内麦系列小麦品种(系)的染色体结构变异分析

为明确内麦系列小麦品种(系)的染色体结构特点,利用荧光原位杂交(fluorescence in situ hybridization, FISH)技术和pTa535、pSc119.2探针对21份内麦系列小麦材料的染色体进行分析。结果显示,5份材料含1RS/1BL易位染色体;探针除3A、7A、2B、3B、4B、2D和4D外的其他染色体在21份材料间存在结构差异;共鉴定出49种染色体多态类型,其中数量最多的是A基因组(22),其次是B基因组(14)和D基因组(13);在5B、6B、3D和7D染色体的同源染色体间存在不对称核型;根据A、B和D基因组的FISH核型,把21份材料分为16类。通过FISH信号模式,发现所有材料在染色体水平上都有变异。多数材料的染色体类型一致,推测这些材料的遗传背景一致或者相似。     

巴西橡胶树MADS-box基因家族6个成员的染色体物理定位

MADS-box基因家族广泛分布于真核生物中,巴西橡胶树的MADS-box基因家族主要参与花形态建成,对生殖生长起到重要的调节作用。目前,MADS-box基因家族的26个相关基因已被克隆分析,但它们在染色体上的具体位置还未确定。本研究以巴西橡胶树‘热研7-33-97’品种为材料,将MADS-box基因家族的6个成员（HbAGL8、HbAG15、HbAGL30、HbTT16、HbAP1和HbSVP1）定位在细胞核染色体上,通过双探针荧光原位杂交技术（FISH）对巴西橡胶树MADS-box基因家族的这6个成员在细胞核染色体上进行物理定位分析。结果表明：MADS-box基因家族的6个基因分别位于不同的染色体上,其中HbAGL15、HbAG8、HbAG30和HbSVP1基因定位在第4、5、7和8号染色体长臂上,其信号位点到着丝粒的平均百分距离是11.85、39.71、48.94和6.70;HbTT16和HbAP1基因定位在第1和13号染色体短臂上,其信号位点到着丝粒的平均百分距离是22.19和18.01。本研究结果揭示了巴西橡胶树MADS-box基因家族的6个成员在细胞核染色体上的实际位置,展现家族基因之间的分布特点和连锁遗传关系,不仅丰富了橡胶树分子细胞遗传学信息,也为橡胶树的分子辅助育种和比较基因组学研究提供了分子细胞遗传学的科学理论依据。     

环境生物新技术在土壤微生态研究中的应用

鉴于传统的微生物分离培养技术已不能满足环境微生物研究的需要。为此,在介绍变性梯度凝胶电泳(DGGE)以及荧光原位杂交(FISH)等分子生物学新技术的基础上,对这些技术在土壤微生态研究中存在的问题和发展前景做了展望;提出将这些技术整合应用到复杂土壤的环境微生态研究中,以了解微生物的时空分布、种群结构及其与功能之间的关系。     

西藏开菲尔粒中酵母菌的分离与初步鉴定

西藏开菲尔粒(俗称藏灵菇)是由多种乳酸菌(间或少量醋酸菌)和酵母菌共生而成一种外形酷似菜花的白色或乳黄色团状聚合体,是目前已知的唯一可用于酸奶制作的天然野生菌种资源。采用扫描电镜、分离培养、26S rRNA基因测序与序列分析、荧光原位杂交(FISH)相结合的技术方法,对西藏开菲尔粒中的酵母菌进行了分离培养、鉴定和形态观察。结果显示:西藏开菲尔粒中酵母菌多为球形(2.0~5.0μm)或椭圆形(1.0~3.0×2.0~4.0μm);从西藏开菲尔粒中分离出的16株酵母菌分属于酿酒酵母属(Saccharomyces)、克鲁维酵母属(Kluyveromyces)、亚罗酵母属(Yarrowia)这3个属中的4个种,其中3株是酿酒酵母菌(Saccharomyces cerevisiae),8株为马克斯克鲁维酵母菌(Kluyveromyces marxianus),3株为解脂亚罗酵母菌(Yarrowia lipolytica)。     

杂交三倍体泥鳅染色体C带及rDNA序列的FISH定位

对杂交三倍体泥鳅(4n♀×2n♂)的胚胎染色体进行了C带及染色体荧光原位杂交(FISH)分析,首次探讨了杂交三倍体泥鳅C带特征,为准确鉴别染色体提供依据,研究了核糖体5.8S+28S r DNA在杂交三倍体泥鳅胚胎中期染色体上的数目和位置。C带结果表明,杂交三倍体泥鳅的染色体C带包括臂端C带和着丝粒C带,没有发现臂间C带。M染色体只有1号染色体既有臂端C带又有着丝粒C带,但臂端C带均比着丝粒C带大,信号也比着丝粒的强;而其他M染色体及SM染色体、T染色体只有着丝粒C带。FISH分析显示,核糖体5.8S+28S r DNA清楚地定位在杂交三倍体泥鳅中期染色体中部着丝粒染色体(M)的端部区域,在杂交三倍体泥鳅中期染色体上可以检测到三簇杂交信号。该结果从分子细胞遗传学层面进一步证实了杂交三倍体泥鳅是含有三套染色体组的遗传三倍体。     

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.     

巴西橡胶树胶乳转化酶HbNIN基因家族物理定位的研究

胶乳转化酶(Invertase,Inv)是控制橡胶树胶乳蔗糖代谢和影响胶乳(橡胶)产量的关键酶。已有研究证实了胶乳转化酶属于中性/碱性转化酶。本研究以热研7-33-97(2n=36)为材料制备叶片细胞染色体标本,采用荧光原位杂交技术(fluorescence in situ hybridization,FISH)对胶乳转化酶基因HbNIN1、HbNIN2和HbNIN3进行了染色体物理定位的初步研究。结合核型分析,初步确定HbNIN1基因位于5号染色体的长臂上,其信号位点到着丝粒的百分距离为33.1;HbNIN2基因位于2号染色体的长臂上,其信号位点到着丝粒的百分距离为35.7;HbNIN3基因位于3号染色体的短臂上,其信号位点到着丝粒的百分距离为40.42。由此揭示了该基因家族在染色体上的位置和分布特点。     

Utilization of flow cytometry for festulolium breeding (Lolium multiflorum (2x) × Festuca arundinacea (6x))

Festulolium is a hybrid between Festuca and Lolium species that has valuable agronomic traits from both grass species. The purpose of our breeding program is to produce hexaploid festulolium that introduces tolerance to summer depression into Italian ryegrass (Lolium multiflorum) by crossing it with tall fescue (Festuca arundinacea). However, we found the DNA ploidy of hexaploids was not stable and was reduced in successive generations. We aimed to find out how to obtain stable high-ploidy festulolium. F1 hybrids of L. multiflorum and F. arundinacea were produced. The F3 generation was produced from putative hexaploid F2 individuals by open pollination. The F4 to F6 generations were obtained by polycrossing. The DNA ploidy levels of F2 to F6 individuals were estimated by flow cytometry. Cytological characteristics of the F5 and F6 individuals were investigated by FISH and GISH. The DNA ploidy level of hexaploid festulolium was reduced and stabilized at almost the same level as a tetraploid. Seed fertility was inversely correlated with an increase in ploidy level. GISH revealed no preferential Lolium transmission. FISH with a telomere probe revealed that counting the exact number of chromosomes in festulolium was difficult. DNA ploidy level was strongly correlated with the number of chromosomes.     

污水处理反硝化除磷-诱导结晶磷回收工艺中除磷微生物特性

为探究反硝化除磷-诱导结晶磷回收工艺中缺氧池污泥释磷、吸磷以及微生物特征,利用荧光原位杂交(fluorescence in situ hybridization,FISH)技术、电子扫描显微镜(scanning electron microscope,SEM)观察了微生物的数量、分布和形态;通过批次试验考察了污泥在厌氧/好氧和厌氧/缺氧2种模式下的释磷和吸磷特征。结果表明:该双污泥系统缺氧池中聚磷菌占总细菌比例的69.7%,明显高于单污泥系统中富集的聚磷菌比例,污泥中的微生物多呈杆状;厌氧/好氧、厌氧/缺氧模式下单位污泥浓度(mixed liquor suspended solids,MLSS)总吸磷量(以PO43--P计)分别为22.84、18.60 mg/g,反硝化聚磷菌(denitrifying polyphosphate-accumulating organisms,DPAO)占聚磷菌(polyphosphate-accumulating organisms,PAO)的比例为81.44%,表明在长期的厌氧/缺氧运行条件下可以富集到以硝酸盐为电子受体的反硝化聚磷菌,同时还存在着仅以氧气为电子受体的聚磷菌;通过pH值和氧化还原电位(oxidation reduction potential,ORP)的实时监测可以快速地了解污水生物处理系统中各类反应的进程,对调控工艺参数有着重要的意义。综上所述,为保证污水生物处理工艺的正常稳定运行,将微生物分析与常规的化学参数分析结合起来考察将是未来发展的必然趋势。     

几种鉴定小麦背景中1BL/1RS易位染色体的分子标记方法比较研究

对 5种鉴定小麦背景中 1BL/ 1RS易位染色体的生化和分子标记方法 ,包括酸性聚丙烯酰胺凝胶电泳 (A PAGE)、荧光原位杂交 (FISH)、RFLP、RAPD和特异性PCR标记 ,进行比较研究。结果表明 ,RAPD标记难以鉴定1BL/ 1RS易位染色体 ,其余均能对 1BL/ 1RS易位染色体进行有效鉴定。FISH和RFLP方法比较费时费力且花费昂贵 ,而特异性PCR标记又在一定程度上受模板DNA浓度的影响。据此认为 ,APAGE方法是一种简单、快速、经济有效的方法 ,最易于在小麦育种选择中应用