哺乳动物卵母细胞非整倍体产生机制的研究进展

雌性生殖细胞进行减数分裂时易发生染色体分离错误而产生非整倍体卵母细胞,其受精后会产生非整倍体胚胎,导致出生缺陷或胚胎致死,是影响哺乳动物繁殖的重要因素。卵母细胞在第一次减数分裂前期发生同源染色体联会,此时DNA双链断裂引发重组。重组时缺乏交叉、重组事件数量的减少及交叉靠近端粒或着丝粒导致染色体发生同向分离或不分离,从而产生非整倍体卵母细胞。减数分裂期间,当染色体的端粒共向于同一极或没有完全附着在纺锤体微管上时,纺锤体组装检查点（spindle assembly checkpoint,SAC）被激活,E3泛素连接酶APC/Cyclome （APC/C）沉默,保护分离酶抑制蛋白（securin）和细胞周期蛋白B （cyclin B）不被降解,从而抑制分离酶和染色体的分离。直到所有染色体与纺锤体实现稳定的双极定向并正确排列到赤道板上,SAC关闭,染色体正确分离。卵母细胞中SAC蛋白缺失,导致SAC不能有效地监测端粒在纺锤体上的正确附着,发生染色体分离错误,从而产生非整倍体卵母细胞。因此,通过现代分子技术手段解析非整倍体卵母细胞所涉及的机制是保护哺乳动物生育的重要目标。作者主要介绍了卵母细胞减数分裂的特点,详细阐述了卵母细胞非整倍体发生的染色体分离错误的分子机制,以期为开发卵母细胞非整倍体的治疗手段提供参考。     

1B/1R易位系的冬小麦单体系统育成

本研究以“中国春”小麦21个单体系为遗传背景,经过7代连续回交,转育成1B/1R易位系的冬小麦73(36)9-1单体系,同时也对转育过程产生的缺体进行了详细的研究。用“中国春”双端体核查结果,除2A单体有19″ 1t″ 1′核型外,其余20个单体系和19个缺体都是20″ t′核型。这表明,后者是正确的,而2A单体已发生“变迁”。3B缺体虽未核查,但依其减数分裂不能正常配对的特征,也可判断是正确的。本文对这20个单体系和相应的缺体植株的形态特征进行了描述,并对1B染色体的异质性以及有关单体和缺体在形态特性上的差异进行了讨论。     

Autosomic 27 Trisomy in a Standardbred Colt

A 19-month-old Standardbred colt was donated to the University of Pennsylvania School of Veterinary Medicine with a suspicion of intersexuality. The anal−genital distance and penis were normal, and there was no evidence of intersexuality, but the colt was bilaterally cryptorchid. Several aspects of the colt's behavior appeared unusual, including general temperament and behavior described as sympathetically dull and affable. With herd mates, the colt appeared slow to perceive or to learn the usual intraspecies social cues. An atypical gait characterized by intermittent unnatural shuffle of the hind limbs, sliding them along in short rhythmic strides for 3 to 10 seconds at a time was noted at times when a horse might normally transition from a slow walk to a fast walk or a slow trot. Occasionally the colt exhibited slight protrusion of the tongue through the teeth and lips with jaw movements and smacking of the tongue against the teeth as if struggling to retract the tongue to the normal position. Evaluation of the karyotype combined with fluorescent in situ hybridization (FISH) revealed an abnormal male karyotype showing trisomy of chromosome 27 (65, XY + 27). The colt was euthanized at 24 months of age, and a necropsy revealed no significant abnormalities. This case of trisomy was not associated with developmental abnormalities described in other rare reports of trisomy in horses; however, some features were strikingly similar to that of humans with trisomy 21. FISH was demonstrated to be an excellent method for correct identification of equine chromosomes.     

Polypolar spindle formation during first cell cycle in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> embryos after heat-shock treatment

ABSTRACT:   Heat shock has been used to inhibit cleavage for the induction of monogenic diploids or tetraploids in animals, but usually the success rate is low. Heat-shocked rainbow trout Oncorhynchus mykiss embryos were used in this histological study to clarify the causes of this low success rate. Embryos treated with hydrostatic pressure were used for comparison. After heat shock had disorganized the spindles, polypolar (tripolar or tetrapolar) spindles in addition to bipolar spindles were often reassembled soon after treatment. The embryos then completed tripolar or tetrapolar division at the first mitosis, and directly turned into three- or four-cell embryos as a result of the first cleavage. During the second mitosis, a monopolar spindle was formed in each blastomere of four-cell embryos and approximately 60% of three-cell embryos. In the remaining three-cell embryos, two of the three blastomeres formed a monopolar spindle, and the third one formed a bipolar spindle. The formation of polypoles is assumed to be caused by insufficient disorganization of daughter centrioles and splitting from the mother centriole by heat shock. Polypolar division is considered to be the cause of aneuploidy and the low success rate of chromosome set doubling. In the case of hydrostatic pressure treatment, the regenerated spindles were bipolar in almost all embryos.     

冬小麦1A缺体单倍体的细胞学研究

对1B/1R易位系冬小麦73(36)9-1的1A缺体单倍体小孢子母细胞和小孢子进行了细胞学观察,结果表明:①减数分裂有双线期,76.7％细胞的全部染色体都有不同形态的双线结构,而中期Ⅰ染色体配对的细胞只有41％;②末端联会较多,并靠联会较少,其分布随每个细胞二价体数目的增加而逐渐减少;③四分体时期有大量的二分体,一分体和三分体,它们进一步发育成大量的败育小孢子和少量异常小孢子;④中期Ⅰ和小孢子中有不同频率的融合细胞。     

Colchine-induced aneuploids from cell culture of sugarcane

Summary Sugarcane (Saccharum spp.) clones are amenable to gross chromosome manipulation due to their high polyploid nature (2n=100–120). This study was conducted to analyze the effects on plant morphology of altering chomosome number via callus culture. Callus cultures from clone H69-9092 were established, and plants were regenerated following colchicine treatment of cultured cells. Cytological analysis showed that variant somaclones were aneuploids with a wide range in chromosome numbers (2n=66–196). Some 22 visually distinct somaclones were planted in 1.35 m² plots with five replications to compare morphological and quality characteristics with H69-9092 at 8 months of growth. Extreme morphological variation was observed between somaclones, but coefficients of variation for quality factors-fibers %, refractometer solids %, pol %, and juice purity-and stomatal length were smaller than those for morphological traits associated with stalk volume and leaf area. Significant negative correlations were found between chromosome number and most morphological traits, e.g., stalk length (r=-0.58), number (r=-0.69), diameter (r=-0.54) and volume (r=-0.65); internode length (r=-0.57); and leaf area (r=-0.48). A positive correlation was found between chromosome number and stomatal length (r=-0.66). No significant correlations were found between chromosome number and quality factors. Aneuploids with higher than parental chromosome number had reduced growth. However, depression in growth was generally not observed in somaclones lower in chromosome number than the parent.Published with the approval of the Director as Paper No. 598 in the Journal Series of the Experiment Station, Hawaiian Sugar Planters' Association.     

Gain or loss of single chromosomes in wheat-rye addition lines and in 6x triticale detected by flow cytometry

Flow cytometric DNA analysis was used to study changes in nuclear DNA content induced by the gain or loss of chromosomes or chromosome arms in wheat-rye addition lines and in triticale. The amount of DNA present was determined by comparison with internal reference values obtained from euploid plants. The smallest difference in the DNA level which could be detected between aneuploid and euploid plants was 1.84%. Thus, it was possible to determine aneuploidy in all cases, except when very short telocentric rye chromosomes were added to wheat (addition line 1RS). The study has proved that flow cytometry is an efficient and simple way to discriminate between euploid and aneuploid plants in wheat and triticale.     

Intergeneric crosses with Fragaria and Potentilla. I. Crosses between Fragaria moschata and Potentilla fruticosa

Summary Crosses between Fragaria moschata (2n=42) and Potentilla fruticosa (2n=14) produced many seeds; about half of these germinated, but only nine plants were obtained from 554 seedlings. Five plants died without flowering, and four were vigorous but sterile. Four plants had the expected chromosome number (2n=28), one plant had 21 chromosomes, and four were aneuploid with 23, 24, 25 and 27 chromosomes respectively. Response to growth substances giving elongated stem internodes and swollen but not succulent receptacles provided evidence that two of the plant were hybrids. It is suggested that all nine plants originated from a normal fertilisation which was followed in some cases by chromosome elimination at an early stage of embryo development.     

Insights on the association between somatic aneuploidy and ostreid herpesvirus 1 detection in the oysters Crassostrea gigas,C. angulata and their F1 hybrids

Cytogenetic abnormalities associated with viral infections, including from viruses of the Herpesvirales order, have been reported in vertebrate species. Ostreid herpesvirus 1 (OsHV‐1) has been detected worldwide during mortality outbreaks of the Pacific oyster Crassostrea gigas. On the other hand, a high proportion of aneuploid cells in somatic tissues have been observed in C. gigas. In this study, we analysed the putative association between aneuploidy levels and the detection of OsHV‐1 in gills of C. gigas, the Portuguese oyster C. angulata and their F1 hybrids cultured in Ria Formosa (Portugal). OsHV‐1 was detected by PCR in 5.4% of the total of oysters analysed (n = 111) namely in 11.1%, 8.0% and 1.7% of C. gigas, C. angulata and F1 hybrid respectively. Sequencing analysis of a viral fragment amplified with the C2/C6 primer pair revealed a high similarity with the OsHV‐1 reference type. Moreover, in situ hybridization confirmed the presence of OsHV‐1 in gill tissue. Oysters where OsHV‐1 was detected had a significantly higher mean percentage of aneuploid cells (25%) than the ones where the virus was not detected (18%). However, the overall low percentage of positive samples contrasted with the high mean percentage of aneuploidy observed, with 50% of the oysters analysed showing a percentage of aneuploid cells between 20% and 30%. We hypothesize that somatic aneuploidy may adversely affect oysters making them more prone to OsHV‐1 infection, but the virus is unlikely to be the cause of somatic aneuploidy.     

Perennial aneuploidy as a potential material for gene introgression between maize and Zea perennis

Hybridization,which allows for gene flow between crops,is difficult between maize and Zea perennis.In this study,we aim to initiate and study gene flow between maize and Z.perennis via a special aneuploid plant(MDT) derived from an interspecific hybrid of the two species.The chromosome constitution and morphological characters of MDT as well as certain backcross progenies were examined.Results from genomic in situ hybridization(GISH) indicate that aneuploid MDT consisted of nine maize chromosomes and 30 Z.perennis chromosomes.The backcross progenies of MDTxmaize displayed significant diversity of vegetative and ear morphology;several unusual plants with specific chromosome constitution were founded in its progenies.Some special perennial progeny with several maize chromosomes were obtained by backcrossing MDT with Z.perennis,and the first whole chromosome introgression from maize to Z.perennis was detected in this study.With this novel material and method,a number of maize-tetraploid teosinte addition or substitution lines can be generated for further study,which has great significance to maize and Z.perennis genetic research,especially for promoting introgression and transferring desirable traits.