Balfourodendron riedelianum (Engl.) Engl. e uma espécie arbórea tropical que ocorre na Mata Atlântica e está em perigo de extinção devido a fragmentação do bioma e a exploração de sua madeira. Atualmente três populações da espécie estão sendo conservadas em um teste de procedências e progênies implantado na Estação Experimental de Luiz Antonio, estado de São Paulo, Brasil. O teste e composto por procedências de Alvorada do Sul, estado do Paraná, Gália e Bauru, ambas do estado de São Paulo. Com o objetivo de verificar o potencial do teste como fonte de sementes para a recomposição florestal, esse estudo avaliou a biometria de frutos, geminação de sementes, mortalidade e sobrevivência de plântulas da espécie. Foram desenvolvidos, marcadores microssatélites para a análise da diversidade genética, endogamia, sistema de reprodução, fluxo e dispersão de pólen e depressão por endogamia (DE). A variação observada entre as matrizes para a biometria de frutos, assim como para germinação, mortalidade e sobrevivência sugerem que estas sejam decorrentes da variabilidade genética, não foi detectada correlação entre a biometria e germinação. Os microssatélites apresentaram alto nível de polimorfismo e ausência de desvios de segregação Mendeliana, ligação genética e desequilíbrio de ligação. A procedência Gália apresentou a maior riqueza alélica (R = 10,3), heterozigosidade esperada (He = 0,60) e índice de fixação (F = 0,11) e a procedência Alvorada do Sul apresentou a maior heterozigosidade observada (Ho = 0,58) e o menor índice de fixação (F = -0,01). As plântulas apresentaram a menor riqueza alélica (R = 9,4) e heterozigosidade observada (Ho = 0,48) e o maior índice de fixação (F = 0,16). A taxa de cruzamento (tm) foi alta (> 0,93), mas foi observado para as plântulas uma variação individual na taxa de cruzamento (tm = 0,68 a 0,99), cruzamento entre parentes (tm - ts = 0 a 0,18) e na correlação de paternidade (rp = 0,041 a 0,182), o que indica que a espécie se reproduz predominantemente por cruzamentos, e autocompatível e que a reprodução não foi aleatória. A distância média de dispersão do pólen (71 m) maior que a mediana (21 m) sugere um padrão de isolamento por distância. Aos 12 meses as plântulas oriundas de autofecundação apresentaram os menores valores de C e Ho e o maior F (2,98 mm; 0,25 e 0,61) do que as de cruzamento entre parentes (3,10 mm, 0,34 e 0,29, respectivamente) e não parentes (3,30 mm, 0,51 e 0,07, respectivamente). A autofecundação resultou em maior DE para circunferência a altura do colo (9,6%) e o cruzamento entre parentes para altura (2,6%). Para fins de conservação, sementes de no mínimo 53 arvores devem ser coletadas para alcançar um Ne(ref) de 150 indivíduos, da mesma forma, devem ser mantidos também pelo menos 53 indivíduos com a mesma fenologia de florescimento a cada ciclo de desbaste. A endogamia decorrente de cruzamentos entre parentes pode ser minimizada se apenas uma planta por família for selecionada dentro das parcelas e pode ser eliminada se apenas uma planta por família for selecionada no teste, sendo possível obter ganhos com a seleção em ambos os casos. / Balfourodendron riedelianum (Engl.) Engl. is a tropical tree species of the Atlantic Forest that is endangered due to exploitation of its wood and biome fragmentation. Currently, three populations of the species are preserved in a provenance and progeny test established in the Luiz Antônio Experimental Station, São Paulo State, Brazil. The test consists of provenances from Alvorada do Sul, Parana State, and Galia and Bauru, both from São Paulo State. In order to verify the potential of the test as a seed source for forest restoration, this study evaluates the species\' fruit biometry, seed germination, mortality and seedling survival. Microsatellite markers were developed to analyze genetic diversity, inbreeding, mating system, pollen flow and dispersal, and inbreeding depression (ID). The variation observed between seed trees for fruit biometry, as well as for germination, mortality and survival, suggests a correlation with genetic variability. No correlation was detected between biometry and germination. Microsatellites presented a high level of polymorphism and an absence of deviation from Mendelian segregation, genetic linkage, and linkage disequilibrium. The Galia population had the highest allelic richness (R = 10.3), expected heterozygosity (He = 0.60), and fixation index (F = 0.11), and Alvorada do Sul had the highest observed heterozygosity (Ho = 0.58) and the lowest fixation index (F = -0.01). Seedlings showed the lowest allelic richness (R = 9.4) and observed heterozygosity (Ho = 0.48) and the highest fixation index (F = 0.16). The outcrossing rate (tm) was high (> 0.93). However, there was individual variation in seedlings for the outcrossing rate (tm = 0.68 to 0.99), mating between relatives (tm - ts = 0 to 0.18), and paternity correlation (rp = 0.041 to 0.182), indicating that the species has a predominantly outcrossed mating system, is selfcompatible, and that reproduction is not random. The mean pollen dispersal distance (71 m) was greater than the median (21 m), suggesting a pattern of isolation by distance. At 12 months, seedlings resulting from selfing had the lowest values of C, Ho, and higher F (2.98 mm, 0.25, and 0.61, respectively) than those from mating between relatives (3.10 mm, 0.34, and 0.29) and unrelated individuals (3.30 mm, 0.51, and 0.07). Selfing resulted in higher ID for root-collar circumference (9.6%) and mating between relatives showed a higher ID for height (2.6%). For conservation purposes, seeds of at least 53 trees must be collected to reach Ne(ref) of 150 individuals and at least 53 individuals with the same flowering phenology must be maintained at each thinning cycle. Inbreeding resulting from crosses between relatives can be minimized if only one plant per family is selected within the plots and can be eliminated if only one plant per family is selected in the test, thus enabling gains with selection in both cases.

Existem poucas informações sobre o uso de intercruzamentos em populações de soja derivadas de retrocruzamento. Os objetivos deste trabalho compreendem a avaliação dos efeitos de uma geração de intercruzamentos nas médias, distribuição de frequências, variâncias genéticas e respostas à seleção em uma população de retrocruzamento de soja. A população básica foi derivada de um cruzamento simples entre duas linhagens contrastantes para a produção de grãos, seguido de um retrocruzamento para a linhagem mais produtiva. Cento e dezessete progênies derivadas da população não intercruzada (progênies RC1F2) e cento e dezoito progênies derivadas da população intercruzada (progênies RC1#F2) foram avaliadas no ano agrícola de 2008/09 na Estação Experimental de Anhumas, do Departamento de Genética da ESALQ/USP, em Piracicaba, SP, em dois experimentos em látice quadruplo 11x11 (quatro repetições). As parcelas foram constituídas de linhas de 2 m, espaçadas de 0,5 m, contendo 30 plantas após o desbaste. Estas foram colhidas em bulk (que correspondem às gerações RC1F3 e RC1#F3) e avaliadas novamente no ano agrícola 2010/11, utilizando o mesmo delineamento experimental, tipo de parcela e local. Foram avaliados os seguintes caracteres: número de dias para florescimento (DF), altura da planta no florescimento (AF), número de dias para maturação (DM), altura da planta na maturação (AM) e produção de grãos (PG). Para os dois tipos de populações, RC1 e RC1#, foram estimados os seguintes parâmetros: média geral, amplitude de variação, distribuição das frequências, variância genética entre progênies (σ2p), variância aditiva (σ2A), variância fenotípica entre médias de progênies (σ2F), e resposta esperada com seleção (Rs). As médias foram similares para a maioria dos caracteres entre as populações RC1 e RC1# dentro de cada ano. Entretanto, houve um acréscimo das variâncias genéticas na população intercruzada para a maioria dos caracteres, o que era esperado com base em um modelo de um loco com dois alelos. Consequentemente, a resposta esperada com seleção para PG foi 39% maior, em média, para as populações intercruzadas (RC1#F2 e RC1#F3). Estes resultados indicam que uma geração de intercruzamento após o retrocruzamento é importante em programas de melhoramento genético de soja. / There is limited information on using random mating after backcrossing in soybeans. This work was carried out to evaluate the effects of one generation of random mating after backcrossing on the means, frequency distributions, genetic variances and responses to selection in a soybean population. The basic population was derived from a two-way cross between two inbred lines contrasting for grain yield and backcrossed to the higher yielding one. One hundred and seventeen progenies derived from a not random-mated backcross population (RC1F2 progenies) and one hundred and eighteen progenies derived from a random-mated backcross population (RC1#F2 progenies) were evaluated in the 2008/09 growing season at Anhumas Experimental Station, of the Department of Genetics, ESALQ/USP, located in Piracicaba, state of São Paulo, Brazil. Evaluation trials were carried out using an 11x11 quadruple lattice design (four replications). Plots consisted of 2 m rows spaced by 0.5 m, with 30 plants after thinning. The entries were harvested in bulks (which correspond to RC1F3 and RC1#F3 progenies) and evaluated again in the 2010/11 growing season, using the same experimental design, plot size and location. The following traits were recorded: number of days to flowering (DF), plant height at flowering (AF), number of days to maturity (DM), plant height at maturity (AM), and grain yield (PG). The following parameters were estimated for both RC1 and RC1#: general mean, amplitude of variation, frequency distribution, genetic variance among progenies (σ2p), additive variance (σ2A), phenotypic variance on a progeny mean basis (σ2F), and expected response to selection (Rs). For most traits general means were similar for RC1 and RC1# populations within years. However, genetic variances increased after one generation of random mating, for most traits, which was expected based on a one-locus two-alleles model. Thus, expected response to selection on a progeny mean basis for grain yield (PG) was 39% higher, on average, for the random-mated populations (RC1#F2 and RC1#F3). Overall, the results indicate that one generation of random mating before selfing in backcross population is useful in soybean breeding programs.

Le sanglier (Sus scrofa scrofa) est une espèce à part entière. C'est une espèce de gibier particulièrement appréciée des chasseurs, un cauchemar pour les agriculteurs et un sujet de débat pour la société en général. La multiplication par dix des populations au cours des dernières décennies en France et dans toute l'Europe, malgré une pression de chasse accrue, a engendré de nombreux conflits entre les humains et la faune sauvage. Le sanglier est responsable de grandes pertes économiques dues aux collisions avec les véhicules, à la transmission de maladies et de dommages aux cultures et écosystèmes. L'amélioration des stratégies de gestion devient un intérêt majeur pour éviter, ou contrôler, de tels conflits. La récolte d'informations sur l'espèce problématique est un premier pas vers de bonnes stratégies de gestion. L'objectif de mon travail est, dans un premier temps, de caractériser le système d’appariement du sanglier et d'identifier certains paramètres influençant les processus de reproduction, notamment la chasse. Dans un deuxième temps, mon travail se concentre sur l'étude de l'influence du système d’appariement sur les traits d'histoire de vie du sanglier. Mes recherches sont basées sur l'étude de plusieurs populations contrastées dans leurs pratiques de chasse et sur des données longitudinales d'une population intensivement suivie. L'étude est basée sur des données recueillies sur des sangliers tués à la chasse. Les génotypes ont été obtenus pour les femelles gestantes et leur portée et des analyses de paternité ont été réalisées pour mesurer le nombre de pères dans une portée et estimer les taux de paternité multiples (proportion de portées engendrées par plus d'un père). J'ai été en mesure de montrer que le système d’appariement est principalement de promiscuite (plusieurs mâles s'accouplent avec plusieurs femelles) contrastant avec la polygynie (un mâle dominant monopolisant un groupe de femelles) habituellement décrite chez cette espèce. De plus, les processus de reproduction, estimés par le nombre de partenaires d'une femelle et les taux de paternité multiples, sont influencés par les variations de chasse dans une population. J'ai aussi montré que le nombre de pères avait un effet positif sur la fécondité des femelles. Des taux élevés de paternité multiple et une grande diversité génétique ont été constatés ensemble dans une population fortement chassée, ce qui suggère que la paternité multiple peut tamponner les goulots d'étranglement annuels. Cependant, l'augmentation du nombre de père n'est pas associée à une augmentation de la variation intraportée / The wild boar (Sus scrofa scrofa) is a peculiar species. It is an appreciated game species for hunters, a nightmare for farmers and a subject of debate for the society in general. The tenfold increase of the population over the last decades in France and all over Europe, despite increased hunting pressure, generated great human-wildlife conflict. The wild boar is responsible for great economic losses due to vehicle collision, diseases transmission and damaged crops and ecosystems. Improving management strategies becomes a prime interest to avoid such conflicts, or at least keep them under control. Obtaining information on the species is a first step toward good management strategies. The objective of my work is, in a first part, to characterize the mating system of the wild boar and to identify some parameters, especially hunting, influencing the reproductive processes. The second part focus on the investigation of the influence of the mating system on wild boar life history traits. My researches are based on the study of several populations contrasting in their hunting practices and on longitudinal data of a highly monitored population. The study is based on data collected on wild boars killed by hunting. Genotypes were obtained for pregnant females and their litter and paternity analyses were realized to measure the number of fathers in a litter and estimate multiple paternity rates (proportion of litter sired by more than one father). I was able to show that the mating system is mainly promiscuous (several males mate with several females) contrasting with the polygyny (a dominant male monopolizing a group of females) usually described in this species. Moreover, reproductive processes, estimated by the number of mates of a female and the multiple paternity rates, are influenced by hunting variations in a population. I also showed that number of fathers has positive effect on female fecundity. High rates of multiple paternity together with high genetic diversity were found in a heavily hunted population, suggesting multiple paternity may buffer yearly bottlenecks. However, the increase of number of fathers is not associated with increase of within-litter variation

Wild Canis species possess a unique suite of reproductive traits including social monogamy, copulatory lock/tie, and biparental care. Females are seasonally monestrous and experience an obligatory pseudopregnancy after spontaneous ovulation. While these characteristics have been ascribed to coyotes, an integrated profile of behavior and physiology has not yet been described. In this study, temporal correlations between steroid hormone levels and socio-sexual mating behaviors were documented, as were changes in vaginal epithelium. Pseudopregnancy was compared to pregnancy by contrasting hormone (progesterone, estradiol, prolactin and relaxin) profiles of unmated females to patterns obtained in alternate years when they bred. Meanwhile, social interactions between pseudopregnant females and their mates appeared similar to pregnant coyotes, suggesting a proximate role of pseudopregnancy in pair-bond enforcement. Finally, out-of-season stimulation of ovarian hormones and estrous behaviors suggested that reproductive seasonality of the coyote may possess some degree of plasticity, providing an adaptive response mechanism to environmental change.

在族群遺傳學中已知有許多因子影響基因型與對偶基因頻率，族群的交配方式即為其中之一。例如，哈溫平衡定律即指族群進行隨機交配，而芮特平衡定律所描述之自交系理論指的是族群進行了某種特定型式的非隨機交配。本文將以往討論過或未討論過之族群基因型頻率會達到哈溫比例及芮特比例的交配系統做一整合性研究。族群基因型頻率會達到哈溫比例或芮特比例之交配系統的整體架構將根據交配群體中雙親二子群體的交配行為、基因型頻率是否在哈溫比例及是否相同、對偶基因頻率是否相同來建立。整體架構中的每一構造點所對應的交配系統將被找出或證明不存在，若存在則會進一步討論其特性。 目　錄 摘要………………………………………………………………….. I ABSTRACT………………………………………………………… II 圖表目錄…………………………………………………………….. VII 第一章 緒論……………………………………………………….. 1 第二章 族群達到Hardy-Weinberg比例及Wright比例的交配系統之整體架構………………………………………….. 3 第三章 族群達到Hardy-Weinberg比例之隨機交配系統……... 6 3.1 基本定義與原理………………………………………………………. 6 3.1.1 基因型頻率與對偶基因頻率…………………………………… 6 3.1.2 隨機交配與非隨機交配………………………………………… 7 3.1.3 基因型隨機交配會導致配子隨機結合原理…………………... 7 3.1.4 平衡族群的交配系統之特性…………………………………… 8 3.2 雌雄二次群體之基因型頻率在相同的Hardy-Weinberg比例且其子代群體基因型頻率會達到Hardy-Weinberg比例的隨機交配系統 ………………………………………………………………… 9 3.3 雌雄二次群體之基因型頻率在不同的Hardy-Weinberg比例的隨機交配系統 其子代群體基因型頻率不會達到Hardy-Weinberg比例……………………………………………………….. 10 3.4 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例但基因型及對偶基因頻率皆相同下其子代群體基因型頻率會達到Hardy-Weinberg比例的隨機交配系統 ……………………………….. 12 3.5 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例且基因型頻率不同但對偶基因頻率相同下其子代群體基因型頻率會達到Hardy-Weinberg比例的隨機交配系統 ………………………. 13 3.6 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例且基因型及對偶基因頻率皆不同的隨機交配系統 之子代群體基因型頻率不會達到Hardy-Weinberg比例………………………………. 14 3.7 隨機交配族群基因型頻率會達到Hardy-Weinberg比例的交配系統之結論與彙整………………………………………………………. 14 第四章 族群達到Hardy-Weinberg比例的隨機交配系統之性質 17 4.1 隨機交配系統 及其子代群體配子結合之相關係數…………... 17 4.1.1 隨機交配系統 之相關係數………………………………... 17 4.1.2 隨機交配系統 之子代群體配子結合的相關係數……….. 18 4.2 ITO方法及其應用……………………………………………………. 19 4.3 隨機交配系統 之親代和子代基因型聯合頻率矩陣及其相關係數. 21 4.3.1 隨機交配系統 之母子基因型聯合頻率矩陣及其相關係數... 20 4.3.2 隨機交配系統 之父子基因型聯合頻率矩陣及其相關係數... 23 4.3.3 隨機交配系統 之親子基因型聯合頻率矩陣及其相關係數... 24 4.4 隨機交配系統 之二同胞基因型聯合頻率矩陣及其相關係數.. 25 4.5 隨機交配族群基因型頻率達到Hardy-Weinberg比例的交配系統之性質比較……………………………………………………………. 27 第五章 母父子基因型的傳遞機率矩陣及其應用……………….. 28 5.1 母父子基因型的傳遞機率矩陣………………………………………. 28 5.2 子代群體基因型頻率之矩陣代數算法………………………………. 31 5.2.1 不區分異質結合基因型的子代群體基因型頻率……………... 32 5.2.2 區分異質結合基因型的子代群體基因型頻率………………... 33 5.3 親代和子代基因型聯合頻率之矩陣代數算法……………………… 34 5.3.1 母子基因型聯合頻率矩陣之矩陣代數算法…………………... 34 5.3.2 父子基因型聯合頻率之矩陣代數算法………………………… 35 5.4 二同胞基因型聯合頻率之矩陣代數算法…………………………… 36 第六章 族群達到Hardy-Weinberg比例之非隨機交配系統…... 41 6.1 雌雄二次群體之基因型頻率在Hardy-Weinberg比例且基因型及對偶基因頻率皆相同下其子代群體基因型頻率會達到Hardy-Weinberg比例的非隨機交配系統 …………………………….. 41 6.2 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例且基因型與對偶基因頻率皆不同但其子代群體基因型頻率達到Hardy-Weinberg比例的非隨機交配系統 …………………………….. 44 6.3 非隨機交配系統 的另一種表示法……………………………… 46 6.4 雌雄二次群體之基因型頻率在Hardy-Weinberg比例但基因型及對偶基因頻率皆不同下其子代群體基因型頻率會達到Hardy-Weinberg比例的非隨機交配系統 …………………………….. 50 6.5 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例但基因型及對偶基因頻率皆相同下其子代群體基因型頻率會達到Hardy-Weinberg比例的非隨機交配系統 …………………………….. 52 6.6 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例且基因型頻率不同但對偶基因頻率相同下其子代群體基因型頻率會達到Hardy-Weinberg比例的非隨機交配系統 …………………… 53 6.7 非隨機交配族群基因型頻率會達到Hardy-Weinberg比例的交配系統之彙整……………………………………………………………. 54 第七章 族群達到Hardy-Weinberg比例的非隨機交配系統之性質……………………………………………………….. 56 7.1 雌雄交配基因型聯合頻率矩陣 之相關係數………………….. 56 7.2 之子代群體配子結合的相關係數……………………………… 58 7.3 之親代和子代基因型聯合頻率矩陣及其相關係數………….. 60 7.3.1 之母子基因型聯合頻率矩陣及其相關係數…………….. 61 7.3.2 之父子基因型聯合頻率矩陣及其相關係數…………….. 64 7.4 之二同胞基因型聯合頻率矩陣及其相關係數……………… 64 7.5 非隨機交配族群之子代群體基因型頻率達到Hardy-Weinberg比例的交配系統之性質比較…………………………………………… 69 第八章 族群達到Wright比例的交配系統及其性質……………. 71 8.1 Wright平衡定律……………………………………………………… 71 8.2 族群基因型頻率達到Wright比例的交配系統之最一般化型式 … 72 8.3 族群基因型頻率會達到Wright比例之交配系統的整體架構中其餘各交配系統之存在性………………………………………………. 77 8.3.1 雌雄二次群體之基因型頻率在Hardy-Weinberg比例且基因型及對偶基因頻率皆相同其子代群體基因型頻率會達到Wright比例的交配系統 …………………………………. 77 8.3.2 雌雄二次群體之基因型頻率不在Hardy-Weinberg比例但基因型及對偶基因頻率皆相同其子代群體基因型頻率會達到Wright比例的交配系統 …………………………………. 78 8.3.3 族群基因型頻率達到Wright比例的交配系統之彙整……….. 80 8.4 族群基因型頻率會達到Wright比例的交配系統之性質………… 82 8.4.1 雌雄交配基因型聯合頻率矩陣 之相關係數……………. 82 8.4.2 之親代和子代基因型聯合頻率矩陣及其相關係數……. 84 8.4.3.1 之母子基因型聯合頻率矩陣及其相關係數……… 84 8.4.3.2 之父子基因型聯合頻率矩陣及其相關係數………. 87 8.4.3 之二同胞基因型聯合頻率矩陣及其相關係數…………. 87 8.4.4 族群基因型頻率會達到Wright比例的交配系統之性質的結論與彙整………………………………………………………… 89 第九章 一般化平衡族群的交配系統及其性質………………….. 91 9.1 一般化平衡定律………………………………………………………. 91 9.2 非隨交配族群基因型頻率達到一般化比例的交配系統之最一般化型式 ……………………………………………………………… 93 9.3 雌雄二次群體之基因型頻率在Hardy-Weinberg比例但基因型與對偶基因頻率皆不同的一般化平衡族群之交配系統 ………… 98 9.4 一般化平衡族群的交配系統之性質………………………………… 100 9.4.1 雌雄交配基因型聯合頻率矩陣 之相關係數……………… 100 9.4.2 之子代群體配子結合之相關係數………………………… 102 9.4.3 之親代和子代基因型聯合頻率矩陣及其相關係數……… 104 9.4.3.1 之母子基因型聯合頻率矩陣及其相關係數……….. 104 9.4.3.2 之父子基因型聯合頻率矩陣及其相關係數………... 106 9.4.4 之二同胞基因型聯合頻率矩陣及其相關係數…………… 106 9.4.5 一般化平衡族群之性質彙整…………………………………… 109 第十章 結論與未來研究方向…………………………………….. 111 10.1 結論……………………………………………………………………. 111 10.2 未來研究方向…………………………………………………………. 115 參考文獻…………………………………………………………….. 117 / In population genetics it is known that there are many factors that may affect the genotypic and gene distributions of a population. The type of mating of a population is one of them. For examples, basically Hardy-Weinberg equilibrium law refers to a population undergoing random mating, and Wright's equilibrium law in inbreeding refers to a special type of nonrandom mating. This study performs a comprehensive investigation of all possible matings that can attain Hardy-Weinberg proportions or Wright proportions that had been or not had been discussed previously. The framework of mating systems attaining the Hardy-Weinberg proportions or Wright proportions will be established on the basis of pooling together factors such as the mating behavior, gene frequencies, genotypic frequencies and the Hardy-Weinberg proportions of the male and female subpopulations. The type and property of each mating system corresponding to each point of the established mating system framework are examined.

The Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), a major pest of horticulture in eastern Australia, is a relatively poor coloniser of new habitat. This thesis examines behavioural properties that might limit the ability of B. tryoni to establish new populations. As the potential for B. tryoni to establish an outbreak population may be most directly limited by mechanisms associated with dispersal and mating behaviour, these two factors were the focus of this research project. The relevance of dispersal and mating behaviour for control of outbreak populations was assessed. Dispersal (i) Dispersal patterns of males and females are not different. Dispersal of post-teneral male B. tryoni from a point within an orchard near Richmond, New South Wales, was monitored following temporally replicated releases. Application of sterile insect technique (SIT) requires knowledge of dispersal from a release point so that effective release rates can be determined. In addition, dispersal following introduction to new habitat can lead to low or negative population growth and an Allee effect. In Spring and Autumn, 2001 � 2003, three different strains of B. tryoni were released: (1) wild flies reared from infested fruit collected in the Sydney Basin; (2) a laboratory-reared strain with a colour mutation (white marks); and (3) sterile flies obtained by gamma-irradiation of a mass-reared strain. Dispersal was monitored using a grid of traps baited with the male attractant, cuelure. During the majority of releases, flies were massmarked using a self-marking technique and fluorescent pigment powder to enable identification of recaptured flies. A preliminary study found that fluorescent pigment marks had no effect on adult survival and marks did not fade significantly in the laboratory over a period of five weeks after eclosion. As cuelure repels inseminated sexually mature female B. tryoni, unbaited, coloured flat sticky traps, and black and yellow sticky sphere traps baited with a food lure (protein autolysate solution) were used to supplement traps baited with cuelure. The effectiveness of these two sticky trap types was assessed, and recaptures used to compare patterns of dispersal from a release point by male and female B. tryoni. Fluorescent yellow (chartreuse), green, and clear unbaited flat sticky traps were relatively ineffective for monitoring dispersal of sterile male and female B. tryoni, recapturing only 0.1% of released sterile flies. Monitoring dispersal with sticky ball traps baited with protein autolysate solution was more successful, with yellow spheres and black spheres recapturing 1.7% and 1.5%, respectively. Trap colour had no effect on recaptures on flat sticky traps or sticky spheres. Equal recapture rates on yellow and black sticky sphere traps suggests that the odour of yeast autolysate solution was more important than colour for attraction of post-teneral flies to traps. Using the results of recaptures on odoriferous black and yellow sticky sphere traps within one week of release, regression equations of male and female recaptures per trap were found to be similar (Figure 4-3). This is the first study to clearly indicate that post-teneral dispersal patterns of male and female B. tryoni released from a point do not differ, enabling the use of existing models to predict density of both sexes of B. tryoni following post-teneral dispersal. (ii) Males disperse further in Spring than in Autumn, but this is not temperature-related. Analysis of replicated recaptures in traps baited with cuelure revealed that dispersal of male B. tryoni in an orchard near Richmond, New South Wales, was higher in Spring than in Autumn (Figure 5-6). As the maximum daily temperature was significantly higher in Spring than in Autumn this result was unexpected, since earlier studies have found that B. tryoni disperse at the onset of cool weather in search of sheltered over-wintering sites. Dispersal of post-teneral B. tryoni may have been affected by habitat suitability; it was found that seasonal trends in dispersal could have been influenced by local habitat variables. Low mean dispersal distances in Autumn may be explained by the presence of fruiting hosts in the orchard, or the availability of resources required by over-wintering flies. There was no significant correlation between temperature and mean dispersal distance, suggesting that higher rates of dispersal cannot be explained by temperature-related increases in activity. Recapture rate per trap was significantly negatively correlated with increasing daily maximum and average temperature. This may have consequences for detection of B. tryoni outbreaks in quarantine areas due to reduced cuelure trap efficiency. (iii) Maturity and source variation affect dispersal and response to cuelure. This research indicated that most male and female B. tryoni do not disperse far from a release point, suggesting that an invading propagule would not spread far in the first generation. However, there is considerable variation in flight capability among individuals. Comparison of wild, laboratory-reared white marks, and gamma-irradiated sterile male B. tryoni indicated that mean dispersal distance and redistribution patterns were not significantly affected by fly origin. Despite no difference in dispersal distance from the release point, recaptures of wild and sterile males per Lynfield trap baited with cuelure were highest within one week after release, whereas recaptures of white marks males per trap increased in the second week. This result may offer evidence to support the hypothesis that sterile male B. tryoni respond to cuelure at an earlier age. Rearing conditions used to produce large quantities of males for sterilisation by gamma-irradiation may select for earlier sexual maturity. Mating Behaviour (i) Density and sex ratio do not affect mating, except at low densities. Demographic stochasticity in the form of sex ratio fluctuations at low population density can lead to an observed Allee effect. The effect of local group density and sex ratio on mating behaviour and male mating success of a laboratory-adapted strain of B. tryoni was examined in laboratory cages. In the laboratory-adapted strain of B. tryoni used in this study, a group of one female and one male was sufficient for a good chance of mating success. The proportion of females mated and male mating success was not significantly affected by density or sex ratio, although variability in male mating success was higher at low density. This could indicate that mating success of B. tryoni can be reduced when local group density is low owing to decreased frequency in encounters between males and females. (ii) Mass-reared males exhibit aberrant mating behaviour, but this does not reduce mating success. Strong artificial selection in mass-rearing facilities may lead to decreased competitiveness of sterile males released in SIT programs as a result of alteration or loss of ecological and behavioural traits required in the field. The effects of domestication and irradiation on the mating behaviour of males of B. tryoni were investigated by caging wild, mass-reared and sterile (mass-reared and gammairradiated) males with wild females. Mating behaviour of mass-reared males was different from that of wild males, but behaviour of wild and sterile males was similar. Mass-reared males were found to engage in mounting of other males much more frequently than wild and sterile males, and began calling significantly earlier before darkness. Male calling did not appear to be associated with female choice of mating partners, although this does not exclude the possibility that calling is a cue used by females to discriminate between mating partners. Conditions used to domesticate and rear large quantities of B. tryoni for SIT may select for an alternative male mating strategy, with mass-reared males calling earlier and exercising less discrimination between potential mating partners. Despite differences in behaviour of wild, mass-reared and sterile males, frequency of successful copulations and mating success were similar. (iii) Pheromone-calling by males was increased in larger aggregations but this did not result in significantly more female visits. Finally, large laboratory cages with artificial leks were used to investigate the importance in B. tryoni of male group size for female visitation at lek sites and initiation of male pheromone-calling. Calling propensity of male B. tryoni was increased by the presence of conspecific males. Females visited the largest lek more frequently than single males, but there was no correlation between lek size and female visitation. Female B. tryoni had a limited capacity to perceive a difference between the number of calling males; female visitation at leks was only weakly associated with male calling, suggesting that lek size and the number of pheromone-calling males may not be the only factor important in locating mates in B. tryoni. The weak, but positive correlation between male calling and female visitation may indicate that passive attraction maintains lek-mating in B. tryoni. Further studies are essential on mating behaviour of B. tryoni, including identification of male mating aggregations in the field, measurement of habitat variables associated with male aggregations, the influence of density on wild B. tryoni mating success, and the role of pheromone-calling, in order to optimise use of SIT for control of this pest.

The cytokine environment of early pregnancy is known to be a key determinant of the development of the pre-implantation embryo, and its subsequent implantation and growth. Factors in male seminal fluid have been identified as regulators of the expression of cytokines in the female tract of mice, humans and other mammalian species, with insemination eliciting a cascade of molecular and cellular events, reminiscent of a classic inflammatory response. In humans, perturbations in seminal fluid signalling have been proposed to predispose to pathologies of pregnancy including implantation failure, recurrent miscarriage and pre-eclampsia. Seminal transforming growth factor-beta (TGFβ) is identified as one key molecule present in seminal fluid responsible for inducing the female post-mating cytokine response in mice. Research in humans however, has shown the seminal TGFβ content of fertile versus infertile couples to be similar, while the content of other known seminal constituents such as interferon-gamma (IFNγ), correlate with reproductive success. This project aimed to investigate the nature of active factors present in seminal fluid in mice, and their interactions in regulating the uterine cytokine environment during early pregnancy, utilising a variety of in vitro and in vivo experimental strategies. Further, the effect of perturbation in the peri-conception cytokine environment on short and long term pregnancy and postnatal outcomes was investigated. Evaluation of uterine fluids from estrous and mated mice showed a marked upregulation of a number of cytokines following mating, including granulocyte macrophage colony stimulating factor (GM-CSF), interleukin-6 (IL-6) and the chemokine KC (rodent IL-8 homologue). Increased production of factors such as GM-CSF and subsequent generation of a receptive uterine environment is thought to be crucial for optimal embryo development and placentation. It has previously been shown that seminal factors such as TGFβ contribute to the uterine post-mating inflammatory response, however other moieties present in seminal fluid, for instance cytokines induced in response to infection such as IFNγ or products from the mucosal microflora, may also play a regulatory role. Using uterine epithelial cells cultured in vitro, it was shown that a variety of immune modulators including the cytokines TGFβ and IFNγ, as well as bacterial products, gram negative lipopolysaccharide (LPS) and gram positive lipoteichoic acid (LTA), can alter basal cytokine production. IFNγ, a pro-inflammatory cytokine secreted by activated natural killer cells and T-cells, is known to interfere with TGFβ signalling in other contexts. Independently TGFβ, LPS and LTA stimulate GM-CSF production while differentially regulating IL-6 and KC production. Conversely IFNγ inhibits GM-CSF production, without effecting IL-6 or KC. Pair wise combinations of TGFβ, LPS and LTA resulted in additive stimulation of GM-CSF, while addition of IFNγ to cultures in conjunction with any of these molecules downregulated GM-CSF and KC stimulation. These in vitro studies indicate factor-specific interactions between seminal fluid constituents and highlight the complex nature of seminal fluid signalling. Consequently we propose that the relative ratio of seminal signalling factors is likely to be more important than the absolute concentration of various regulators, in determining the optimal female reproductive tract response. Using the mouse as an in vivo model, I have in addition demonstrated that LPS and LTA instilled into an estrous uterus can elicit cytokine production comparable to that observed following insemination. Further, these studies have shown that IFNγ instilled into the uterus of a recently mated mouse can reduce the post-copulatory GM-CSF and KC surge. However administration of IFNγ had no effect on near term pregnancy outcomes including fetal or placental weights, fetal crown-rump length, or implantation or resorption rates. The ‘developmental origins of adult disease hypothesis’ proposes the idea that the early uterine environment encountered by the conceptus contributes toward the risk of metabolic disorders in adulthood, hence a long term study of progeny conceived after IFNγ administration was also undertaken. Neo-natal outcomes, such as birth weight, litter size and gestation length were unaltered, as was growth trajectory to 22 weeks of age. Adult metabolic markers, glucose tolerance, organ weight, muscle weight, adiposity and systolic blood pressure were not affected by the perturbation of peri-conceptual cytokine parameters. This work has examined the potential regulatory role of a number of seminal fluid signalling agents in directing the post-mating cytokine response, and has furthermore shown the relatively resilient nature of the early cytokine environment to subtle perturbation. Delineating the identity and roles of seminal fluid factors in early pregnancy brings us closer to an understanding of the key physiological events of early pregnancy and assists in identifying potential risk factors for human pregnancy pathologies. / Thesis (Ph.D.) -- University of Adelaide, School of Paediatrics and Reproductive Health, 2008

This thesis investigates aspects of the ecology, morphology, bioacoustics, genetic relatedness, biogeography and taxonomy of cicadas in the Pauropsalta annulata Goding and Froggatt species complex, across their entire eastern Australian geographical distribution. These cicadas seem to emerge at the same sites annually, although the exact duration of their life cycle is not known. They are wary and mobile insects, with individuals generally taking flight at the first sign of disturbance, which makes them challenging to record, capture and observe. The males produce distinctive calling songs that have a consistent rhythmic structure, which is critically important for attracting females. In most cases, the songs are biphasic, with a penetrating introductory or "buzzing" component that apparently functions in mate attraction and a strongly structured cueing or "lilting" component for mate localisation. Initially Pauropsalta annulata was thought to comprise a single species with some degree of geographically structured variation in its calling song. Consequently, various "song types" had been recognised, but their precise relationship to one another had never been investigated. Therefore the structure of their calling songs were compared statistically across individuals of three song types, and this revealed four discrete clusters that were demonstrated to be independent of one another and consistent in their calling song structure. Plotting the geographical distributions of these cicadas revealed that each of the P. annulata song types is independently distributed geographically, with areas of overlap that are relatively small. Calling song structure is consistent for each song type across extensive geographical space and this structure holds even into areas of sympatry. One song type showed consistent differences from the others in male genitalia structure, and female ovipositor length differs significantly among three of the other song types. Each song type was found to be strongly associated with a small number of tree species and these associations are maintained into areas of sympatry. The spatial ecological and morphological comparisons in song structure, plant associations and morphology made in this study demonstrate that the P. annulata song types actually represent a complex of cryptic species. Two of the song types appear to intergrade in areas of sympatric overlap in terms of calling songs, plant associations and morphology, and thus represent subspecies as defined in population genetics terms. The components of the male calling song were then investigated to determine their specific functions and thus how they could delimit species boundaries. Significant differences were found in dominant song frequency between three of the four species, and may contribute to differential mate attraction. Both components of the song of each species have the same dominant frequency. Consistent differences were also evident, among species and subspecies, in the rhythmic structure of the "lilting" component of the male calling song. This component contains repeated phrases and each one of these provides a cue to which the conspecific female may respond. Her response is timed for the brief silent interval between the phrases. At this point the calling song becomes a duet, which enables the male to locate the female, as he actively searches for her on the surrounding branches at this stage. The calling song is discussed in terms of random mating within gene pools of these cicadas and in terms of its role as part of their broader fertilisation mechanism. A molecular analysis of the P. annulata species complex was performed to examine the phylogenetic relationships across 12 species and four subspecies defined in this study, and estimate divergence times within the group. Individual specimens were sampled widely across the geographical distributions of the species and subspecies where possible to account for genetic variation across space. DNA sequences from two loci were amplified: mitochondrial CO1 ("barcoding region") and a large intron from the dynamin nuclear gene. Separate phylogenies were reconstructed for each locus using maximum parsimony procedures and Bayesian posterior sampling with implementation of a relaxed molecular clock. The phylogenies from both genes provided strong support for the monophyly of the P. annulata species complex, and nine of the species were monophyletic based on the CO1 gene. The remaining three emerged non-monophyletic. Based on a clock calibration of 0.0165s/s/myr, the monophyletic clades represented by extant P. annulata species diverged about 4.5-8.0 million years ago. Those species that emerged non-monophyletic had shallower divergences, with the exception of one species, which exhibited haplotype diversity that conferred up to 13.2% sequence divergence between allopatric populations in CO1. Dynamin produced a broadly similar phylogenetic pattern to that of CO1, but the relationships among individuals across the species and subspecies that emerged non-monophyletic differed substantially. This lack of congruence between the two genomes, in combination with the dominance of internal haplotypes in both loci, indicates an overall pattern of deep coalescence rather than interspecific hybridization. Therefore the molecular data do not provide an alternative definition of species limits in the P. annulata species complex, despite some emerging non-monophyletic in this analysis. Sound interpretation of the phylogenetic pattern discovered here would not have been possible without the acoustic, ecological and geographical investigations on species limits that preceded this work. To determine what biological and climatic factors influence the present day distributions of these cicadas, the distributions of two of the most closely related species in the P. annulata species complex were compared with the distributions of the tree species with which each is mainly associated. Because a large part of their life cycle is subterranean, soil texture, pH, electrical conductivity and force required for surface penetration were compared across sites where each of the cicadas occur in sympatry and allopatry. Finally, the influence of temperature and rainfall variables were investigated by testing 'predicted distribution' models (formed using positive distribution records) against negative records for both sets of variables, individually and in combination. The results show that the extent of the distribution of the cicada species is substantially less than that of the associated tree species. The geographical distributions of one of the species may be influenced more by rainfall, or a combination of temperature and rainfall, whereas the other species appears to be more influenced by temperature alone. Both species tolerate soils with a wide range of pH levels, electrical conductivity and forces required for surface penetration. They both showed a strong association with soils that had a silt loam texture, with only few records from sandy soils. However, none of the soils sampled where the cicadas occurred were heavy clays, which suggests that the physical properties of such soils may provide an unsuitable environment for the nymphal stages of the life cycle of these particular cicadas. The resolution of species limits within the P. annulata species complex allowed the redescription of Pauropsalta annulata Goding and Froggatt sensu stricto and the description of 11 new species belonging to the P. annulata species group, all from eastern Australia. Two of these species comprise two subspecies each, also all new. New distribution records and calling song data are documented for the allied species, P. ayrensis Ewart, which is redescribed to include the characters newly recognised in the present study as significant taxonomically with respect to Pauropsalta cicadas. The treatment includes comprehensive descriptions of the morphology and calling songs of the species and subspecies, and separate descriptive keys are provided for both sets of characters. The new taxa comprise P. artatus sp. nov., P. corymbiae sp. nov., P. decorus sp. nov., P. graniticus sp. nov., P. inversus inversus subsp. nov., P. i. laboris subsp. nov., P. notialis notialis subsp. nov., P. notialis incitatus subsp. nov., P. simplex sp. nov., P. subtropicus sp. nov, P. torrensis sp. nov. and P. tremulus sp. nov. Areas of hybridization between P. n. notialis subsp. nov. and P. n. incitatus subsp. nov. are also documented, together with their calling songs and morphology, which justifies their subspecific status. The P. inversus subspecies are allopatric, but consistently differ in the duration between phrases of the calling song. Finally, the results and conclusions are amalgamated into a critical reassessment of what defines species limits and the most appropriate approaches to investigating species limits in sexual organisms. Some historical discussions are revisited, such as the question of the reality of species and how species are perceived under the premises of neoDarwinism. The realism of species demands that species limits are most realistically defined in terms of their fertilisation mechanism, for this delimits the gene pool and thus the distribution of adaptations (the calling song of cicadas, for example).

<p>This thesis relates modern speciation theory to the effects of sudden changes in the range and size of populations. Special reference is made to the climatic oscillations during the Quaternary ice ages. A meta-analysis of laboratory experiments showed that support for allopatric speciation is weak, especially for the peripatric type of allopatric speciation. Furthermore, factors traditionally believed to increase the likelihood of speciation have had little effect on the generation of reproductive isolation in speciation experiments. However, the method of testing reproductive isolation appeared important, in the sense that experimentally derived sister populations were likely to demonstrate reproductive isolation from each other but not from the unaffected mother population. Raw data from mating tests showed that the poor isolation between mother and daughter populations was an effect of asymmetric mate preferences towards males from the mother population. This suggests that peripatric speciation can be effective in generating reproductive isolation between sister populations. The proposed mechanism is that males become less attractive to females by losing certain secondary sexual traits during population bottlenecks, and that females shift their preferences towards other male traits. Support for this mode of speciation is found in the widespread bird genus <i>Motacilla</i> (wagtails). This genus is characterised by extensive plumage variation and contains a large number of widely distributed taxa in the northern parts of its distribution. This thesis shows that taxonomic diversity of wagtails is inversely related to complexity in song and to diversity in molecular and mitochondrial markers. The northern taxa seem to be descendants of southern populations, which were subjected to bottlenecks during expansions into re-opened habitats after the last ice age. The bottlenecks would not only reduce genetic diversity but also inhibit cultural transmission of song to the leading edge of colonisers, allowing sexual selection on other traits, such as plumage. Rapid plumage differentiation among wagtail taxa appears to be a recurrent process and has lead to convergent evolution, making the currently recognised species <i>Motacilla flava</i> (Yellow Wagtail) polyphyletic.</p>

<p>Genetic structuring is common in natural populations. It is important to identify and consider population structure when studying evolutionary processes. Recently, the discovery of genetic structuring in some lekking bird species has opened up new perspectives on our understanding of the evolution of lek mating systems.</p><p>This thesis uses molecular data to identify patterns of broad and fine scale genetic structuring in the lekking white-bearded manakin Manacus manacus. Additionally, data on male mating success, female visiting patterns and behavioural, morphological and territorial characteristics of individual males are used to identify variables that may influence the distribution of matings in this species.</p><p>Analysis of genetic divergence within the genus Manacus revealed genetic sub-structuring and limited gene flow between species/subspecies. There was no significant isolation by distance relationship. Factors such as physical barriers to gene flow may play a role in shaping the genetic structure of the bearded manakin genus.</p><p>White-bearded manakin leks on Trinidad were composed of groups of related males. More than one such kin group existed on each lek. That related males gather in groups suggests that genetic structuring is not simply a consequence of limited dispersal. Active choices must take place by both residential and newly arrived birds.</p><p>Female visits to, and matings with, males were non-random. Centrality of male display court was the only measured variable that consistently correlated with male mating success. More aggressive displays were made as the distance between courts decreased. There was no significant relationship between the number of aggressive displays made between males and relatedness levels. Males with high mating success spent more time in aggressive behaviours.</p><p>In conclusion, white-bearded manakin lek formation and display court acquisition is likely to be influenced by genetic relatedness levels and male-male interactions. Centrality of court seemed important in mating success and may be an indicator of male dominance. However, a variety of other factors may also influence mating success and may be variable over time.</p>