Tochtergenerationen von heterologen Implantaten beiAcetabularia

Zusammenfassung Zellkerne wurden ausAcetabularia mediterranea undAcetabularia crenulata isoliert und in das Cytoplasma kernloserAcetabularia (Polyphysa) cliftonii implantiert. Die so gewonnenen Kombinationen wurden bis zur Hut- und Zystenbildung gebracht. Die Morphogenese der Zellen, die nach Freisetzen der Gameten und nach Kopulation entstanden, entsprach der Art, von der die Kerne der Elterngeneration stammten. Auch das Isozymmuster der Malatdehydrogenase der F₁-Generation zeigte die Kernkontrolle der Elterngeneration.

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F₁-generation of heterologous implants inAcetabularia

Summary Cell nuclei were isolated fromAcetabularia mediterranea andAcetabularia crenulata and were implanted into the cytoplasm of enucleatedAcetabularia (Polyphysa) cliftonii. Such combinations formed caps, crysts, and gametes which were capable of copulation. The cells of the F₁-generation corresponded well to the species from which the nuclei of the parent generation originated with respect to morphogenesis as well as to the isozyme pattern of the malic dehydrogenase.

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Herrn H.Kretschmer danken wir für die Mithilfe bei der Herstellung der Implantate und bei der Aufzucht der Zellen.     

Changes of peroxidase,esterase isozyme activities and some cell inclusions in regenerated vascular tissues after girdling inBroussonetia papyrifera (L.) Vent

The isozyme zymogram of peroxidase and esterase, and some cell inclusion contents were changed with the differentiation of regenerated vascular stem tissues after girdling inBroussnetia papyrifera (L.). Vent. Presence or disappearance of some peroxidase and esterase isozyme bands was related to wounding. Some isoperoxidase bands disappeared at the time of vascular tissue formation, but some esterase isozyme bands appeared in phloem or cambial regions as sieve-like elements or mature xylem were formed. The inclusion grains progressively disappeared with the formation of callus and initiation of vascular meristems. The cell inclusions reappeared during the formation of regenrated vascular tissues. Histochemical study indicated that the inclusion grains could be a complex compound of a protein mass encircling polysaccharide in the center with a proteinous nucleus.     

Genetic and biochemical aspects of lactate dehydrogenase isozymes in the salmonid eye

Polyacrylamide gel electrophoresis reveals similar differences between the retinal-specific lactate dehydrogenase (LDH) isozymes of the salmon (Salmo salar L.) and the sea-trout (Salmo trutta forma trutta L.) as those previously described for the salmon and the brown trout (Salmo trutta L.). F₁ salmon × sea-trout hybrids give a classic hybrid isozyme pattern, but the F₂ hybrids all possess the parental sea-trout type pattern. Loss of part of the salmon genome in these latter hybrids is the most likely explanation. It was observed that when the individual eye isozymes of the salmon, the sea-trout, and the rainbow trout (Salmo gairdneri Richardson) were eluted from preparative polyacrylamide gels and re-electrophoresed, an apparent interconversion of certain isozyme bands occurred. This phenomenon was also evident using starch gel. However, the major cathodally migrating isozyme in each case (presumably the E4 isozyme) re-electrophoresed pure. The reasons for these interconversions are, as yet, unclear. Attempts to produce in vitro hybridization between the various isolated individual isozymes were unsuccessful. K_m pyruvate values for the different salmon isozymes were of the order expected from results already published for other teleosts.     

Phytohemagglutinin-enhanced hybrid colony formation by cells from aged mice: Expression of th 4mod-1 mouse locus in human-mouse hybrids

Summary Cells from a continuous human line and freshly isolated cells from old adult mice heterozygous at theMod-1 locus were fused in the presence of polyethylene glycol (PEG). The production of hybrid cells, as a function of PEG concentration in the presence and absence of phytohemagglutining (PHA), was measured by cell survival and proliferation on selective medium. The incorporation of PHA into the fusion mixture allowed cell fusion to take place at nontoxic concentrations of PEG. PHA increased the frequency of cell fusion and increased the production of viable hybrid cells from 138- to over 2800-fold depending on cell type. The results suggest that the procedure may have broad application in promoting the fusion of cells sensitive to PEG. Clones were analyzed for isozymes of malic enzyme and glucose-6-phosphate dehydrogenase. The expression of the gene encoding X-linked mouse glucose-6-phosphate dehydrogenase confirmed that the cells were hybrids. These cells lost other mouse isozymes rapidly. In those clones in which the mouse malic enzyme gene was expressed, the product ofMod-1 ^α was detected significantly more frequently than that ofMod-1 ^b.     

Characterization of clonal populations of theHeliothis zea cell line IPLB-HZ 1075

Summary A total of 24 clones (HZ 1075/UND-A through X) were initially isolated by dilution plating from the established IPLB-HZ 1075 cell line. Many of the isolates with highly vacuolated cytoplasms eventually died during subculturing. The cloned cell strains differed in their predominant morphology, cell doubling times, and relative ability to support replication of the singly encapsulated nuclear polyhedrosis virus ofHeliothis zea (HzSNPV). The origin of the cloned cell strains was confirmed by comparing their isozyme profiles with those of the parental IPLB-HZ 1075 cell line andH. zea larvae using stains for fumerate hydratase, lactate dehydrogenase (LDH), and malic dehydrogenase (MDH). One dipteran and several lepidopteran cell lines maintained in our lab were also separable using stains for LDH and MDH. This research was supported in part by USDA grant number GAM 8400211, and by a Jesse Jones Faculty Research Award from the University of Notre Dame to M. J. F.     

Isozymes in nutrient medium of suspension-cultured apple cells (Malus sylvestris Mill.)

The time course of the activities of esterase, -galactosidase, and -glucosidase in cell sap and nutrient medium in in vitro cultured apple cells (Malus sylvestris Mill.) was studied. The corresponding isozyme patterns and the intracellular and extracellular isozyme patterns of acid phosphatase and polyphenol oxidase were compared using isozyme visualization methods adapted to ultra-thin-layer isoelectric focusing. Neither quantitative (total activity) nor qualitative (isozyme pattern) data were congruent for cell saps and nutrient media. Malate dehydrogenase, malic enzyme, and glutamate dehydrogenase occurred in cell sap only. The extracellular activities probably originate to a great part from a programmed release by intact cells. Nutrient media of plant cell cultures constitute a rich source of active plant isozymes.     

Somatic hybridization in the gramineae: Pennisetum americanum (L.) K. Schum. (Pearl millet) +Panicum maximum Jacq. (Guinea grass)

Summary Protoplasts from Pennisetum americanum resistant to S-2-amino-ethyl-l-cysteine (AEC) were fused with protoplasts of Panicum maximum utilizing polyethylene glycol-dimethylsulfoxide after inactivation of the Pennisetum protoplasts with 1 mM iodoacetic acid. The iodoacetate treatment prevented division of Pennisetum protoplasts; therefore, only Panicum protoplasts and heterokaryons potentially could give rise to colonies. A second level of selection was imposed by plating 3–4-week-old colonies on AEC medium. Putative somatic hybrid calli were analyzed for alcohol dehydrogenase, 6-phosphogluconate dehydrogenase, aminopeptidase, and shikimate dehydrogenase isozymes. Three somatic hybrid cell lines (lines 2, 3, and 67) were identified which showed two bands of alcohol dehydrogenase activity representing homodimers of P. maximum and P. americanum as well as a novel intermediate band of activity where Panicum-Pennisetum heterodimers would be expected. Aminopeptidase and shikimate dehydrogenase were useful for identifying presumptive hybrid calli but the isozyme patterns were additive-evidence which would not preclude the selection of chimeric callus. A more complex isozyme pattern which varied among the somatic hybrids was observed for 6-phosphogluconate dehydrogenase. In the hybrid calli, the presence of DNA sequences homologous to both P. maximum and P. americanum sequences was confirmed by hybridization of a maize ribosomal DNA probe to XbaI and EcoRI restriction fragments. Growth of hybrid lines on various concentrations of AEC was either similar to the AEC-resistant parent (hybrid line 2) or intermediate between the resistant and sensitive parents (hybrid lines 3, 67).     

Analysis of the Esterase Isozymes in Three Lines and F1 in Oryza sativa and Prediction of Heterosis

Esterase isozyme patterns in the embryos of dry seeds of 114 combinations of steriles, maintainers, restorers and their F1 hybrids were analyzed with acrylamide gel eleetrophoresis. Usually six major bands were found and named 1A, 3A, 4A, 5A, 6A and 7A. The isoesterase zymograms in three lines--sterile, maintainer and restorer were diffcrent. There were seven types of zymograms in F1 hybrids. The eomplementary bands were shown in F1 hybrids when sterile with 6A band and restorer with 3A or 5A band were used as parents. F1 hybrids with 3A and 6A complementary each other were more vigorous in vegetative growth and only those 5A and 6A complemontary each other displayed economic superiority. It was shown that the pattern of esterase zymograms of F1 hybrids was influenced by both cytoplasm and nucleus of their parents. It was concluded that esterase isozyme patterns could be used as one of the biochemical markers for the predicting hybrid vigor in heterosis breeding.     

趋磁细菌WD-1同工酶分析

趋磁细菌在微好氧和好氧条件下生长时,它们在酯酶、乙醇脱氢酶、过氧化物酶、苹果酸脱氢酶、苹果酸酶、乳酸脱氢酶、谷草转氨酶、谷氨酸脱氢酶和异柠檬酸脱氢酶等同工酶中具有明显不同的酶带或酶活性,呈现酶的多分子形态。     

Autosomal determination of erythrocyte glucose 6-phosphate dehydrogenase in domestic chickens and ring-necked pheasants

Erythrocyte glucose 6-phosphate dehydrogenase isozymes of domestic chickens, ring-necked pheasants, and their hybrids were studied, using the starch gel zone electrophoresis technique. In domestic chickens G6PD isozymes were represented by two fast-moving bands and an indistinct third band, whereas in ring-necked pheasants a slow-moving broad band which seemed to consist of two closely apposed G6PD isozymes was observed. The F₁ hybrids showed three distinct bands combining the characteristic mobility pattern of the two parents, which seemed to indicate that both parental alleles are expressed in F₁ hybrids. Since both male and female hybrids exhibited strikingly similar isozyme patterns representing both sire and dam, it was assumed that the genes controlling the production of G6PD in chicken and pheasant red blood cells are located on the autosomes.This study was supported in part by a research grant from the National Research Council of Canada.     

Arrangement of cortical microtubules at the surface of the shoot apex inVinca major L.: Observations by immunofluorescence microscopy

Arrangements of cortical microtubules (MTs) and of cellulose microfibrils at the surface of the vegetative shoot apex ofVinca major L. were examined by immunofluorescence microscopy and polarizing microscopy, respectively. Cortical MTs adjacent to the outermost walls of the apex were arranged more or less randomly in individual cells: especially in cells in the central region of the apex the arrangement was almost completely random. However, in the peripheral region MTs tended to show parallel alignment in individual cells, and an overall pattern that was roughly concentric around the apical dome was discerned. Observations of birefringence of cell walls indicated that cellulose microfibrils in the peripheral region of the apex were also arranged in a pattern which was roughly concentric around the apical dome. These patterns of arrangements of MTs and microfibrils are understood to be perpendicular to the radial cell files observed in the peripheral region of the apex, and can be related to the radial expansion of the surface of the apex.     

Division of the primary nucleus ofAcetabularia

Summary The division of the primary nucleus ofAcetabularia mediterranea, andAcetabularia cliftonii was studied by light microscopical observation of living cells. Nuclear and nucleolar volumes are reduced when the caps of the cells have reached their maximum diameter. When the nucleus has reached a size of about 30–50 m in diameter, condensed chromosomes are formed which are separated by an intranuclear spindle.     

Somatic hybridization between selected Lycopersicon and Solanum species

Summary Mesophyll protoplasts of an interspecific Lycopersicon esculentum Mill, (tomato) x Lycopersicon pennellii hybrid plant (EP) were fused with callus-derived protoplasts of Solanum lycopersicoides Dun. using a modified PEG/DMSO procedure. The EP plant was previously transformed by Agrobacterium tumefaciens which carried the NPTII and nopaline synthase genes. Protoplasts were plated at 10⁵/ml in modified KM medium and 16 days post-fusion 25 ug/ml kanamycin was added to the culture medium. During shoot regeneration, 212 morphologically similar putative somatic hybrids were delineated visually from kanamycin resistant EP's. Forty-eight shoots, randomly selected among the 212, were further verified as somatic hybrids by their leaf phosphoglucoisomerase heterodimer isozyme pattern. However, the resulting plants were virtually pollen sterile. In a second fusion, mesophyll protoplasts of Solanum melongena (eggplant) were fused with EP callus-derived protoplasts. Using the same fusion and culture procedure, only two dark green calli were visually selected among the pale green parental EP and verified as somatic cell hybrids by several isozyme patterns. These two calli have produced only leaf primordia in one and half years on regeneration medium.Abbreviations ABA abscisic acid - BAP 6 benzylaminopurine - 2,4-D 2,4 dichlorophenoxy acetic acid - DMSO dimethyl sulfoxide - GA₃ gibberellic acid - GOT glutamate oxaloacetate - IAA indoleacetic acid - IBA indolebutyric acid - IDH isocitrate dehydrogenase - MDH malate dehydrogenase - MES morpholinoethane-sulfonic acid - PEG polyethylene glycol - 6-PGDH 6 phosphogluconate dehydrogenase - PGI phosphoglucoisomerase     

Relationships between cell surface protease and acid phosphatase activities ofLeishmania promastigote

A correlation between the ratio of the cell surface protease activity to phosphatase activity and the complexity of the pattern of cell surface exposed polypeptides ofLeishmania promastigotes was demonstrated for various strains grown under similar conditions. The ratio of the cell surface protease activity to acid phosphatase activity was high forL. major andL.b. panamensis and it correlates with the expression of a single polypeptide of 63 KDa on their cell surface. Intermediate and lower ratios of these enzymatic activites relate with more complex radio-iodinated patterns: two main bands inL.b. guyanensis (70 and 58 KDa) andL.b. braziliensis (72 and 60 KDa) and three main bands 65, 50, 27 KDa in allL.m. mexicana strains tested. Evidence is presented that the acid phosphatase located on theL.m. mexicana cell surface is not an artifact due to a secondary absorption of the secreted acid phosphatase from the culture medium. These results confirm theLeishmania antigen cell surface heterogeneity. The implications on the biology ofLeishmania and the clinical manifestation of leishmaniasis are discussed.     

A Sequential Study of Cell Divisions and Expansion Patterns on a Single Developing Shoot Apex of Vinca major

During the growth of a single developing vegetative apex ofVinca major, both the orientation and frequency of cell divisions,and the pattern of cell expansion, were observed using a non-destructivereplica technique. Micrographs taken at daily intervals illustratethat the central region of the apical dome remains relativelyinactive, except for a phase of cell division which occurs after2 d of growth. The majority of growth takes place at the proximalregions of the dome from which develop the successive pairsof leaves. The developing leaf primordia are initiated by aseries of divisions which occur at the periphery of the centraldome and are oriented parallel to the axis of the subsequentleaves. The cells which develop into the outer leaf surfaceof the new leaves undergo expansion and these cells divide allowingfor the formation of the new leaf. This paper describes thefirst high-resolution sequential study of cell patterns in asingle developing plant apex. Sequential development, cell division, expansion patterns, SEM, Vinca major, apical dome, leaf primordium, leaf initiation     

Branch formation induced by microbeam irradiation ofAdiantum protonemata

Branches were induced in centrifugedAdiantum protonemal cells by partial irradiation with polarized red light. Nuclear behavior and microtubule pattern change during branch formation were investigated. A branch formed at any part where a red microbeam was focused along a long apical cell. The nucleus moved towards the irradiated area and remained there until a branch developed. The pattern of microtubules changed from parallel to oblique at the irradiated area and then a transverse arrangement of microtubules appeared on both sides of the area. It appeared as if the nucleus was suspended between two microtubule rings. This nuclear behavior and the changes in microtubule pattern were different from those observed during branch formation under whole cell irradiation. From the results of this work we suggest that there is an importance for precise control of experimental conditions.     

Alcohol dehydrogenase isozymes in grain sorghum (Sorghum bicolor): Evidence for a gene duplication

Two sets of alcohol dehydrogenase (ADH) bands are regularly observed in grain sorghum (Sorghum bicolor): set I is a permanent triplet; set II is variable, as either two or three bands. A faint set III is detected only when extracts from seeds subjected to anerobiosis are run in neutralpH gels. Dissociation-reassociation experiments reveal that the central band of the set I triplet is a heterodimer of the other two. Full-sib progeny analysis from selfed plants shows that the set II bands are doublets, with heterozygotes having only three apparent bands instead of four because of the similar mobilities of the fast-migrating isozyme specified by the slow allele and the slow isozyme specified by the fast allele. We propose a three-locus model as the best explanation of these patterns. Set I consists of the products of two loci and their intergenic heterodimer. Set III is specified by a third locus. Set II isozymes are the intergenic heterodimers of the two set I loci and the set III locus. This explanation is similar to that of Schwartz and Freeling for maize but suggests that the evolution ofSorghum includes a gene duplication of the homologue of theAdh-1 locus inZea. Supported by USDA Grant 59-2063-01522 to NCE and KWF.     

Transfer of kanamycin resistance fromNicotiana tabacum toNicotiana plumbaginifolia by fusion of x-irradiated protoplasts

Summary Nuclear hybrids have been obtained by fusion of mesophyll protoplasts ofNicotiana plumbaginifolia and x-irradiated or iodoacetate-treated mesophyll protoplasts of a kanamycin-resistant line ofN. tabacum. The effect of irradiation on the recovery of asymmetric hybrids was evaluated by analysis of their morphology, fertility, chromosome number, isozyme patterns, restriction patterns in their organelle DNAs, and presence of the kanamycin-resistance gene. The results presented in this paper show that x-ray irradiation leads to a significant reduction in the amount ofN. tabacum genome present in the hybrids and demonstrates, once more, the power of this technique to induce directional loss of genomic traits of the irradiated parent.     

从烟草与龙葵细胞杂交获得杂种植株(简报)

烟草与龙葵是茄科不同属的植物,经原生质体融合可能将龙葵某些性状转入烟草。本文简要报道这一组合细胞杂交的结果。实验通过对诱导融合后再生植株的选择和鉴定,确定了两个株系(TS-28,TS-33)为烟草与龙葵的细胞杂种。材料为烟草(Nicotiana tabacum L.)品种革新一号,龙葵(Solanum nigrum L.)野生种。均取全展幼叶,用酶法制备原生质体。诱导融合方法按PEG和高pH高Ca~(++)法。培养基为原生质体培养用Du培养基(NAA0.186 mg/1,ZT 0.11 mg/1,2,4-D 0.5mg/1,KT 0.125 mg/1)。愈伤组织用MS 1 D(2,4-D 1 mg/1)。芽分化用MS_2(IAA l mg/1,KT2 mg/1)。根分化用MS0(无激素)。     

Isozyme Variability in Plants Regenerated from Calli of Cereus peruvianus (Cactaceae)

Morphological and isozyme variation was observed among plants regenerated from callus cultures of Cereus peruvianus. Different morphological types of shoots (68%) were observed in 4-year-old regenerated plants, while no distinct morphological variants were observed in plants grown from germinated seeds. Isozyme patterns of 633 plants regenerated from calli and of 261 plants grown from germinated seeds showed no variation in isocitrate dehydrogenase isozyme, and the differential sorbitol dehydrogenase, alcohol dehydrogenase, malate dehydrogenase, acid phosphatase, and peroxidase isozyme patterns observed in regenerated plants were attributed to nonallelic variation. Allelic variation was detected at three isoesterase loci. The proportion of polymorphic loci for both populations was 13.6% and the deviation from Hardy–Weinberg equilibrium for the Est-1 and Est-7 loci observed in somaclones was attributed to the manner in which the regenerant population was established. The high values for genetic identity among regenerant and seed-grown plant populations are in accordance with the low levels of interpopulation genetic divergence. In somaclones of C. peruvianus, morphological divergence was achieved within a short time but was not associated with any isozyme changes and also was not accompanied by biochemical genetic divergence.