Molecular cytogenetics of the equidae. II. purification and cytological localization of a (G + C)-rich satellite DNA from Equus hemionus onager and cross-species hybridization to E. asinus chromosomes

A (G + C)-rich satellite DNA component (p = 1.716 g/ml) has been fractionated from the total DNA of the Iranian subspecies of the Asiatic wild ass, Equus hemionus onager, by successive dactinomycin-CsCl and netropsin sulfate-CsCl isopycnic gradients. Complementary 3H-RNA (cRNA) transcribed from the satellite DNA hybridized predominantly to the centromeric and telomeric constitutive heterochromatic regions of onager chromosomes. These studies have suggested that satellite DNA's with similar sequences are present in the centromeric, as well as telomeric, heterochromatic regions of some onager chromosomes. The centromeric region of the fusion metacentric t(23;24) of the onager is deficient in sequences homologous to the onager 1.716 g/ml satellite DNA, indicating a loss of satellite DNA during fusion or an amplification of the satellite DNA in the centromeric regions of the acrocentric chromosomes 23 and 24 subsequent to fission. Sequences complementary to onager 1.716 g/ml satellite DNA show extensive hybridization to the constitutive heterochromatin of the feral donkey (E. asinus) karyotype, consistent with a view of conservation and amplification of similar or identical sequences in the two species.     

Homologous fission event(s) implicated for chromosomal polymorphisms among five species in the genus Equus

The genus Equus is unusual in that five of the ten extant species have documented centric fission (Robertsonian translocation) polymorphisms within their populations, namely E. hemionus onager, E. hemionus kulan, E. kiang, E. africanus somaliensis, and E. quagga burchelli. Here we report evidence that the polymorphism involves the same homologous chromosome segments in each species, and that these chromosome segments have homology to human chromosome 4 (HSA4). Bacterial artificial chromosome clones containing equine genes SMARCA5 (ECA2q21 homologue to HSA4q31. 21) and UCHL1 (ECA3q22 homologue to HSA4p13) were mapped to a single metacentric chromosome and two unpaired acrocentrics by FISH mapping for individuals possessing odd numbers of chromosomes. These data suggest that the polymorphism is either ancient and conserved within the genus or has occurred recently and independently within each species. Since these species are separated by 1-3 million years of evolution, this polymorphism is remarkable and worthy of further investigations.     

Mitochondrial DNA evolution in the genus Equus

Employing mitochondrial DNA (mtDNA) restriction-endonuclease maps as the basis of comparison, we have investigated the evolutionary affinities of the seven species generally recognized as the genus Equus. Individual species' cleavage maps contained an average of 60 cleavage sites for 16 enzymes, of which 29 were invariant for all species. Based on an average divergence rate of 2%/Myr, the variation between species supports a divergence of extant lineages from a common ancestor approximately 3.9 Myr before the present. Comparisons of cleavage maps between Equus przewalskii (Mongolian wild horse) and E. caballus (domestic horse) yielded estimates of nucleotide sequence divergence ranging from 0.27% to 0.41%. This range was due to intraspecific variation, which was noted only for E. caballus. For pairwise comparisons within this family, estimates of sequence divergence ranged from 0% (E. hemionus onager vs. E. h. kulan) to 7.8% (E. przewalskii vs. E. h. onager). Trees constructed according to the parsimony principle, on the basis of 31 phylogenetically informative restriction sites, indicate that the three extant zebra species represent a monophyletic group with E. grevyi and E. burchelli antiquorum diverging most recently. The phylogenetic relationships of E. africanus and E. hemionus remain enigmatic on the basis of the mtDNA analysis, although a recent divergence is unsupported.      

Chromosomal and molecular evolution in Asiatic wild asses

Chromosomal studies of Tibetan wild ass,Equus kiang holdereri are reported for the first time. A Robertsonian polymorphism resulting in diploid numbers of 2n=51 and 2n=52 was identified. This polymorphism involves the same elements that participate in the polymorphic Robertsonian rearrangement producing diploid numbers of 54–56 inE. hemionus kulan andE. hemionus onager. Comparison of mitochondrial DNA restriction fragment patterns among three unrelated kiang individuals and between kiangs and a Transcaspian kulan,E. hemionus kulan, indicates an approximately 1% nucleotide sequence divergence of mitochondrial DNAs between the two taxa. The chromosomal divergence betweenE. hemionus andE. kiang probably originated less than 500,000 years ago. Studies of additional Asiatic wild ass taxa are indicated in order to gain further insights into the speciation and systematics of this threatened group of mammals.     

Genetic characterization of free-ranging Asiatic wild ass in Central Asia as a basis for future conservation strategies

Loss of genetic diversity due to drift and inbreeding reduces a population’s ability to respond to environmental change and may result in inbreeding depression. The Asiatic wild ass (Equus hemionus), regionally also known as Gobi khulan, Turkmen kulan, or Persian onager, has become confined to less than 3% of its historic distribution range. Remaining populations in Central Asia outside of the Mongolian Gobi are small and fragmented. Questions concerning subpopulation status remain disputed and concerns over the viability of these populations have been raised because of small size, past bottlenecks, or recent founder events. We used non-invasive faecal samples to assess the genetic diversity and divergence among Turkmen kulan and Persian onager from five free-ranging and one captive population from Turkmenistan, Kazakhstan and Iran and compared their genetic constitution to the large autochthonous population in the Mongolian Gobi. We observed loss of genetic diversity (drift and inbreeding) in the captive and reintroduced populations as well as in one rapidly declining autochthonous population. Population differentiation and structure using microsatellites and mtDNA based phylogenetic analysis do not support the current separation of the autochthonous populations of Turkmen kulan and Persian onager into different subspecies, but rather suggest a cline with the Iranian population in Bahram-e-Goor at the southern end and the Turkmen population in Badhyz at the northern end falling into two distinct clusters, and the northern Iranian population in Touran being intermediate. We compare our findings to other population genetics studies of equids and discuss the implications of our findings for the future conservation of the Asiatic wild ass in the region.     

Variation in skin surface lipid composition among the Equidae

Skin surface lipids from Equus caballus, E. przewalskii, E. asinus, E. grevyi, E. hemionus onager and a mule (E. asinus/E. caballus) were analyzed in detail. In all species the surface lipid mixtures consisted of giant-ring lactones, cholesterol, cholesteryl esters and minor amounts of wax diesters. In E. caballus, the lactone hydroxyacids were entirely branched chained, while in E. asinus and E. grevyi they were almost exclusively straight chained. In E. przewalskii, the onager and the mule there were both straight and branched chain hydroxyacid lactones. These results are in harmony with published interpretations of the evolutionary relationships among Equus species.     

Chromosomal evolution in Cervidae

On the basis of chromosome data obtained on 30 species and 20 subspecies of Cervidae, a report is submitted on the karyosystematics of this family. The primitive karyotype of Cervidae may be inferred to be composed of 35 acrocentric pairs (2n = 70 FN = 70). During the phyletic evolution of this family different types of chromosome rearrangements were probably selected and the group may have differentiated karyologically into three branches: (1) the Cervinae that fixed a centric fusion resulting in a metacentric pair of autosomes (2n = 68, FN = 70), as shown by the basic karyotype of Cervus elaphus, and where Robertsonian fusions are the preeminent type of chromosome rearrangement; (2) the Odocoileinae, in which pericentric inversions and Robertsonian fusions were favored, yielding first a submetacentric X and then a submetacentric autosome pair. The most representative karyotype is 2n = 70, FN = 74--as in Odocoileus hemionus; and (3) the Muntiacinae, in which centric and tandem fusions were the most common chromosome rearrangements. While Muntiacus reevesi has a karyotype 2n = 46, FN = 46, the chromosome number drops down to 2n = 6 in the females of the M. muntjak vaginalis subspecies group and M. rooseveltorum. Therefore, while the karyotypes are conserved within the subfamilies Cervinae and Odocoileinae; the subfamily Muntiacinae appears to be the most chromosomally diversified group. The few karyological data on the Moschus berezovskii suggest that the Moschinae should be placed in a separate family, the Moschidae.     

Genetic and cytological characterisation of fusion chromosomes of Dictyostelium discoideum

Abnormally large chromosomes which appear to result from the fusion of 2 chromosomes of the normal karyotype have been found in diploids of Dictyostelium discoideum formed by parasexual fusion of haploid strains HU483 (n=7) and HU245 (n=7). These fusion chromosomes appear to be the products of the tandem translocation of most, if not all, of one acrocentric chromosome to the telomere of a second acrocentric. Thus the chromosome number of the diploids is reduced from the normal 2n=14 to 2n=13 with the formation of an abnormally large acrocentric fusion chromosome. Experimental haploidisation of such diploids results in two types of products, those with a normal 7 chromosome karyotype and those with an abnormal 6 chromosome karyotype which contains the fusion chromosome. Genetic analysis of haploid segregants indicates that linkage groups II and VII are involved in this fusion. Phenotypes of recombinant diploids obtained following mitotic crossing-over establishes that linkage group II is proximal to linkage group VII. Cytological examination of the karyotypes of haploid strains bearing the fusion chromosome suggest that chromosome 2 may correspond to linkage group II and chromosome 3 to linkage group VII. Haploid strains bearing the fusion chromosome grow and develop normally so little or no genetic information can have been lost in the fusion event. While the nature of this event is unknown it may have involved aberrant recombinational DNA repair since the parental haploid strain HU483 bears the radB13 DNA repair mutation.     

新疆独尾草属植物的核型分析

对新疆独尾草属（Eremurus）植物的核型进行了研究。核型公式如下：阿尔泰独尾草[E.altaicus（Pall.）Stev.]2n=2x=14=4m＋8sm＋2st;异翅独尾草[E.anisopterus（Kar.et Kit）Regel]2n=4x=28=4m＋4sm＋20st;粗柄独尾草[E.inderiensis （M.Bieb）Regel]2n=2x=14=10sm＋4st,首次发现古尔班通古特沙漠南缘所产的异翅独尾草2n=4x=28,与前人报道其为二倍体2n=2x=14的结果不一致。     

The use of endobiotic ciliates from old collections in transmission electron microscopic investigations

The opportunity of the use of formalin-fixed endobiotic ciliates from old collections in transmission electron microscopic investigations (TEM) has been studied. Ciliates from the following species were examined: Ditoxum funinucleum Gassovsky, 1919 from the hindgut of Equus hemionus kulan Groves et Mazak, 1967 preserved in a collection during 19 years, Blepharoprosthium pireum Bundle, 1895 and Cochliatoxum periachtum Gassovsky, 1919 from the hindgut of the Yakut horse Equus caballus L. stored during 1.5 years, and Triplumaria heterofasciculata Timoshenko et Imai, 1995 from faeces of the Asian elephant Elephas maximus L. stored during 2.5 years. It is shown, that the main taxonomically important characters of the cortex ultrastructure, ciliature, and internal fibril structure of the cell of Trichostomatia keep well during a long-term storage in formalin.     

The discovery of triploid Lycoris sprengeri Comes ex Baker from Anhui,China

Lycoris sprengeri Comes ex Baker is endemic to China. Reported in the present paper are the chromosomes number and karyotypes for two wild populations of the species from Anhui. ( 1 )Caishi population has a karyotype 2n=33=9st+21t+3T. The length of chromosomes ranges from 5.58～9.15μm. The karyotype belongs to Stebbin’s (1971) “4A”. (2)Longyashan populations have two karyotypes. The karyotype formula of the type I is 2n=22=8st+14t, with chromosomes ranging from 6.88～9.15μm. The karyotype belongs to “4A”. The karyotype formula of the type Ⅱ is 2n＝22＝1m+1sm+14st+6t, with chromosomes ranging from 7.20～15.80μm. The karyotype belongs to “3B”. The triploid type of L. sprengeri was discovered in Anhui for the first time. The karyotype 2n=22 =1m+1sm+14st+6t in diploid type of this species is here reported for the first time.The Robertsonian change plays a key role in karyotype evolution of Lycoris.     

The karyotype of Cepaea sylvatica (Pulmonata: Helicidae) and its relationship to those of C. hortensis and C. nemoralis

The karyotypes of Cepaea nemoralis (L.) and C. hortensis (Müller), with 2n=44 and a conspicuously large pair of chromosomes, are described and compared with that of C. sylvatica (Draparnaud) which has 2n=50. The karyotype of C. sylvatica also has a conspicuously large pair of chromosomes but the comparison suggests that these have an independent origin from those in the 2n = 44 species. There is no evidence that the large chromosomes in C. nemoralis and C. hortensis have originated from simple fusion of chromosomes from a 2n=50 karyotype with chromosomes all sub-equal such as is reported for C. vindobonensis. It may be that such a karyotype with little size differentiation amongst the chromosomes is not a primitive feature in the Helicinae. The relationship of shell colour and banding polymorphism to the chromosome architecture is discussed.     

中国4种蝙蝠的G-带和C-带

首次报道了中国4种蝙蝠的G-带和C-带核型。大长舌果蝠(Eonycteris spelaea)二倍染色体数目(2n)为36,常染色体臂数(FN)为56;马来假吸血蝠(Megaderma spasma)2n=38,FN=70;黑髯墓蝠(Taphozous melanopogon)2n=42,FN=64;皱唇蝠(Chaerephon plicata)2n=48,FN=54。通过C-带显示,除着丝粒异染色质外,在皱唇蝠的许多染色体臂内和马来假吸血蝠染色体的端粒处也有较多的插入异染色质,大长舌果蝠的基因组中既有臂内异染色质也有端粒异染色质。     

蒙古野驴、鹅喉羚和家畜的食物重叠

采用粪样显微分析技术研究了新疆卡拉麦里山有蹄类自然保护区及周边区域蒙古野驴、鹅喉羚及家羊、家马和家骆驼3种家养有蹄类春季、秋季和冬季食性组成及食物生态位。结果发现：（1）蒙古野驴、鹅喉羚和家畜主要采食针茅、驼绒藜、蒿和梭梭,但是,它们采食的植物科数和种数都不相同,各植物种类在食谱中所占的比例不同;（2）各个季节有蹄类动物两两之间的食物生态位重叠均在0．8以上,最低为0．832（冬季鹅喉羚和家马）,最高达到0．986（秋季蒙古野驴和家羊;秋季家马和家羊）,五种有蹄类之间的食物生态位重叠度也达到了0．3以上。表明在卡拉麦里山有蹄类自然保护区,上述野生动物及季节性进入该自然保护区的家畜之间均存在食物竞争;冬季积雪深,食物短缺,荒漠有蹄类易因冻饿及疾病等造成死亡。因此,应采取限制秋冬季进入该自然保护区,家畜数量及调整放牧区域等保护管理措施,对该区域荒漠有蹄类动物实施有效保护[动物学报54（6）：941-954,2008]。     

Cytogenetics studies in Brazilian species of Pseudophyllinae (Orthoptera: Tettigoniidae): 2n(♂)=35 and fn=35 the probable basic and ancestral karyotype of the family Tettigoniidae

The karyotypes of five species of Brazilian Pseudophyllinae belonging to four tribes were here studied. The data available in the literature altogether with those obtained with species in here studied allowed us to infer that 2n(♂)=35 is the highest chromosome number found in the family Tettigoniidae and that it is present in species belonging to Pseudophyllinae, Zaprochilinae and in one species of Tettigoniinae. In spite of that all five species exhibit secondary karyotypes arisen surely by a mechanism of chromosomal rearrangement of centric fusion, tandem fusion and centric inversion types from those with 2n(♂)=35 and FN=35, they share some common traits. The X chromosome is submetacentric (FN=36), heteropicnotic during the first prophase, the largest of the set but its size is rather variable among the species and the sex chromosomal mechanism is of the XO( ♂ ), XX( ♀ ) type. The chromosomal rearrangements involved in the karyotype evolution of the Pseudophyllinae and its relationship with those of the family Tettigoniidae are discussed and we propose that the basic and the ancestral karyotype of the Tettigoniidae is formed by 2n(♂)=35, FN=35 and not by 2n(♂)=31, FN= 31, as usually accepted.     

Chromosomal characteristics and karyotype evolution of Oxyopidae spiders (Araneae, Entelegynae)

We made a cytogenetic analysis of four species of Oxyopidae and compared it with the karyotype data of all species of this family. In Hamataliwa sp, the mitotic cells showed 2n♂ = 26+X(1)X(2) and telocentric chromosomes. The 2n♂ = 28, which has been described for only one oxyopid spider, is the highest diploid number reported for this family. Peucetia species exhibited distinct karyotype characteristics, i.e., 2n♂ = 20+X(1)X(2) in P. flava and 2n♂ = 20+X in P. rubrolineata, revealing interspecific chromosome variability within this genus. However, both Peucetia species exhibited telocentric chromosomes. The most unexpected karyotype was encountered in Oxyopes salticus, which presented 2n♂ = 10+X in most individuals and a predominance of biarmed chromosomes. Additionally, one male of the sample of O. salticus was heterozygous for a centric fusion that originated the first chromosomal pair and exhibited one supernumerary chromosome in some cells. Testicular nuclei of Hamataliwa sp and O. salticus revealed NORs on autosomal pairs, after silver impregnation. The majority of Oxyopidae spiders have their karyotype differentiated by both reduction in diploid number chromosome number and change of the sex chromosome system to X type; however, certain species retain the ancestral chromosome constitution 2n = 26+X1X2. The most remarkable karyotype differentiation occurred in O. salticus studied here, which showed the lowest diploid number ever observed in Oxyopidae and the second lowest registered for Entelegynae spiders.     

A Study on Karyotypes of 6 Species of Elymus in China

The present paper deals with the karyotype analysis of 6 species of Elymus which are native to Qinghai-Tibetan plateau. The number of somatic chromosomes in roottip cells of the 6 species and their karyotypes are reportad here for the first time, and they are all hexaploid, with 2n=6x=42. The karyotype formulae are as follows: E. melantherus, 2n=6x=42=32m+10sm, E. kengii, 2n=6x=42=34m+8sm, E grandiglumis, 2n=6x=42= 30 m+12 sm, E. laxiflorus, 2n=6x=42=32 m +10 sm, E. kokonoricus, 2n+6x=42=34 m +8 sm, E. longiglumis, 2n+6x=42=34 m+8 sm. No satellites have been founded in the 6 species and all the karyotypes belong to 1B or 2B type:     

Karyotypic study of Echinops (Asteraceae) in Fars Province, Iran

A karyotypic study was performed on 19 Echinops species in Fars Province, Iran. The taxa revealed chromosome counts of 2 n = 28, 30, 32 and 34. Somatic chromosome numbers of 18 species are reported for the first time. Using somatic chromosome number as a criterion for section delimitation, the transfer of E. gedrosiacus, E. cyanocephalm and E. sojakii from Sect. Oligolepis to Sect. Ritropsis is suggested. Differences in basic chromosome numbers point towards the possible role played by centric fusion/fission in the karyotypic evolution of the genus. The Karl Pearson coefficient of correlation and principal components analysis indicated the occurrence of structural changes in chromosomes. The species occupied classes 1A, 2A & 2B of Stebbins' karyotype classification, indicating the presence of a primitive symmetrical karyotype in the genus.     

Defining the orientation of the tandem fusions that occurred during the evolution of Indian muntjac chromosomes by BAC mapping

The Indian muntjac (Muntiacus muntjak vaginalis) has a karyotype of 2n=6 in the female and 7 in the male, the karyotypic evolution of which through extensive tandem fusions and several centric fusions has been well-documented by recent molecular cytogenetic studies. In an attempt to define the fusion orientations of conserved chromosomal segments and the molecular mechanisms underlying the tandem fusions, we have constructed a highly redundant (more than six times of whole genome coverage) bacterial artificial chromosome (BAC) library of Indian muntjac. The BAC library contains 124,800 clones with no chromosome bias and has an average insert DNA size of 120 kb. A total of 223 clones have been mapped by fluorescent in situ hybridization onto the chromosomes of both Indian muntjac and Chinese muntjac and a high-resolution comparative map has been established. Our mapping results demonstrate that all tandem fusions that occurred during the evolution of Indian muntjac karyotype from the acrocentric 2n=70 hypothetical ancestral karyotype are centromere–telomere (head–tail) fusions.     

淫羊藿属十二个种的核型

用细胞压片法对12个淫羊藿属(Epimedium)植物的根尖有丝分裂进行了观察,并对其核型进行了比较研究。结果表明:这12种淫羊藿在核型上基本相似,都有1对中间随体染色体,而且都为对称核型。粗毛淫羊藿(E.acuminatum)、印江淫羊藿(E.yinjiangense)、单叶淫羊藿(E.simplicifloum)、巫山淫羊藿(E.wushanense)、光叶淫羊藿(E.myrianthum)、宝兴淫羊藿(E.davidi)、罗甸淫羊藿(E.luodianense)、木鱼坪淫羊藿(E.franchetii)、箭叶淫羊藿(E.sagittatum)、E.pubigerum、高山淫羊藿(E.alpinum)等11个种的核型公式均为2n=2x=6m(2SAT) 6sm,而黔岭淫羊藿(E.leptorrhizum)的核型公式为2n=2x=8m(2SAT) 4sm。结果显示12种淫羊藿的体细胞染色体数目均为2n=12,基数X=6;都属于对称核型、都有1对中间随体;染色体均为m、sm两种类型。