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1.
《Plant science》1988,55(2):159-167
Red clover (Trifolium pratense L.) cvs ‘Altaswede’ (2n = 2x = 14) and ‘Norseman’ (2n = 4x = 28) have been used to investigate tissue culture initiation, plant regeneration and the occurrence of somaclonal variation. After callus induction shoots were induced both when calli on L2 medium containing 2 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D), 2 mg l−1 6-benzylaminopurine (BA) and 2 mg l−1 6-amino-purine (AP) were subcultured on media containing naphthalene acetic acid (NAA) (0.05 mg l−1) and kinetin (KIN) (0.05 or 0.5 mg l−1) and when embryogenic calli were cultured and subcultured on L2 medium containing 0.002 mg l−1 4-amino-3,5,6-trichloropicolinic acid (PIC) and 0.2 mg l−1 BA. Shoot tip cultures were also established to induce multiple shoots for regeneration of plants via organogenesis.Regenerants from different regeneration pathways were evaluated for chromosome number stability, morphology and several biochemical traits. Regenerated plants showed stable isozyme banding patterns for malate dehydrogenase, 6-phosphogluconate dehydrogenase, phosphoglucose isomerase, phosphoglucomutase and shikimate dehydrogenase, as well as their nodule leghemoglobin profiles. Variations were detected in the chromosome number of some regenerants as well as in leaflet length-to-width ratio and leaflet number. Factors related to the incidence of somaclonal variation are discussed.  相似文献   

2.
Thirty-six sour (Prunus cerasus L.), sweet (P. avium L.), and ground cherry (P. fruticosa Pall.) selections were evaluated for seven enzyme systems and principal coordinate analysis was used to examine isozyme divergence among these cherry species. The enzyme systems studied were phosphoglucose isomerase (PGI), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphogluconate dehydrogenase (6-PGD), leucine aminopeptidase (LAP), shikimate dehydrogenase (SKDH), and malate dehydrogenase (MDH). The first principal coordinate, which accounted for 41% of the total variation, separated the diploid sweet cherry selections from the sour, ground, and sour x ground cherry tetraploids. An additional 86 selections were evaluated for up to six of the enzyme systems to determine the polymorphisms at the enzyme loci and the level of heterozygosity between the diploid sweet cherry and the tetraploid species and interspecific hybrids. 6-PGD was the most polymorphic enzyme exhibiting 16 patterns. The tetraploid cherry species were more heterozygous than the diploid sweet cherry with an average heterozygosity of 78% compared to 19% for the diploids.  相似文献   

3.
An electrophoretic study of isozyme number for seven soluble enzymes revealed extensive gene duplications in eight diploid species of American Eupatorium belonging to three morphological groups. The enzymes isocitrate dehydrogenase, phosphoglucomutase, phosphoglucose isomerase, 6-phosphogluconate dehydrogenase, and shikimate dehydrogenase occur as three to six isozymes in all species, whereas the minimal conserved number typical of diploid plants is two isozymes for each. Fructose 1, 6-biphosphate aldolase is expressed as multibanded pattern suggesting fixed heterozygosity in all examined species. It was not possible to document gene duplication for triosephosphate isomerase from the electrophoretic patterns. All species examined have a chromosome number of 2n = 20, which has been regarded as the basic diploid number for Eupatorium. However, the detection of extensive duplications suggests that 2n = 10 may be the original diploid chromosome number in Eupatorium and that plants with 2n = 20 are of polyploid origin. This hypothesis would mean that extensive duplications at isozyme gene loci have been maintained since the origin of the genus, despite chromosomal diploidization having occurred.  相似文献   

4.
A method for long-term plant regeneration of Phaseolus coccineus L, is described. Shoot-tips and cotyledonary nodes cultured on a Murashige and Skoog medium supplemented with N6-benzylaminopurine, 10 M, and -naphthaleneacetic acid, 1M, formed multiple bud-shoots. These shoots were transferred to medium containing BAP 1 M, NAA 0.1 M, and gibberellic acid 3 M to promote shoot growth and further shoot multiplication. Rooting was achieved in medium with 11 M indole-3-acetic acid. Rooted plants grew to maturity and were fertile. Cultures have maintained their ability to regenerate plants for more than two years. A sample of 30 regenerated plants (R0) was tested for chromosome number, all of them being diploid; seven isozymatic systems were electrophpretically analyzed in 82 R0 regenerated plants. No differences were observed in their electrophoretic patterns in comparison with those shown by seedlings. Histological studies revealed the origin of buds from calluses via organogenesis.Abbreviations BAP N6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA3 gibberellic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic acid - ADH alcohol dehydrogenase - GOT glutamic-oxaloacetic transaminase - MDH malate dehydrogenase - 6PGD 6-phosphogluconate dehydrogenase - PGI Phosphoglucose isomerase - PGM phosphoglucose mutase - SK shikimate dehydrogenase  相似文献   

5.
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  相似文献   

6.
Summary Callus ofNicotiana tabacum SRI, a mutant with maternally inherited streptomycin resistance, was induced from leaf sections. Callus pieces were mutagenised with N-ethyl-N-nitrosourea and inoculated onto a shoot-induction medium on which calli are normally green. White callus sectors were observed in the mutagenised cultures, and white and variegated shoots were regenerated from these sectored calli. The SR1-A10 line regenerated a chimeric shoot with white leaf margins. The chimeric shoot was grafted onto a normal green rootstock, grown into a flowering plant in the greenhouse, and crosses were made. The SRI-A15 line was crossed using flowers formed on albino plants grown in sterile culture. Pigment deficiency was maternally inherited in both lines. Physical mapping of the chloroplast genome of the SR1-A15 mutant by SalI, PstI and BamHI restriction endonucleases did not reveal any difference between the SR1-A15 and the parental SRI chloroplast genomes.  相似文献   

7.
Phosphoglucose isomerase negative mutant of mucoid Pseudomonas aeruginosa accumulated relatively higher concentration of fructose 1,6-bisphosphate (Fru-1,6-P2) when mannitol induced cells were incubated with this sugar alcohol. Also the toluene-treated cells of fructose 1,6-bisphosphate aldolase negative mutant of this organism produced Fru-1,6-P2 from fructose 6-phosphate in presence of ATP, but not from 6-phosphogluconate. The results together suggested the presence of an ATP-dependent fructose 6-phosphate kinase (EC 2.7.1.11) in mucoid P. aeruginosa.Abbreviations ALD Fru-1,6-P2 aldolse - DHAP dihydroxyacetone phosphate - F6P fructose 6-phosphate - G6P glucose 6-phosphate - Gly3P glyceraldehyde 3-phosphate - KDPG 2-keto 3-deoxy 6-phosphogluconate - PFK fructose 6-phosphate kinase - PGI phosphoglucose isomerase - 6PG 6-phosphogluconate  相似文献   

8.
Summary Allotetraploid intergeneric somatic hybrid plants between Citrus reticulata Blanco cv. Cleopatra mandarin and Citropsis gilletiana Swing. & M. Kell. (common name Gillet's cherry orange) were regenerated following protoplast fusion. Cleopatra protoplasts were isolated from an ovule-derived embryogenic suspension culture and fused chemically with leaf-derived protoplasts of Citropsis gilletiana. Cleopatra mandarin and somatic hybrid plants were regenerated via somatic embryogenesis. Hybrid plant identification was based on differential leaf morphology, root-tip cell chromosome number, and electrophoretic analyses of phosphoglucose mutase (PGM) and phosphohexose isomerase (PHI) isozyme banding patterns. This is the first somatic hybrid within the Rutaceae reported that does not have Citrus sinensis (sweet orange) as a parent, and the first produced with a commercially important citrus rootstock and a complementary but sexually incompatible, related species.Abbreviations PGM phosphoglucose mutase - PHI phosphohexose isomerase - MES 2[N-morpholino] ethane sulfonic acid - BH3 protoplast culture medium (Grosser and Chandler, 1987) - PEG polyethylene glycol - MT Murashige and Tucker (1969) basal medium - NAA 1-naphthaleneacetic acid - GA3 gibberellic acid - H+H and EME citrus embryogenic cell culture media (Grosser and Gmitter, 1990b) - B embryo germination medium - RMAN rooting medium Florida Agricultural Experiment Station Journal Series No. R-00298.  相似文献   

9.
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.  相似文献   

10.
The activity of phosphoglucose isomerase, its kinetic properties, and the effect of 6-phosphogluconate on its activity in the forward (glucose 6-phosphate----fructose 6-phosphate) and the reverse (fructose 6-phosphate----glucose 6-phosphate) reactions were determined in adult rat brain in vitro. The activity of phosphoglucose isomerase (in nmol/min/mg of whole brain protein) was 1,865 +/- 20 in the forward reaction and 1,756 +/- 32 in the reverse reaction at pH 7.5. It was 1,992 +/- 28 and 2,620 +/- 46, respectively, at pH 8.5. The apparent Km and Vmax of phosphoglucose isomerase were 0.593 +/- 0.031 mM and 2,291 +/- 61 nmol/min/mg of protein, respectively, for glucose 6-phosphate and 0.095 +/- 0.013 mM and 2,035 +/- 98 nmol/min/mg of protein, respectively, for fructose 6-phosphate. The activity of phosphoglucose isomerase was inhibited intensely and competitively by 6-phosphogluconate, with an apparent Ki of 0.048 +/- 0.005 mM for glucose 6-phosphate and 0.042 +/- 0.004 mM for fructose 6-phosphate as the substrate. With glucose 6-phosphate as the substrate, at concentrations from 0.05 to 0.5 mM, the activity of the enzyme was inhibited completely in the presence of 0.5-2.0 mM 6-phosphogluconate. With 0.05-0.2 mM fructose 6-phosphate as the substrate, it was inhibited greater than or equal to 85% at the same concentrations of the inhibitor. No significant changes were observed in the values of Km, Vmax, and Ki for phosphoglucose isomerase in the brain of 6-aminonicotinamide-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
Isozymes of hexose-phosphate isomerase (HPI; EC 5.3.1.9), pyruvate kinase (PK; EC 2.7.1.40) and 6-phosphogluconate dehydrogenase (6PGDH; EC 1.1.1.44) have been detected in the developing cotyledons of soybean (Glycine max (L.) Merr.), safflower (Carthamnus tinctorius L.) and sunflower (Helianthus annuus L.). In each seed there are two isozymes each of PK and HPI. The isozyme patterns of 6PGDH are more complex: soybean has two forms of the enzyme, safflower three, and sunflower six. In each tissue, at least 25% of the activity of each of the three enzymes is in the plastids. This supports the proposal that the glycolytic and pentose-phosphate pathways are operating in the plastids and that the plastids are the site of long-chain fatty-acid biosynthesis in developing oilseeds.Abbreviations HPI hexose-phosphate isomerase - 6PGDH 6-phosphogluconate dehydrogenase - PK pyruvate kinase  相似文献   

12.
Pupal ovaries of the wild oak silkworm Antheraea yamamai Guerin-Meneville were cultured in MGM-448 (Modified Grace Medium-448) medium containing 10% fetal bovine serum. After the primary culture was set up in 1988, a continuous cell line was obtained in 1991, designated as NISES-Anya-0611 (Anya-0611). The population doubling time was 54 hrs. and 19 min. at 96 passages and 88 hrs. and 29 min. at 387 passages. Spindle-shaped and spherical cells coexisted in the cell group. The cell line karyotype line was typical of lepidopteran cell lines, consisting of numerous small chromosomes. The cell line was distinguished from other lepidopteran cell lines by comparing malic enzyme, phosphoglucose isomerase, phosphoglucose mutase, and isocitric dehydrogenase isozyme patterns. The cell line was highly infected to the Antheraea yamamai nuclear polyhedrosis virus (Anya NPV). The luciferase gene of recombinant Bm NPV (BmNPVP6ETL) was able to express in the cell line, too, so that luciferase recombinant products were able to be detected in the cell body and in supernatant. The Anya NPV clone group was isolated on the cell seat using plaque purification.  相似文献   

13.
A quantitative analysis of malate dehydrogenase isozymes has been carried out in a hexaploid wheat Triticum aestivum variety Holdfast, a diploid rye Secale cereale variety King II, a series of seven addition lines each having the Holdfast wheat chromosome complement, and also a different homologous pair of King II rye chromosomes. In young shoots of three of these addition lines grown in a defined salts medium lacking sucrose, at least one isozyme activity was elevated. This did not occur in shoots grown in a medium containing 0.5% sucrose or in the Triticale possessing the full wheat and rye chromosomal complements grown in the absence of exogenous sucrose. On the basis of cellular localization and substrate inhibition studies, the particular isozyme activities enhanced by the rye chromosomes were indistinguishable from isozyme activities in Holdfast wheat and dissimilar to all malate dehydrogenase isozyme activities observed in King II rye. These results suggest that three different rye chromosomes produce gene products which can interact with the wheat malate dehydrogenase regulatory system.  相似文献   

14.
Twenty genotypes (individual plants) of Kentucky 31 tall fescue (Festuca arundinacea Schreb., 2n=6x=42) were evaluated to determine regeneration response, and meiotic and isozyme changes in the regenerants. Six genotypes (K8, K16, K25, K27, P3 and P13) were selected for study of efficiency in producing calli and regenerating plants. Panicle pieces (3569) were plated on Schenk & Hildebrandt medium with one of three auxins, 2,4-D (2.0 mg 1-1), pCPA (3.8 mg 1-1) or 2,4,5-T (2.5 mg 1-1). Square-root transformed data were analysed as a completely randomized 6×3 factorial with genotype and auxin as the main effects. F-protected means were compared by the Waller-Duncan K-ratio T test. Genotype K8 produced significantly more calli per panicle piece (88/877), whereas genotype K25 regenerated significantly more plantlets per panicle piece (58/214) than the other genotypes. Callus production was significantly higher using 2,4-D (102/1244 calli per piece) than pCPA or 2,4,5-T. The type of auxin did not have a statistically significant effect on plant regeneration, which suggested that genotype was the most important variable. Twelve of the 210 regenerants were albino. Cytological evaluation of 95 green regenerants showed that all plants had 21 bivalents and pollen stainability ranged from 43.0–98.4%, which suggested all plants were male fertile. Zymograms of these 95 regenerants for ACPH, ADH, GOT, MDH, 6-PGD, PGI and SOD showed no differences from that of the parental genotype, which suggested that the green plantlets regenerated from somatic tissue.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - pCPA parachlorophenoxyacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - ACPH acid phosphatase - ADH alcohol dehydrogenase - GOT glutamate oxaloacetate transaminase - MDH malate dehydrogenase - 6-PGD 6-phosphogluconate dehydrogenase - PGI phosphoglucoisomerase - SOD sulfoxide dismutase  相似文献   

15.
Summary Extracts from phylloclads of Asparagus officinails were electrophoretically analyzed for isozyme polymorphism. Fourteen enzyme systems were examined using four buffer systems: seven enzymes (acid phosphatase, catalase, glutamate-oxaloacetate transaminase, isocitrate dehydrogenase, malate dehydrogenase, peroxidase, and 6-phosphogluconate dehydrogenase) exhibited clear and consistent banding patterns. Isozyme polymorphism was studied in seven pairs of male and female doubled haploids and in their male F1s. Segregation of polymorphic loci was examined in the backcross progenies and was found to be consistent with a simple Mendelian inheritance in all cases, except for three anodical peroxidases, where two factors have been hypothesized. No linkage could be found between isozyme markers that were segregating in the same cross, but association was demonstrated between one malate dehydrogenase locus and the sex determining genes. The availability of isozyme markers may be useful in breeding and, in particular, the localization of one malate dehydrogenase locus on the sex chromosomes may be helpful in mapping the sex genes.  相似文献   

16.
Six ecologically central (old field) and marginal (strip mine) populations of the hexaploid Rumex acetosella were collected, grown under uniform conditions, and examined for genetic and morphological variation. Extensive electrophoretic variation was found in alcohol dehydrogenase and phosphoglucose isomerase, while other enzymes surveyed showed little or no variation. Hedrick's genotypic measure of identity revealed mean values of 0.506 for central populations, 0.836 for marginal populations, and 0.633 for comparisons of central with marginal populations. Alcohol dehydrogenase phenotypes had significantly fewer electromorphs per individual in marginal populations. Clones of individuals from both environments were subjected to different watering regimes. No significant differences in root/shoot ratio, leaf number, total leaf area, or relative growth rate were found between strip mine and old-field individuals within each watering treatment, although significant differences were found between watering treatments. There are small, but significant amounts of isozyme differentiation between central and marginal populations, while there was no such differentation for morphological characters.  相似文献   

17.
For the characterization of null mutants identified in Drosophila populations, several Drosophila enzymes including alcohol dehydrogenase, cytoplasmic malate dehydrogenase, alpha-glycerol phosphate dehydrogenase, and phosphoglucose isomerase were co-purified to homogeneity using an 8-(6-aminohexyl)-amino-ATP-Sepharose affinity column followed by DEAE-Sepharose column chromatography. Mitochondrial malate dehydrogenase was purified by the use of CM-Sepharose and the same ATP affinity column. Alcohol dehydrogenase and alpha-glycerol phosphate dehydrogenase were mapped on a two-dimensional gel. Antiserum was raised in rabbits against these Drosophila enzymes. The presence of cross-reacting material in null mutants was characterized by double immunodiffusion, immunoelectrophoresis, and two-dimensional gel electrophoresis. By immunological techniques, two natural null variants of malic enzyme and one of phosphoglucose isomerase were shown to be negative to cross-reacting material. Two low-dose-rate gamma-radiation-induced null mutants of cytoplasmic malate dehydrogenase were shown to be positive to cross-reacting material. Two-dimensional gel analyses enabled the characterization of three natural null variants of alpha-glycerol phosphate dehydrogenase. The viability of some null mutants with homozygous null or null/deficiency genotypes is discussed in terms of the in vivo metabolic roles of the related enzymes.  相似文献   

18.
Genetic variation in 12 populations of the terrestrial planarian Artioposthia triangulata from Northern Ireland and one in the Republic of Ireland were assessed using starch-gel electrophoresis. Seven enzymes - adenylate kinase, aspartate aminotransferase, enolase, malate dehydrogenase, phosphoglucose isomerase, phosphomannose isomerase and superoxide dismutase - provided consistent, resolved stains. Coefficients of genetic identity were similar for all comparisons suggesting that the populations did not belong to distinct taxa. There was no relationship between genetic distance and geographic distance for populations, thus confirming the role of passive dispersal in the spread of A. triangulata. Average percentage polymorphic loci and mean heterozygosities were higher in the east of the Province (41.4% and 0.240), than the west (37.5% and 0.185) suggesting that the planarian had spread westwards after an initial colonisation in the east.  相似文献   

19.
Summary Plant regeneration in Kentucky bluegrass (Poa pratensis L. cv. Touchdown) via culture of seedling tissues was investigated. When coleoptile, leaf, and stem sections of dark-germinated seedlings were cultured on Murashige and Skoog (MS) medium, different types of callus were produced, depending on the expiant source and growth regulator combinations. Only compact-friable callus (type 3) and moderately compact, friable callus (type 2) produced shoots upon subculture. The nonstructured watery callus (type 4) produced roots without shoots. Shoot differentiation from callus tissues was highest when the culture medium contained 0.2 mgL–1 picloram + 0.01 mgL–1 -naphthaleneacetic acid (NAA). Calli grown from coleoptiles had higher shoot regeneration frequency (32%) than that obtained from either stem sections (12%) or young leaf tissues (2%) of the same seedlings. Some organogenic callus lines produced exclusively green plants, while others produced albino shoots or a mixture of green and albino shoots. The green plants were multiplied in a medium containing 0.1 mgL–1 BAP plus either 0.2 mgL–1 picloram or 0.1 mgL–1 indole-3-acetic acid (IAA). Over 90% of the cultures in the shoot proliferation medium produced roots in 4 weeks. The rooted plants were successfully established in soil medium and grown in the greenhouse.Abbreviations BAP 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - MS Murashige and Skoog (1962) medium - NAA -naphthaleneacetic acid - picloram 4-amino-3,5,6-trichloropicolinic acid - TDZ thidiazuron  相似文献   

20.
Analysis of genetic variation between the banana and the citrus races of Radopholus similis by starch gel eleclrophoresis demonstrated that 7 of 16 enzyme-encoding loci could be used for their diagnostic separation. The two races are closely related arid share approximately 75% of the enzymes evaluated. The level of dissimilarities o1 inherited bands indicates that no gene flow occurs between the races. Aldolase, α + β esterase, glucose-6-phosphate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, and phosphoglucose isomerase are diagnostic markers of the races.  相似文献   

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