No description available.

No description available.

<p> The ɣ-radiosensitivity of haploid and diploid Oedogonium cardiacum cells was measured and compared to other cell lines. With the doubling of the chromosome complement, the Do value doubled, but the extrapolation number decreased four-fold. A general conclusion was drawn from the results that at all doses of ɣ-radiation, the diploids were more resistant than the haploids. A new radiation technique was used and compared to that used routinely in the laboratory. The further use of the technique was not recommended since the data obtained with the diploid line only was not as reliable as one would like.</p> <p> The degree of synchrony of Oe. cardiacum zoospore cultures was measured using cell division as the biological end-point and a mathematical expression, the percent phasing, as the index of synchrony. It was intended that this research problem be secondary to the radiation studies. The percent phasing values were determined for cells growing in two inorganic media and in the presence of an inhibitor, hydroxyurea. However the degree of synchrony was not improved beyond that of the routine laboratory procedure. Attempts to improve the size of the synchronous populations collected also proved unsuccessful.</p> / Thesis / Master of Science (MSc)

This thesis aimed to study three main aspects: (i) the mechanisms underlying unreduced pollen gamete formation in the diploid `CSO¿ tangor hybrid used as male parent in 4x x 2x triploid breeding programs, (ii) the frequencies and mechanisms involved in the unreduced 2n female gametes production for `Eureka Frost¿ and `Fino¿ lemon genotypes, and (iii) the interspecific recombination and the resulting diploid gamete structures of doubled-diploid `Mexican¿ lime to evaluate the possibility that natural interploid hybridization maybe the origin of C. latifolia (`Tahiti¿ lime type) and C. aurantifolia (`Tanepao¿ lime type) triploid varieties. The production of 54 tetraploid hybrids from 4x x 2x sexual hybridizations allowed the analysis of the mechanisms underlying unreduced pollen gamete formation. SSR and SNP molecular markers revealed that the majority of these plants were obtained from 2n pollen of the diploid tangor parent. Then, the maximum-likelihood method based on parental heterozygosity restitution (PHR) of centromeric loci revealed that both FDR (First Division Restitution ) with predominant occurrence and to a lower extend SDR (Second Division Restitution) were the mechanisms leading to unreduced male gamete formation in the tangor studied. These observations were confirmed by the analysis of PHR pattern along the linkage group (LG) 2. To our knowledge, this is the first report of tetraploid citrus progenies arising from unreduced pollen and the first description of the coexistence of two meiotic restitution mechanisms (SDR and FDR) producing unreduced pollen in citrus. In order to study the frequencies and the mechanisms involved in the unreduced 2n female gametes production in two different genotypes of lemon, we produced 43 triploid and tetraploid hybrids from 2x x 2x and 2x x 4x sexual hybridizations using `Eureka Frost¿ and `Fino¿ as female parents. The frequencies of 2n gamete production were respectively 4.9% and 8.3%. The maximum-likelihood analysis and pattern of PHR along LG1 revealed that SDR is the main mechanism of unreduced female lemon gametes (88%), followed by FDR or pre-meiotic doubling (PRD) (7%) and post-meiotic genome doubling (PMD) mechanisms (5%). This is the first report of the production of a large number of lemon progenies from 2n gametes and the first identification of a new mechanism, PMD that has never been observed in citrus and has rarely been described in other herbaceous or woody species. Across both studies, we demonstrated at the methodological level the effectiveness of using two complementary approaches, the analysis of the PHR pattern in one LG and the maximum-likelihood method based on centromeric loci for distinguishing between the different mechanisms of unreduced gamete production. We analyzed the meiotic mechanisms of a doubled diploid `Mexican¿ lime, the interspecific recombination and the resulting diploid gamete structures combining a segregation analysis of SSR and SNPs markers, a cytogenetic study and pollen viability evaluation. We concluded that the doubled-diploid `Mexican¿ lime had a predominantly disomic segregation for three LGs, intermediate inheritance with disomic tendency was found for five LGs and intermediate models for one LG. The resulting interspecific diploid gamete structures displayed high C. medica / C. micrantha heterozygosity. The revealed genetic structures of the diploid gametes produced by the doubled-diploid `Mexican¿ lime are compatible with the hypothesis that `Tahiti¿ and `Tanepao¿ triploid varieties results from interploid hybridization involving a doubled-diploid `Mexican¿ like lime. This disomic tendency limits the recombination and the diversity of the diploid gamete population; however the observed pollen viability restoration at tetraploid level could be advantageous for intensive breeding projects. / Esta tesis estudia tres aspectos principales: (i) los mecanismos de la formación de gametos 2n de polen originados por un híbrido diploide (tangor `CSO¿) utilizado como parental masculino en hibridaciones sexuales 4x x 2x, (ii) las frecuencias y los mecanismos de la producción de gametos 2n femeninos en dos genotipos de limón, `Eureka Frost¿ y `Fino¿, y (iii) el análisis de la recombinación interespecífica y las estructuras de los gametos diploides originados por la lima `Mejicana¿ doble diploide para evaluar el origen de las las variedades triploides de lima C. latifolia (`Tahiti¿) y C. aurantifolia (`Tanepao¿). La obtención de 54 híbridos tetraploides a partir de hibridaciones 4x x 2x permitió analizar los mecanismos de la formación de gametos 2n de polen. El análisis molecular reveló que la mayoría de estas plantas se obtuvieron a partir de gametos 2n de polen del parental `CSO¿. A continuación, el análisis mediante el método de máxima verosimilitud indicó que FDR (Restitución de la Primera División meiótica) y SDR (Restitución de la Segunda División meiótica) son los mecanismos implicados con una mayor dominancia de FDR respecto SDR. Estos resultados se confirmaron mediante el análisis de la restitución de la heterocigosidad en el grupo de ligamiento (LG) 2. Con los datos publicados hasta la fecha, es la primera vez que se han obtenido progenies tetraploides de cítricos mediante gametos 2n de polen y es la primera descripción en cítricos de la coexistencia de dos mecanismos SDR y FDR. Para estudiar las frecuencias y los mecanismos de la producción de gametos 2n en dos genotipos de limón, se obtuvieron 43 híbridos triploides y tetraploides a partir de hibridaciones sexuales 2x x 2x y 2x x 4x utilizando los limones `Eureka Frost¿ y `Fino¿ como parentales femeninos. Las frecuencias de producción de gametos 2n fueron respectivamente 4,9% y 8,3%. El análisis de máxima verosimilitud y el patrón de PHR a lo largo del LG1 reveló que SDR es el mecanismo principal de formación de gametos 2n femeninos (88%), seguido por FDR o duplicación del genoma pre-meiosis (PRD) (7%) y se identificó un nuevo mecanismo originado a partir de la duplicación del genoma post-meiosis (PMD) (5%). En este trabajo se describe por primera vez en cítricos la producción de un elevado número de híbridos de limón a partir de gametos 2n y es la primera vez que se identifica un nuevo mecanismo PMD que nunca se ha observado en cítricos. En ambos estudios se demostró la efectividad del uso de dos métodos complementarios, el análisis del patrón de PHR a lo largo de un LG y el método de máxima verosimilitud para distinguir entre los diferentes mecanismos implicados en la formación de gametos 2n. También se ha analizado el modelo de segregación cromosómica de la lima `Mejicana¿ doble diploide (DD) así como la recombinación interespecífica y las estructuras de los gametos diploides resultantes. Se ha realizado análisis de la viabilidad del polen junto con un análisis citogenético y con marcadores SSRs y SNPs. Estos trabajos nos han permitido concluir que la lima `Mejicana¿ DD presenta una segregación predominantemente disómica para tres LGs, herencia intermedia con tendencia disómica para cinco LGs y un tipo de segregación intermedia para un LG. Las estructuras de los gametos diploides mostraron una alta heterocigosis en C. medica/C. micrantha, parentales de la lima `Mejicana¿. Las estructuras genéticas de los gametos diploides de la lima `Mejicana¿ DD son compatibles con la hipótesis de que las variedades triploides `Tahiti¿ y `Tanepao¿ se obtuvieran a partir de una hibridación interploide a partir de la lima `Mejicana¿ DD. El tipo de segregación disómico conlleva una limitación de la recombinación y la diversidad genética de la población de gametos 2n. Sin embargo la viabilidad del polen de la lima `Mejicana¿ DD en comparación con la lima `Mejicana¿ diploide permite / Aquesta tesi ha tingut com a objectius l'estudi de (i) els mecanismes subjacents a la formació de pol·len de gàmetes 2n al tangor híbrid diploide `CSO', utilitzat com a progenitor masculí en els programes de millora de triploides 4x x 2x (ii) les freqüències i els mecanismes implicats en la producció gàmetes 2n en la femella als genotips de llimona `Eureka Frost¿ i Fina' (iii) la recombinació interespecífica i les estructures resultants de gàmetes diploides del doble-diploide (DD) de `llima Mèxicana' per avaluar la possibilitat que la hibridació interploid natural potser l'origen de les varietats triploides C. latifolia (llima tipus `Tahiti') i C. aurantifolia (llima tipus `Tanepao'). La producció de 54 híbrids tetraploides obtinguts d¿hibridacions sexuals 4x x 2x va permetre l'anàlisi dels mecanismes de formació de gàmetes 2n de pol·len. Els marcadors moleculars SSR i SNP van revelar que la majoria d'aquestes plantes es van obtenir de pol·len 2n del parental diploide tangor. Llavors, el mètode de màxima probabilitat va revelar que tant FDR (Restitució en la Primera Divisió), amb ocurrència predominant, com SDR (Restitució en la Segona Divisió) van ser els mecanismes que condueixen a la formació de gàmetes masculins 2n en aquest tangor. Aquestes observacions van ser confirmades per l'anàlisi de patró de RHP al llarg del cromosoma 2. Des del nostre coneixement, aquest és el primer estudi de progènies de cítrics tetraploides derivats de pol·len no reduït i la primera descripció de la coexistència de dos mecanismes de restitució meiòtiques (FDR i SDR) produint pol·len no reduït en els cítrics. Per tal d'estudiar les freqüències i els mecanismes implicats en la producció de gàmetes 2n sense reduir de la femella, en dos genotips diferents de llimona, vam obtenir 43 híbrids triploides i tetraploides d¿hibridacions sexuals 2x x 2x i 4x x 2x utilitzant `Eureka Frost ' i `Fino' com progenitors femenins. Les freqüències de la producció de gàmetes 2n van ser, respectivament, 4,9% i 8,3%. L'anàlisi de màxima probabilitat i el patró de RHP al llarg del cromosoma 1 van revelar que SDR és el principal mecanisme de gàmetes 2n en la llimona utilitzada com parent femení (88%), seguit pels mecanismes FDR o duplicació pre-meiòtica (PRD) (7%) i la duplicació del genoma post-meiòtica (PMD) (5%). Per primera volta en els cítrics s¿ha obtingut un gran nombre de progènie de llimona a partir de gàmetes 2n i s¿ha identificat un nou mecanisme, el PMD que poques vegades s'ha descrit en altres espècies herbàcies o llenyoses. A través dels dos treballs, hem demostrat, a nivell metodològic, l'eficàcia d'utilitzar dos enfocaments complementaris, és a dir, l'anàlisi del patró de RHP en un cromosoma amb el mètode de màxima probabilitat basat en loci centromèrics per distingir entre els diferents mecanismes de la producció de gàmetes 2n . Es van analitzar els mecanismes meiòtics d'un DD `llima Mèxicana', la recombinació interespecífica i les estructures resultants de gàmetes diploides combinant una anàlisi de segregació de marcadors SSR i SNP, un estudi citogenètic i l'avaluació de la viabilitat del pol·len. Hem arribat a la conclusió que el DD de `llima Mèxicana' tenia una segregació predominantment disómica en tres cromosomes, herència intermèdia amb tendència disómica en cinc cromosomes i els models intermedis per a un altre. Les estructures resultants de gàmetes diploides interespecífiques mostren alta heterozigositat C. medica / C. micrantha. Les estructures genètiques revelades dels gàmetes diploides produïts pel DD de `llima Mèxicana' són compatibles amb la hipòtesi que les varietats triploides `Tahiti¿ i `Tanepao¿ resulten de la hibridació interploide que impliquen un DD de tipus `llima Mèxicana'. Aquesta tendència disómica limita la recombinació i la diversitat de la població de gàmetes diploides, però, la restauració de la viabilitat / Houssem, R. (2017). Genetic Structure of Diploid Gametes for the Production of Triploid Citrus Hybrids [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/90634 / TESIS

Triploid (sterile) trout potentially offer a more risk-averse option for stocking popular non-native sport fish; however the relative performance (e.g., survival and growth) of triploid versus diploid fish in natural settings is not well understood. I evaluated the relative performance of triploid versus diploid brook trout (Salvelinus fontinalis) stocked in high mountain Uinta lakes in response to food availability and lake morphology. I chose a set of 9 lakes that included a range of elevation and lake morphology. I observed no difference in CPUE or relative weight (Wr) of both types of trout in all lakes. Food availability (e.g., zooplankton and macroinvertebrates) varied substantially among lakes; however I observed no discernible difference between diploid and triploid diets, diet preference, or isotopic trophic signatures. Physical lake characteristics (e.g., dissolved oxygen [DO, mg/L] and temperature [°C]) were within or near optimal brook trout conditions (metabolically beneficial range) during the summer, but over-winter DO reached near lethal levels under the ice. In sum, between the two strains, I did not observe any significant differences in relative performance measured across a number of indices; however the size distribution of diploid fish was marginally (20 mm) skewed towards larger fish (a difference likely not great enough to be detectable by the average angler). In contrast to the similarity in performance between strains, I did observe considerable variability in performance of brook trout across lakes as a function of lake productivity, food availability, and most importantly fish density. Fish performance was greater in lakes with a lower density of stocked fish. Overall, the results from this study indicate triploid brook trout offer a viable and risk-averse alternative to stocking diploid fish in Uinta mountain lakes. Stocking triploid fish should decrease the threat of uncontrolled expansion into adjacent water bodies, while still allowing managers to maintain a popular non-native sport fishery.

An exceptionally effective transformation procedure has been established by using class I embryo-derived rice callus. Every treated callus clump yielded multiple independently transformed plants (average 40 plantlets). Analysis of genomic DNA blots and pollen expressing green fluorescent protein (GFP) from T0 plants revealed that 64% bore a single locus T-DNA insertion in which half had one T-DNA copy. Additive transgene expression was observed fromT0 plants with GFP driven by mUbi1 promoter. Transgenic plants could be rapidly characterized by analyzing GFP pollen from T0 plants without the need for further generations or genomic DNA blot analysis. Agrobacterium tumefaciens-mediated transformation of microspore-derived callus for generating large numbers of T-DNA haploid and doubled haploid(DH) plants has also been investigated. The established transformation procedure resulted in 100% transformation frequency for class I microspore-derived rice callus. Each callus typically yields multiple independent transgenic plants. Genomic DNA blot analysis suggested 98% of the transgenic plants are independent events. About half of the transgenic plants were identified as haploid plants, whereas half are DH hemizygous or homozygous transgenic plants. DH homozygous transgenic plants were obtained from T0plants and confirmed by pollen GFP expression and genomic blot analysis in T0transgenic DH plants. In this study, about 60% ofT0transgenic DH plants had a single locus T-DNA insertion of which 45% bore one T-DNA copy. Furthermore, in a population of over 2,000 haploid and doubled haploid T-DNA plants , about 25% showed phenotypic differences from non-transformed haploid plants. Approximately 5% were seriously phenotypically abnormal including lethal or semi-lethal mutants. This highly efficient transformation procedure using microspore-derived callus could be valuable in speeding up plant breeding and in new gene discovery. Diversification of the mUbi1 promoter led to a minimal promoter that has a similar function as the original mUbi1. Transient and stable transformation as measured from gene expression driven by the minimal promoter suggested that it has a similar function as the original wild type promoter.

Annual ryegrass (Lolium multiflorum Lam.) can be divided into two cytotypes: diploid (2n = 2x = 14) or tetraploid (2n = 4x = 28). Transforming annual ryegrass from diploid to tetraploid often results in increased seedling vigor, more robust growth and, thus, supposedly greater yield. There has been conflicting research that suggests no difference in yield between diploid and tetraploid cultivars, but livestock producers’ question whether benefits exist to utilizing tetraploid cultivars for grazing cattle. This study sought to determine if annual ryegrass cytotype affected cattle average daily gain. Although there were no differences in average daily gain between cytotypes, there were differences among cultivars. In 2017, Jumbo (4x) produced the greatest weight gain, while in 2018, Tam-90 (2x) generated the greatest gain. This study suggests that there are no advantages attributed to average daily gain by utilizing tetraploid, as opposed to diploid cultivars.

No description available.

Bean pod mottle virus (BPMV), a member of the genus Comovirus in the family Comoviridae, is widespread in the major soybean-growing areas in the United States. Soybean yield losses of 10-40% have been reported as a consequence of BPMV infection. The complete nucleotide sequences of two strains, K-Ha1 and K-Ho1, were determined. Field isolates of BPMV were classified into two distinct subgroups (I and II) based on slot blot hybridization and sequence analyses. Full-length cDNA clones from which infectious transcripts can be produced were constructed for strains K-G7, K-Ho1 and K-Ha1. Whereas strains K-Ha1 and K-G7 induced mild or moderate symptoms in infected soybean plants, strain K-Ho1 produced very severe symptoms. Symptom severity was mapped to RNA1. Chimeric RNA1 constructs were generated by exchanging full or partial coding regions of the five RNA1-encoded mature proteins between the full-length cDNA clones of the three RNA1s and the resultant transcripts were inoculated onto soybean. The results showed that the coding regions of the protease co-factor (Co-pro) and the putative helicase (Hel) are determinants of symptom severity. Although symptom severity correlated well with accumulation of viral RNA, neither the Co-pro nor Hel protein could be demonstrated as a suppressor of RNA silencing. Furthermore, separate expression of the Co-pro or Hel proteins from a PVX vector induced necrosis on the inoculated leaves of Nicotiana benthamiana. Characterization of BPMV K-Ho1 indicated that it is a diploid reassortant, containing two distinct types of RNA1s and one type of RNA2. Examination of field isolates from various locations in the United States and Canada revealed that diploid reassortants are of frequent occurrence in natural populations of BPMV. The vary severe symptoms induced by BPMV K-Ho1 can be mimicked by inoculation of plants with a mixture of RNA1 transcripts from two distinct strain subgroups and RNA2 transcript from either subgroup. Plants inoculated with a mixture of transcripts containing two types of RNA1 from the same strain subgroup did not produce very severe symptoms. These are due to interactions between two distinct types of RNA1s. At present, no soybean cultivars with resistance to BPMV are commercially available. Therefore, the feasibility of cross protection as an alternative disease management strategy was studied. Two mild strains of BPMV (K-Da1 and K-Ha1), belonging to subgroup II, were tested for their ability to protect infected plants against a severe strain (K-Ho1). Inoculation of the soybean cultivar Essex on the primary leaves with either of the two mild strains conferred complete protection against challenge inoculation with the severe strain K-Ho1, regardless of the timing of challenge inoculation. Cross-protection was evident regardless of whether virions or BPMV-RNA were used as inocula. Cross protection was independent of the soybean cultivar used and method of virus inoculation, sap-inoculation or by the bean leaf beetle, vector of BPMV. Protection was complete and durable.

As abelhas sem ferrão exercem importante papel ecológico como polinizadores de muitas espécies vegetais das regiões tropicais e tem significativo potencial para uso na polinização de culturas agrícolas. Contudo, com a contínua degradação de habitats, as populações de inúmeras espécies tem se tornado cada vez menores e separadas umas das outras por grandes distâncias. A criação de espécies de abelhas é um componente essencial para a conservação da biodiversidade, além de uma alternativa de fonte de renda. Para tanto, esforços de conservação e programas de criação em escala comercial requerem uma combinação de fatores, como o conhecimento biológico mais amplo, principalmente os relacionados à produção de sexuados e à diversidade genética necessária para manter a viabilidade de pequenas populações destas abelhas. Nesse contexto, os principais objetivos desta tese foram avaliar a variabilidade genética em populações manejadas, sob condições de isolamento genético ou não, e a produção de machos e rainhas nessas populações e o papel na reprodução. Para isso estudamos duas populações de Melipona scutellaris mantidas em regiões geográficas distintas, uma no município de Igarassu (PE; 7°50S 34°54W), onde a espécie ocorre naturalmente e outra no município de São Simão (SP; 21°26 47°34W), onde a população foi iniciada com duas colônias e criada por mais de 10 anos, quando chegou, a partir de sucessivas multiplicações, a 55 ninhos. Assim, embora a população de S. Simão tivesse maior redução na diversidade alélica e maior frequência de machos diplóides, quando comparada à mantida em Igarassu, ela foi criada com sucesso por um extenso período (ca. 10 anos). Provavelmente o baixo número de alelos sexuais em S. Simão, e a conseqüente produção de machos diplóides, foi a principal explicação para a freqüência significativamente maior de sexuados criados nessa população. Como contraponto à alta produção de machos diplóides, as substituições das rainhas-mãe foram mais frequentes e as colônias produziram mais rainhas. Além disso, a alta produção de machos e rainhas também pode ser entendida em termos de benefícios reprodutivos individuais. Tanto as rainhas fisogástricas como as operárias poedeiras foram responsáveis pela maternidade de machos haplóides. Contudo, 80% dos machos filhos de operárias foram produzidos por operárias filhas da rainha-mãe substituída, indicando que essas operárias especiais tem sobrevida maior que as demais e parasitam reprodutivamente a força de trabalho da geração seguinte, que são menos relacionadas a elas. Quanto à super-produção de rainhas, detectou-se que 25% das colônias órfãs, após a perda da rainha-mãe, eram invadidas por rainhas que foram produzidas e vieram de colônias próximas. Nessas colônias não-natais, elas iniciaram suas atividades de postura. Este importante resultado muda as bases para melhoramento genético destas abelhas estabelecidas até o momento. Outro estudo relacionado à alta produção de rainhas foi realizado em colônia poligínica de M. quadrifasciata, em que oito rainhas fisogástricas co-existiam. Ao contrário da hipótese de que alguma das rainhas poderia ter vindo de outro ninho, todas eram irmãs completas. Isto sugere novas estratégias reprodutivas ainda desconhecidas para as abelhas do gênero Melipona. / Stingless bees play an important ecological role as pollinators of many wild plant species in the tropics and have significant potential for the pollination of agricultural crops. However, as a consequence of habit degradation, populations of a number of bee species have became increasingly small and separated from one another by large distances. Thus, stingless beekeeping is an essential component of biodiversity conservation, as well as a profitable business. Therefore, conservation efforts and breeding programme on a large scale require a combination of factors, including a broader biological knowledge, especially those related to production of sexuals and to the genetic diversity needed to maintain the viability of small population. In this context, the main goals were to evaluate the genetic variability in managed populations under or not genetic isolation and the production of males and queens. Two Melipona scutellaris populations were studied and they were kept in different geographic regions, one in Igarassu (PE; 7°50\'S 34°54\'W), in the species natural area of occurrence, and the other in São Simão (SP; 21°26 47°34W), where the population was started from only two foundress colonies and which after a breeding programme of ten years increased to about 55. Despite a great reduction in the number of alleles and an increased frequency of diploid males in the S. Simão population, it could be successfully bred and maintained for a prolonged period (ca. 10 years). Probably the low number of sex alleles in S. Simão population, leading to production of diploid males, was the main reason for the highest level of sexual production. To counter-balance the high production of diploid males, the replacement of mother queen was more frequent leading to higher levels of queen production by the colonies. Furthermore, the high production of males and queens can also be explained by the individual fitness benefits. Queen and reproductive workers were haploid males mothers. However, 80% of the workers sons had genotypes that were compatible with them being the sons of workers that were the offspring of a superseded queen, indicating that these workers greatly outlive all other workers and reproductively parasitize the next-generation workforce, that are less related individuals. Related to queen overproduction, 25% of all queenless colonies were invaded by unrelated queens that fly in from unrelated hives nearby. In these non-natal colonies, the alien queens started their egg laying activities. Another study related to the high queen production was conducted in a polygyne colony of M. quadrifasciata, where eight physogastric queens coexisted. Contrary to the hypothesis that some of these queens could be an alien queen, it was confirmed that they were full-sisters. This suggests new reproductive strategies that are unknown for Melipona bees.