Development and characterization of interspecific hybrids between <Emphasis Type="Italic">Oryza sativa</Emphasis> and <Subscript>O. latifolia</Subscript> by <Subscript>in situ</Subscript> hybridization

Oryza sativa and O. latifolia belong to the AA and CCDD genomes of Oryza, respectively. In this study, interspecific hybrids of these species were obtained using the embryo rescue technique. Hybrid panicle traits, such as long awns, small grain, exoteric large purple stigma, grain shattering and dispersed panicles, resemble that of the paternal parent, O. latifolia, whereas there is obvious heterosis in such respects as plant height, tillering ability and vegetative vigor. Chromosome pairing and the genomic components of the hybrid were subsequently investigated using genomic in situ hybridization （GISH） and fluorescent in situ hybridization （FISH） analysis. Based on the mitotic metaphase chromosome numbers of the root tips investigated, the hybrid is a triploid with 36 chromosomes. The genomic constitution of the hybrid is ACD. In the meiotic metaphase Ⅰ of the hybrid pollen mother cell, poor chromosome pairing was identified and most of the chromosomes were univalent, which resulted in complete male sterility in the hybrid.     

甘蓝-白芥单体异附加系自交后代的GISH分析

将甘蓝-白芥单体异附加系自交,获得了其自交后代.利用基因组原位杂交(genomicin situhy-bridization,GISH),结合双色荧光原位杂交(dual-colour fluorescencein situhybridization,dcFISH)技术,从这些自交后代中鉴定出了纯合的甘蓝-白芥二体异附加系植株.GISH分析结果表明,甘蓝-白芥二体异附加系有丝分裂中期具有18条甘蓝染色体及2条白芥染色体,减数分裂中期I表现为9个C染色体二价体及1个S染色体二价体,减数分裂后期I会出现落后的1对S染色体,有时落后的1对S染色体形成染色体桥.     

利用粳稻基因组DNA和C_ot-1 DNA探针对普通野生稻和亚洲栽培稻的比较分析

以粳稻日本晴基因组DNA和Cot-1 DNA为探针,分别对日本晴、籼稻广陆矮4号和普通野生稻的染色体组进行了基因组原位杂交(GISH)和Cot-1 DNA荧光原位杂交(FISH)分析,并对3种染色体组进行了同源聚类和比较研究.结果表明:粳稻基因组DNA和Cot-1 DNA探针信号在3种水稻染色体组中的分布状况和覆盖率相似,Cot-1 DNA的覆盖率分别为(47.13±0.18)%、(45.89±0.22)%、(44.24±0.21)%,3种水稻基因组同源性高,亲缘关系接近.Cot-1 DNA在3种水稻染色体上的杂交信号分布各有特点,中高度重复序列的变异在普通野生稻向栽培稻进化和亚洲栽培稻籼、粳分化过程中具有重要意义,中高度重复序列含量较低的2、5、8号染色体是水稻染色体组进化过程中相对活跃的成分.     

着丝粒串联重复序列RCS2对亚洲栽培稻和非洲栽培稻的比较分析

为了揭示中高度重复序列在同为AA基因组的亚洲栽培稻和非洲栽培稻基因组中的差异以及重复序列在.栽培稻种的分化过程中可能起到的作用,利用水稻着丝粒串联重复序列RCS2作为探针分别对籼稻广陆矮4号、粳稻日本晴和非洲栽培稻的体细胞染色体进行荧光原位杂交(FISH)实验,并对其核型进行同源性聚类和比较分析,杂交结果显示:RCS2序列位于在3种栽培稻染色体组中,RCS2序列位于每条染色体的着丝粒位置,但有不同的分布特点,表明该3种栽培稻基因组的RCS2序列有不同的进化方向.探讨了RCS2序列结合Cot-1 DNA FISH方法对水稻染色体组进行核型分析的可行性和优势.     

Comparative analysis of A, B, C and D genomes in the genus Oryza with Cot-1 DNA of C genome

Fluorescence in situ hybridization （FISH） was applied to somatic chromosomes preparations of Oryza officinalis Wall. （CC）, O. sativa L. （AA）xO. officinalis F1 hybrid （AC）, backcross progenies BC1 （AAC and ACC）, O. latifolia Desv. （CCDD）, O. alta Swallen （CCDD） and O. punctata Kotschy （BBCC） with a labelled probe of Cot-1 DNA from O. officinalis. In O. officinalis, the homologous chromosomes showed similar signal bands probed by Cot-1 DNA and karyotype analysis was conducted based on the band patterns. Using no blocking DNA, the probe identified the chromosomes of C genome clearly, but detected few signals on chromosomes of A genome in the F1 hybrid and two backcross progenies of BC1. It is obvious that the highly and moderately repetitive DNA sequences were considerably different between C and A genomes. The chromosomes of C genome were also discriminated from the chromosomes of Dand B-genome in the tetraploid species O. latifolia, O. alta and O. punctata by Cot-1 DNA-FISH. Comparison of the fluorescence intensity on the chromosomes of B, C and D genomes in O. latifolia, O. alta, and O. punctata indicated that the differentiations between C and D genomes are less than that between C and B genomes. The relationship between C and D genomes in O. alta is closer than that of C and D genomes in O. latifolia. This would be one of the causes for the fact that both the genomes are of the same karyotype （CCDD） but belong to different species. The above results showed that the Cot-1 DNA had a high specificity of genome and species. In this paper, the origin of allotetraploid in genus Oryza is also discussed.     

Comparative physical mapping of rice BAC clones linked to resistance genes Glh,Bph-3 and xa-5 in Oryza sativa L. and O. granulata Nees et Arn. ex Watt.

Oryza granulata Nees et Arn. ex Watt. is one of the three wild relatives of rice,which are the most valuable for study and utilization in China.In this study,the homology and physical locations of three rice resistance genes,Glh,Bph-3 and xa-5 are comparatively analyzed between O.sativa and O. granulata by Southern blotting and fluorescence in situ hybridization (FISH).The results of Southern blotting indicate that there exist homologous sequences of the tested RFLP markers in O. granulata.By using three bacterial artificial chromosome (BAC) clones scanned by the tested RFLP as probes, FISH signals are detected on both mitotic and pachytene chromosomes in O.sativa and O.granulata.Dual-color FISH demonstrates that two of the three BAC clones (14E16 and 38J9) are located on the short arm of the same chromosome pair in O. granulata. Additionally, colinearity is shown for the two clones between O.sativa and O.granulata. Another BAC clone 44B4 is located on the end of the short arm of other chromosome pair in these two species.Although the phylogenetic relationship between O.sativa and O.granulata is the most distinct in Oryza and these two species have evidently different biological features and ecological habits, the relative lengths and arm ratios of the detected chromosomes and the relative positions of the tested clone signals on chromosomes in O. granulata are quite similar to those in O. sativa.     

电镜原位杂交对淡水三角头涡虫中期染色体RNA的研究

本文用ＲＮａｓｅ－ｇｏｌｄ标记法定位了淡水三角头涡虫中期染色体内ＲＮＡ，发现ＲＮＡ在染色单体切面内部及边缘均有分布。此外，以玉米１８ｓｒＲＮＡ的ｃＤＮＡ和水稻５ｓｒＲＮＡ的ＤＮＡ为探针，经电镜原位杂交后观察了涡虫１８ｓｒＲＮＡ（也包括４５ｓｐｒｅ－ｒＲＮＡ）及５ｓｒＲＮＡ在中期染色单体切面上的分布特点，证明涡虫染色单体边缘及内部ＲＮＡ的两个来源是核仁核糖体ＲＮＡ和核基质内的５ｓｒＲＮＡ。     

一种改进的重复多色荧光原位杂交技术

多色荧光原位杂交技术是在常规荧光原位杂交技术基础上建立的一种染色体分析手段,已广泛应用于肿瘤遗传学的研究,尤其是用于染色体隐匿性重排及复杂核型分析。本研究旨在利用现有实验条件,建立一种可靠的人类全染色体分析方法——重复多色荧光原位杂交技术（repetitive multicolour fluorescence in situ hybridization,RM-FISH）。采用特异的全染色体涂染探针,分别用C3、Cy5、FITC3种荧光素直接标记,组合制备4组六色的探针池。对同一染色体片先后进行4次杂交,通过装备了4组滤镜的荧光显微镜捕获信号。RM-FISH技术结合反相DAPI显带对1例原发性食管鳞癌进行核型分析,获得了全部染色体的特异信号。本研究建立的RM-FISH简便、易行,是光谱核型分析的一种较理想的替代技术。     

应用基因组原位杂交鉴定杂交后代中的簇毛麦染色体

用地高辛（Digoxigenin-11-dTup)标记的簇毛麦染色体组DNA为探针，以普通小麦“中国春”总DNA作封阻进行基因组荧光原位杂交，对小麦和簇毛麦杂交和回交后代进行检测。结果显示，在杂交后代的28条染色体中有7条簇毛麦染色体，在发生部分染色体加倍的回交代后鉴定出含7条簇毛麦染色体重的易位系。GISH的准确鉴定以及易位系的获得为向小麦导入簇志麦的有用基因提供了宝贵材料。     

Breeding and molecular cytogenetic identification of wheat-Thinopyrum intermedium addition lines resistant to powdery mildew

Wheat-related species Th. intermedium was used to cross with common wheat Yannong 15. In the self progenies of the hybrid, two addition lines, Ⅱ-1-7-1 and Ⅱ-3-3-2, stable in cytology, were developed by cytology and powdery mildew resistance identification. Their chromosome number were 2n = 44 and formed 22 bivalents at PMC MI. In F1 of the two addition lines crossing with Yannong 15, there appeared about one univalent at PMC MI, respectively. Resistance identification in greenhouse and field using the No. 15 and mixed strains of E. gramnis f. sp. tritici showed that they were immune to powdery mildew. Chromosome number and resistance identification using the F2 single plants of the addition line crossing with Yannong 15 indicated that the resistant gene was located on the alien chromosomes. In situ hybridization using St and E genomic DNA as probe showed that the added chromosome in the two addition lines probably came from the E genome of Th. intermedium, which indicated that a pair of E genome chromosomes carried a new resistant gene to powdery mildew.     

PVDF/Al2O3杂化膜的结构及性能表征

为了改善超滤膜的使用性能,采用溶胶-凝胶法,在聚偏氟乙烯(PVDF)注膜液中直接添加异丙醇铝(AIP),制备了不同Al2O3含量的PVDF/Al2O3杂化超滤膜.并用扫描电镜、红外光谱、超滤实验、接触角和机械性能测试等手段对膜的结构和性能进行了表征.结果表明: 由于AIP发生水解聚合反应,生成铝氧聚合物掺杂在杂化膜中,并与PVDF发生键联.当AIP添加量为12%时,杂化膜的通量比纯PVDF膜提高了83.36%,截留率由88.27%提高到96.64%,同时膜的亲水性、机械性能和抗污染性都得到改善.     

Comparative genome research between maize and rice using genomicin situ hybridization

Using the genomic DNAs of maize and rice as probes respectively, the homology of maize and rice genomes was assessed by genomic in situ hybridization. When rice genomic DNAs were hybridized to maize, all chromosomes displayed many multiple discrete regions, while each rice chromosome delineated a single consecutive chromosomal region after they were hybridized with maize genomic DNAs. The results indicate that the genomes of maize and rice share high homology, and confirm the proposal that maize and rice are diverged from a common ancestor.     

一个药用野生稻异源单体附加系在减数分裂时期的GISH分析鉴定

异源单体附加系是从亲缘关系较远或属间的一个物种单条染色体附加到另一个物种中．栽培稻珍籼97B与药用野生稻Hy18杂交与连续回交，在BC2后代中得到一个药用野生稻单体附加系．生物素标屺的药用野生稻总DNA作为探针，未标记的栽培稻总DNA封阻，对其异源单体系减数分裂染色体进行基因组原位杂交．FISH结果表明，在栽培稻（AA，2n=24）基因组中附加了一条药用野生稻染色体，并鉴定为第8号染色体．研究表明，药用野生稻异源单体附加系的建立为药用野生稻的基因组学和遗传学的研究提供一个新的操作平台．而GISH技术在水稻远缘杂交育种中是最准确有效的染色体鉴定方法．在水稻育种改良中具有重要应用前景．     

Cytological mechanisms of interspecific incrossability and hybrid sterility between Oryza sativa L. and O. alta Swallen

Oryza sativa and O. alta belong to AA and CCDD genomes in Oryza, respectively. Interspecific repro-ductive isolation limits the transfer of favorable genes from O. alta into O. sativa. The cytological mechanisms of interspecific incrossability and hybrid sterility between O. sativa and O. alta were studied systematically in this paper. We indentified two cytological causes of interspecific incrossabil-ity. First, we observed embryo sac incompatibility that caused fertilization barriers of variable severity such as non-fertilization, fertilization stagnation and egg cell single-fertilization. Second, we observed hybrid inviability, the major cause for incrossability, apparent from hybrid embryo developmental stagnation and embryo abortion. Hybrid sterility included both embryo sac sterility and pollen sterility. The hybrid embryo sac was completely sterile and exhibited mainly embryo sac degeneration. Hybrid pollen was also sterile and mainly typical abortive. Hybrid sterility was mainly caused by severely ab-normal meioses of megasporocytes and pollen mother cells; it is the most important abnormality, being chromosome sterility. Several methods are suggested to overcome the interspecific reproductive iso-lation between O. sativa and O. alta.     

褐家鼠染色体减数分裂联会复合体的研究

以界面铺张－硝酸银和磷钨酸（ＰＴＡ）染色技术，对褐家鼠（Ｒａｔｔｕｓｎｏｒｖｅｇｉｃｕｓ）减数分裂前期精母细胞联会复合体（ＳＣ）进行了显微和亚显微结构的观察．银染和ＰＴＡ染色都清楚地显示出褐家鼠ＳＣ的结构和减数分裂的行为，但ＰＴＡ染色的ＳＣ标本，着丝位区域较为明显．ＳＣｓ核型与有丝分裂染色体核型有较好的一致性．     

Integration of cytogenetic landmarks into the draft sequence of the human genome

We have placed 7,600 cytogenetically defined landmarks on the draft sequence of the human genome to help with the characterization of genes altered by gross chromosomal aberrations that cause human disease. The landmarks are large-insert clones mapped to chromosome bands by fluorescence in situ hybridization. Each clone contains a sequence tag that is positioned on the genomic sequence. This genome-wide set of sequence-anchored clones allows structural and functional analyses of the genome. This resource represents the first comprehensive integration of cytogenetic, radiation hybrid, linkage and sequence maps of the human genome; provides an independent validation of the sequence map and framework for contig order and orientation; surveys the genome for large-scale duplications, which are likely to require special attention during sequence assembly; and allows a stringent assessment of sequence differences between the dark and light bands of chromosomes. It also provides insight into large-scale chromatin structure and the evolution of chromosomes and gene families and will accelerate our understanding of the molecular bases of human disease and cancer.     

七星瓢虫精母细胞减数分裂观察研究

研究昆虫染色体一般选择在生殖细胞减数分裂阶段.七星瓢虫初级精母细胞的染色体计数为2n=16+X,减数分裂中期,n=8、n=8+X,染色体形态多为短棒状(参见照片).其性染色体是属XD型,雌性XX,为同配性别,雄性为异配性别,即X0(2n—1).昆虫染色体数量、组型,以及减数分裂的配对情况均与分类学亲缘关系有相关性,是昆虫正确鉴定和系统分类的重要参考.     

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L

Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O.sativa and O.meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.     

Hybridization and adaptive mate choice in flycatchers

Hybridization in natural populations is strongly selected against when hybrid offspring have reduced fitness. Here we show that, paradoxically, pairing with another species may offer the best fitness return for an individual, despite reduced fitness of hybrid offspring. Two mechanisms reduce the costs to female collared flycatchers of pairing with male pied flycatchers. A large proportion of young are sired by conspecific male collared flycatchers through extra-pair copulations, and there is a bias in favour of male offspring (which, unlike females, are fertile) within hybrid pairs. In combination with temporal variation in breeding success, these cost-reducing mechanisms yield quantitative predictions about when female collared flycatchers should accept a male pied flycatcher as a mate; empirical data agree with these predictions. Apparent hybridization may thus represent adaptive mate choice under some circumstances.     

Molecular cytogenetic analysis of Triticum aestivum-Leymus racemosus reciprocal chromosomal translocation T7DS·5LrL/T5LrS·7DL

In order to induce chromosome translocation between wheat chromosomes and chromosome 5Lr of Leymus racemosus, the mi- crosporocytes during meiosis of T. aestivum-L. racemosus disomic addition line DA5Lr were irradiated by 60Co γ-rays 800 R (100 R/min). Before flowering, the treated spikes were emasculated and bagged. After 2-3 d, the emasculated flowerets were pollinated using pollens from T. aestivum cv. Chinese Spring. One plant with two translocation chromosomes involved in both the long and short arm of...