Dorota Juchno

Ploidy-dependent survival of progeny arising from crosses between natural allotriploid Cobitis females and diploid C. taenia males (Pisces, Cobitidae)

Crosses between 21 triploid hybrid Cobitis females and 19 C. taenia (2n = 48) males led to viable progeny; whereas no embryonic development was observed in crosses with tetraploid males (4n = 98). The ploidy status of 491 progenies randomly selected with flow cytometry (316) or chromosome analysis (175) revealed an average of 55.2 % triploids and 44.8 % tetraploids, but the ratio of 3n versus 4n fish did change during development. In the first 2 days after hatching, approximately 65.1 % of tetraploid larvae were observed. Their number decreased significantly to 30.8 and 6.2 % on average during 2–5 and 10–15 months of life, respectively. The karyotype of tetraploid progeny (4n = 98) included 3n = 74 chromosomes of the parental female and n = 24 of C. taenia male. The number of tetraploid progeny indicated indirectly that about 66 % of eggs from 3n females were fertilized with C. taenia. The rest of the eggs developed clonally via gynogenesis or hemiclonally via hybridogenesis into triploids of the same karyotype structure as parental females. We have documented for the first time that (at least under experimental conditions) tetraploids are commonly formed, but are less viable than triploids, and a ratio similar to what is found under natural conditions is finally attained. The current explanation concerning the ploidy and karyotype structure of the progeny confirms that the eggs of 3n Cobitis females are not only capable of maintaining all chromosomes but are also capable of incorporating the sperm genome, thus creating the potential to produce tetraploids.

Fecundity of the spined loach, Cobitis taenia (Pisces, Cobitidae) and natural allopolyploids of Cobitis from a diploid-polyploid population

Abstract. A comparison was made between the absolute and actual fecundity of C. taenia and allotriploid Cobitis females from a diploid-polyploid population inhabiting the Bug River, of Vistula River basin in Poland. All specimens were measured, weighed and aged. The absolute fecundity was determined by a gravimetric method using 29 ovaries (three of C. taenia, 25 of triploids and one of tetraploid). The actual fecundity was estimated according to the method adopted by Halačka et al. (2000) and used five ovaries of C. taenia and 40 of Cobitis triploids. Absolute fecundity of C. taenia under analysis ranged from 1 819 to 3 302 eggs and equaled 2 487 on average. It was significantly lower than absolute fecundity of an exclusively diploid population in Klawój Lake described previously (Juchno & Boroń 2006a). Absolute fecundity of triploid females ranged from 285 to 3 710, with an average of 1 577 eggs, whereas the fecundity of tetraploid Cobitis female was low, with only 882 eggs. The highest actual fecundity of C. taenia as well as of triploids was observed after they laid their first batch of eggs.

Molecular cytogenetic analysis of the crucian carp, Carassius carassius (Linnaeus, 1758) (Teleostei, Cyprinidae), using chromosome staining and fluorescence in situ hybridisation with rDNA probes

Abstract The crucian carp Carassius carassius (Linnaeus, 1758) is a species with restricted and decreasing distribution in Europe. Six males and six females of the species from the Baltic Sea basin in Poland were examined to show sequentially CMA3/AgNO3 staining pattern, DAPI staining, and, for the first time in literature, molecular cytogenetic analysis using double-colour fluorescence in situ hybridisation (FISH) with 28S and 5S rDNA probes. The karyotype consisted of 20 m, 36 sm and 44 sta chromosomes, NF=156. The AgNO3 stained NORs were most frequently located terminally in the short arms of two sm and two sta elements, and CMA3-positive sites were also observed suggesting abundant GC-rich repetitive DNA in the regions. Other CMA3-positive sites in the short arms of six to ten sm and sta chromosomes were detected. The results based on 28S rDNA FISH confirmed the location of rDNA sites. DAPI-negative staining of NORs suggested the scarcity of AT-rich DNA in the regions. FISH with 5S rDNA probe revealed 8–14 loci (ten and 12 in respectively 49 and 29% of metaphases). They were located in two sm and eight to ten sta chromosomes and six of them were larger than others. Simultaneously, mapping of the two rDNA families on the chromosomes of C. carassius revealed that both 28S and 5S rDNA probes were located in different chromosomes. Molecular cytogenetic data of C. carassius presented here for the first time give an important insight into the structure of chromosomes of this polyploid and declining species and may be useful in its systematics.

Comparison of egg and offspring size of karyologically identified spined loach, Cobitis taenia L., and hybrid triploid Cobitis females (Pisces, Cobitidae)

Abstract The aim of this study was to determine and compare for the first time the size of eggs laid by C. taenia (2n=48) and its naturally occurring triploid (3n=74) hybrid females and the total length of offspring of these females obtained from experimental crosses, in the period from hatch to 22 days post hatch. The eggs laid by triploid females had an average diameter of 1.18mmand were significantly larger than the eggs spawned by C. taenia females, which averaged 0.93 mm in diameter. The size of the triploid female eggs was associated with their level of ploidy. Similarly, the average total length of triploid female progeny from one to ten days post hatch was statistically significantly higher than the average total length of C. taenia offspring in the same period of life. After two weeks, the offspring of diploid and triploid females reached average total lengths of 13.27 mm and 13.60 mm, respectively, which was not a significant difference. The possible causes of triploid Cobitis female domination in diploid-polyploid populations in terms of egg size and other traits associated with polyploid functioning was also investigated.

Evidence of the sterility of allotetraploid Cobitis loaches (Teleostei, Cobitidae) using testes ultrastructure

The diploid-polyploid populations of Cobitis distributed in Poland are usually composed of the spined loach Cobitis taenia or, less often, the Danubian loach C. elongatoides and their triploid (females) and tetraploid hybrids (females and males). The aim of this study was to determine whether tetraploid males participate in the reproduction process by analyzing their testis ultrastructure and the process of spermatogenesis in comparison with diploid males of both parental species. Tetraploid loaches were obtained from three different diploid-polyploid populations distributed in Poland. The structure of Cobitis testes are typical for most Teleostei fish with cystic-type spermatogenesis. The successive stages of developing germ cells are enclosed within cysts formed by the Sertoli cells. This paper morphologically describes the different germ cell stages of spermatogenesis (spermatogonia, spermatocytes, spermatids, and spermatozoa) of C. taenia and C. elongatoides and provides a pioneering ultrastructural analysis of tetraploid Cobitis testes which reveals their unusual structure for the first time. Thus, cysts with normal spermatogonia and spermatocytes (pachyten or leptoten stages) containing synaptonemal complexes were present and no spermatids or spermatozoa were observed. Moreover, in contrast to previously analyzed diploid species, single cells or all of the cells within the cysts displayed chromatin condensation and/or chromatin fragmentation. The obtained results clearly demonstrated that tetraploid males are sterile and diploids are fertile and are the only sperm donors in the reproduction processes of diploid-polyploid Cobitis populations.

Reproduction and fecundity of the golden loach, Sabanejewia baltica Witkowski, 1994 from Bug River in Poland

In this paper we report for the first time the results of histology of the golden loach (S. baltica) gonads (25 females and 8 males) and absolute fecundity of females from the Bug River during the reproductive season. The golden loach has an asynchronous ovary and spawns in batches. The absolute fecundity of the golden loach ranged from 1507 to 7220 eggs (3050±1377). We hypothesize that the golden loach spawns twice a year.

Histological evidence that diploid hybrids of Cobitis taenia and C. elongatoides (Teleostei, Cobitidae) develop into fertile females and sterile males

In Europe, two bisexual fish species, Cobitis taenia (TT) and Cobitis elongatoides (EE), hybridized, producing diploid and polyploid clonal lineages. This study compared, for the first time, embryonic development, hatching success, sex ratio, body size and as well as reproductive ability based on the gonad histology of F1 reciprocal diploid hybrids (TE, ET) of both species. Hybrid F1 progeny showed the same proper pattern of embryonic and larval development. Among TE and ET offspring, slightly more females and males, respectively, occurred, but sex parity among 18-month-old hybrids was observed. Two- and three-year-old F1 hybrid females were mature, possessing all stages of oogenesis in their ovaries. Females (TE) back-crossed with C. taenia males had properly developing progeny. In testes of two- and three-year-old F1 hybrids, only early stages of spermatogenesis and pyknotic cells indicating the degeneration process were observed, but they exhibited an external feature (lamina circularis) of maturation. The results were confirmed by the structure of gonads that two sexual species, C. taenia and C. elongatoides, hybridize, producing F1 progeny of sterile males and fertile females. Hybrid females may participate in subsequent steps of speciation via hybridization and polyploidy or, on the contrary, represent an element of new species isolation.