A Brassica campestris-alboglabra addition line and its use for gene mapping,intergenomic gene transfer and generation of trisomics

Summary Brassica campestris-alboglabra monosomic addition lines were developed from a trigenomic Brassica hybrid (2 n=3 x=29, AAC) obtained by backcrossing a resynthesized B. napus (2 n=4 x=38, AACC) line to its parental B. campestris (2 n=2 x=20, AA) line. One addition line was characterized genetically with three loci specific for the alien chromosome and cytologically by meiotic analysis. The following results were obtained. (1) The same chromosome in the B. alboglabra (2 n= 2 x=18, CC) genome carried the three loci, E _c, W _c and Lap-1 C ^c, which control the biosynthesis of erucic acid, white flower colour and the faster migrating band of leucine aminopeptidase, respectively. The linear order and possible positions of the three loci were inferred. The meiotic behaviour of the alien chromosome was documented and its transmission frequency was assessed. (2) Intergenomic recombination frequently occurred in the monosomic addition line, resulting in the introgression of one or two loci from the alien chromosome into the B. campestris genome. (3) B. campestris trisomics were found in the progeny of the monosomic addition line. (4) The removal of the other eight C-genome chromosomes from the trigenomic Brassica hybrid led to a dramatic increase in the erucic acid content of the monosomic addition line. (5) No offspring of the trigenomic Brassica hybrid showed evidence of intergenomic recombination and introgression of the W _c locus into the B. campestris genome. It is questioned whether such a difference might be due to a possible regulating mechanism for homoeologous chromosome pairing.     

Herbicide-resistant Indica rice plants from IRRI breeding line IR72 after PEG-mediated transformation of protoplasts

The commercially important Indica rice cultivar Oryza sativa cv. IR72 has been transformed using direct gene transfer to protoplasts. PEG-mediated transformation was done with two plasmid constructs containing either a CaMV 35S promoter/HPH chimaeric gene conferring resistance to hygromycin (Hg) or a CaMV 35S promoter/BAR chimaeric gene conferring resistance to a commercial herbicide (Basta) containing phosphinothricin (PPT). We have obtained so far 92 Hg^r and 170 PPT^r IR72 plants from protoplasts through selection. 31 Hg^r and 70 PPT^r plants are being grown in the greenhouse to maturity. Data from Southern analysis and enzyme assays proved that the transgene was stably integrated into the host genome and expressed. Transgenic plants showed complete resistance to high doses of the commercial formulations of PPT.     

Methionine adenosyltransferase activity in cultured cells and in human tissues

We have investigated methionine adenosyltransferase activity (MAT) in extracts of a variety of normal and malignant human tissues and cultured cell lines. MAT activity assayed from 17 different cultured cell lines varied to a great extent. Ramos (human, Burkitt's lymphoma) and EL4 (mouse, T cell lymphoma) cell showed MAT activity near 300 pmol/mg per min. Daudi (human, Burkitt's lymphoma) and almost all monolayer cells had MAT activity below 100 pmol/mg per min. Human peripheral blood lymphocytes had MAT activity of 36 pmol/mg permin. The MAT activity of the cell lines can be related to doubling time: cell lines with short doubling times have much higher MAT activity than other cell lines. A large variation in MAT activity in different human tissues was observed. In autopsy samples MAT activity was highest in the brain and in the colon. Malignant tissue samples gave much higher MAT activity than normal tissues. Lung cancer (carcinoma squamocellulare pulmonis) had MAT activity of 30.7 pmol/mg per min, while in normal lung it was 2.4 pmol/mg per min.     

粘、易型1B/1R小麦雄性不育系产生单倍体的遗传机理及育性恢复性能的研究

调查了ms(Ae.kotschyi)-77(2)和ms(Ae.variabilis)-77(2)低、高世代和在转育、组配中单倍体频率的变化趋势及在不同胞质间、核型间存在的变异。结果表明:(1)粘、易型1B/1R小麦雄性不育系产生单倍体的遗传机理是由于1B/1R卵细胞与粘、易胞质的专一互作,并在花粉蒙导下而导致孤雌生殖的结果;(2)1B·1B/1R杂合核型比1B/1R·1B/1R纯合核型产生的单倍体频率高,1B/1R·1B/1R纯合核型世代间单倍体诱导频率相对稳定;(3)在同一核背景下,诱导单倍体频率粘质高于易质;(4)用不同来源的1B/1R易位系来转育粘、易型不育系及用不同核型的父本与其组配杂种,诱导单倍体频率明显不同;依此差异进行亲本选择,分别可选出与组配出不产生或很少产生单倍体的粘、易型1B/1R不育系和F_1杂种。此外,分析了粘、易型1B/1R不育系一般恢复度不高的内在原由,认为与1B·1B/1R杂合核型中的易位染色体在减数分裂中能否正常联会配对直接相关。     

Development of the jamming avoidance response and its morphological correlates in the gymnotiform electric fish,Eigenmannia

The electric fish, Eigenmannia, will smoothly shift the frequency of its electric organ discharge away from an interfering electric signal. This shift in frequency is called the jamming avoidance response (JAR). In this article, we analyze the behavioral development of the JAR and the anatomical development of structures critical for the performance of the JAR. The JAR first appears when juvenile Eigenmannia are approximately 1 month old, at a total length of 13–18 mm. We have found that the establishment of much of the sensory periphery and of central connections precedes the onset of the JAR. We describe three aspects of the behavioral development of the JAR: (a) the onset and development of the behavior is closely correlated with size, not age; (b) the magnitude (in Hz) of the JAR increases with size until the juveniles display values within the adult range (10–20 Hz) at a total length of 25–30 mm; and (3) the JAR does not require prior experience or exposure to electrical signals. Raised in total electrical isolation from the egg stage, animals tested at a total length of 25 mm performed a correct JAR when first exposed to the stimulus. We examine the development of anatomical areas important for the performance of the JAR: the peripheral electrosensory system (mechano- and electroreceptors and peripheral nerves); and central electrosensory pathways and nuclei [the electrosensory lateral line lobe (ELL), the lateral lemniscus, the torus semicircularis, and the pacemaker nucleus]. The first recognizable structures in the developing electrosensory system are the peripheral neurites of the anterior lateral line nerve. The afferent nerves are established by day 2, which is prior to the formation of receptors in the epidermis. Thus, the neurites wait for their targets. This sequence of events suggests that receptor formation may be induced by innervation of primordial cells within the epidermis. Mechanoreceptors are first formed between day 3 and 4, while electroreceptors are first formed on day 7. Electroreceptor multiplication is observed for the first time at an age of 25 days and correlates with the onset of the JAR. The somata of the anterior lateral line nerve ganglion project afferents out to peripheral electroreceptors and also send axons centrally into the ELL. The first electroreceptive axons invade the ELL by day 6, and presumably a rough somatotopic organization and segmentation within the ELL may arise as early as day 7. Axonal projections from the ELL to the torus develop after day 18. Within the torus semicircularis, giant cells are necessary for the performance of the JAR. Giant cell numbers increase exponentially during development and the onset of the JAR coincides with a minimum of at least 150 giant cells and the attainment of a total length of at least 15 mm and at least 150 giant cells. Pacemaker and relay cells comprise the adult Eigenmannia pacemaker nucleus. The growth and differentiation of these cell types also correlates with the onset of the JAR in developing animals. We describe a gradual improvement of sensory abilities, as opposed to an explosive onset of the mature JAR. We further suggest that this may be a rule common in most developing behavioral systems. © 1992 John Wiley & Sons, Inc.     

Formation of Hassall's corpuscles in vitro by the thymic epithelial cell line IT-26 R 21 of the rat

Summary In the monolayer of an established epithelial cell line from the rat thymus, IT-26R21, characteristic cell aggregates quite similar to Hassall's corpuscles were formed. These aggregates were examined by light and electron microscopy, and immunohistochemically. Their interpretation as Hassall's corpuscles is based on the following observations: (1) The aggregates are formed in the monolayer of cells that greatly resemble medullary epithelial cells of the thymus. (2) They consist of flattened epithelial cells in a concentric pattern with one or more degenerating cells in the center. (3) Loss of microvilli suggests that these cells are keratinizing. (4) The aggregates show strongly positive reactions in immunofluorescent staining with antikeratin and antiprekeratin.When Hassall's corpuscles increase in size, cellular proliferation is somewhat suppressed. Both in vivo and in vitro, they may be interpreted as an expression of a changing growth pattern in confined spaces and thus seem to have little immunological function.     

Predominance of gonadotropin α-subunit resulting from preferential loss of hCGβ production in an established cell line

Summary The BeWo line of trophoblastic cells, maintained in continuous culture since 1966, was employed to investigate the phenomenon of gonadotropin α-subunit predominance that exists in several cell lines. The secretion of complete human chorionic gonadotropin (hCG) relative to α-subunit was compared in several different BeWo sublines, all of which were derived from BeWo stock roller tube colonies. In all of the BeWo sublines, secretion of hCG originally exceeded secretion of α-subunit. With time in culture, however, there was a marked decline in production of hCG/hCGβ, but not in α-subunit. Thus it appears that the production of hCGβ by BeWo choriocarcinoma cells is more labile than the production of the α-subunit.     

Pokeweed mitogen inhibition of protein synthesis in cultured lymphoblastoid lines

Summary Pokeweed mitogen (PWM) and ricin are both lectins derived from plant seeds. They are glycoproteins and share the ability to agglutinate a variety of animal cells including erythrocytes. The effect of these two lectins on protein synthesis was studied in four longterm lymphoblastoid lines (8866 and GM1531, which are B cell lines; and CCRF/CEM and MOLT 4, which are T-cell lines). Ricin (50 μg/ml) completely inhibited protein synthesis by 2 hr in both B-cell and T-cell lines as measured by the uptake to [³H]leucine. The PWM appeared more specific and at a concentration of 500 μg/ml inhibited protein synthesis only in B-cell lines (8866 and GM 1531). This effect was maximal at 5 hr. To investigate the reason for the differential effect of PWM on T and B cells,¹²⁵I-labeled PWM was incubated with 8866, MOLT 4, and CCRF/CEM to see if a significant difference in binding to B cells and T cells could be demonstrated. It does not appear that the differential effect on T and B cells is due to a difference in the amount of PWM bound. On the other hand it is possible that the B cells may bind some toxic subcomponent of the PWM preparation that the T cells do not bind because of a difference in composition or arrangement of cell surface glycoproteins.     

Two relative efficiencies of polymorphic enzymes for characterizing cell lines,detecting contaminations,and monitoring transplants

Summary A new calculation of the relative efficiency of polymorphic enzyme markers, called the REB, was determined and compared with one of Fisher's determinations of the relative efficiency called REA here. The REA estimates the chance of failing, and 1-REA of succeeding, to show a phenotypic difference between two randomly selected persons or cultured cell lines (Case 1). In this study it was shown that the REA also estimates the chance of detecting a cell line mislabeling or similar mixup (Case 2) and a cell line cross-contamination leading to the complete replacement of an original line by contaminating line (Case 3). The new REB determines the probability of failing, and 1-REB of succeeding, to detect a contamination of an original line by another line leading to their coexistence, or at least a sufficiently long period of transitional coexistence before one overgrows the other. The REA and REB also apply to determining the efficiency of polymorphic markers in detecting donor and recipient cells in tissue transplants. This work was developed from the author's involvement in the human tumor cell-line characterization project at Sloan-Kettering Institute and he acknowledges this opportunity and the benefits of his association with Dr. J?rgen Fogh and colleagues in the Human Tumor Cell Laboratory.