远缘杂交形成的二倍体鱼和多倍体鱼生殖细胞染色体研究

本文采用性腺染色体制片及组织学切片方法,系统地研究了不同发育时期的鲫鲤杂交第二代(F2) (2n=100)、异源四倍体鲫鲤(4n=200)、三倍体鲫鱼(3n=150))、雌核发育二倍体鲫鲤第二代(G2)(2n=100)及鲤鱼(Cypninus carpio L)(2n=100)(对照组)生殖细胞的染色体特征.研究结果表明,对照组中鲤鱼精原细胞染色体数与体细胞染色体数一致,为二倍体精原细胞(2n=100),而远缘杂交形成的二倍体鱼和多倍体鱼的生殖细胞中则观察到明显的染色体数加倍现象,其中,鲫鲤杂交第二代(F2)精巢生殖细胞染色体数加倍现象特别丰富,占检测的染色体分裂相的21.6%,为其产生不减半的二倍体配子提供了直接的细胞学证据,同时也说明远缘杂交是导致生殖细胞染色体数加倍的一个重要因素.该研究在探讨多倍体鱼的发生及鱼类遗传育种方面具有重要意义.     

四倍体鲫鲤、三倍体湘云鲫染色体减数分裂观察

用精巢细胞直接制片法观察了异源四倍体鲫鲤、三倍体湘云鲫和二倍体红鲫、湘江野鲤精母细胞染色体第一次减数分裂中期配对情况 ;作为对照 ,观察了上述四种鱼肾细胞的有丝分裂中期染色体。在精母细胞第一次减数分裂中 ,异源四倍体鲫鲤同源染色体两两配对 ,形成 10 0个二价体 ,没有观察到单价体、三价体和四价体 ;三倍体湘云鲫精母细胞形成 5 0个二价体和 5 0个单价体 ;红鲫和湘江野鲤精母细胞分别形成 5 0个二价体。肾细胞检测表明异源四倍体的染色体数目为 4n =2 0 0 ;湘云鲫为 3n =15 0 ;红鲫和湘江野鲤分别为 2n =10 0。减数分裂时染色体分布情况与肾细胞染色体检测结果相吻合。具有四套染色体的异源四倍体鲫鲤在减数分裂中只形成 10 0个二价体 ,而不形成 2 5个四价体或其它形式 ,为产生稳定一致的二倍体配子提供了重要的遗传保障 ,也为人工培育的异源四倍体鲫鲤群体能够世世代代自身繁衍下去提供了重要的遗传学证据。三倍体湘云鲫在减数分裂过程中出现二价体、单价体共存 ,同源染色体在配对和分离中出现紊乱 ,导致非整倍体生殖细胞的产生 ,为湘云鲫的不育性提供了染色体水平上的证据     

Massive production of all-female diploids and triploids in the crucian carp

In many species of aquaculture importance, all-female and sterile populations possess superior productivity due to faster growth and a relatively homogenous size of individuals. However, the production of all-female and sterile fish in a large scale for aquaculture is a challenge in practice, because treatments necessary for gynogenesis induction usually cause massive embryonic and larval mortality, and the number of induced gynogens is too small for their direct use in aquaculture. Here we report the massive production of all-female triploid crucian carp by combining artificial gynogenesis, sex reversal and diploid-tetraploid hybridization. Previously, we have obtained an allotetraploid carp population (4n = 200) by hybridization between red crucian carp (Carassius auratus red var; ♀) and common carp (Cyprinus carpio; ♂). We induced all-female diploid gynogens of the Japanese crucian carp (Carassius cuvieri; 2n = 100). We also generated male diploid gynogens of the same species treated gynogenetic fry with 17-α-methyltestosterone, leading to the production of sex-revered gynogenetic males. Finally, these males were used to cross with the female diploid Japanese crucian carp gynogens and the allotetraploid females, resulting in the production of fertile all-female diploid Japanese crucian carp (2n=100) and sterile all-female triploid hybrids (3n = 150), respectively. Therefore, diploid crucian carp gynogenetic females and sex-reversed male together with an allotetraploid line provide an opportunity to produce all-female triploid populations in a large scale to meet demands in aquaculture industry.     

Development of 10 tri- and tetranucleotide microsatellite loci for population studies in the common carp (Cyprinus carpio L.)

Ten tri- and tetranucleotide microsatellite DNA markers were isolated and characterized from common carp (Cyprinus carpio L.) to estimate genetic potential. These markers were tested in the samples from two closely related carp populations (Cyprinus carpio var. xingguonensis and Cyprinus carpio var. wananensis). The number of the alleles ranged from three to nine, and observed and expected hererozygosities varied from 0.207 to 1.000 and from 0.499 to 0.900 in each population, respectively. No evidence for linkage disequilibrium was found, indicating that these markers will be useful for population studies.     

草鱼和鲤杂交的细胞学研究—鱼类远缘杂交核质不同步现象

以草鱼(Ctenopharyngodon idellus)为母本、鲤(Cyprinus carpio)为父本进行人工杂交,杂种胚胎发育至孵化期全部死亡;同时获得了少数雌核发育草鱼和雄核发育的鲤。分析比较了草鲤杂种胚胎染色体变化及胚胎发育情况。发现杂种胚胎染色体数目变化较大,一般在24—73之间,绝大部分细胞染色体在发育过程中不断丢失而出现非整倍体;极少数细胞在受精后雌性原核和雄性原核不结合而引起雌核发育和雄核发育;草鱼和鲤胚胎发育时序有较大差别;因此细胞分裂不能同步。可能是杂种胚胎染色体不断丢失的主要原因。     

Reproduction mode of an artificial allotetraploid carp (Pisces; Cyprinidae)

Mature eggs of allotetraploid carp were activated by inactive sperm or crossed with normal sperms of common carp (Cyprinus carpio), crucian carp (Carassius auratus), Chinese blunt snout bream (Megalobrama amblycephala), Hemiculter leucisculus and Pseudorasbora parva. Chromosome counts showed that all offspring of these crosses presented a mode number of 200 chromosomes (4n = 200), and their morphological traits are much like maternal. Microsatelite marker and RAPD patterns between allotetraploid maternal and its offspring, reproduced from different paternal species, were identical. Cytological, morphological and molecular evidences suggested that allotetraploid carp female nucleus would not fuse with any male nucleus and its reproduction mode might be gynogenesis and therefore their offspring are retaining their tetraploidy and give origin to clonal individuals.     

Genome evolution trend of common carp （Cyprinus carpio L.） as revealed by the analysis of microsatellite loci in a gynogentic family

Genome evolution arises from two main ways of duplication and reduction. Fish specific genome duplication （FSGD） may have occurred before the radiation of the teleosts. Common carp （Cyprinus carpio L.） has been considered to be a tetraploid species, because of its chromosome numbers （2n=100） and its high DNA content. Using 69 microsatellite primer pairs, the variations were studied to better understand the genome evolution （genome duplication and diploidization） of common carp from a gynogenetic family. About 48% of primer pairs were estimated to amplify duplicates based on the number of PCR amplification per individual. Segregation patterns in the family suggested a partially duplicated genome structure and disomic inheritance. This indicates that the common carp is tetraploid and polyploidy occurred by allotetraploidy. Two primer pairs （HLJ021 and HLJ332） were estimated to amplify reduction based on the number of PCR amplification per individual. One allele in HLJ002 locus and HLJ332 locus was clearly lost in the gynogenetic family and the same as in six wild populations. Segregation patterns in the family suggested a partially diplodization genome structure. A hypothesis transition （dynamic） and equilibrium （static） were proposed to explain the common carp genome evolution between genome duplication and diploidization.     

用PCR-DGGE对岩原鲤×锦鲤杂交F1代进行分子鉴定

运用变性梯度凝胶电泳(DGGE)技术,选用核基因光间受体视黄类物质结合蛋白(IRBP)作为分子标记,成功鉴定了岩原鲤Procypris rabaudi×锦鲤Cyprinus carpio的杂交F1代.DGGE胶图显示,杂交个体IRBP基因的等位基因条带分别与岩原鲤和锦鲤的IRBP基因条带相对应.对各条带切胶回收后的测序表明,岩原鲤等位基因的相似性为99.8％,锦鲤等位基因的相似性为98.8％,而杂交个体等位基因的相似性为95.8％ ～96.7％,与岩原鲤和锦鲤间基因的相似性(95.5％～96.9％)接近.这一方法将有助于鱼类的杂种鉴定和种质研究.     

Aquarium studies on the selectivity of 16 aquatic plants as food by fingerling hybrids of the cross between Ctenopharyngodon idella ♂ and Cyprinus carpio ♀

Fish hybrids of the cross Ctenopharyngodon idella ♂ (grass carp) and Cyprinus carpio ♀ (Israeli carp), averaging 124 mm total length and 20 g in weight were stocked in aquaria along with 16 different aquatic plants to determine which plants they would eat. Observations showed that one group of plants was highly utilized (Azolla caroliniana, Lemna spp., Cladophora sp., Spirogyra sp., Nymphaea odorata); another group was moderately utilized (Pistia stratiotes, Ruppia maritima, Najas flexilis, Limnobium spongia, Potamogeton pecti-natus). Two other groups were only poorly utilized (Ceratophyllum demersum, Eichhornia crassipes, Elodea sp.) , or not utilized (Cabomba caroliniana, Sigittaria sp., Myriophyllum pinnatum).     

Modelling energetic costs of fish swimming

The oxygen consumption rates of two cyprinid fishes, carp (Cyprinus carpio L.) and roach (Rutilus rutilus (L.)), were analysed for a wide range of body mass and swimming speed by computerized intermittent-flow respirometry. Bioenergetic models were derived, based on fish mass (M) and swimming speed (U), to predict the minimal speed and mass-specific active metabolic rate (AMR) in these fishes (AMR=aMbUc). Mass and speed together explained more than 90% of the variance in total swimming costs in both cases. The derived models show that carp consume far more oxygen at a specific speed and body mass, thus being less efficient in energy use during swimming than roach. It was further found that in carp (AMR=0.02M0.8U0.95) the metabolic increment during swimming is more strongly effected by speed, whereas in roach (AMR=0.02M0.93U0.6) it is more strongly effected by body mass. The different swimming traits of carp and roach are suitable for their respective lifestyles and ecological demands.     

人工诱导雌核发育日本白鲫

分别用遗传失活的散鳞镜鲤(Cyprinus carpio L.)、团头鲂(Megalobrama amblycephala)精子诱导日本白鲫(Carassius cuvieri)进行雌核发育.未经冷休克处理,用UV照射过的镜鲤的精子(其最佳UV照射剂量为4 200 mJ/cm2)与日本白鲫卵子受精获得(32.4±3.3)%雌核发育单倍体和(0.7±0.3)%杂交二倍体,而照射过的团头鲂精子(其最佳UV照射剂量为3 600mJ/cm2)与日本白鲫卵子受精得到了(33.8±1.4)%雌核发育单倍体和(0.5-±0.3)%杂交二倍体.日本白鲫卵子分别经灭活的镜鲤、团头鲂的精子激活(精子经各自最佳UV剂量处理),施以冷休克处理(受精后2min,0-4℃,40min),其孵化率分别为(27.8±2.1)%和(29.4±3.3)%,发育到摄食期,其正常的雌核发育二倍体鱼苗分别为(15.7±3.4)%和(23.6±4.1)%,在镜鲤实验组中,其正常的雌核发育二倍体鱼苗占孵化鱼苗总数的56%,这比团头鲂实验组中正常雌核发育二倍体鱼苗的比率(达到80%)低,结果表明诱导日本白鲫雌核发育,与母本遗传关系远的团头鲂精子较镜鲤的更有效.从染色体数目、外形特征和性腺结构等方面证实雌核发育后代的生物学特性.雌核发育日本白鲫全为雌性,这表明其雌性是同配XX型.雌核发育日本白鲫在生产上将有着潜在的价值,如比普通日本白鲫长得更快,抗病能力强等.所有的雌核发育日本白鲫的染色体数目都是100,而所有的团头鲂与日本白鲫的杂交后代都是三倍体(3n=124),新三倍体鱼在鱼类养殖和理论研究中将存在潜在的价值.     

Effect of photoperiod on growth and feed conversion of juvenile wild carp, Cyprinus carpio

Because of the effect of photoperiod on physiological and biochemical processes in fish, this study aimed to evaluate the effect of manipulated photoperiod on growth, feed conversion and survival of wild carp, Cyprinus carpio. Fish received six photoperiod regimes (light:dark cycle) including: natural photoperiod (control), 24L:0D, 16L:8D, 12L:12D, 8L:16D and 0L:24D by the three replications. Regulated photoperiods as a 16L:8D or 12L:12D light/dark cycle significantly improved growth rate and food conversion ratio of wild carp.     

Identification of a new granulocyte type in the skin of carp Cyprinus carpio (L.)

The skin of carp Cyprinus carpio (L.) was investigated by electron microscopy. A new granulocyte cell type was identified, and is described as being similar to the eosinophilic granular cell (EGC) of salmonids.     

Genetic differences between the Chinese and the European races of the common carp III. Gonad abnormalities in hybrids

Abnormalities in the gonads of carp, Cyprinus carpio L., were present in a relatively high proportion of hybrids derived from the Chinese and European strains. The aberrations involved were the absence or marked reduction of the gonads in mature fish.     

Karyotypes of Cyprinus carpio and Leuciscus cephalus

Karyological investigations on the common carp (Cyprinus carpio) and chub (Leuciscus cephalus) are described. The diploid chromosome complement in the carp was found to be 98, with 50 meta- to submetacentric chromosomes, 48 subtelo- to telocentric chromosomes, and an arm number of 148. Chromosomal studies on the chub revealed the diploid chromosome complement to be 50, with 34 meta- to submetacentric chromosomes, 16 subtelo- to telocentric chromosomes, and an arm number of 84.     

Purification and characterization of Cu,Zn-superoxide dismutase from common carp liver

1. Common carp (Cyprinus carpio L.) liver Cu,Zn-superoxide dismutase (Cu,Zn-SOD) was purified and characterized. 2. Its molecular weight, isoelectric point, electrophoretic mobility, amino acid pattern and some other characteristics were determined.     

Introgression in the genus Salmo via allotriploids

Hybridization between sympatric species is not uncommon in the wild. Wild allotriploids (individuals with two chromosome sets from a species + one chromosome set from another species) are generally the result of a backcross between interspecific hybrids that produce unreduced gametes and one of the parental species. In animals, allotriploids are commonly sterile, except for some vertebrate species complexes in which allotriploids reproduce by parthenogenesis, gynogenesis and/or hybridogenesis, producing generally clonal or hemiclonal gametes; nuclear DNA introgression between hybridizing species is considered to be extremely rare. Employing species-specific molecular markers, we show genetic introgression between the chromosomally well-differentiated salmonids Atlantic salmon (2n = 58) and brown trout (2n = 80) through spontaneous bisexual reproduction of allotriploids leading to salmon-like offspring bearing some brown trout genes. Although introgression between these Salmo species can occur via allotriploids, we hypothesize that extinction of parental species can be discarded based on very low survival of allotriploid offspring.     

异育淇鲫及其双亲同工酶的比较研究

用4.5％聚丙烯酰胺凝胶平板电泳研究了异育淇鲫及其母本淇鲫和父本兴国红鲤的肌可溶性蛋白以及肾、肝、眼、背白肌和心等五种组织的乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)和酯酶(EST)。结果发现:异育淇鲫的肌可溶性蛋白以及同工酶的电泳图谱与母本淇鲫相同而与父本兴国红鲤显著不同,因而认为异育淇鲫是淇鲫雌核发育的产物,父本基因对子代基本无影响。在此基础上,本文对异源精子在雌核发育中所起的生物学作用进行了初步探讨。     

Structural and immunochemical studies of the immunoglobulins of carp (Cyprinus carpio L.) II. Analysis of the subunits]

The 14 S immunoglobulin of the carp (Cyprinus carpio L.) was split into subunits with 0,01 M dithioerythritol. These 5,7 S subunits have a molecular weight of 104 000 and a hexose and hexosamine content of 6.2%. It is likely that the subunits represent HL-half-molecules and not H2L2-monomeres. The values for the molecular of H- and L-chains were 77 000 and 24 000, respectively.