Intragenic recombination in maize: pollen analysis methods and the effect of parental adh1 isoalleles

The ability to stain mature pollen grains for the presence of alcohol dehydrogenase (ADH) activity permits the quantitation of ADH( +) gametophytes at frequencies below 10(-6). This resolution allows reversion and genetic fine structure analyses. The rationale of pollen analysis follows Nelson's prototype studies with waxy. As with the waxy gene, revertant frequencies for seven Adh1-deficient ( Adh1(-)) alleles appear to be in excess of microbially derived expectations. Each of the seven Adh1(-) alleles were derived from one of three naturally occurring isoalleles. Based on Schwartz's protein level characterizations of the mutants' products, it was anticipated that the seven Adh1(-) alleles should recombine to yield ADH(+) cistrons in certain pairwise combinations. This expectation was not met. The parental "wild-type" isoalleles from which the mutants were derived appear to be structurally divergent. The discussion interprets these data in view of understanding naturally occurring cistronic variation.     

The effects of light intensity and spectral quality on growth and shoot initiation in tobacco callus

The effects of eight different narrow band-emitting fluorescent lamps (371-750 nm) and four commercial broad band-emitting fluorescent sources upon growth and shoot initiation in tobacco callus (Nicotiana tabacum var. Wisconsin 38) have been characterized. Wavelength and intensity are equally important parameters in determining morphogenic changes. Near ultraviolet light (371 nm) was found to stimulate (0.024 mw/cm²) or inhibit (above 0.15 mw/cm²) callus growth and shoot initiation, depending on the light intensity. Stimulation of growth and shoot production occurs also in blue light region, but at higher intensity than in the near ultraviolet. Red and far red light (up to 1.7 mw/cm²) do not appear to affect callus growth or stimulate shoot initiation. The enhancement of callus growth and the stimulation of shoot initiation are controlled by the same near ultraviolet-absorbing photoreceptor system present in a small enough concentration that it cannot be recognized in the absorption spectrum of the intact tissue. Carotenoids, porphyrins, and phytochrome associated with the high irradiance response do not appear to qualify as the photoreceptor. Flavonoids are possible candidates. Radiation emitted by fluorescent lamps outside the near visible region was determined, and we concluded that energy levels were not sufficient to affect the reported results. The spectral output of several commercial lamps in the visible and near visible regions is such that there could be different effects on growth and development of tissue cultures.     

l-Ornithine:2-Oxoacid Aminotransferase from Squash (Cucurbita pepo, L.) Cotyledons: Purification and Properties

ORNITHINE: 2-oxoacid aminotransferase (EC 2.6.1.13) has been purified over 400-fold with a total recovery of 14% from acetone powders of cotyledons of germinating squash (Cucurbita pepo, L.) seedlings. The pH optimum of the transamination between l-ornithine and alpha-ketoglutarate is 8 and the Michaelis constants are 4.7 mm and 6.3 mm, respectively. The enzyme has a molecular weight of 48,000 as determined by gel filtration. The reaction is essentially specific for alpha-ketoglutarate as the amino group acceptor. The enzyme is inhibited very strongly by hydroxylamine, and less severely by NaCN and isonicotinylhydrazide. No inhibition is observed in the presence of 10 mml-cysteine. The energy of activation is 7.6 kcal/mole. The stability of the enzyme preparation is enhanced by the presence of dithioerythritol and glycerol. The enzyme activity of the most purified fraction is stimulated 30% by the addition of pyridoxal phosphate; however, the evidence for the unequivocal involvement of pyridoxal phosphate was inconclusive.     

Perithecial Formation in Gelasinospora reticulispora: IV. Action Spectra for the Photoinduction

Action spectra for photoinduction of perithecia after different lengths of dark period were determined with apically growing mycelium of a sordariaceous fungus Gelasinospora reticulispora. When hyphae were exposed to monochromatic light in near ultraviolet and visible regions, reciprocity between intensity and exposure time was observed within the range of incident energy used. The resulting action spectrum determined after a dark period of 48 hours showed a peak at 460 nm with shoulders at 420 and 480 nm and another peak at 370 nm, indicating minima at 410, 430, and 470 nm. After 72 hours darkness the spectrum was very similar to the above, except that the major peak shifted to 450 nm and the near ultraviolet region was somewhat less effective. In both cases, wavelengths longer than 520 nm showed no effect.     

Ribulose Diphosphate Carboxylase from Autotrophic Euglena gracilis

Ribulose 1,5-diphosphate carboxylase (RUDPcase) from autotrophically grown Euglena gracilis was purified to homogeneity as measured by analytical ultracentrifugation, polyacrylamide gel electrophoresis, and immunoprecipitation reactions. The enzyme represented about 9% of total protein and 24% of soluble protein in the autotrophic cell. Light-grown, heterotrophic cells seemed to contain considerably less RUDPcase. Native carboxylase from autotrophic Euglena showed an s_{20, w} at low protein concentrations of 17 to 17.5, suggesting a molecular weight of >500,000 daltons. Upon denaturation, the enzyme dissociated into two subunits having different amino acid compositions and molecular weights of 59,000 and 12,000 daltons. Based upon the amino acid mass ratios, a quaternary organization of 7 to 8 large and 8 to 10 small subunits per native enzyme molecule was indicated.     

Effect of glucose on the induction of nitrate reductase in corn roots

In Zea mays L., addition of glucose to the induction medium has no effect on the induction of nitrate reductase during the initial 3 hours either in root tips (0-10 mm) or mature root sections (25-35 mm). With longer times, higher levels of enzyme activity are recovered from both root segments when glucose is present in the incubation medium. The induction in root tips is saturated by 10 mm NO(3) (-). Higher concentrations of NO(3) (-) are required for saturation in mature root sections. The response to glucose is seen over a wide range of external NO(3) (-) concentrations.Nitrate reductase activity is lost rapidly when nitrate is withdrawn from the induction medium. Additions of glucose do not prevent this loss. Additions of glucose have no effect on total uptake of NO(3) (-) by the root segments but they increase the anaerobic NO(2) (-) production in both root tips and mature root segments. This latter measurement is considered to be an estimate of an active NO(3) (-) pool in the cytoplasm. Thus the results show that glucose alters the distribution of NO(3) (-) within the root sections. This may be an important factor in controlling the in vivo stability of the enzyme or its rate of synthesis.     

Benzyladenine reversal of abscisic Acid inhibition of growth and RNA synthesis in germinating bean axes

The effect of benzyladenine on growth, ATP pool size and specific radioactivity, and the rate and amount of RNA synthesis in aseptically cultured axes of Phaseolus vulgaris during the first 24 hours of germination were measured in experiments where the duration of benzyladenine application and its concentration were varied. Maximum promotion of growth (25%) occurs at 10(-5)m benzyladenine. Maximum promotion of RNA synthesis (44%) occurs at 10(-5)m benzyladenine. Benzyladenine has little effect on the size or specific radioactivity of the ATP pool. Benzyladenine can completely counteract abscisic acid inhibition of growth and RNA synthesis, and these reversals are measurable in 2 hours. GA(1) and GA(3) do not promote growth or counteract abscisic acid inhibition of growth in germinating bean axes in these experiments.