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Most apomictic plants are facultative, maintaining the ability to reproduce sexually at different frequencies depending on the taxa, ploidy, and reproductive stage. In this context, Paspalum species are good model systems for studies evaluating the varying levels of apomixis expression. We aimed to identify, in apomictic tetraploid Paspalum species, the degree of apomixis and residual sexuality in three stages of reproductive development, and if their expression varies along them in order to predict their realized impact on the genetic diversity of future generations. Three main stages in the reproductive development (i.e., ovule, seed, and progeny) were studied in tetraploids from populations of P. cromyorhizon and P. maculosum. Mature ovules were studied using cytoembryological analysis, seeds by flow cytometry, and progeny tests with molecular markers. The expression of sexuality and apomixis was compared in each stage. We observed a decline in expression of sexual reproduction through the consecutive stages, jointly with an increase of apomixis expression. Both species showed at least one tetraploid plant capable of producing progeny by sexual means. These small rates of sexually originated progeny prove the ability of apomictic plants to produce low levels of genetic variation through rare events of sexuality. This study also demonstrates the importance of analyzing different reproductive stages in order to get a whole picture of the reproductive outcomes in plant evolution.

Introduction and objectives: The members of the genus Pilosella are native in Europe and Asia, but they are successful invasive species on most continents. These species form an agamic complex with common apomixis. Apomictic species hybridize, they have different degree of residual sexuality. Main aim of this paper was to determine if the interspecific hybridization already occurred in Patagonia. M&M: This study is based on analysis of seed progeny collected at thirteen populations of Pilosella in southern Argentina and Chile. The plants were examined for their taxonomic identity, DNA ploidy level (using flow cytometry), chromosome number, reproduction, formation of parthenogenetic seeds and clonal identity (using isozyme phenotypes). Results: No mixed-species population was recorded. Two apomictic clones of P. officinarum (one pentaploid and the other hexaploid) were found in populations: eight were hexaploid and one was mixed in cytotype composition. A new species for Patagonia, the apomictic pentaploid P. caespitosa, was represented by plants from two populations in Argentina. Some of the progeny plants cultivated from seeds sampled at three localities represented seed-fertile aneuploids the morphology of which implied a hybrid origin and indicated P. officinarum as one of the parents. Conclusions: The presence of seed-fertile, aneuploid and parthenogenetic hybrids among the cultivated plants signifies an increased risk of the formation of new hybridogeneous genotypes of Pilosella in southern Patagonia.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior FAPEMIG - Fundação de Amparo a Pesquisa do Estado de Minas Gerais Apomixia, reprodução assexuada a partir de sementes, é considerada como uma alternativa reprodutiva para muitas angiospermas. Em Melastomataceae, este modo de reprodução tem sido esclarecido em Miconieae, embora seja reportado em outras tribos, como Microlicieae. Assim, este estudo utilizou três espécies de Microlicia D. Don como modelo, Microlicia fasciculata Mart. ex Naudin, Microlicia polystemma Naudin e Microlicia sp. nov. a fim de (i) verificar a ocorrência de apomixia nas espécies estudadas; (ii) verificar se este modo de reprodução leva a formação de sementes poliembriônicas; (iii) investigar se as espécies apomíticas apresentam alterações durante o desenvolvimento dos grãos de pólen, com consequente esterilidade polínica, e (iv) averiguar se a apomixia pode ocorrer em paralelo ao processo sexuado no grupo. Para tal, realizamos um teste para detecção de apomixia autônoma, outro para verificar a presença de poliembrionia, análise de inviabilidade polínica, análise de crescimento de tubos polínicos e estudo da esporogênese e gametogênese por meio de técnicas utilizadas em estudos anatômicos. As três espécies são apomíticas autônomas e apresentam sementes poliembriônicas. A inviabilidade polínica foi alta em M. fasciculata e M. polystemma e total em M. sp. nov, decorrente de anormalidades meióticas, mitose simétrica e atraso no desenvolvimento dos grãos de pólen. A pequena formação de grãos de pólen viáveis e possível ocorrência de polinização natural em M. fasciculata e M. polystemma permite que haja eventos de reprodução sexuada, caracterizando-as como apomíticas facultativas. Em M. sp. nov., a completa esterilidade polínica e ausência de indícios de polinização natural indicam apomixia obrigatória. Apesar de apomíticas, todas as espécies retêm etapas de desenvolvimento que levou a formação de sacos embrionários reduzidos. A independência de polinizadores para formação de frutos, reprodução uniparental e sementes poliembriônicas com embriões de diferentes origens, pode conferir a tais táxons flexibilidade reprodutiva e diversidade genética. Apomixis, asexual reproduction through seeds, is considered as a reproductive alternative for many angiosperms. In Melastomataceae, this mode of reproduction has been elucidated in Miconieae, although it is reported in other tribes, such as Microlicieae. Thus, this study used three species of Microlicia D. Don as model, Microlicia fasciculata Mart. ex Naudin, Microlicia polystemma Naudin and Microlicia sp. nov. in order to (i) verify the occurrence of apomixis in the studied species; (ii) verify if this mode of reproduction leads to the formation of polyembryonic seeds; (iii) investigate whether the apomic species present alterations during pollen grain development, with consequent pollen sterility, and (iv) investigate whether apomixis can occur in parallel to sexual process in the studied species. For this, we performed a test to detect autonomic apomixis, other to verify the presence of polyembryony, pollen viability analysis, pollen tubes growth analysis and the study of sporogenesis and gametogenesis by techniques used in anatomical studies. The three species were autonomous apomictic and presented polyembryonic seeds. The percentage of inviable pollen grains was high in M. fasciculata and M. polystemma, and total in M. sp. nov., due to meiotic abnormalities, symmetric mitosis and to the delay in the pollen development. The small formation of viable pollen grains and possible occurrence of natural pollination in M. fasciculata and M. polystemma allows sexual reproduction events, characterizing them as facultative apomictics. In M. sp. nov., the complete pollen sterility and the absence of evidence of natural pollination indicate obligatory apomixis. Although apomictic, all species retain stages of development that led to the formation of reduced embryo sacs. The independence of pollinators for fruit set, unipariental reproduction and polyembryonic seeds with embryos of different origins, can confer to these taxa reproductive flexibility and genetic diversity. Dissertação (Mestrado)

Buffel grass (Cenchrus ciliaris L. syn. Pennisetum ciliare (L.) Link) is a species that is highly tolerant to drought and is used primarily as forage in drier regions throughout the subtropics and tropics. It reproduces mainly by apomixis and the acquisition of obligate sexual genotypes or facultative apomicts with high levels of sexuality is required for performing crosses and plant improvement. The aim of this study was to obtain sexual genotypes from controlled crosses using obligate apomictic cultivars and a sexual line. Twelve putative hybrid F1 plants were selected morphologically and two of them were identified as sexual genotypes by PCR using specific primers for reproductive mechanism. Cytoembryological analysis showed 65.5 and 71.3% meiotic embryo sacs in these plants and their hybrid nature was corroborated by AFLP. Both highly sexual genotypes could be used as female parents in crosses for obtaining improved cultivars of buffel grass. Fil: Quiroga, M.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina Fil: Grunberg, Karina Alejandra. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ribotta, A.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina Fil: Lopez Colomba, Eliana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Carloni, E.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina Fil: Tomassino, E.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina Fil: Luna, Celina Mercedes. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Griffa, S.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Fisiología y Recursos Geneticos Vegetales; Argentina

Traditionally, tetraploid Paspalum notatum was considered an obligate or a facultative apomict according to cytoembryological analyses. The degree of facultativeness was usually determined by the relative amount of mature ovules bearing aposporous or meiotic (sexual) embryo sacs, or both together. We established, through progeny tests conducted with the aid of AFLP markers, the degree of residual sexuality expressed in four selected biotypes. The results showed it to be substantially and significantly lower than predicted by previous embryological analyses for the same biotypes. Moreover, the lowest expression of residual sexuality was coincident with maximum flowering period. Seed development in facultative apomictic P. notatum shows a definite bias against meiotic embryo sacs Fil: Rebozzio, Romina Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina Fil: Sartor, Maria Esperanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina Fil: Quarin, Camilo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina Fil: Espinoza, Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina

In flowering plants, apomixis is asexual reproduction by seeds. Apomixis allows the production of offspring with the same genetic characteristics as the mother plant. Fertilization is not required. Apomixis could become a tool for naturally cloning high-yielding crop hybrids through their own seed. However, apomixis does not occur in major crop plants, except for citrus. In the present study, genes that might cause apomixis in naturally occurring apomictic plants were investigated. Sexual and apomictic species of the genus Boechera were exposed to stressed and non-stressed conditions. Effects of these treatments on the expression of apomixis was then measured. Stress triggered an increase in the frequency of sexual development in apomictic plants, but continuation of sexual development to form sexual seeds did not occur. Stress also triggered alterations in the expression of stress-related genes.

Cenchrus ciliaris L. is a C4 perennial grass of arid lands which is under the focus of different ecological issues such as response to desertification, quality of forage grass and impacts of invasions. Here, molecular and morphological analyses of the genetic diversity of several Tunisian provenances of C. ciliaris were performed to better understand the phenotypic polymorphism of this agamospermous and polyploid grass. Ten phenotypic traits associated with productivity were measured in a common garden environment. Amplified Fragment Length Polymorphism (AFLP) markers were developed to investigate the structure of genetic diversity among and within provenances and between the three ploidy levels. Heritable phenotypic traits showed considerable differences within provenances. Surprisingly, AFLP markers revealed the existence of genotypic variations between individuals of the same sibship and a high G/N value (0.55). A neighbour-joining tree based on AFLP markers revealed three major groups; tetraploid, pentaploid and a mix of pentaploid and hexaploids. These groups do not correspond completely to the geographical origin of samples. The results underline the possibility of sexual reproduction, recombination and gene flow within and between populations of C. ciliaris. In respect with the well known dynamic nature of polyploid genomes, these results should have strong consequences for the future management of this grass for both conservation and invasion issues.

Some citrus varieties express a form of apomixis termed nucellar embryony in which the adventive embryos develop from nucellus tissue surrounding the embryo sac. This trait results in many seeds containing multiple embryos (polyembryony). Inheritance of the frequency of polyembryony was studied in 88 progeny from a cross of Citrus maxima (monoembryonic) × Poncirus trifoliata (polyembryonic). The frequency of polyembryonic seed produced by each progeny was determined by scoring 100–500 seeds for the number of seedlings to emerge from each seed. Two groups of eight individuals from each extreme of the population were chosen for bulked segregant analysis with amplified fragment length polymorphism markers amplified with 256 primer combinations. Candidate markers identified in the bulks as linked to the trait were tested on the 32 individuals used to create the bulks and then on the remaining plants in the population. Five candidate markers tightly linked to polyembryony in P. trifoliata were identified. Specific marker alleles were present in nearly all progeny that produced polyembryonic seed, and alternate alleles were present in nearly all progeny that produced only monoembryonic seed. The region defined by these markers very likely contains a gene that is essential for the production of polyembryonic seeds by apomixis, but also shows segregation distortion. The proportion of polyembryonic seeds varied widely among the hybrid progeny, probably due to other genes. Scoring 119 progeny of a P. trifoliata selfed population for the closely linked markers and the proportion of polyembryonic seeds confirmed close linkage between these markers and polyembryony.