Department of Genetics & CytologyDepartment of Genetics & Cytology

Laboratory of Plant CytologyLaboratory of Plant Cytology & & Embryology Embryology

Laboratory of Laboratory of GeneticsGenetics

FACULTY OF BIOLOGY, GEOGRAPHY AND OCEANOLOGYFACULTY OF BIOLOGY, GEOGRAPHY AND OCEANOLOGY

Living Matter

Laboratory of Laboratory of GeneticsGenetics

Head: Tadeusz Sywula Head: Tadeusz Sywula PhD, DSc,PhD, DSc, ProfessorProfessor  

Research InterestsResearch InterestsResearch in this laboratory is concentrated on assessing intraspecific Research in this laboratory is concentrated on assessing intraspecific genetic polymorphism of hydrobionts, especially crustaceans and fishes. genetic polymorphism of hydrobionts, especially crustaceans and fishes. We use various types of markers such as allozymes and mitochondrial and We use various types of markers such as allozymes and mitochondrial and nuclear DNA to estimate levels of genetic polymorphism of natural nuclear DNA to estimate levels of genetic polymorphism of natural populations of studied species, population range and subdivision, populations of studied species, population range and subdivision, characteristics of gene flowcharacteristics of gene flow.. The research activities contribute to our The research activities contribute to our understanding of microevolution of species representing various biological understanding of microevolution of species representing various biological groups, e.g. relict species, invading species, eurytopic species and, groups, e.g. relict species, invading species, eurytopic species and, contrarily, species specialized with respect to type of environment such as contrarily, species specialized with respect to type of environment such as astatic water–bodies, lake profundal, lenitic littoral, upwelling waters, astatic water–bodies, lake profundal, lenitic littoral, upwelling waters, inland saline waters.inland saline waters.

Calanoides carinatus (KRØYER) (Crustacea, Copepoda), a planktonic

species inhabiting South Atlantic Central Waters below 200 m depth as

diapausing stage V copepodites, reproducing rapidly in upwelling

waters

It was shown that individuals of C. carinatus arrive with the upwelling waters on the shelf off Ghana in two genetically different stocks. This result is indicative of the non-random (geographic, or perhaps behavioural) segregation of the C. carinatus population in the South Atlantic Central Waters off West African coast. (Sywula T., Wiafe G., Sell J., Głażewska I. 2002. Genetic subdivision of the upwelling copepod Calanoides carinatus (Krøyer, 1849) off the continental shelf of Ghana. J. Plankton Res. 24: 523-525).

Candona neglecta SARS (Crustacea, Ostracoda), a fresh-water, Palaearctic, eurytopic species, reproducing amphigonically, unable to swim

Our results unexpectedly suggest lack of genetic isolation betweenpopulations inhabiting as extremely different environments as profundal of post-glacial lakes from Pomeranian and Masurian Lake Districts and the deep muddy bottom of the Baltic Sea (it might be probably a matter of birds and fish as mediators of so highly effective passive migration of C. neglecta between quite old populations connected with large water-bodies).On the other hand a very distinct founder effect can be noted in the case of young populations from small astatic basins. We was able to show that a population inhabiting a large lake may be genetically subdivided due to differentiated eutrophication. (Wysocka A., Sell J., Sywula T. 2000. Genetic variability in natural populations of eurytopic ostracod Candona neglecta Sars. Zool. Sci. 17: 55-59).

Saduria entomon L. (Crustacea, Isopoda), the glacial relict crustacean

The distribution pattern of mtDNA haplotypes in populations from the Baltic, the White Sea and the Barents Sea was examined to assess phylogeographic relationships among them. Significant differences in haplotype frequency distributions among populations were found. The isolated populations of S. entomon have diverged since the retreat of the last glaciation. The geographical pattern of variation is most likely the result of stochastic (founder effect, genetic drift) mechanisms and suggests that the haplotype differentiation observed is probably older than the isolation of the Baltic and Arctic seas. (Sell J. 2003. Haplotype Frequency Distribution in Northeastern European Saduria entomon (Crustacea: Isopoda) Populations. A Phylogeographic Approach. Int. Rev. Hydr. 88: 584-595).

Salmo letnica (KARAMAN) the endemic trout species

from the Balkan Lake Ohrid

Mitochondrial DNA diversity in sympatric, reproductively isolated populations of Ohrid trout, Salmo letnica was investigated. Results obtained allowed us to: (i) provide insight into the distribution of mtDNA diversity in the typicus and aestivalis populations, (ii) test the level of their reproductive isolation and (iii) provide clues regarding their evolutionary origin, (iv) present new data concerning evolution of the salmonid mitochondrial control region. (Sell J, Spirkovski Z. 2003. A new insight into genetic diversity of trout Salmo letnica from the Balkan Lake Ohrid: tandemly repeated sequences in mtDNA control region ( in press)).

Genetic polymorphism of the Baltic Sea invading species:

American mud crab Rhithropanopeus harrisii (GOULD) (Brachyura,

Decapoda)Ponto-Caspian goby Neogobius melanostomus (PALLAS) (Pisces)

Despite of expected consequences of the recurrent founder effect and at least one bottleneck effect two geographically isolated populations from the Vistula River and the Vistula Lagoon of the mud crab show unexpectedly high level of genetic polymorphism. (Laszczuk J. 1998. Impact of colonisation process on gene pool of Rhithropanopeus harrisii (Gould) ssp. tridentatus (Maitland) (Brachyura, Decapoda). Pol. Arch. Hydrobiol. 45: 65-75).

No signs of the founder effects in a newly established population of the fishy invader Neogobius melanostomus may indicate very intensive colonisation of the Baltic Sea. (Szybkowska J. 2003. Genetic diversity of the invading fish species Neogobius melanostomus (Pallas, 1811) (Gobiidae: Perciformes) from the Baltic Sea. Annal. Zool. 53: 339-346).

Dendrocoelum ØRSTED and Phagocata LEIDY (Tricladida) endemic species flocks from

the relict Lake Ohrid

As many as 23 endemic species of Tricladida were found in the relict Balkan Lake Ohrid. We were able to show that the separation of particular lineages within two mentioned species flocks was widely dispersed over time. (Sywula T., Krstanovski Z., Tasevska O., Sell J., Kretowicz T. 2003. Genetic differences among several species of Tricladida from the relict Lake Ohrid as revealed by enzyme electrophoresis. Folia Biol. 51: 105-109).

Equus caballus L. (Mammalia) - Polish

Arabian horses

Genetic structure of this non-randomly mating pedigreed population was studied with innovative use of components of pedigree analysis. By this means it was possible to show reasons of quite high level of genetic polymorphism of Polish Arabian horses despite of seemingly significant influences of directional selection and inbreeding (Głażewska I. 2000. The founder contribution analysis in currently living Polish Arabian brood mares. Animal Sci. Pap. Rep. 18: 19-31).

Ostracode Research Group Rich collection of European ostracodes including type-specimens

of several taxa is located at the Department of Genetics and Cytology. On the basis of this comparative material numerous papers were published, concerning taxonomy, biology and ecology of contemporary and extinct species and assemblages of species.

Achievements useful for the BioMoBiL activity: evidenced by subfossil and Recent Ostracoda concept of the coherent succession of lacustrine meiofauna in the profundal zone of postglacial lakes in relation to the progressing eutrophication and pollution as a tool for the bio-indication of sequencing changes in the lake environment. (Namiotko T. 1998. Changes in the profundal lacustrine ostracod fauna as an indicator of environmental perturbations in Polish lakes undergoing eutrophication. Bull. Centres Rech., Explor.-Prod. Elf-Aquitaine, Mém. 20: 117-124).

Values of the selected trophy indices combined with established profundal Ostracod assemblages.

Recorded directions of stratigraphic succesion of the established profundal ostracoda assemblages A, B, b, C, D, E. Size of arrows shows in a semiquantitative way frequency of the sequential alternations and size of the figured species corresponds to their domination in assemblages.

Four season life cycle. Points show the stages of life cycle of C. rectangulata present in samples.

 

 

Stage of life cycle years of life

Five season life cycle. Points show the stages of life cycle of C. rectangulata present in samples.

Stage of life cycle years of life

Candona rectangulata ALM, an Arctic, fresh-water species inhabiting shallow tundra water-bodies,reproducing

parthenogenetically

Peculiar life strategy was discovered in Spitsbergen populations of this tiny ostracode resulting in extremely prolonged development, which can take as many as five years. (Wiśniewska B. The life cycle of Candona rectangulata Alm, 1914 – submitted).

Laboratory of Plant Cytology Laboratory of Plant Cytology and Embryologyand Embryology

Head: Head: Jerzy BohdanowiczJerzy Bohdanowicz PhDPhD

Research InterestsResearch Interests::• biology of sexual reproduction in vascular plants:biology of sexual reproduction in vascular plants:

• development, ultrastructure and cytochemistry of embryodevelopment, ultrastructure and cytochemistry of embryo sac, embryo and endosperm sac, embryo and endosperm

• development, ultrastructure and immunocytochemistry of development, ultrastructure and immunocytochemistry of pollen and pollen tubepollen and pollen tube

• differentiationdifferentiation of stigma and pistil transmitting tract of stigma and pistil transmitting tract• experimental embryology of vascular plantsexperimental embryology of vascular plants

Laboratory of Plant Cytology Laboratory of Plant Cytology and Embryologyand Embryology

Techniques and methods we use routinely include:Techniques and methods we use routinely include:

• transmission and scanning electron microscopy transmission and scanning electron microscopy • light microscopy, epifluorescence and confocal laser scanning light microscopy, epifluorescence and confocal laser scanning

microscopymicroscopy• cytophotometrycytophotometry• cytochemistry and immunodetectioncytochemistry and immunodetection• morphometry and stereologymorphometry and stereology

DDevelopment, ultrastructure and cytochemistry of evelopment, ultrastructure and cytochemistry of embryoembryo sac, embryo and endosperm sac, embryo and endosperm

Research of our group focuses on the development and functional Research of our group focuses on the development and functional ultrastructure of embryo-suspensor and endosperm haustoria – ultrastructure of embryo-suspensor and endosperm haustoria – highly polyploid cells engaged in the nutrition of the young embryo highly polyploid cells engaged in the nutrition of the young embryo and seed.and seed.

Sagittaria sagittifolia

DDevelopment, ultrastructure and cytochemistry of evelopment, ultrastructure and cytochemistry of embryoembryo sac, embryo and endosperm sac, embryo and endosperm

The processes of suspensor differentiation in The processes of suspensor differentiation in Sedum acreSedum acre (Crassula- (Crassula-ceae), ceae), Triglochin palustreTriglochin palustre and and T.maritimumT.maritimum (Juncaginaceae), (Juncaginaceae), Gagea Gagea lutealutea (Liliaceae), and (Liliaceae), and Luronium natansLuronium natans (Alismataceae) have been (Alismataceae) have been studied in our lab. studied in our lab.

Kozieradzka-Kiszkurno M, Bohdanowicz J (2003)Kozieradzka-Kiszkurno M, Bohdanowicz J (2003).. Sedum acreSedum acre embryogenesis: poly embryogenesis: poly--ploidization in the suspensor. Acta Biol Crac Ser Bot 45(2): ploidization in the suspensor. Acta Biol Crac Ser Bot 45(2): in pressin press; ; Kozieradzka-Kiszkurno Kozieradzka-Kiszkurno M, Świerczyńska J, Bohdanowicz J (2002)M, Świerczyńska J, Bohdanowicz J (2002).. Polyploidization in the suspensor of Polyploidization in the suspensor of Triglochin Triglochin palustrepalustre L. (Juncaginaceae). Acta Biol Crac Ser Bot 44: 189-193 L. (Juncaginaceae). Acta Biol Crac Ser Bot 44: 189-193; ; Kozieradzka-Kiszkurno M, Kozieradzka-Kiszkurno M, Bohdanowicz J (2000)Bohdanowicz J (2000).. Ultrastructure of the suspensor in Ultrastructure of the suspensor in Triglochin palustreTriglochin palustre L. L. (Juncaginaceae). Acta Biol Crac Ser Bot 42 (1): 24(Juncaginaceae). Acta Biol Crac Ser Bot 42 (1): 24.......etc........etc.

Sedum acre

The suspensor basal cell (BC) of all the studied species is highly The suspensor basal cell (BC) of all the studied species is highly polyploid, contains dense cytoplasm with numerous organelles, and polyploid, contains dense cytoplasm with numerous organelles, and develops special wall ingrowths facilitating the transfer of solutes develops special wall ingrowths facilitating the transfer of solutes from the surrounding cells of the ovule into the embryo. from the surrounding cells of the ovule into the embryo.

Usually, the plasmodesmata present in the common cell wall Usually, the plasmodesmata present in the common cell wall allowallowthe symplastic flow of nutrients between the BC and the rest of the symplastic flow of nutrients between the BC and the rest of embryo. embryo.

Gagea lutea Sedum acreGagea lutea Sedum acre

Recently, in the suspensor BC of Recently, in the suspensor BC of Sedum acreSedum acre, we have discovered a , we have discovered a new type of compound plasmodesmata, not described earlier. The new type of compound plasmodesmata, not described earlier. The BC forms special haustorial protrusionsBC forms special haustorial protrusions (MH) (MH), penetrating the , penetrating the ovular tissues, and is apparently the main source of nutritive ovular tissues, and is apparently the main source of nutritive substances for the embryo-proper. The unusual structure of these substances for the embryo-proper. The unusual structure of these plasmodesmata evolved pplasmodesmata evolved probablyrobably to facilitate and/or control the to facilitate and/or control the mass flow of nutrients from the basal cell mass flow of nutrients from the basal cell

Kozieradzka-Kiszkurno MKozieradzka-Kiszkurno M (2003). Development, ultrastructure and cytochemistry of the (2003). Development, ultrastructure and cytochemistry of the embryo-suspensor in embryo-suspensor in Sedum acreSedum acre (Crassulaceae). Doctoral thesis, University of Gdańsk. (Crassulaceae). Doctoral thesis, University of Gdańsk.

Unexpectedly, we have found no plasmodesmata in the basal cell Unexpectedly, we have found no plasmodesmata in the basal cell wall of wall of Gagea lutea Gagea lutea – the cell is symplastically isolated from the – the cell is symplastically isolated from the other embryo cells. In the BC a peculiar labyrinth of wall ingrowths other embryo cells. In the BC a peculiar labyrinth of wall ingrowths (WI) (WI) forms on the common cell wall, probably to facilitate the forms on the common cell wall, probably to facilitate the transfer of nutrients to the embryo proper through the wall apoplast. transfer of nutrients to the embryo proper through the wall apoplast.

Bohdanowicz JBohdanowicz J. . Development and ultrastructure of suspensor basal cell in Development and ultrastructure of suspensor basal cell in Gagea luteaGagea lutea. . Presented atPresented at Xth Intern Xth Internatiational Conference on Plant Embryology. Nitra, Slovak Republic 2001.onal Conference on Plant Embryology. Nitra, Slovak Republic 2001.(paper in preparation)(paper in preparation)..

DDevelopment, ultrastructure andevelopment, ultrastructure and immunocytochemistry of pollen and pollen tubeimmunocytochemistry of pollen and pollen tube

Del Casino C, Bohdanowicz J, LewandowskaDel Casino C, Bohdanowicz J, Lewandowska B, Cresti M (1999):B, Cresti M (1999): The The organization of microtubules during generative-cell division in organization of microtubules during generative-cell division in Convallaria Convallaria majalis. majalis. Protoplasma 207: 147-153.Protoplasma 207: 147-153. [in cooperation with [in cooperation with Dipartimento Biologia Ambientale, Università degli Studi di Siena, ItalDipartimento Biologia Ambientale, Università degli Studi di Siena, Italy]y]

Analysis by confocal laser scanning microscopy after tubulin Analysis by confocal laser scanning microscopy after tubulin immunostaining was used to investigate changes of the microtubule immunostaining was used to investigate changes of the microtubule cytoskeleton during generative-cell (GC) migration through the cytoskeleton during generative-cell (GC) migration through the pollen tube and its division into the two sperm cells (SC).pollen tube and its division into the two sperm cells (SC).

DifferentiationDifferentiation of stigma and pistil transmitting tract of stigma and pistil transmitting tract

The function of the stigma is collection, hydration and germination The function of the stigma is collection, hydration and germination of pollen. The stigma develops a highly specialized receptive of pollen. The stigma develops a highly specialized receptive surface which fulfills these functions.surface which fulfills these functions.

Świerczyńska J, Bohdanowicz J (2000): Endopolyploidization of stigmatic Świerczyńska J, Bohdanowicz J (2000): Endopolyploidization of stigmatic papillae in papillae in Triglochin palustreTriglochin palustre L. (Juncaginaceae). Acta Biol Crac Ser Bot 42 L. (Juncaginaceae). Acta Biol Crac Ser Bot 42/1//1/: : 62.62.; ; Bohdanowicz J, Dąbrowska D (1997): Polyploidization of stigmatic papillae Bohdanowicz J, Dąbrowska D (1997): Polyploidization of stigmatic papillae in in Triglochin maritimumTriglochin maritimum L. (Juncaginaceae). Acta Biol Crac Ser Bot 39: 63-67. L. (Juncaginaceae). Acta Biol Crac Ser Bot 39: 63-67.

Our cytophotometric studies on the dry papillate stigma of Our cytophotometric studies on the dry papillate stigma of Triglochin maritimumTriglochin maritimum and and T. palustreT. palustre revealed that all papillae revealed that all papillae constituting the receptive surface underwent regular endoredupli-constituting the receptive surface underwent regular endoredupli-cation during maturation. The highest ploidy level determined in the cation during maturation. The highest ploidy level determined in the nuclei of the studied cells was 64C DNA content. nuclei of the studied cells was 64C DNA content.

Świerczyńska J, Bohdanowicz J (2000): Endopolyploidization of stigmatic Świerczyńska J, Bohdanowicz J (2000): Endopolyploidization of stigmatic papillae in papillae in Triglochin palustreTriglochin palustre L. (Juncaginaceae). Acta Biol Crac Ser Bot 42 L. (Juncaginaceae). Acta Biol Crac Ser Bot 42/1//1/: : 62.62.; ; Bohdanowicz J, Dąbrowska D (1997): Polyploidization of stigmatic papillae Bohdanowicz J, Dąbrowska D (1997): Polyploidization of stigmatic papillae in in Triglochin maritimumTriglochin maritimum L. (Juncaginaceae). Acta Biol Crac Ser Bot 39: 63-67. L. (Juncaginaceae). Acta Biol Crac Ser Bot 39: 63-67.

The process of multiplication of basic nuclear DNA content in The process of multiplication of basic nuclear DNA content in secretory cells of the stigma has not been described previously.secretory cells of the stigma has not been described previously.

Experimental embryology: Experimental embryology: Induction of autonomous endosperm developmentInduction of autonomous endosperm development

Experimental embryology: Experimental embryology: Induction of autonomous endosperm developmentInduction of autonomous endosperm development

Endosperm is a very important Endosperm is a very important plant tissue, the main source of plant tissue, the main source of nutrition for the developing nutrition for the developing embryo and germinating seed.embryo and germinating seed.

The endosperm usually The endosperm usually develops from the fertilized develops from the fertilized central cell of the embryo sac.central cell of the embryo sac.

Experimental embryology: Experimental embryology: Induction of autonomous endosperm developmentInduction of autonomous endosperm development

Rojek J, Kuta E, Przywara L (2002): Autonomous endosperm development in Rojek J, Kuta E, Przywara L (2002): Autonomous endosperm development in unpollinated ovaries of unpollinated ovaries of Brassica napus Brassica napus L. cv. Topas cultured in vitro. Acta Biol L. cv. Topas cultured in vitro. Acta Biol Crac Ser Bot 44: 195-202.Crac Ser Bot 44: 195-202. [in cooperation with Dept. of Plant Cytology and Embryology, Jagiellonian University, Cracow][in cooperation with Dept. of Plant Cytology and Embryology, Jagiellonian University, Cracow]

In this studyIn this study, , using phytohormousing phytohormo--nes in different concentrations nes in different concentrations and combinationsand combinations, we induced , we induced the development of autonomous the development of autonomous endosperm in culture of unpolli-endosperm in culture of unpolli-nated ovaries of nated ovaries of Brassica napusBrassica napusL. cv. TopasL. cv. Topas

Rojek J, Kuta E, Przywara L (2002): Autonomous endosperm development in Rojek J, Kuta E, Przywara L (2002): Autonomous endosperm development in unpollinated ovaries of unpollinated ovaries of Brassica napus Brassica napus L. cv. Topas cultured in vitro. Acta Biol L. cv. Topas cultured in vitro. Acta Biol Crac Ser Bot 44: 195-202.Crac Ser Bot 44: 195-202. [in cooperation with Dept. of Plant Cytology and Embryology, Jagiellonian University, Cracow][in cooperation with Dept. of Plant Cytology and Embryology, Jagiellonian University, Cracow]

In this studyIn this study, , using phytohormousing phytohormo--nes in different concentrations nes in different concentrations and combinationsand combinations, we induced , we induced the development of autonomous the development of autonomous endosperm in culture of unpolli-endosperm in culture of unpolli-nated ovaries of nated ovaries of Brassica napusBrassica napusL. cv. TopasL. cv. Topas