via neurotransmitter liberation. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Histochemical and ultrastructural evidence of lipid secretion by the silk gland of the sugarcane borer Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae)

The silk gland in Lepidoptera larvae is responsible for the silk production used for shelter or cocoon construction. The secretion of fibroin and sericin by the different silk gland regions are well established. There are few attempts to detect lipid components in the insect silk secretion, although the presence of such element may contribute to the resistance of the shelter to wet environment. This study characterizes the glandular region and detects the presence of lipid components in the secretion of the silk gland of Diatraea saccharalis (Fabricius). The silk gland was submitted to histochemical procedure for lipid detection or conventionally prepared for ultrastructural analyses. Lipid droplets were histochemically detected in both the apical cytoplasm of cell of the anterior region and in the lumen among the microvilli. Ultrastructural analyses of the anterior region showed lipid material, visualized as myelin-like structures within the vesicular Golgi complex and in the apical secretory globules, mixed up with the sericin; similar material was observed into the lumen, adjacent to the microvilli. Lipids were not detected in the cells neither in the lumen of the posterior region. Our results suggest that the silk produced by D. saccharalis has a minor lipid content that is secreted by the anterior region together with the sericin.

Characterization and impact of the sugarcane borer on sugarcane yield and quality

The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), is a key pest of sugarcane (Saccharum spp.). While damage caused by this pest has increased in the past 20 yr, studies investigating the insect-plant interactions are still lacking. Moreover, there is no information about the consequences of borer damage on the parameters of sugar quality. Therefore, two field experiments were performed during the 2010 and 2011 growing seasons in Brazil to compare the raw material and sugar quality of SP80-3280 sugarcane plants with and without the sugarcane borer. Plants were protected within screen cages and infested weekly during the 2010 and 2011 seasons, using egg masses starting at the second and third internode stage. At harvest, 25.77 and 19.01% of the internodes were bored by larvae (infestation intensity, II) in the first and second seasons, respectively. There was no correlation between the borer gallery total volume and II. The fiber content significantly increased with increasing II. The stalk biometric parameters, such as length, diameter, and yield, were not correlated with II. The sucrose yield significantly decreased with increasing II. Consequently, sugar yield losses were estimated at 8.83 and 19.80% per 1% bored internode for the first and second seasons, respectively. The concentration of phenolic compounds increased, and unclarified juice color quality decreased, with increasing II. Significant differences were detected in the quality of the sugar. These results should be confirmed for other sugarcane cultivars and incorporated into an economic injury level to enhance decision-making strategies for borer management. © 2013 by the American Society of Agronomy, 5585 Guilford Road, Madison, WI 53711. All rights reserved.

Binding analysis of Bacillus thuringiensis Cry1 proteins in the sugarcane borer, Diatraea saccharalis (Lepidoptera: Crambidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Testis morphology and dichotomic spermatogenesis in the sugarcane borer Diatraea saccharalis (F.) (Lepidoptera: Crambidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Resistance of sugarcane cultivars to Diatraea saccharalis

The objective of this work was to evaluate the oviposition preference of Diatraea saccharalis and the effect of ten sugarcane cultivars on larval development. Oviposition preference was assessed under greenhouse conditions by three releases of couples of moths, with subsequent counting of egg masses and eggs per plant. In order to evaluate the effect of the cultivars on larval development, each plant was infected with about 150 eggs, and, 29 days later, the total number of internodes, number of bored internodes, number of life forms found, larval and pupal weight and length, and the width of larval head capsule were evaluated. The cultivars IACSP94-2101 and IACSP96-2042, the least preferred by D. saccharalis for oviposition, and IACSP94-2094, the most unfavorable for larvae entrance and development, show resistance to the pest.