A nuclear protein associated with cell divisions in plants

A nuclear protein, present in carrot meristems and rapidly proliferating cultured cells of carrot (Daucus carota L.) has been identified by the use of a monoclonal antibody (MAb 21D7). By combining the techniques of two-dimensional polyacrylamide gel analysis and blotting separated proteins onto nitrocellulose sheets, it was shown that the antibody detected a single polypeptide of apparent molecular mass (M _r) of 45000 and an isoelectric focusing point (pI) of 6.7. This protein was found by subcellular fractionation and immunofluorescence to be highly concentrated in the nucleoli of somatic and zygotic embryos of a wide range of plants. It was not detectable in logarthmically growing cells ofEscherichia coli, yeast, embryos ofDrosophila melanogaster or cultured C3H mouse cells. These data indicate that this protein is a highly conserved non-histone protein associated with nuclei of rapidly dividing plant cells.Abbreviations M _r apparent molecular mass - Da dalton - Ig immunoglobulins - MAb monoclonal antibody - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - 2-D gel two-dimensional gel electrophoresis - 2,4-D 2,4-dichlorophenoxyacetic acid     

Monensin inhibits the secretion of α-amylase but not polysaccharide slime from seedling tissues ofZea mays

Summary The effect of monensin on polysaccharide slime secretion by root tips of corn (Zea mays) was studied. Various treatment times and ionophore concentrations were tested: none resulted in inhibition of slime secretion. Because monensin changes the pH of the medium, its effect was also monitored in strongly buffered media and at different pH's. Even in such media, monensin did not inhibit slime secretion. We also measured the effect of the drug after a pulse with [³H]fucose or a pulse followed by a chase. The amount of labeled slimed secreted was not altered by the ionophore. However, 10M monensin affected the development of root tips and drastically reduced their growth. We showed that monensin inhibits the secretion of -amylase by the scutellum of the same plantlet. The importance of the nature of the secretory compound in relation to monensin inhibition of its secretion is discussed.Abbrevations Hepes N-2-hydroxyethylpiperazine-N-2-ethane-sul-fonic acid - Mes 2-(N-morpholino)ethane-sulfonic acid     

Characterization of the pyrenoid isolated from unicellular green algaChlamydomonas reinhardtii: Particulate form of RuBisCO protein

Summary The pyrenoid is a protein complex in the chloroplast stroma of eukaryotic algae. After the treatment with mercury chloride, pyrenoids were isolated by sucrose density gradient centrifugation from cell-wall less mutant cells, CW-15, as well as wild type cells, C-9, of unicellular green algaChlamydomonas reinhardtii. Pyrenoids were characterized as a fraction whose protein/chlorophyll ratio was very high, and also examined by Nomarski differential interference microscopy. Most of the components consisted of 55 kDa and 16 kDa polypeptides (11) which were immunologically identified as the large and small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) protein, respectively. Some minor polypeptides were also detected. Substantial amount of RuBisCO protein is present as a particulate form in the pyrenoid in addition to the soluble form in algal chloroplast stroma.Abbreviations BPB bromophenol blue - DAB 3,3-diaminobenzidine - DTT dithiothreitol - ELISA enzyme-linked immunosorbent assay - High-CO₂ cells cells grown under air enriched with 4% CO₂ - Low-CO₂ cells cells grown under ordinary air (containing 0.04% CO₂) - NP-40 nonionic detergent (Nonidet) P-40 - PAGE polyacrylamide gel electrophoresis - PAP peroxidase-antiperoxidase conjugate - RuBisCO ribulose-1,5-bisphosphate carboxylase/oxygenase - RuBP ribulose-1,5-bisphosphate - SDS sodium dodecylsulfate     

Detection of linkage between marker loci and loci affecting quantitative traits in crosses between segregating populations

Summary By making use of pedigree information and information on marker-genotypes of the parent and F-1 individuals crossed to form an F-2 population, it is possible to carry out a linkage analysis between marker loci and loci affecting quantitative traits in a cross between segregating parent populations that are at fixation for alternative alleles at the QTL, but share the same alleles at the marker loci. For two-allele systems, depending on marker allele frequencies in the parent populations, 2–4 times as many F-2 offspring will have to be raised and scored for markers and quantitative traits in order to provide power equivalent to that obtained in a cross between fully inbred lines. Major savings in number of F-2 offspring raised can be achieved by scoring each parent pair for a large number of markers in each chromosomal region and scoring F-1 and F-2 offspring only for those markers for which the parents were homozygous for alternative alleles. For multiple allele systems, particularly when dealing with hypervariable loci, only 10%–20% additional F-2 offspring will have to be raised and scored to provide power equivalent to that obtained in a cross between inbred lines. When a resource population contains novel favorable alleles at quantitative trait loci that are not present (or rare) in a commercial population, analyses of this sort will enable the loci of interest to be identified, mapped and manipulated effectively in breeding programs.Contribution no. 2124-E, 1987 series from The Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel     

大剂量~(60)Co照射后造血干细胞潜在的残留损伤

本文对受到临界全致死剂量——8.5Gy~(60)Co照射后的小鼠造血干细胞(HSC)的自我更新力进行了测试,并对照射后4个月,造血功能业已恢复的小鼠HSC的造血重建功能进行了研究。结果表明:应用骨髓连续移植实验所测得的照后3个月中骨髓CFU_s的自我更新潜能明显衰退了。用照射后造血恢复小鼠的CFU_s给全致死剂量照射的受体进行移植治疗,发现其移植效力比正常的显著减弱。在受体存活30天时,CFU_s的再生速率只有正常的1/17。通过对性染色体追踪观察的资料分析,此种小鼠的??造血细胞在受体中难以形成长期稳定的嵌合体。以上事实反映出,照射后小鼠的HSC的造血重建功能大大削弱了,揭示出残存干细胞质量上的缺陷。对于这种潜在的残留损伤的机制,从“干细胞增殖力耗竭学说”和“基因自我修整假说”的角度进行了讨论。     

核微管组织中心的研究

本文利用间接免疫荧光手段对间期核微管组织中心的性质进行了初步探讨。当分离核与6S微管蛋白保温后,用抗管蛋白抗体进行间接免疫荧光染色,在群体中只有极少数核上有微管生长。Triton X-100或溶血卵磷脂去膜核的实验证实能长微管的核是核膜缺失或破损核。用秋水仙素解聚核上长出的微管,可见核内荧光亮点数多于染色体数,说明核内微管组织中心数目多于着丝粒数,很可能包括着丝粒和一部分染色质颗粒。将秋水仙素和6S微管蛋白混合后与去膜核保温,结果6S微管蛋白与核内微管组织中心有亲和力。本文就这些结果进行了讨论。     

Responses of Cultured Melanocytes to Defined Growth Factors

To proliferate in vitro, normal melanocytes, unlike normal fibroblasts, require specific growth factors in addition to those supplied in serum. The substances that promote melanocyte proliferation, such as 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and stimulators of cyclic adenosine monophosphate (cAMP), also promote pigmentation. Consequently, cell division and expression of at least some differentiated functions are not mutually exclusive for melanocytes. At present, the only known natural growth factor that can replace TPA in normal human melanocyte cultures is basic fibroblast growth factor (bFGF). Like TPA, bFGF is effective, most of the time, only in the presence of added cAMP. Some preparations of bFGF, however, may have a highly labile, intrinsinc cAMP stimulatory activity. It is thus possible that bFGF can assume two forms, dependent on and independent of cAMP stimulatory activity. Alternatively, a second factor may exist in pituitary glands that co-purifies with bFGF but deteriorates with storage. Abnormal melanocytes in culture, such as those derived from dysplastic nevi and primary melanomas, depend on the specific factors (bFGF and cAMP), whereas melanocytes from metastatic melanomas do not     

白毛新木姜子[Neolitsea aurata var.glauca]胚胎学研究

应用光学显微镜对白毛新木姜子的胚胎学特征进行了研究,首次在非寄生性樟科植物中发现了细胞型胚乳。对樟科8属进行了胚胎学特征的比较。花药四室,药室壁的发育属于"基本型",周原质团型绒毡层。小孢子母细胞连续型分裂。等四面体型四分体。二细胞成熟花粉,无孔沟。雌孢原多个,一般仅一个能继续发育。蓼型胚囊。助细胞具丝状器。反足细胞宿存。大孢子母细胞和合子具极性。宿存的一个助细胞具有吸器功能。细胞型胚乳,胚胎发育属于柳叶菜型的三叶变型。种皮源于外珠被,内表皮细胞壁螺旋状加厚。胚胎学特征表明,新木姜子属与木姜子属有密切的亲缘关系。较多的双胚囊异常现象,支持樟科与Monimiaceae具有紧密关系的推测。胚胎学特征不支持将无根藤属独立为科的观点。     

泡状饶氏藻营养细胞的超微结构研究

本文报道我国特有的一种绿藻门植物－泡状饶氏藻营养细胞的超微结构特征,植物体由3层细胞组成,外层细胞最小,细胞质比较丰富,含有较多的各种细胞器,液泡体积较小,中层细胞具有很大的中央液泡,细胞质成为贴壁的薄层,各种细胞器结构仍清晰可见,内层细胞极度液泡化,细胞质呈现退化状态,周生的片状叶绿体上有许多大小不等的穿孔,使叶绿体呈网孔状外貌,叶绿体主要由许多成对的类囊体组成,叶绿体上往往有几个蛋白核,蛋白核经常被1或2条类囊体穿过,呈现分隔状,本文也报道了泡状饶氏藻的线粒体,质体,内质网,高尔基体和核内微管的结构特征,根据泡状饶氏藻的类囊体形态与Ulva mutabilis非常相似以及蛋白核的超微结构特征,它与石莼科植物可能有较密切的亲缘关系,属于绿藻门中进化的类群。     

Correlation of immunomodulatory and therapeutic activities of interferon and interferon inducers in metastatic disease

The mechanism of therapeutic activity of recombinant murine interferon-gamma (rMu IFN-gamma) and the IFN inducer polyinosinic-polycytidylic acid solubilized with poly-L-lysine in carboxy methyl cellulose (pICLC) in treating metastatic disease was investigated by comparing effector cell augmentation with therapeutic activity in mice bearing experimental lung metastases (B16-BL6 melanoma). Effector cell functions in spleen, peripheral blood, and lung (the organ with tumor) were tested after 1 and 3 weeks of rMu IFN-gamma or pICLC administration (intravenous, three times a week). In these studies, natural killer (NK), lymphokine-activated killer (LAK), cytolytic T lymphocytes (CTL) (against specific and nonspecific targets), and macrophage tumoricidal and tumoristatic activities were measured. rM IFN-gamma and pICLC had therapeutic activity and immunomodulatory activity in most assays of immune function examined. Specific CTL activity of pulmonary parenchymal mononuclear cells (PPMC), but not in splenocytes or peripheral blood lymphocytes (PBL), during week 3 and not during week 1, correlated with the therapeutic activity of rMu IFN-gamma and of pICLC. Macrophage tumoricidal activity in PPMC, but not in alveolar macrophages, also correlated with the therapeutic activity of rMu IFN-gamma, but the opposite was true for the therapeutic activity of pICLC. NK activity of PPMC, but not of splenocytes or PBL, during week 1 correlated with the therapeutic activity of pICLC; in contrast, NK activity at any site did not correlate with the therapeutic activity of rMu IFN-gamma. LAK activity at any site did not correlate with the therapeutic activity of either agent.