The aim of the study was to develop a method of laparoscopic embryo transfer in pigs and to compare different variants of this method. Two catheter diameters (1.6 mm and 1.0 mm), the method and site of embryo deposition (oviduct or uterus), the embryo development stage (2 – 4 cell or blastocyst), the method for oviduct or uterus stabilization, the potential for cryopreserved embryo transfer, the developmental potential of the embryos after transfer to the oviduct, patomorphology of the oviduct after transfer and possible clinical complications were taken into consideration. Two studies compared two variants of transfer to the uterus, and five variants of transfer to the fallopian tube. The transfer of embryos by the infundibulum may be of limited use due to handling problems and very low efficiency (pregnancy was not achieved). Very low efficiency was shown after transfer of vitrified embryos. Transfer to the fallopian tube by puncture of the fallopian tube, regardless of the developmental stage of the embryo, is the recommended method of embryo transfer. The histopathological examination of the fallopian tube revealed possible changes within the puncture site. The numerous clinical complications observed did not affect the effectiveness of the method.

In this study, female gametophytes of Silene muradica, which is a gynodioecious species, were examined histologically. Buds and blossoms of S. muradica were used as the research material. They were collected in the Sivas province (Turkey) in July 2019, and fixed with ethanol:acetic acid solution (3:1, v/v). Flower parts were dissected under a stereo microscope. They were dehydrated in rising alcohol series and then embedded in Historesin. The sections were taken by a rotary microtome and stained with 0.5% Toluidine blue O. The ovary of S. muradica has three carpels and a single chamber, the ovules are arranged on a central column. The mature ovule is of the campylotropous type, crassinucellate and bitegmig. The megaspore mother cell undergoes regular meiotic division and forms a linear megaspore tetrad after meiosis. The development of the embryo sac is monosporic. The chalazal megaspore is functional and the others degenerate. The mature embryo sac is eight-nucleated and of the Polygonum type. The synergid cells and the egg cell are completely surrounded by the cell wall. Antipodal cells are temporary cells, which degenerate immediately after fertilization. Before fertilization, polar nuclei are fused in the central cell and form the secondary nucleus. The endosperm development is of the nuclear type. Nucellar tissue is permanent and forms perisperm in mature seeds. The embryo development is of the Caryophyllad type. In this study, the development of the female gametophyte of S. muradica, which was determined to be a gynodioecious species, was reported for the first time.

The article presents the main discoveries of Prof. Andrzej K. Tarkowski, which proved to be fundamental for modern mammalian developmental biology and also for progress in animal breeding and assisted reproduction. Among his achievements the most important are: the demonstration of regulative abilities of blastomeres isolated from early mammalian embryos, generation of first chimaeric mice, studies on mammalian parthenogenesis and establishment of blastomere electrofusion technique for production of tetraploid embryos. Studies on nucleocytoplasmic interactions in germ cells and early embryos contributed substantially to the development of mammalian cloning. Prof. Tarkowski’s work and discoveries provided a tremendous input to the contemporary developmental biology of mammals.

In this study, the effects of oleic (18:1 cis-9-octadecenoic acid) and linoleic (18:2 (n-6), 9,12-octadecadienoic acid) acids added to the embryo culture media for bovine embryonic development after vitrification were investigated in cattle. Following maturation and fertilization, the oocytes were placed in Charles Rosencrans (CR1aa) culture drops containing 10, 100, 500, and 1000 μM of oleic or linoleic acids. On day 7 or 8 of the culture, the blastocysts and expanded blastocysts were vitrified and warmed to evaluate the viability and development. High doses of oleic acid (1000 μM) in the culture media increased the viability of embryos after vitrification. Similarly, linoleic acid at 1000 μM increased the viability compared to the other linoleic acid doses. It was observed that the addition of essential fatty acids improved the development of embryos. Increasing the concentration of linoleic and oleic acid concentrations in the media proportionally advanced the embryonic development and hatching capability after vitrification/warming. Specifically, the addition of high doses of oleic acid had dramatic effects on the embryonic development after vitrification/warming probably due to the increased lipid storage. In conclusion, the present results suggest that the ratio of unsaturated fatty acids in the culture media affects significantly the embryonic development in vitro.

We used cytological and embryological methods to study reproductive cycle stages in Cerasus fruticosa Pall., Cerasus × eminens (Beck) Buia and Cerasus × mohacsyana (Kárpáti) Janchen from SW Slovakia, focusing on development of the male and female reproductive organs, fertilization processes and embryo formation. We found that reproductive potential was reduced by synergistic effects of negative biotic and abiotic factors. Despite the presence of degenerated, deformed pollen grains and their great variability of shape and size, a sufficient amount of normally developed viable pollen grains developed in anthers of C. fruticosa and C. × mohacsyana. Disturbed microsporogenesis in C. × eminens led to significantly lower production of viable pollen grains. We did not observe serious disturbances during megasporogenesis and megagametogenesis. Lower fruit set was caused by degeneration of ovules as a result of unsuccessful pollination, fertilization failure, or embryo degeneration during its initial development.

An efficient system of micropropagation via somatic embryogenesis from root-derived callus was established in

Arabica coffee (Coffea arabica L.). Twenty-six callus lines were induced on MS (Murashige and Skoog, 1962)

medium supplemented with combinations of NAA (0, 0.1, 0.5, 1 and 2 mg/L) plus BA (0, 1 and 2 mg/L), or 2,4-D

(0, 0.1, 0.5, 1 and 2 mg/L) plus TDZ (0, 1 and 2 mg/L). Subsequently, two types of somatic embryos were obtained

from callus cultures and named S-type and I-type embryos. The S-type embryos were obtained from an 18-monthold

callus line which was induced and maintained at 2 mg/L TDZ and 0.1 mg/L 2,4-D near the end of each period

of the subculture. These embryos have a developmental barrier, which did not pass through the torpedo stage

and could be overcome by a supplement of 2 or 5 mg/L BA. The I-type embryos were induced from 3-month-old

callus when transferred onto induction media, i.e., MS supplemented with TDZ (2 and 5 mg/L) plus 2,4-D (0 and

0.1 mg/L). The significantly highest response, i.e., 13.3 embryos per callus clump was obtained at 2 mg/L TDZ.

In this study, the results reveal that TDZ has a crucial effect on embryogenic callus induction, proliferation and

subsequent somatic embryogenesis.

Our study involved the first-ever evaluation of the performance of anther culture and wheat × maize hybridization techniques in producing haploids or doubled haploids as a result of spontaneous doubling of the chromosome number during androgenesis in plants from 30 wheat genotypes including ancient, local and modern types. The results indicated that the best induction rates of androgenic structures and haploid embryos for the hexaploid and tetraploid wheat genotypes were obtained with anther culture and wheat × maize hybridization, respectively. Whereas only one regenerated plant from 15 genotypes of tetraploid wheat was obtained, 13 plants were regenerated from 15 genotypes of hexaploid wheat. Moreover, haploid embryos obtained in wheat × maize hybridization 60 and 100% green plants regenerated in relation to the number of the cultured haploid embryos. Genotypes with high induction capacity to produce androgenic structure or haploid embryos did not have desired haploid plantlets regeneration capacity and vice-versa. However, with both methods, hexaploid wheat genotypes had a considerable ability to produce green plants. Doubled haploid plants were obtained from ancient and local wheat genotypes by both methods, but not from modern wheat. Those genotypes can be used as parents in future wheat breeding programs and new varieties may be obtained by selecting pure lines in wheat populations

We studied the embryology of Withania somnifera (L.) Dunal by light microscopy in order to reveal specific embryological features of the genus, and compared the results with embryological data on other members of the family Solanaceae. The key embryological characters of W. somnifera include dicotyledonous-type anther wall formation, simultaneous cytokinesis in pollen mother cells, binucleate tapetal cells, 2-celled mature pollen, anatropous, tenuinucellate and unitegmic ovules, polygonum-type embryo sac formation, the presence of an endothelium, and cellular endosperm formation. We give the first report of the dicotyledonous mode of anther wall formation (previously described as basic type) for the species. Comparative study suggests that anther wall formation, number of nuclei in tapetal cells, number of cells in mature pollen, mode of embryo sac formation and endosperm development are the most variable embryological features in Solanaceae. Some of these embryological features of W. somnifera should be of value for comparative study of related species and their phylogenetic relationships within the family.

Poor seed set is a limiting factor in alfalfa breeding, as it slows the selection response. One strategy used to overcome this problem is to search for mutations of inflorescence morphology. Long-peduncle (lp), branched-raceme (br) and top-flowering (tf) inflorescence mutations increase the number of flowers per inflorescence, but they do not improve seed set per flower. Here we assessed pollen tube growth in styles of those inflorescence mutants and we observed embryo and endosperm development in seeds 1 to 16 days after pollination (DAP). The number of pollen tubes penetrating the style and the ovary was similar in all tested mutants and in the reference cultivar Radius. At 2 DAP, fertilized ovules were 2.7-3.9 times less numerous in certain inflorescence mutants than in the short-raceme cv. Radius. Ovule degeneration progressed at 2-4 DAP in all analyzed plants. Most ovules were not properly developed in the control cultivar (62%), nor in the forms with mutated inflorescence morphology (69-86%). The number of seeds per pod was lowest in the tf form despite its having the highest number of ovules per ovary. It appears that the number of ovules per pistil is not a crucial factor in seed set in alfalfa when fertilization efficiency is very low. Both poor fertilization and gradual ovule degeneration were factors causing poor seed set in the investigated alfalfa genotypes.

We made interspecific crosses to facilitate the introgression of desirable traits of Allium roylei into the Alliumcepa genome. After hand-pollination, 906 interspecific F1Allium cepa × A. roylei plants were obtained by in vitro culture via embryo rescue. Nuclear DNA analysis showed that 97.6% of the regenerants were interspecific F1Allium cepa × A. roylei hybrids. Genomic in situ hybridization (GISH) showed that each hybrid had 16 chromosomes, eight of which were identified as A. cepa and eight as A. roylei chromosomes.

The embryology of two species, Deschampsia antarctica , a native species, and Poa annua , an alien species in the Antarctic we studied. Flowering buds of plants growing in their natural habitats on King George Island and generative tissues of both plant species grown in a greenhouse were analyzed. Adaptations to autogamy and anemogamy were observed in the flower anatomy of both species. The microsporangia of the evaluated grasses produce a small number of three−celled pollen grains. Numerous pollen grains do not leave the microsporangium and germinate in the thecae. Deschampsia antarctica and P. annua plants harvested in Antarctica developed a particularly small number of microspores in pollen chambers. In D. antarctica , male gametophytes were produced at a faster rate: generative cells in pollen did not become detached from the wall of the pollen grain, they were not embedded in the cytoplasm of vegetative cells, and they divided into two sperm cells situated close to the wall. The monosporous Polygonum type of embryo sac development was observed in the studied species. The egg apparatus had typical polarization, and the filiform apparatus did not develop in synergids. Large antipodals with polyploidal nuclei were formed in the embryo sacs of D. antarctica and P. annua . Poa annua was characterized by numerous antipodal cells which formed antipodal tissue in the chalazal region of the embryo sac. Three distinct antipodals with atypical, lateral position in the vicinity of the egg apparatus were observed in D. antarctica. The diaspores of the investigated grass species were characterized by small size, low weight and species−specific primary and secondary sculpture of the testa and caryopsis coat.