Krystyna Musiał

Anatomy of ovary and ovule in dandelions (Taraxacum, Asteraceae)

The genus Taraxacum Wigg. (Asteraceae) forms a polyploid complex within which there are strong links between the ploidy level and the mode of reproduction. Diploids are obligate sexual, whereas polyploids are usually apomictic. The paper reports on a comparative study of the ovary and especially the ovule anatomy in the diploid dandelion T. linearisquameum and the triploid T. gentile. Observations with light and electron microscopy revealed no essential differences in the anatomy of both the ovary and ovule in the examined species. Dandelion ovules are anatropous, unitegmic and tenuinucellate. In both sexual and apomictic species, a zonal differentiation of the integument is characteristic of the ovule. In the integumentary layers situated next to the endothelium, the cell walls are extremely thick and PAS positive. Data obtained from TEM indicate that these special walls have an open spongy structure and their cytoplasm shows evidence of gradual degeneration. Increased deposition of wall material in the integumentary cells surrounding the endothelium takes place especially around the chalazal pole of the embryo sac as well as around the central cell. In contrast, the integumentary cells surrounding the micropylar region have thin walls and exhibit a high metabolic activity. The role of the thick-walled integumentary layers in the dandelion ovule is discussed. We also consider whether this may be a feature of taxonomic importance.

Embryological study on gynogenesis in onion (Allium cepa L.)

Abstract  Haploid induction in onion can, to date, be induced only via gynogenesis by culturing unfertilized flowers, ovaries or ovules. The process of haploid embryo induction has been macroscopically well studied, but only limited data exist from microscopic examination of ovule development status at the inoculation stage and of the origin of gynogenic embryos. Microscopic studies were carried out using individual donor plants with relatively high embryo induction frequencies (45.9 embryos formed per 100 flowers, on average, for 2 years). Ovaries from flower bud culture were fixed at 1 week intervals up to the 7th week of culture. These were compared with pollinated ovaries at 1 or 2 weeks after pollination. In total, 1428 unfertilized embryo sacs were examined. The results indicate that, at the time of inoculation, ovules within ovaries 2.0–3.0 mm in diameter contained two- or four-nucleate embryo sacs in the smallest ovaries to mature embryo sacs in the largest ovaries. It seems likely that the embryos are actually induced from ovaries cultured at the immature stage. After 1 or 2 weeks in culture, the egg apparatus primarily consisted of distinctly enlarged synergids and the egg cell, which was often detached from the micropylar pole. But free nuclear endosperm was also formed. From the 2nd to 7th week in culture, formation of haploid embryos (from globular to the almost mature cylindrical stage) was detected in 5.7% of the ovules. Their origin, for several reasons, was most likely the egg cell. In addition, ovules containing endosperm only (3.6%) and ovules containing the egg apparatus (0.5%) or both endosperm and embryo (0.4%) were detected. This observation is probably unique and has not yet been reported in other species studied.

The development of onion (Allium cepa L.) Embryo sacs in vitro and gynogenesis induction in relation to flower size

SummaryThe development of embryo sacs (ES) in vitro and induction of gynogenesis were studied in onion flower bud culture. Explants were divided into three groups according to their size at inoculation: (a) small flower buds (2.3–3.0 mm in diameter); (b) medium flower buds (3.1–3.7 mm); and (c) large flower buds (3.8–4.4 mm). For histological study, excised ovaries were fixed at inoculation and then at 3-d intervals until day 12, and after 2 and 3 wk of culture. Some explants were cultured until embryo emergence, i.e., 3–5 mo. In total, 2592 ovules were examined histologically. At inoculation, 83% of ovules in small flower buds contained a megaspore mother cell; in 17% of ovules, two-nucleate ES occurred. In medium flower buds two-nucleate, four-nucleate, and mature ES were present at frequencies of 15%, 46%, and 40%, respectively. In large flower buds, only mature ES occurred. In vitro conditions did not disturb meiosis and megagametophyte development in non-degenerated ovules. Regardless of the developmental stage at inoculation, only mature ES occurred on day 12. Gynogenic embryos were found after 2 wk of culture, indicating that embryos developed in mature ES exclusively. Embryos were detected in 5.4% of histological studied ovules; however, the number of embryos after 3–5 mo. was higher (12.4%). The parthenogenetic origin of the embryos is discussed. In addition, ES containing endosperm only (6.5%) and both endosperm and embryo (0.4%) were observed.

Taraxacum zajacii (Asteraceae), a New Species from Poland

A description of Taraxacum zajacii J. & P. Marciniuk, a new species of T. sect. Palustria in Poland is given. Taraxacum zajacii is a pentaploid (2n = 40). Morphologically, the new species is closest to the T. subalpinum/T. neterophilum group.

Deposition of callose in young ovules of two Taraxacum species varying in the mode of reproduction

Although callose occurs during megasporogenesis in most flowering plants, the knowledge about its general function and the mechanisms by which the callose layer is formed in particular places is still not sufficient. The results of previous studies suggest a total lack of callose in the ovules of diplosporous plants in which meiosis is omitted or disturbed. This report is the first documentation of callose events in dandelions ovules. We demonstrated the pattern of callose deposition during the formation of megaspores through diplospory of Taraxacum type and during normal meiotic megasporogenesis in apomictic triploid Taraxacum atricapillum and amphimictic diploid Taraxacum linearisquameum. We found the presence of callose in the megasporocyte wall of both diplosporous and sexual dandelions. However, in a diplosporous dandelion, callose predominated at the micropylar pole of megaspore mother cell (MMC) which may be correlated with abnormal asynaptic meiosis and may indicate diplospory of the Taraxacum type. After meiotic division, callose is mainly deposited in the walls between megaspores in tetrads and in diplodyads. In subsequent stages, callose gradually disappears around the chalazal functional megaspore. However, some variations in the pattern of callose deposition within tetrad may reflect variable positioning of the functional megaspore (FM) observed in the ovules of T. linearisquameum.

Synergids and filiform apparatus in the sexual and apomictic dandelions from section Palustria (Taraxacum, Asteraceae)

An evolutionary trend to reduce “unnecessary costs” associated with the sexual reproduction of their amphimictic ancestors, which may result in greater reproductive success, has been observed among the obligatory apomicts. However, in the case of the female gametophyte, knowledge about this trend in apomicts is not sufficient because most of the ultrastructural studies of the female gametophyte have dealt with amphimictic angiosperms. In this paper, we tested the hypothesis that, in contrast to amphimictic plants, synergids in apomictic embryo sacs do not form a filiform apparatus. We compared the synergid structure in two dandelions from sect. Palustria: the amphimictic diploid Taraxacum tenuifolium and the apomictic tetraploid, male-sterile Taraxacum brandenburgicum. Synergids in both species possessed a filiform apparatus. In T. brandenburgicum, both synergids persisted for a long time without any degeneration, in spite of the presence of an embryo and endosperm. We propose that the persistent synergids in apomicts may play a role in the transport of nutrients to the embryo.

Pattern of callose deposition during the course of meiotic diplospory in Chondrilla juncea (Asteraceae, Cichorioideae).

Total absence of callose in the ovules of diplosporous species has been previously suggested. This paper is the first description of callose events in the ovules of Chondrilla juncea, which exhibits meiotic diplospory of the Taraxacum type. We found the presence of callose in the megasporocyte wall and stated that the pattern of callose deposition is dynamically changing during megasporogenesis. At the premeiotic stage, no callose was observed in the ovules. Callose appeared at the micropylar pole of the cell entering prophase of the first meioticdivision restitution but did not surround the megasporocyte. After the formation of a restitution nucleus, a conspicuous callose micropylar cap and dispersed deposits of callose were detected in the megasporocyte wall. During the formation of a diplodyad, the micropylar callose cap decreased and the walls of a newly formed megaspores showed scattered distribution of callose. Within the older diplodyad, callose was mainly accumulated in the wall between megaspores, as well as in the wall of the micropylar cell; however, a dotted fluorescence of callose was also visible in the wall of the chalazal megaspore. Gradual degradation of callose in the wall of the chalazal cell and intense callose accumulation in the wall of the micropylar cell were related to the selection of the functional megaspore. Thus, our findings may suggest that callose fulfills a similar role both during megasporogenesis in sexual angiosperms and in the course of meiotic diplospory in apomicts and seems to form a regulatory interface between reproductive and somatic cells.

Insights into developmental processes in anthers, ovaries, and ovules of Taraxacum belorussicum (Asteraceae-Cichorioideae) using DIC optics

This study represents the first report on the embryological characteristics of triploid male-sterile dandelion Taraxacum belorussicum (section Palustria) from Poland. While this taxon is considered to be a male-sterile species, we found that the investigated individuals produced pollen. Irregular tetrads, triads and diads with microspores of unequal size were observed in the pollen loculi as a result of disturbed meiotic division, while anthers’ tapetum did not show structural disorders. Possible reasons for the plasticity in the expression of male sterility, as well as the role of pollen in apomicts, are discussed. Flowers of the examined individuals contained well-developed nectaries. The course of embryological processes in the ovules indicated an apomictic mode of reproduction in T. belorussicum. We observed meiotic diplospory of the Taraxacum type, in which first meiotic division starts but results in nuclear restitution, while undisturbed second meiotic division gives rise to a dyad of unreduced megaspores (diplodyad). After three mitotic divisions of the chalazal megaspore, a seven-celled unreduced female gametophyte developed. The features of ovule anatomy and characteristics of a mature female gametophyte corresponded to these described in sexually reproducing dandelions.