Comparative cytogenetics and chromosomal characteristics of ribosomal DNA in the fish genus Vimba (Cyprinidae)

Karyotypic and cytogenetic characteristics of Vimba vimba and V. elongata were investigated using differential staining techniques (sequential C-banding, Ag- and CMA₃-staining) and fluorescent in situ hybridization (FISH) with 28S rDNA probe. The diploid chromosome number in both species was 2n = 50 with 8 pairs of metacentrics, 14 pairs of submetacentrics to subtelocentrics and 3 pairs of subtelo- to acrocentrics. The largest chromosome pair of the complements was characteristically subtelo- to acrocentric. The nucleolar organizer regions (NORs) in both species were detected in the telomeres of a single, middle-sized subtelocentric chromosome pair, a pattern common in a number of other Leuciscinae. FISH with rDNA probe produced consistently positive hybridization signals detected in the same regions indicated by Ag-staining and CMA₃-fluorescence. The distribution of C-positive heterochromatin was identical in both species, including a conspicuous size polymorphism of heterochromatic blocks in the largest metacentric and subtelo- to acrocentric chromosomal pairs. No heteromorphic sex chromosomes were detected. A single analyzed individual of V. melanops possessed the same karyotype and NOR phenotype as V. vimba and V. elongata. The apparent karyotype homogeneity and chromosomal characteristics of ribosomal DNA in all three species of the genus Vimba is consistent to that found in most other representatives of the European leuciscine cyprinid fishes.     

Cytogenetic characterization of six species of flatfishes with comments to karyotype differentiation patterns in Pleuronectiformes (Teleostei)

The karyotypes and cytogenetic characteristics of flatfishes species Paralichthys orbignyanus , Paralichthys patagonicus , Citarichthys spilopterus and Etropus crossotus (Paralichthyidae), Bothus ocellatus (Bothidae) and Symphurus tessellatus (Cynoglossidae) were investigated by conventional [Giemsa staining, C-banding, Ag- and chromomycin (CMA₃)-stainings] and molecular [ in situ hybridization (ISH)] cytogenetic techniques. The results showed 2n = 46 and FN = 48 (2msm + 46sta) in P. orbignyanus , 2n = 46 and FN = 46 (46sta) in P. patagonicus , 2n = 26 and FN = 44 (18msm + 8sta) in C. spilopterus , 2n = 38 and FN = 64 (26msm + 12sta) in E. crossotus , 2n = 32 and FN = 50 (18msm + 14sta) in B. ocellatus , and 2n = 46 and FN = 62 (46msm + 62sta) in S. tessellatus . All species exhibited weak C-band positive segments in terminal and centromeric positions of some chromosome pairs. Silver staining of the nucleolus organizer regions (Ag-NOR) technique showed a single Ag-NOR-bearing chromosome pair in all species except E. crossotus . All these sites were CMA₃ positive and showed clear ISH signals after probing with a 18S rRNA probe. Etropus crossotus presented until seven chromosomes with Ag-NORs and CMA₃ positively stained segments in five chromosome pairs. Conversely only one chromosome pair was identified with the ISH experiments in this species. The available results show that the fishes of the order Pleuronectiformes experienced a marked chromosome evolution that included reduction in diploid number, mainly due to Robertsonian rearrangements, and several chromosome inversions.     

Extensive polymorphism and chromosomal characteristics of ribosomal DNA in a loach fish, Cobitis vardarensis (Ostariophysi, Cobitidae) detected by different banding techniques and fluorescence in situ hybridization (FISH)

When surveying the karyotype diversity of European loaches of the genus Cobitis to identify species involved in hybrid polyploid complexes, an extensive polymorphism in number and location of NORs was discovered in C. vardarensis using Ag-staining, C-banding, CMA₃-fluorescence and fluorescence in situ hybridization (FISH). This species had 2n=50, the karyotype contained 13 pairs of metacentric, 10 pairs of submetacentric and two pairs of subtelocentric chromosomes. The NOR-bearing chromosomes included one medium-sized metacentric pair with a large CMA₃-positive heterochromatic pericentromeric block, one small metacentric as well as one large submetacentric pairs. Ribosomal sites were always located in telomeres of these chromosomes. Each of the pair of NOR-bearing chromosomes occurred in three variants – (1) presence and/or (2) absence of NORs on both homologues and (3) heterozygous combination where only one of the homologues bears NORs. Altogether, 10 different NOR cytotypes from 27 theoretically possible ones were discovered among 20 indviduals examined. The number of NORs ranged from two to five per specimen. The results regarding the number and locations of NORs as revealed by banding techniques were confirmed using FISH with rDNA probe. NOR sites were of CMA₃-positive, suggesting that ribosomal sites are associated with GC-rich DNA. Very similar structural polymorphism with multiple NORs is expressed in the Danubian loach C. elongatoides indicating a close relationship between both species.     

Developmental morphology of the Indian cyprinid fish Barilius canarensis

Embryonic and larval development of an Indian cyprinid fish, Barilius canarensis, is described from laboratory-reared specimens. The eggs, measuring 2.1–2.4 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk without oil globules. Hatching occurred 39–45 h after fertilization at 26.8°–27.4°C. The newly hatched larvae, measuring 4.8–5.1 mm in body length (BL) with 22 + 17 = 39 myomeres, were characterized by melanophores already deposited on the eyes. The eggs of B. canarensis resembled those of the related danionin species Candidia barbatus, Opsariichthys uncirostris uncirostris, Zacco platypus, Z. sieboldii, and Z. temminckii. Although the larvae of B. canarensis were also similar to those of the foregoing species in general morphology, they differed in having a straight notochord tip and pigmentation on the eyes at hatching and the almost entire absence of melanophores on the ventral body surface from the yolk sac to postflexion larval stages. Conversely, melanophores occurred on the anterior abdominal and pericardial cavities from the preflexion to postflexion larval stages.     

Developmental morphology of the cyprinid fish Tanichthys albonubes

Embryonic, larval, and juvenile development of a small cyprinid species, Tanichthys albonubes, is described from laboratory-reared specimens. The eggs, measuring 1.0–1.2 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yolk without oil globules. Hatching occurred 45–53 h after fertilization at 25.5°–26.9°C. The newly hatched larvae, measuring 2.2–2.6 mm in body length (BL), had melanophores on the head and body. In particular, a dark vertical streak occurring posterior to the otic capsule and melanophores above the eyes were distinctive. The yolk was completely absorbed at 3.4 mm BL. Notochord flexion was initiated at 5.0 mm BL and finished at 6.0 mm BL. Aggregate numbers of all fin rays were completed at 11 mm BL. Squamation was initiated at 8.4 mm BL and completed at 13 mm BL. Although the eggs of T. albonubes resembled those of other small danionin species, including Aphyocypris chinensis, Chela dadiburjori, Danio rerio, Devario malabaricus, Gobiocypris rarus, Hemigrammocypris rasborella, and Horadandia atukorali, they differed from those of A. chinensis, C. dadiburjori, G. rarus, and Horadandia atukorali in having a wider perivitelline space. The larvae and juveniles of T. albonubes were similar to those of the aforementioned seven species plus Danio albolineatus, Danio kerri, and Devario sp. (cf. D. aequipinnatus) in general morphology. However, the early life stage morphology of T. albonubes differed from them in having a dark vertical streak posterior to the otic capsule and melanophores above the eyes in the yolk sac larval stage, and a dark lateral streak with an unpigmented area just above the former on the body, a dark blotch on the caudal fin, and reddish dorsal, anal, and caudal fins during the postflexion larval and juvenile stages.     

铁线莲属21个类群的染色体核型分析

铁线莲属(Clematis L.)为毛茛科大属之一,具有重要的园艺与药用价值。为探讨铁线莲属植物染色体组演化规律,揭示属下组间及种间的亲缘关系,该研究采用常规压片法对铁线莲属21个类群的根尖进行处理、压片,对染色体的形态特征进行观察及核型分析,同时利用Ward联接法进行聚类分析研究。狭裂太行铁线莲、毛果扬子铁线莲、卷萼铁线莲、中印铁线莲、钝萼铁线莲的染色体形态为首次报道。结果表明:铁线莲属21个类群均为二倍体,染色体数均为16(2n=2x=16),除中印铁线莲外其他类群均具随体;长瓣铁线莲、钝萼铁线莲、芹叶铁线莲、褐毛铁线莲、C.flammula、毛果扬子铁线莲的染色体为“2B”型,其他类群染色体为“2A”型;铁线莲属核型不对称系数在60.29%~63.79%之间;铁线莲属植物染色体组表现的较为原始,种间核型存在广泛变异。综上结果表明,铁线莲属植物染色体数目应由二倍体向多倍体演化后通过多倍体的二倍化过程产生非整倍体方向演化。铁线莲属染色体的演化主要在二倍体水平上进行,通过产生染色体结构变异的方式实现,通过产生杂合染色体、加强核型不对称性、染色体类型改变以及随体染色体的变化四种途径进化。同时,核型特征在分组水平与物种水平上的划分与传统分类基本一致,说明核型分析可为铁线莲属下组一级分类提供一定的线索。该研究结果为铁线莲属植物系统分类、遗传演化与资源利用等研究领域提供了新的参考资料。     

Developmental morphology of the cyprinid fish Chela dadiburjori

Embryonic, larval, and juvenile development of an Indian cyprinid fish, Chela dadiburjori, is described from laboratory-reared specimens. The eggs, measuring 0.7–0.9mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk and no oil globule. Hatching occurred 50–61h after fertilization at ca. 27°C. The newly hatched larvae, measuring 2.4–2.6mm in body length (BL), had melanophores on the body with 14–16+14–17=29–31 myomeres. Two dark transverse bands on the ventral body surface and one melanophore on the lower margin of the eye in newly hatched larvae were diagnostic. Additionally, a cement organ for adhering to objects was present on the forehead of yolk sac larvae <3.1mm BL. The yolk was completely absorbed at 3.5mm BL. Notochord flexion was initiated at 5.0mm BL and finished at 6.0mm BL. Aggregate numbers of all fin rays were completed at 9.2mm BL. Squamation was initiated on the caudal peduncle at 8.0mm BL and completed at 10mm BL. The eggs of C. dadiburjori resembled those of the closely related species Devario malabaricus and Danio rerio. The larvae and juveniles of C. dadiburjori were also similar to those of the latter species in general morphology, especially the presence of body melanophores in newly hatched individuals and a distinctive lateral streak on the head during the period from yolk sac to postflexion larvae. However, early yolk sac larvae of C. dadiburjori were more similar to those of Devario malabaricus than Danio rerio in having a cement organ on the forehead. Larvae and juveniles of C. dadiburjori differed from those of the latter two species in pigmentation on the ventral body surface at hatching and around the mouth during the period from preflexion to early postflexion larvae and in having a dark lateral streak or band on the body in postflexion larvae and juveniles.     

Note on the banded karyotype of the enigmatic South Asian loach <Emphasis Type="Italic">Vaillantella </Emphasis><Emphasis Type="Italic">maassi</Emphasis> (Cypriniformes: Vaillantellidae)

The karyotype of Vaillantella maassi, a loach with an unusually long dorsal fin, was investigated using sequential CMA₃ and AgNO₃ staining to test if this character evolved in parallel with Vaillantella’s morphological distinctness. The karyotype was 2n = 50 with 13 pairs of metacentric-submetacentric and 12 pairs of subtelocentric chromosomes. Sequential CMA₃ banding and AgNO₃ staining evidenced NORs in the short arms of pair no. 2. Additional CMA₃-positive, Ag-negative signals were present in the short arms of pairs 14 and 15. The karyotype paralleled those of most other cobitoid loaches studied so far and is apparently not associated with the morphological distinctness of Vaillantella.     

Developmental morphology of the cyprinid fish Horadandia atukorali

Embryonic, larval, and juvenile development of a small Indian cyprinid, Horadandia atukorali, is described from laboratory-reared specimens. The eggs, measuring 0.7–0.8mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk without oil globules. Hatching occurred 47–54h after fertilization at 26.3–27.5°C. The newly hatched larvae, measuring 2.3–2.6mm in body length (BL) with 16+13=29 myomeres, had no melanophores, except on the eye, a single melanophore occurring on the lower margin, and xanthophores surrounding the pupil. The yolk was completely absorbed at 3.0mm BL. Notochord flexion was initiated at 4.0mm BL and finished at 4.4mm BL. Aggregate numbers of all fin rays were completed at 8.0mm BL. Squamation was initiated at 6.4mm BL and completed at 9.5mm BL. Although the eggs of Horadandia atukorali resembled those of other small danionin species, including Aphyocypris chinensis, Chela dadiburjori, Danio rerio, Devario malabaricus, and Hemigrammocypris rasborella, they differed from those of Danio rerio and Devario malabaricus in having a narrower perivitelline space. The larvae and juveniles of Horadandia atukorali were also similar to those of the latter five species in general morphology, especially in the presence of a melanophore on the lower margin of the eye at hatching, as in C. dadiburjori. However, the early life stage morphology of Horadandia atukorali differed from the other danionin species in having a conical yolk sac at hatching, no cement organ on the forehead in the yolk-sac larval stage, a divided gas bladder in the flexion larval stage, two dark lateral streaks on the head and chevron-like melanophores on the ventral body surface from the preflexion to postflexion larval stages, and xanthophores on the eyes at hatching.     

Developmental morphology of the cyprinid fish Inlecypris auropurpureus

Embryonic, larval, and juvenile development of a Myanmarese cyprinid fish, Inlecypris auropurpureus, is described from laboratory-reared specimens. The eggs, measuring 0.9–1.0 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk without oil globules. Hatching occurred 49–56 h after fertilization at 26.2°–27.3°C. The newly hatched larvae, measuring 2.9–3.1 mm in body length (BL) with 17 + 19–20 = 36–37 myomeres, had melanophores on the head and body. A cement organ on the forehead for adhering to objects during the yolk sac and early preflexion larval stages was distinctive. The yolk was completely absorbed at 3.6–4.0 mm BL. Notochord flexion was initiated at 5.1–5.6 mm BL and finished at 7.1 mm BL. Aggregate numbers of all fin rays were completed at 14 mm BL. Squamation was initiated midlaterally on the anterior trunk at 14 mm BL and completed at 27 mm BL. Although the eggs of I. auropurpureus resembled those of the closely related species Chela dadiburjori, Danio rerio, and Devario malabaricus, they differed from those of Danio rerio and Devario malabaricus in having a narrower perivitelline space. The larvae and juveniles of I. auropurpureus were also similar to those of C. dadiburjori, Danio rerio, and Devario malabaricus in general morphology, but they differed from the latter three species in having a series of dark blotches laterally on the body in the juvenile stage. Moreover, I. auropurpureus differed from C. dadiburjori in having more myomeres and a near-single row of melanophores on the body along the dorsal midline from the yolk-sac to early postflexion larval stages, from Danio rerio in having a cement organ on the forehead during the yolk-sac and early preflexion larvae, and a single melanophore on the lower eye margin in the early yolk-sac larvae, and from Devario malabaricus in having a single melanophore on the lower eye margin in the early yolk-sac larvae. The presence of a cement organ on the forehead indicates a close relationship among the genera Inlecypris, Chela, and Devario.     

The phylogenetic position of Apterosperma (Theaceae) based on morphological and karyotype characters

Classifications of Theaceae have usually placed the endangered monotypic genus Apterosperma in tribe Schimeae (x=18), whereas recent molecular phylogenetic evidence supports its transfer to tribe Theeae (x=15). Molecular data have not resolved the phylogenetic position of Apterosperma within Theeae. We investigated the chromosome number and karyotype of Apterosperma in the context of molecular and morphological phylogenetic evidence to provide further insight into the placement of Apterosperma within Theaceae. The chromosome number and karyotype was found to be 2n = 30 = 26m + 4sm, consistent with the transfer of Apterosperma to tribe Theeae. When the chromosome data were incorporated into a data set of 46 other nonmolecular characters, Apterosperma was placed as the first-diverging lineage within the clade comprising tribe Theeae. This supports its placement based on molecular data. The low intrachromosomal asymmetry (type 1A) of Apterosperma, presumably ancestral for the family, is also consistent with this placement. Character optimization strongly supports a base chromosome number of x=15 for tribe Theeae. Because of variable and sometimes conflicting chromosome count reports of species in tribes Schimeae and Stewartieae, the base chromosome number of Theaceae could be either x=15 or 17.     

Molecular cytogenetics of <Emphasis Type="Italic">Oligosarcus hepsetus</Emphasis> (Teleostei,Characiformes) from two Brazilian locations

Karyotype and cytogenetic markers of Oligosarcus hepsetus from two Brazilian locations in the Paraíba do Sul River Basin (Brazil) were investigated using differential staining techniques (C-banding, silver (Ag)- and chromomycin A₃ (CMA₃)-staining) and fluorescent in situ hybridization (FISH) using 18 S rDNA and 5 S rDNA probes. The diploid chromosome number was invariably 2n = 50 with 3 pairs of metacentric, 5 pairs of submetacentric, 8 pairs of subtelocentric and 9 pairs of acrocentric chromosomes. No heteromorphic sex chromosomes were observed. The nucleolar organizer regions (NORs) were detected in the short arms of the largest acrocentric pair using Ag-, CMA₃- stainings and FISH with 18 S rDNA probe, the latter showing also positive labeling in the short arms of a small acrocentric pair, not visualized by the former methods. FISH with 5 S rDNA probe showed positive labeling in the two chromosome pairs. While the CMA₃-staining exhibited GC-rich heterochromatin segments in two pairs of chromosomes, including those coincided with Ag-NORs, the DAPI staining did not reveal any signal, indicating the absence of AT-rich heterochromatin. FISH with an As-51 satellite DNA probe derived from the closely related Astyanax scabripinnis did not reveal any positive signal, demonstrating the absence of this class of DNA in the genome of the specimens under study.     

Meiosis,SC-formation,and karyotype structure in diploidPaeonia tenuifolia and tetraploidP. officinalis

The meiosis of the diploidPaeonia tenuifola and the allotetraploidP. officinalis was studied after conventional methanol/acetic acid-fixation and synaptonemal complex (SC) spreading. Meiosis inP. tenuifolia (2n = 10) is normal with five bivalents in metaphase I, and the SCs in pachytene show regular features. InP. officinalis (2n = 4x = 20) univalents, bivalents and multivalents are found in metaphase I. The SCs reveal several abnormalities: a high number of unpaired lateral elements, partner exchanges between three and four lateral elements, loops and lateral element thickenings. These characteristics are compared with the situations found in other polyploid and hybrid species. It is noteworthy that the abnormalities in meiosis ofP. officinalis are not reflected in its somatic karyotype. Its features were analysed after silver staining and fluorescent staining with chromomycin and compared with those ofP. tenuifolia. Synaptonemal Complex Spreading in Plants2; for part1 see Pl. Syst. Evol.154, 129–136 (1986).     

The karyotype ofLimnobium spongia (Hydrocharitaceae)

The somatic chromosomes ofLimnobium spongia have been studied for the first time. Its number is 2n = 24; the karyotype is composed of 5 m pairs + 3 sm pairs + 4 st pairs. Pairs 3 and 11 bear microsatellites. Results are compared with previous chromosome reports for the family, mainly as regards the taxonomic status ofLimnobium versusHydromystria; our findings seem to support the point of view of those who favour the independence of both generic entities.The first author is member of the Carrera del Investigador (CONICET, Argentina) and the second one fellowship from the same institution.     

Banded karyotype of spined loach Cobitis taenia and triploid Cobitis from Poland

The present work provides new data on the banding pattern of diploid Cobitis taenia and its triploid hybrid females, which belong to the diploid–polyploid complex in the Vistula River tributary. C-banding, silver-staining (Ag), and fluorescent staining with chromomycin A₃ techniques were used to describe the diploid and triploid karyotype. The karyotype of Cobitis taenia of 2n=48 was characterised by one pair of NOR-bearing subtelocentric chromosomes and at least four chromosomes with CMA₃-positive sites. The C-positive heterochromatin was present in the centromeres of almost all chromosomes and the pericentromeric regions of several metacentric and submetacentric chromosomes. The triploid females of 3n=74 had two pairs of chromosomes with active NORs. The NORs-sites were located terminally on two biarmed and two uniarmed chromosomes. The CMA₃-staining revealed at least six A₃-positive sites. The C-banded and A₃-stained triploid karyotype was composed of haploid set of Cobitis taenia and diploid set of unidentified species, so heterochromatin pattern confirmed the possibility of their hybrid origin. The characteristics of banded diploid and triploid karyotype, and the hypothetical karyotype of an unknown species of 2n=50 is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Replication banding patterns in the spined loach,Cobitis taenia L. (Pisces,Cobitidae)

The chromosomal complement of Cobitis taenia was analysed by replication banding techniques to determine whether there were specific patterns that could allow distinction of the different chromosomes. The diploid chromosome number of 2n = 48 is diagnostic of this species. In vivo 5-bromodeoxyuridine (5-BrdU) incorporation induced highly reproducible replication bands. Most of the chromosome pairs were distinguishable on the base of their banding patterns. The karyotype, consisting of five pairs of metacentrics, nine pairs of submetacentrics and 10 pairs of subtelocentrics and acrocentrics, was confirmed. C-banding and replication banding patterns were compared, and heterochromatin was both early and later replicating. C-positive heterochromatin in centromeric regions was mainly early replicating, but that located in pericentromeric regions was late replicating. Most of the late-replicating regions found interstitially were C-band negative. The results obtained so far for combined chromosomal staining methods of C. taenia and other Cobitis fish species are discussed.     

铺道蚁的染色体核型分析

以商丘地区的铺道蚁Tetramorium caespitum(L.)早期胚胎为材料,采用低渗处理—烘干法制备染色体标本,对其染色体组型进行研究。结果表明,铺道蚁染色体组型为n=22,2n=44,是由10条中央着丝粒染色体,3条亚中着丝粒染色体和9条亚端着丝粒染色体组成。     

A contribution to the study of the food of six cyprinid fishes in three areas of the Lake Victoria basin,East Africa

An account of the food of six species of cyprinid fish from the Lake Victoria basin is given. All the species studied possess tubular stomachs and analysis was carried out on entire guts, the relative importance of the various dietary items being assessed by the frequency occurrence method. Results indicate that these species are omnivorous, and their food ranges from debris, diatoms through algae, insect larvae to molluscs. Comparison of the food of Barbus spp. from the lake and river shows that riverine fish ingest more plant material than lake fish.