Elements Of Apomixis Research Articles

METHODOLOGICAL IMPROVEMENT OF THE APPLICATION OF APOMIXIS IN TOBACCO BREEDING FOR CONFIRMATION OF HETEROSIS

The source material for the research was collection samples bred by breeders of the station and introduced from European countries, registered in the National Genetic Fund of Ukraine. Hybridization was performed according to the diallel scheme and F1 seeds from 36 hybrid combinations were obtained. The best hybrids with a high effect of heterosis are transferred to the apomictic basis in order to consolidate heterosis. The obtained apomicts were sown and 7 with the best indicators were selected according to the set of traits and checked for the presence of apomictic properties for sowing in the future to obtain hypomites-apomict with high productivity. The best hybrid combinations were selected by quantitative characteristics, which showed a positive dominance and are of practical interest for the consolidation of heterosis through apomixis and further testing in a competitive nursery. As a result of a detailed analysis of the specific splitting of forms in A1, it should be noted that each hybrid combination has its own specific splitting and there is no pattern of manifestation, as observed in the second hybrid generation. Criteria for selection of possible apomites according to the ability to form plants in A1 with maternal traits in about 25% of plants and selection of seeds for further sowing only after castration of flowers and removal to the isolator. In the second generation A2 there is a high level of constancy, which allows the selection of plants without castration, but the selection under the insulator. The main marker for the identification of genotypes with elements of apomixis are morphological features of the maternal type, which requires planting next to hybrid seeds of the first generation. In the breeding practice of tobacco to improve the source material and consolidate heterosis through apomixis should use an improved method of evaluation, which consists in collecting seeds for further study of A1 through castration of flowers and growing seeds under the insulator, selection of A1 plants by maternal type, which requires placement next to the plant F1 and formation of seed stock for re-seeding. Keywords: tobacco, apomixis, ecological stability and plasticity, heterosis, consolidation of heterosis effect

Is apomixis occurring in walnut (Juglans regia L.)? New data from progeny molecular tests and cytological investigations shed light on its reproductive system.

Persian walnut (Juglans regia) is an economically important nut fruit species cultivated worldwide for its nutritious kernel and timber quality wood. Walnut trees are mostly hetero-dichogamous and, depending on the genotype, some cultivars are protogynous, while others are protandrous. Although selfing is possible when male and female blooms overlap, the dichogamy of the species promotes outcrossing. In addition to sexual reproduction, some reports indicate that elements of apomixis may occur in commercial orchards of walnut varieties and in the last two decades, nut production by apomixis has been reported in walnut. However, there are no reliable studies on the occurrence of apomictic reproduction based on cytoembryological observations and/or molecular marker-progeny tests. This study addresses the combined use of molecular and cytological analyses to gain new insights into the population genetics and reproduction systems of J. regia. We systematically analyzed the reproductive origin of individual progeny plants from 8 different cultivated walnut genotypes using microsatellite genotyping and carried out cytohistological investigations of 5 cultivated walnut genotypes arising seed sets from isolated flowers, to shed light on the mode of reproduction. These cytometric and genotyping analyses did not support any asexual mode of reproduction or asexual propagation by seed and all individuals studied were identified as zygotic plants produced by crossing. Likewise, the cytological findings did not confirm completely the first component of apomixis, namely apomeiosis. On the other hand, according to histological evidence, adventitious embryony seems to take place at low frequency. Overall, our findings suggest that the occurrence of gametophytic apomixis is unlikely in J. regia, but sporophytic apomixis cannot be completely ruled out.

Evolution of an Apomixis-Specific Allele Class in Supernumerary Chromatin of Apomictic Boechera.

Asexual reproduction through seeds in plants (i.e., apomixis) is a heritable trait, and apomixis- linked loci have been identified in multiple species. However, direct identification of genomic elements is typically hindered as apomixis-linked loci and are commonly found in recombination-suppressed and repetitive regions. Heterochromatinized elements, such as B chromosomes and other supernumerary chromosomal DNA fragments have long been known to be associated with asexuality in both plants and animals and are prime candidate regions for the evolution of multiple apomixis factors controlling the individual elements of apomixis. Here, we examined molecular evolution, gene regulation, and chromosomal location of a male apomeiosis factor (UPG2), a long noncoding RNA gene, in sexual and apomictic Boechera with and without male apomeiosis (i.e., balanced and unbalanced apomicts). We revealed the origin of the gene in the apomixis genome on an apomixis-specific, supernumerary heterochromatic Boechera chromosome (Boe1). The UPG2 is active in the tapetum at male meiosis. We found allele classes specific to apomictic and sexual Boechera accessions and a third class that shares the features of both and points to a convergent transition state. Sex alleles are found only in some of the sexual accessions and have higher nucleotide divergence and lower transcriptional activity compared to apo alleles. These data demonstrate selective pressure to maintain the function of UPG2 for unreduced pollen formation in apomicts as the occasional transmission of the allele from unbalanced apomicts into sexual organisms that lead to pseudogenization and functional decay of copies in sexual organisms.

RH17 restricts reproductive fate and represses autonomous seed coat development in sexual Arabidopsis.

Plant sexual and asexual reproduction through seeds (apomixis) is tightly controlled by complex gene regulatory programs, which are not yet fully understood. Recent findings suggest that RNA helicases are required for plant germline development. This resembles their crucial roles in animals, where they are involved in controlling gene activity and the maintenance of genome integrity. Here, we identified previously unknown roles of Arabidopsis RH17 during reproductive development. Interestingly, RH17 is involved in repression of reproductive fate and of elements of seed development in the absence of fertilization. In lines carrying a mutant rh17 allele, development of supernumerary reproductive cell lineages in the female flower tissues (ovules) was observed, occasionally leading to formation of two embryos per seed. Furthermore, seed coat, and putatively also endosperm development, frequently initiated autonomously. Such induction of several features phenocopying distinct elements of apomixis by a single mutation is unusual and suggests that RH17 acts in regulatory control of plant reproductive development. Furthermore, an in-depth understanding of its action might be of use for agricultural applications.

The spread of infectious asexuality through haploid pollen.

The mechanisms of initiation and transmission of apomixis (asexual reproduction through seeds) in natural plant populations are important for understanding the evolution of reproductive variation. Here, we used the phylogenetic diversity of the genus Boechera (Brassicaceae), together with natural diversity in pollen types produced by apomictic lines, to test whether hybridization triggers the transition to asexuality, and whether a 'triploid bridge' is required for the formation of polyploid apomicts. We performed crosses between diploid sexual recipient and diploid apomictic donor lines and tested whether the mating system (interspecific hybridization vs intraspecific outcrossing) or pollen type (haploid vs diploid) influenced the transmission of apomixis from diploid apomictic donors into sexual recipients. We used genetic markers and flow cytometric analyses of embryo and endosperm in seeds to infer the reproductive mode. Within a single generation, initiation of both diploid and polyploid apomixis in sexual Boechera can occur. Diploid apomixis is transmitted through haploid pollen (infectious asexuality) and polyploids can form through multiple pathways. The three functional elements of apomixis occasionally segregate. Variation in pollen ploidy and the segregation of apomixis elements drive reproductive diversity of hybrids and outcrosses and can be utilized for apomixis initiation in crop breeding programs.

Viewpoints on apornictic and sexual reproduction in angiosperms

Gametophytic apomixis implies different changes of the reproductive cycle of sexual plants. Each of these "elements of apomixis" occurs in sexual plants, which may even display functional diploid parthenogenesis. Thus, apomixis, like vegetative reproduction, is part of the reproductive behaviour of sexual plants, becoming important when sexual reproduction is impaired. The elements of apomixis are probably to a large extent under polygenic control.

The intriguing complexity of parthenogenesis inheritance in Pilosella rubra (Asteraceae, Lactuceae)

Neither the genetic basis nor the inheritance of apomixis is fully understood in plants. The present study is focused on the inheritance of parthenogenesis, one of the basic elements of apomixis, in Pilosella (Asteraceae). A complex pattern of inheritance was recorded in the segregating F(1) progeny recovered from reciprocal crosses between the facultatively apomictic hexaploid P. rubra and the sexual tetraploid P. officinarum. Although both female and male reduced gametes of P. rubra transmitted parthenogenesis at the same rate in the reciprocal crosses, the resulting segregating F(1) progeny inherited parthenogenesis at different rates. The actual transmission rates of parthenogenesis were significantly correlated with the mode of origin of the respective F(1) progeny class. The inheritance of parthenogenesis was significantly reduced in F(1) n + n hybrid progeny from the cross where parthenogenesis was transmitted by female gametes. In F(1) n + 0 polyhaploid progeny from the same cross, however, the transmission rate of parthenogenesis was high; all fertile polyhaploids were parthenogenetic. It appeared that reduced female gametes transmitting parthenogenesis preferentially developed parthenogenetically and only rarely were fertilized in P. rubra. The fact that the determinant for parthenogenesis acts gametophytically in Pilosella and the precocious embryogenesis in parthenogenesis-transmitting megagametophytes was suggested as the most probable explanations for this observation. Furthermore, we observed the different expression of complete apomixis in the non-segregating F(1) 2n + n hybrids as compared to their apomictic maternal parent P. rubra. We suggest that this difference is a result of unspecified interactions between the parental genomes.

Expression of the apomictic potential and selection for apomixis in sorghum line AS-1a

Expression of elements of apomixis was studied for ten seasons in sorghum line AS-la and its backcross hybrids on the 9E and A3 sterile cytoplasms. Cytoembryological analysis revealed aposporous embryo sacks (apo-ESs), their initial cells, and, rare, parthenogeneic proembryos in ovules of line AS-la and its BC2 and BC3 hybrids on the 9E cytoplasm. The A3 sterile cytoplasm suppressed the development of parthenogenetic proembryos, but did not affect the apo-ES formation. The frequency of apomictic elements increased in seasons with high daily temperatures and total precipitation deficiency in the period when the ovule and megagametophyte developed (r = -0.805, P < 0.01). Selection was used to isolate the families where the frequency of ovules with apo-ESs was 28% and the frequency of parthenogenetic proembryos was 14%. Emasculated panicles of line AS-la were pollinated with pollen of line Volzhskoe-4v, which carried the Rs marker dominant gene, responsible for the anthocyan color of coleoptyles and leaves in seedlings. Plants of the maternal type were found in the progenies of these crosses at a frequency of 1.4-28.1%. The genetic structure of the endosperm in grains with maternal-type seedlings was inferred from the electrophoretic patterns of storage proteins (kafirins). The kafirin spectra of grains producing maternal-type seedlings was similar to the spectrum of line AS-la and differed from the spectra of grains producing hybrid seedlings, indicating that the endosperm developed independently when apomictic grains formed in line AS-1a. The results showed that lines with facultative apomixis can be constructed in functionally diploid plants.

Chromosome control of apomixis in maize-gamagrass hybrids

The results of long-term studies on the transmission of the mode of asexual reproduction through seeds to maize from gamagrass, a closely related wild plant, performed in the Laboratory of Plant Cytology and Apomixis are summarized. The first apomictic hybrids between Zea mays and Tripsacum dactyloides were obtained in this laboratory more than 40 years ago and have been maintained until the present time. Cytogenetic studies on the hybrids have shown that at least nine chromosomes of the wild parent are necessary for the expression of asexual reproduction through seeds. In addition, the genes controlling two elements of apomixis (apomeiosis and parthenogenesis) have been found to be inherited independently from each other.

Gametophytic apomixis: elements and genetic regulation

Gametophytic apomixis implies formation of unreduced embryo-sacs and capacity of their egg cells for parthenogenetic development. Each of these processes, as well as their union to give matromorphous offspring, are known from several species with sexual reproduction. A comparative survey is given of meiotic modifications, occurrence of apospory, female and male parthenogenesis and aberrant endosperm formation in sexual and apomictic species. Knowledge and models concerning the genetic basis of apomictic reproduction are summarized, as well as effects upon apomixis of changes of chromosome number and of hybridization. In the author's opinion, the constituents or elements of apomixis are — with the possible exception of apospory—to a large extent quantitative traits under polygenic control.

The role of tetraploids in the sexual-asexual cycle in dandelions (Taraxacum).

Apomictic plants often produce pollen that can function in crosses with related sexuals. Moreover, facultative apomicts can produce some sexual offspring. In dandelions, Taraxacum, a sexual-asexual cycle between diploid sexuals and triploid apomicts, has been described, based on experimental crosses and population genetic studies. Little is known about the actual hybridization processes in nature. We therefore studied the sexual-asexual cycle in a mixed dandelion population in the Netherlands. In this population, the frequencies of sexual diploids and triploids were 0.31 and 0.68, respectively. In addition, less than 1% tetraploids were detected. Diploids were strict sexuals, triploids were obligate apomicts, but tetraploids were most often only partly apomictic, lacking certain elements of apomixis. Tetraploid seed fertility in the field was significantly lower than that of apomictic triploids. Field-pollinated sexual diploids produced on average less than 2% polyploid offspring, implying that the effect of hybridization in the 2x-3x cycle in Taraxacum will be low. Until now, 2x-3x crosses were assumed to be the main pathway of new formation of triploid apomicts in the sexual-asexual cycle in Taraxacum. However, tetraploid pollen donors produced 28 times more triploid offspring in experimental crosses with diploid sexuals than triploid pollen donors. Rare tetraploids may therefore act as an important bridge in the formation of new triploid apomicts.

The occurrence of phenotypically complementary apomixis-recombinants in crosses between sexual and apomictic dandelions (Taraxacum officinale)

Apomictic seed development in dandelion (Taraxacum officinale) involves (1) restitutional meiosis (diplospory), (2) egg cell parthenogenesis, and (3) autonomous endosperm development. The question is whether these elements of apomixis are controlled by one single gene or by several independent genes. Five triploid non-apomictic hybrids, obtained in diploid sexual × triploid apomict crosses were characterized using cyto-embryological and genetic methods. Nomarski-differential interference contrast microscopy and the transmission of microsatellite markers and ploidy levels indicated that the hybrids combined elements of the apomictic and the sexual developmental pathway. Hybrids form two complementary groups with respect to the presence or absence of parthenogenesis and autonomous endosperm development. The occurrence of complementary apomixis-recombinants suggests that parthenogenesis and autonomous endosperm development in Taraxacum are regulated independently by different genes. This study also indicates that early embryo development is independent of endosperm formation, but that endosperm is essential for later embryo growth.

Apomixis in agriculture: the quest for clonal seeds

Apomixis, or asexual reproduction through seeds, is a natural trait that could have an immense positive impact on crop production. Apomictic breeding strategies could allow the fixation and indefinite propagation of any desired genotype, however complex. Apomicts display a wide variety of developmental mechanisms, which can be viewed as a short-circuiting of sexual development. Gametophytic and sporophytic apomixis are distinguished by the developmental origin of apomictically derived embryos. Genetic studies suggest that individual elements of gametophytic apomixis, such as apomeiosis and parthenogenesis, are either controlled by one or two dominant Mendelian factors. As recombination around apomeiosis loci is suppressed, it is currently not known how complex these loci are. Much less is known regarding the genetic control of sporophytic apomixis but initial studies suggest a complex genetic control. Genetic analyses of sexual reproduction in plant model systems have identified genes that, when mutated, display elements of apomixis. Such studies help in the identification of candidate genes and promoters that can be used for the de novo engineering of apomixis through biotechnology. Molecular genetic studies in apomictic and sexual systems will generate the knowledge necessary for the engineering of conditional apomixis technology. Approaches encouraging collaboration and widespread dissemination of the acquired knowledge will constitute the most innovative route to the development, deployment and acceptance of apomixis technology in agriculture.

Crosses between sexual and apomictic dandelions (Taraxacum). II. The breakdown of apomixis.

Some dandelions are diplosporous gametophytic apomicts. In order to study the inheritance and breakdown of apomixis, crosses were made between diploid sexuals and triploid apomicts. To investigate their breeding system, four nonapomictic diploid and 10 nonapomictic triploid hybrids were pollinated with diploids and the progenies were analysed. Seed fertility was significantly reduced in two diploid hybrids. Nine triploid hybrids were fertile and could be classified into three types, with respect to the composition of their progenies. Type A produced n+n hybrids. Type B produced either a mixture of n + n and 2n + n hybrids, or a mixture of pseudogamous 2n + 0 apomicts and 2n + n hybrids. Type C produced exclusively 2n + n hybrids. Inheritance of a microsatellite marker strongly suggested that 2n egg cells in type C plants were produced by a first division restitution mechanism. As in apomicts, microsporogenesis in type C plants was reductional. This suggests that type C plants are diplosporous plants that lack parthenogenesis. Such plants are very rare in other apomictic plant species. It is concluded that 'elements of apomixis', diplospory and parthenogenesis, can be uncoupled. This is inconsistent with the single-locus model for apomixis in Taraxacum as suggested by Mogie (1992). Instead, our results suggest that several loci are involved in the genetic control of apomixis in Taraxacum.

Crosses between sexual and apomictic dandelions (Taraxacum). II. The breakdown of apomixis

Some dandelions are diplosporous gametophytic apomicts. In order to study the inheritance and breakdown of apomixis, crosses were made between diploid sexuals and triploid apomicts. To investigate their breeding system, four nonapomictic diploid and 10 nonapomictic triploid hybrids were pollinated with diploids and the progenies were analysed. Seed fertility was significantly reduced in two diploid hybrids. Nine triploid hybrids were fertile and could be classified into three types, with respect to the composition of their progenies. Type A produced n+n hybrids. Type B produced either a mixture of n + n and 2n + n hybrids, or a mixture of pseudogamous 2n + 0 apomicts and 2n + n hybrids. Type C produced exclusively 2n + n hybrids. Inheritance of a microsatellite marker strongly suggested that 2n egg cells in type C plants were produced by a first division restitution mechanism. As in apomicts, microsporogenesis in type C plants was reductional. This suggests that type C plants are diplosporous plants that lack parthenogenesis. Such plants are very rare in other apomictic plant species. It is concluded that ‘elements of apomixis’, diplospory and parthenogenesis, can be uncoupled. This is inconsistent with the single-locus model for apomixis in Taraxacum as suggested by Mogie (1992). Instead, our results suggest that several loci are involved in the genetic control of apomixis in Taraxacum.

Breeding for apomixis in potato: Pursuing a utopian scheme

Apomictic propagation in potato holds a great promise for the new technology of growing potatoes from botanical seeds in stead of tubers. It is explained that apomixis results from the combination of abnormal meiosis, abnormal fertilization and parthenogenesis. An ideal scheme is given for growing potatoes from true seeds. This scheme remains utopian when autonomous apomixis is not available. Autonomous apomixis occurs when the reduction division is lacking (no gene recombination, no reduction of chromosome number) and the resulting 2n egg cell develops parthenogenetically without fertilization of the central nucleus. The first best approach is pseudogamous apomixis, which differs from autonomous apomixis in the necessity of fertilization of the central nucleus. Second best is the parthenogenetic development of 2n eggs, since these have originated through an abnormal type of reduction division. This type is characterized by the lack of formation of a reduction cell wall and fusion of the two spindles at metaphase II of meiosis (first division restitution). Apomixis in the narrow sense, either pseudogamous or autonomous, is defined. It has not been found in potato species or species hybrids. On the basis of our knowledge about characteristic features of existing apomictic species, about genetics and evolution of apomixis and about the occurrence of particular elements of apomixis in potato the possibility of introducing apomixis into potato from existing elements is discussed along with methods to induce missing elements. Some tentative approaches are suggested which in the long run may lead to the detection of potato genotypes with apomictic propagation.

Chromosomal and embryological analyses in sexual x apomictic hybrids ofPanicum maximum Jacq.

Cytological analyses in series of crosses between 7 sexual pistillate and 8 apomictic staminate parents of speciesPanicum maximum (Gramineae) are reported. Although these 15 progenitors were tetraploid (2n = 32), 2 dihaploids (2n = 16), 45 hexaploids (2n = 48) and 5 octoploids (2n = 64) were observed among 333 progeny plants. The role of unreduced gametes as the originators of polyploidy is discussed in relation to the so-called 'elements of apomixis'. The 2 dihaploids appeared to be sexual while the hexaploids and octoploids were all apomictic. At the tetraploid level sexual and apomictic hybrids segregated in a ratio close to 1∶1. These results were then compared to those already obtained from studies on other tropical grasses and indicate a simple genetic determinism for gametophytic apomixis.