棉花体细胞染色体rDNA-FISH技术

介绍了棉花体细胞染色体为靶、r DNA为探针的荧光原位杂交实验技术 ,并着重分析和讨论了棉花 r DNA- FISH技术的关键因素 ,包括染色体制片、探针与染色体共变性等。作为靶染色体的棉种包括陆地棉、海岛棉、草棉、亚洲棉、克劳茨基棉和比克氏棉等 ,作为封阻的是鲑鱼精 DNA。     

Quantitative impact of CO2 enriched atmosphere on abundances of methanotrophic bacteria in a meadow soil

In the Giessen free-air CO₂ enrichment (GiFACE) experiment, 5 years of CO₂ enrichment led to decreased CH₄ uptake rates of the investigated meadow soil. In soils, CH₄ is mainly oxidised by methanotrophic bacteria. In the present study, abundances of methanotrophic bacteria and total bacteria in soil samples from the GiFACE experiment were quantified by applying pmoA- and 16S rRNA gene-targeted real-time PCR and fluorescence in situ hybridisation (FISH). Methanotrophic bacteria of the Methylosinus group (Alphaproteobacteria) and the Methylobacter/Methylosarcina group (Gammaproteobacteria) were detectable by real-time PCR as well as by FISH. Both quantitative analytical approaches revealed that abundances of either bacteria or methanotrophic bacteria in soil samples from sites under CO₂-enriched atmosphere were decreased. Compared to ambient site, only 46 and 30.5% of methanotrophic bacteria and 38 and 63.2% of total bacterial cell numbers could be detected under CO₂-enriched atmosphere by FISH and real-time PCR, respectively. These results suggest that significantly decreased abundances of methanotrophic bacteria could explain reduced CH₄ uptake rates.     

薄片牡蛎的核型及18S-28S核糖体rRNA基因的染色体定位

以薄片牡蛎（Dendostrea folium）成体鳃组织为材料制备有丝分裂中期染色体标本,对其染色体核型进行了分析,并运用荧光原位杂交技术（FISH）将18S-28S核糖体RNA基因定位于中期染色体上。FISH探针是通过PCR扩增介于18S-28S rRNA基因之间的ITS和5.8S rRNA基因序列,并在PCR扩增过程中掺入了Biotin-11-dUTP进行生物素标记。结果显示,薄片牡蛎的单倍染色体数目为n=10,全部为中部着丝粒染色体。与大多数已知巨蛎属牡蛎的染色体核型相似。ITS探针在薄片牡蛎中期分裂体相上产生两簇FISH信号,分别杂交于2号染色体短臂的近端粒区域。本研究首次报道了薄片牡蛎的中期染色体核型以及18S-28S核糖体RNA基因在染色体上的定位。     

玉米和类玉米杂交后代异染色质纽重复序列的遗传稳定性

【目的】探究异染色质纽在玉米、类玉米及其杂种后代染色体上分布的特点及遗传稳定性。【方法】利用组成玉米异染色质纽的180-bp重复序列和TR-1元件对二倍体多年生类玉米（Zea diploperennis, DP）、玉米自交系330及其远缘杂交后代异源种质纯系540的有丝分裂中期染色体进行荧光原位杂交,观察杂交信号在3个材料染色体上的位置、强弱及分布数量。【结果】玉米自交系330的第2、3、5、6、7号染色体长臂的近末端区显示较强杂交信号。DP的第2、3、5、6、7、8、9号染色体的长臂末端检测出异染色质纽,其中2号染色体在短臂的末端也检测到异染色质纽杂交信号。杂交后代540的第2、5、7染色体长臂近末端区检测到较强的杂交信号;3个材料的第6染色体短臂末端的随体上均显示强杂交信号。【结论】玉米大部分异染色质纽成分不能稳定遗传,其表现出来的多态性可以作为鉴定玉米和类玉米杂种后代的细胞学标记。     

应用荧光原位杂交技术进行哺乳动物性别鉴定的研究进展

荧光原位杂交（FISH）技术应用于哺乳动物性别鉴定,近年来取得了长足进展。本文阐述了荧光原位杂交技术的原理及其在胚胎性别鉴定方面和精细胞筛选鉴定方面较PCR法和LAMP法所具有的优势,并着重概述了该技术在人类医学和家畜性别鉴定方面的研究进展和应用前景。     

顶芒和无芒山羊草育种价值及细胞学标记

为了评价顶芒山羊草和无芒山羊草在小麦育种中的利用价值,本研究利用人工接种和近红外二极管阵列分析仪分别对圆锥小麦-顶芒山羊草双二倍体和中国春-无芒山羊草双二倍体的抗病性及籽粒品质进行了鉴/测定。结果表明,二者均近免疫小麦条锈病和白粉病,且其籽粒蛋白和湿面筋含量均极显著高于对照小麦,因此,这2个种质值得向小麦进行回交转育。为了建立可用于鉴定小麦背景中2种山羊草的细胞遗传标记辅助鉴定相应回交群体,利用寡聚核苷酸原位杂交(FISH)方法对2份双二倍体进行分析。结果发现,探针(GAA)8可有效鉴定小麦背景中的顶芒山羊草染色体;探针Oligop Sc119.2-1、Oligo-p Ta-535-1和(GAA)8结合使用可有效鉴定小麦背景中的无芒山羊草染色体。本研究首次建立的2种山羊草细胞遗传学标记可用于相应杂种后代的筛选与鉴定工作,为辅助选育含有2种山羊草优异基因的小麦新种质奠定了基础。     

Acute myeloblastic leukemia with associated BCR-ABL translocation in a dog

An 8-year-old male neutered Labrador Retriever was referred to the University of Wisconsin Veterinary Medical Teaching Hospital with a presumptive diagnosis of leukemia. Hematologic abnormalities included normal neutrophil count with a left shift, monocytosis, eosinophilia, thrombocytopenia, and circulating immature mononuclear cells. Bone marrow was effaced by immature hematopoietic cells of various morphologic appearances. In addition, large multinucleated cells were observed frequently. Flow cytometric analysis of nucleated cells in blood revealed 34% CD34(+) cells, consistent with acute leukemia. By immunocytochemical analysis of cells in blood and bone marrow, some mononuclear cells expressed CD18, myeloperoxidase, and CD11b, indicating myeloid origin; some, but not all, large multinucleated cells expressed CD117 and CD42b, the latter supporting megakaryocytic lineage. The diagnosis was acute myeloblastic leukemia without maturation (AML-M1). To identify genetic aberrations associated with this malignancy, cells from formalin-fixed paraffin-embedded bone marrow were analyzed cytogenetically by multicolor fluorescence in situ hybridization (FISH). Co-localization of bacterial artificial chromosome (BAC) containing BCR and ABL was evident in 32% of cells. This confirmed the presence of the canine BCR-ABL translocation or Raleigh chromosome. In people, the analogous translocation or Philadelphia chromosome is characteristic of chronic myelogenous leukemia (CML) and is rarely reported in AML. BCR-ABL translocation also has been identified in dogs with CML; however, to our knowledge this is the first report of AML with a BCR-ABL translocation in a domestic animal.     

六倍体小黑麦染色体的荧光原位杂交分析

采用基因组原位杂交(GISH)和荧光原位杂交(FISH)技术对六倍体小黑麦进行了研究,以鉴定其含有的黑麦染色体的数目和结构特点,了解小黑麦基因组组成,为其在小麦遗传育种中的应用提供参考.分别以黑麦基因组DNA及pSc200和pSc250探针进行顺序GISH-FISH分析,结果显示该小黑麦含有21对染色体,来源于黑麦的2对染色体的长臂端部出现pSc200和pSc250信号,而另外5对染色体的两端部都有该信号.以Oligo-pSc119.2-1,pSc200和pSc250探针对小黑麦进行了双色FISH分析,发现小黑麦的Oligo-pSc119.2-1信号与黑麦基本相似,而pSc200和pSc250信号与黑麦的差异较大.以黑麦基因组DNA及Oligo-pAs1-1和Oligo-pSc119.2-1为探针对小黑麦进行顺序GISH-FISH分析,鉴定其基因组组成为AABBRR;同时发现小黑麦染色体的一些FISH信号发生了变化,其原因可能是小黑麦形成过程中染色体结构发生了改变,从而导致小黑麦的DNA重复序列呈现多态性.     

45S rDNA在不同倍性沙田柚染色体上的荧光原位杂交分析

 应用荧光原位杂交技术进行45S rDNA在不同倍性沙田柚中期染色体上的定位研究。结果表明: 不同材料的NOR位点表现出明显的多态性和杂合性, 即二倍体具4个45S rDNA位点, 四倍体单株t₁和t₂ 则分别具有10个和8个位点。位点在染色体上的位置包括短臂末端、着丝粒区域、着丝粒至短臂区域、着丝粒至长臂区域、长臂末端共5种不同类型。不同材料中包含的位置类型不完全相同。不同位置类型的位点数也不相同, 以着丝粒及着丝粒至短臂区域的类型位点数最多, 其次是短臂末端。同源四倍体和相应的二倍体相比, 出现了DNA水平的变异。对用于荧光原位杂交的柑橘染色体制片方法和rDNA在染色体上的分布位置、同源四倍体和相应二倍体的遗传差异分析、重复序列的染色体原位杂交用于染色体识别和物种演化研究等进行了讨论。