This thesis reports the effects of disturbance and harbourage on the fitness of Tribolium confusum and T. castaneum, as well as the the efficacy of pheromone monitoring traps for monitoring for populations of Tribolium in laboratory and mill settings. Behavioural studies were also carried out on mill and laboratory-reared beetles and the distributions of both species in a mill were examined. Twenty-four hour sieving disturbance decreased the rates of dispersal for both species, and decreased T. castaneum fecundity. Rolling disturbance decreased T. confusum dispersal rate while shaking disturbance decreased T. castaneum dispersal rate. When undisturbed beetles were given differing amounts of flour in the presence or absence of harbourage, beetles laid more eggs in larger amounts of flour, but harbourage only affected T. castaneum at one level of flour (2 g). Throughout disturbance and harbourage experiments, T. castaneum laid more eggs than T. confusum. Pheromone monitoring traps placed in three Canadian flour mills were not useful in predicting the degree of infestation inside Simons rollstands. Pheromone monitoring traps also showed low efficacy (i.e. caught few beetles) in both mill and laboratory settings, and T. confusum was caught less often than T. castaneum in both mills and in a warehouse. Mill-strain beetles of both species were caught less often than laboratory-strain beetles in a warehouse. In one Canadian flour mill, both T. castaneum and T. confusum were found inside rollstands but the two species were spatially segregated from one another, rarely being found together in the same rollstand. In contrast to this, both species were consistently found together in samples taken from the same mill less than a year beforehand. In behavioural laboratory studies, beetles collected directly from a mill moved slower than beetles collected from a laboratory culture and this response was shown to be phenotypic. Mill-strain and laboratory-strain beetles also differed in burrowing tendencies, with T. confusum from the laboratory strain burrowing less than T. confusum from a mill and T. castaneum from different mills sometimes burrowing more and sometimes less than T. castaneum from the laboratory strain.

This thesis reports the effects of disturbance and harbourage on the fitness of Tribolium confusum and T. castaneum, as well as the the efficacy of pheromone monitoring traps for monitoring for populations of Tribolium in laboratory and mill settings. Behavioural studies were also carried out on mill and laboratory-reared beetles and the distributions of both species in a mill were examined. Twenty-four hour sieving disturbance decreased the rates of dispersal for both species, and decreased T. castaneum fecundity. Rolling disturbance decreased T. confusum dispersal rate while shaking disturbance decreased T. castaneum dispersal rate. When undisturbed beetles were given differing amounts of flour in the presence or absence of harbourage, beetles laid more eggs in larger amounts of flour, but harbourage only affected T. castaneum at one level of flour (2 g). Throughout disturbance and harbourage experiments, T. castaneum laid more eggs than T. confusum. Pheromone monitoring traps placed in three Canadian flour mills were not useful in predicting the degree of infestation inside Simons rollstands. Pheromone monitoring traps also showed low efficacy (i.e. caught few beetles) in both mill and laboratory settings, and T. confusum was caught less often than T. castaneum in both mills and in a warehouse. Mill-strain beetles of both species were caught less often than laboratory-strain beetles in a warehouse. In one Canadian flour mill, both T. castaneum and T. confusum were found inside rollstands but the two species were spatially segregated from one another, rarely being found together in the same rollstand. In contrast to this, both species were consistently found together in samples taken from the same mill less than a year beforehand. In behavioural laboratory studies, beetles collected directly from a mill moved slower than beetles collected from a laboratory culture and this response was shown to be phenotypic. Mill-strain and laboratory-strain beetles also differed in burrowing tendencies, with T. confusum from the laboratory strain burrowing less than T. confusum from a mill and T. castaneum from different mills sometimes burrowing more and sometimes less than T. castaneum from the laboratory strain.

Orientador: Carminda da Cruz Landim / Banca: Flávio Henrique Caetano / Banca: Betina Blochtein / Resumo: As abelhas eussociais apresentam uma variedade de glândulas exócrinas distribuídas por todo o corpo, cujos produtos são usados no desempenho de suas tarefas. Em todos os insetos, pares de glândulas estão associados às peças bucais e, nas abelhas, as principais destas glândulas estão ligadas ao lábio (glândulas labiais ou salivares), às mandíbulas (glândulas mandibulares) e ao assoalho da cavidade bucal (glândulas hipofaríngeas). O par de glândulas labiais ou salivares dos insetos adultos localiza-se no tórax, porém ramos cefálicos se desenvolvem em algumas espécies de abelhas, formando as glândulas salivares cefálicas, as quais são plenamente desenvolvidas em algumas espécies da subfamília Apinae. O presente trabalho estudou a histologia e a histoquímica das glândulas salivares cefálicas de fêmeas e machos, em diferentes idades, de Apis mellifera e Scaptotrigona postica. Os resultados mostram que a histologia das glândulas salivares cefálicas difere entre as espécies. Enquanto as células alveolares e dos ductos são predominantemente cúbicas nas operárias e rainhas de A. mellifera, em S. postica passam de cúbicas a escamosas conforme a operária envelhece, e os ductos, nesta espécie, são invariavelmente constituídos por células escamosas. Machos recém-emergidos de A. mellifera apresentam a glândula com grau de desenvolvimento semelhante ao das operárias recém-emergidas de sua espécie, porém conforme atingem a maturidade sexual, ocorre regressão glandular de modo que nem sempre foi possível encontrá-la. Por outro lado, machos velhos de S. postica são dotados de glândulas tão desenvolvidas quanto as fêmeas de sua espécie... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Eusocial bees present a variety exocrine glands disperse on body whose products are used in tasks performance. Pairs of glands are attached at mouthparts in whole insects and, in bees, main glands are attached to labium (labial or salivary glands), mandibules (mandibular glands) and floor mouth cavity (hiphofaringeal glands). Adult insects labial or salivary pair glands are located in torax. However cephalic branchs develop in some bees species, composing the cephalic salivary glands which are completely develop in some species of Apinae subfamily. The present work studied the histology and histochemic of females and males cephalic salivary glands of Apis mellifera and Scaptotrigona postica in different ages. The histology results showed cephalic salivary glands differs between species. While cells of alveoli and ducts are primarily cuboidal in workers and queens of A. mellifera, in workers of S. postica, cells undergo of cuboidal to squamous with age and ducts invariably consist of squamous cells. Newly emerged males of A. mellifera exhibit glands in developmental stages similar to that of newly emerged workers of their species. However, as males become sexually mature, the gland degenerates difficulting its localization. Nevertheless, old males of S. postica present glands as developed as those of females of their species. Histochemical techniques demonstrate no protein inclusions and indicated the secretion of glands of workers and queens of both species were neutral lipids. The secretion gradually accumulates in the lumen of alveoli, and as a result, foragers and egg-laying queens exhibit more turgid alveoli that younger individuals... (Complete abstract click electronic access below) / Mestre

Reproductive isolation is an essential stage in speciation. In Ascomycetes, the ubiquitous distribution of many species suggests that sympatric speciation through assertive mating should be an important factor. The MAT locus and the pheromone/receptor system could both potentially contribute to the development of such sexual isolation. Alterations at the MAT loci could lead to distinct reproductive habits or a change in mating system, both of which can reduce gene-flow between species. However, if deliberate pre-mating sexual preferences exist, they are more likely to be determined by the pheromone/receptor system. This study of Fusarium reproductive genes, and comparisons with other Ascomycetes, has yielded numerous interesting findings regarding the evolution of these mate-recognition mechanisms and the implications thereof. The G. fujikuroi and F. graminearum species complexes have offered an interesting comparison between heterothallic and homothallic MAT locus evolution. The value of comparative sequence analysis has been demonstrated in the discovery of a previously unknown gene, MAT1-2-3, which may be specific to members of the Order Hypocreales. While all MAT genes share similar regulatory elements, this is the first report of evidence that a transition to homothallism can be accompanied by the recruitment of distinct elements that could facilitate alternate expression of MAT genes. The MAT genes are also highly divergent between Fusarium spp., largely due to relaxed selective constraint, particularly in homothallic species. However, inter-specific gene-flow could curb MAT gene divergence among homothallic species. Despite strong reproductive barriers in the G. fujikuroi complex, the F. sacchari MAT1-1 sequence appears to have been acquired through lateral transfer from a distant relative. Analytical analysis of the MAT locus novelties reported here, including the new MAT gene, will be necessary to determine their biological significance. To investigate the extent of pheromone diversity in the Ascomycetes, and to gain clues as to its biological importance, pheromone peptides from seventy ascomycete species were compared. A number of reproductively incompatible species, such as those in the G. fujikuroi complex, share identical pheromones; which implies that another mechanism must be responsible for the observed reproductive barriers. However, on the whole, pheromones are highly divergent among species. Both adaptive and non-adaptive evolution could have contributed to this pattern. In fact the structure of the á-class pheromone precursor gene, which consists of multiple repeats of the pheromone module, could facilitate rapid diversification through “birth-and-death” evolution. Within species, selection maintains pheromone peptides, implying that much of the inter-specific variation is functionally relevant. This further suggests that pheromone evolution could contribute to the generation of reproductive isolation between species. The most general trend in the findings of this study is that ascomycete reproductive genes are highly divergent. This is in agreement with findings in other Kingdoms. A number of evolutionary forces are probably involved but weaker selective constraint, resulting from the fact that reproduction is not essential in these fungi, appears to be a common factor. This reproductive gene variability could be directly linked to speciation and, therefore, the great diversity in Ascomycetes. Additional information on the appendices is available on a CD, stored at the Merensky Library on Level 3 / Dissertation (MSc)--University of Pretoria, 2011. / Genetics / Unrestricted

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Stinkbugs (Pentatomidae) and other agricultural pests such as bark beetles and flea beetles are known to synthesize terpenoids as aggregation pheromones. Knowledge of the genes and enzymes involved in pheromone biosynthesis may allow engineering of pheromone biosynthetic pathways in plants to develop new forms of trap crops and agricultural practices for pest management. The harlequin bug, Murgantia histrionica, a specialist pest on crucifer crops, produces the sesquiterpene, murgantiol, as a male-specific aggregation pheromone. Similarly, the southern green stink bug, Nezara viridula, a generalist pest worldwide on soybean and other crops, releases sesquiterpene cis-/trans-(Z)-α-bisabolene epoxides as male-specific aggregation pheromone. In both species, enzymes called terpene synthases (TPSs) synthesize precursors of the aggregation pheromones, which are sesquipiperitol and (Z)-α-bisabolene as the precursor of murgantiol and cis-/trans-(Z)-α-bisabolene epoxide, respectively. We hypothesized that enzymes in the family of cytochrome P450 monooxygenases are involved in the conversion of these precursors to the final epoxide products. This study investigated the tissue specificity and sequence of these conversions by performing crude enzyme assays with protein extracts from male tissues. Furthermore, candidate P450 genes were selected by RNA-sequencing and co-expression analysis and the corresponding recombinant proteins tested for enzyme activity. To engineer the pheromone biosynthetic enzymes in plants, transient expression of the TPSs of both stink bugs was performed in Nicotiana benthamiana leaves. Both sesquipiperitol and (Z)-α-bisabolene were found to be produced and emitted from inoculated N. benthamiana leaves. Future work will implement stable transformation to engineer murgantiol biosynthesis in crucifer trap crops and develop similar approaches for pheromone engineering of other agricultural pests. / Master of Science / Stinkbugs including the harlequin bug, Murgantia histrionica and southern green stinkbug, Nezara viridula, are major agricultural pests in the US and worldwide. To control these pests with alternative pest management strategies, we have proposed to develop trap crops that emit pheromones to lure the insects away from crop fields. To establish pheromone biosynthesis in plants, we investigated the corresponding enzymatic steps in both stink bugs. We show that terpene synthase enzyme from both stink bugs can be transformed into plants for the engineering of pheromones in trap crops. With identification of P450 genes in pheromone biosynthesis enhanced trap crops can be made.

The pink hibiscus mealybug (PHM), Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) was investigated with regard to damage caused to hibiscus by feeding, dispersal of nymphs, evaluation of management tactics, and the use of sex pheromone based monitoring in southern Florida from 2005 to 2008. Understanding the ability of PHM to locate and colonize new hosts, and the response of hosts is essential to optimized monitoring and management strategies. Investigation of the onset and severity of PHM feeding symptoms by Hibiscus rosa-sinensis L. showed that severity of symptoms differed among cultivars and that PHM were found on plants that did not exhibit feeding symptoms. Aerially dispersing PHM were predominantly first instars. Dispersal occurred with a diel periodicity that peaked between 14:00 and 18:00 h and was significantly influenced by mean wind speed. Initial infestation with 5, 10, or 20 PHM adult females had no significant affect on the number of dispersing individuals captured from hibiscus plants and PHM were captured at 50 m from infested source plants. The effects of mating disruption, the insecticide (dinotefuran), the parasitoid, Anagyrus kamali (Moursi), and the predator, Cryptolaemus montrouzieri (Mulsant) on PHM on hibiscus plants in screened field cages were evaluated. The total number of mealybugs captured in sticky band and pheromone traps during the study was reduced by dinotefuran and the predator. At the end of the study, the number of nymphs recovered from hibiscus terminals was reduced by the dinotefuran, predator and parasitoid treatments. Field experiments showed that the time of day during which male PHM were captured in pheromone traps in May and September was crepuscular, with most captures occurring from 18:00 to 21:00 h. Significantly more males were captured in traps placed in non-host trees at an elevation of 2 m above the ground than 6 m, and more males were captured in traps placed within host plants than in those 3 m upwind. Pheromone traps placed in hibiscus treated with soil applied dinotefuran or left untreated captured equal numbers of males during the 3 wk prior to treatment and during the 12 wk after treatment. Release of parasitoids at residential sites did not have a significant effect on the total number of males captured in sex pheromone traps over 18 mo. The number of mealybugs found at both parasitoid release and untreated sites were highly variable and corresponded with males captured in sex pheromone traps, as high and low levels of mealybugs corresponded with high and low levels of males captured. The number of males captured in pheromone traps during a two week survey at residential sites in May were a strong predictor of subsequent captures in 2006 (r2 = 0. 712), but not 2007 (r2 = 0.019). The relationship between PHM populations and males captured in sex pheromone traps was influenced by a multitude of factors that can impact the ability of traps to accurately reflect populations at a given location. Pheromone traps have the potential to provide meaningful information towards monitoring and mitigating risk from PHM. The contributions of this dissertation towards optimizing PHM sex pheromone monitoring, as well as facets of PHM monitoring that have yet to be resolved are discussed. / Ph. D.

My research addressed pheromone lure design and the activity of the grape berry moth, Paralobesia viteana, flight and infestation across three years of study. In my lure evaluations, I found all commercial lures contained impurities and inconsistencies that have implications for management. First, sex pheromone concentration in lures affected both target and non-target attraction to traps, while the blend of sex pheromones impacted attraction to P. viteana. Second, over the duration of study, 54 vineyard blocks were sampled for the pest in and around cultivated wine grape in Virginia. The trapping studies indicated earliest and sustained emergence of the spring generation in sex pheromone traps placed in a wooded periphery. Later, moths were detected most often in the vineyard, which indicated that P. viteana emerged and aggregated in woods prior to flying and egg-laying in vineyards. My research supports use of woods and vineyard trap monitoring at both the height of 2 meters and in the periphery of respective environments. These conditions should improve grower efficiency when using trapping as a tool. Cluster infestation was assessed relative to vineyard growth stage and location in the vineyard. My results are in agreement with historical studies that suggest infestation is highest in the periphery of vineyards. However, I found that infestation peaked at veraison before falling somewhat around pre-harvest. This indicated that growers should manage this pest prior to veraison, perhaps as early as budbreak, to prevent the insect from reaching damaging levels when the crop is more susceptible to loss. Lastly, by combining field-based data collection with remote climate and landscape monitoring, growers can achieve a greater level of surveillance of P. viteana. A mash up of remote and vineyard-based data can provide substantive pertinent information for management at both local and regional scales. If one grower identifies moth emergence, then growers nearby potentially can use that same information to initiate models to monitor development of the moth population in their own vineyard. This research complements the notion that the more a community shares information, the broader its applicability becomes to neighbors. / Ph. D.

The stink bugs, (Pentatomidae) harlequin bug (Murgantia histrionica), brown marmorated stink bug (Halyomorpha halys), and southern green stink bug (Nezara viridula) are significant agricultural pests both in the United States and globally. The aggregation or sex pheromones produced by these insects are known to be bisabolene-type sesquiterpenoids; however, the biosynthetic pathways in the formation of these pheromones are unknown. Here we provide evidence that Pentatomidae produce sesquiterpene aggregation pheromones de novo and discuss the evolution of terpene biosynthesis in stink bugs. According to transcriptome analyses, the investigated stink bug species express at least two isoprenyl diphosphate synthases (IDSs), one of which makes (E,E)-farnesyl diphosphate (FPP) as the general precursor in sesquiterpene synthesis, whereas other IDS-type proteins function as terpene synthases (TPSs) generating intermediates in sesquiterpene pheromone formation. The TPS genes are expressed in a sex- and tissue-specific manner. Based on phylogenetic analysis, these IDS-type TPSs arose from trans-IDS progenitors in divergence from bona fide IDS proteins. Compared to microbes and plants, the evolution of TPS function from IDS progenitors in insects appears to have occurred more recently. The discovery of TPS genes in stink bugs provides valuable insight into pentatomid and insect terpene biosynthesis. Moreover, the identified genes may be used in developing alternative management strategies for stink bug pests. / PHD

Diss. (sammanfattning) Alnarp : Sveriges lantbruksuniv. / Härtill 6 uppsatser.