Induction of embryogenicTriticum aestivum L. calli. I. Quantification of genotype and culture medium effects

Somatic embryo (embryoid) formation from immature-embryo-derived calli was quantified in replicated experiments involving 10Triticum aestivum L. genotypes. Several published media formulations, which had previously been optimized for wheat tissue culture, were tested for each genotype. Embryos from each plant were randomly assigned to each medium. Percentage precocious germination of immature embryos and mean percentage scutellar callus per explant were recorded. Embryoids per callus were determined by microscopic examination at 28 and 56 days. There were highly significant differences among genotypes, media, and individual plants from which explants were taken. A medium based on double the Murashige and Skoog (MS) inorganic salt concentration was significantly better than other media. Inclusion of all MS vitamins appeared essential for optimal response. Two genotypes were tested in a second experiment where both 3,6-dichloro-o-anisic acid (9.05 M) and 6-furfurylaminopurine (0.46 M) were substituted for 2,4-dichlorophenoxyacetic acid (4.52 M) in either double or normal MS medium. This substitution significantly increased embryoid formation at 28 days. Additions of either 6-furfurylaminopurine or coconut water increased precocious germination of both embryo explants and embryoids.This study was supported in part by NASA-Ames Cooperative Agreement No. NCC2-139. Contribution of the Utah Agricultural Experiment Station, Utah State University, Logan, UT, Journal Paper No. 3358.     

Ac transposition in transgenic tomato plants

Summary As an initial step towards developing a transposon mutagenesis system in tomato, the maize transposable element Ac was transformed into tomato plants via Agrobacterium tumefaciens. Southern analysis of leaf tissue indicated that in nine out of eleven transgenic plants, Ac excised from the T-DNA and reintegrated into new chromosomal locations. The comparison of Ac banding pattern in different leaves of the same primary transformant provided evidnece for transposition during later stages of transgenic plant development. There was no evidence of Ds mobilization in tomato transformants.     

Nitrate reductase and its role in nitrate assimilation in plants

Nitrate reductase (EC 1.6.6.1) is an enzyme found in most higher plants and appears to be a key regulator of nitrate assimilation as a result of enzyme induction by nitrate. The biochemistry of nitrate reductase has been elucidated to a great extent and the role that nitrate reductase plays in regulation of nitrate assimilation is becoming understood.     

Phytochrome-mediated light regulation of nitrate reductase expression in squash cotyledons

In etiolated squash (Cucurbita maxima L.) cotyledons, nitrate-inducible NADH:nitrate reductase activity and protein were increased in darkness by red light pulses with red/far-red photoreversibility. Continuous far-red light also led to increased levels of nitrate reductase activity and protein. Poly(A)+RNA, which hybridizes to squash nitrate reductase cDNA, was also increased by light treatments. Thus, we found that after nitrate triggering, nitrate reductase expression appears to be regulated by light via phytochrome.     

Biochemical and ultrastructural characterization of the 1,4-dihydropyridine receptor from rabbit skeletal muscle. Evidence for a 52,000 Da subunit

The 1,4-dihydropyridine receptor purified from rabbit skeletal muscle contains four polypeptide components of 175,000 Da (nonreduced)/150,000 Da (reduced), 170,000, 52,000, and 32,000 Da (Leung, A. T., Imagawa, T., and Campbell, K. P. (1987) J. Biol. Chem. 262, 7943-7946). A monoclonal antibody specific to the 52,000-Da polypeptide component of the dihydropyridine receptor has been produced and used in immunoprecipitation and immunoblotting experiments to demonstrate that the 52,000-Da polypeptide is an integral subunit of the purified dihydropyridine receptor. Peptide mapping experiments with 32P-labeled dihydropyridine receptor have also demonstrated that the 52,000-Da polypeptide is distinct from and not a proteolytic fragment of the 170,000-Da subunit. Densitometric scanning of Coomassie Blue-stained sodium dodecyl sulfate-polyacrylamide gels of the purified dihydropyridine receptor has demonstrated that the 52,000-Da polypeptide exists in a 1:1 stoichiometric ratio with the 170,000-, 175,000/150,000-, and 32,000-Da subunits of the dihydropyridine receptor. Electron microscopy of the freeze-dried, rotary-shadowed dihydropyridine receptor has shown that the preparation contains a homogeneous population of 16 x 22-nm ovoidal particles large enough to contain all four polypeptides of the dihydropyridine receptor. The particles have two distinct components of similar size which may represent the location in the molecule of the two larger subunits.     

Aortic velocity contours at abdominal branches in anesthetized dogs

Pulsed Doppler velocimetry was used to record a grid of velocity waveforms 0.5 cm downstream of the cephalic mesenteric and left renal branches of the aorta in anesthetized dogs. Aortic velocity contours at different phases of the pulse cycle were developed from the grid of waveforms. Transient flow recirculation occurred in the post-systolic phase of the pulse cycle at the aortic wall opposite the branch artery. There was no recirculation at the systolic peak but there was asymmetry of the velocity profile. In contrast, in the in vivo post-stenotic velocity field recirculation persists throughout most of the cycle. These results compare well with similar results from pulsatile model studies. Spectral width of the Doppler signal was not increased in any of the velocity waveforms immediately downstream of the branches, this suggests that flow is relatively undisturbed in these locations.     

Mating structure and the cost of deleterious mutation: I. Postponing inbreeding

Mating structure governs the distribution of alleles in populations and thus the extent to which the phenotypes associated with the alleles are manifested. A mating system which initially achieves more genetic identity within individuals than between individuals enhances the probability that a finite population without reproductive excess will become extinct from a recessive lethal or semidominant lethal mutation; however, such a mating system decreases the number of deaths that will ensue if the population size is maintained by replacement of inviable progeny with individuals engendered from the entire mating pool. This is illustrated with Markov chain models for half-sib and double-first-cousin mating in populations of four individuals and by various techniques for analogous large populations. An appropriate choice of mating strategy can mitigate the effect of deleterious mutations, but the determination of which strategy is appropriate depends on how much reproductive excess is available and on the relative costs assigned to individual deaths and the extinction of a population.     

Heterogeneity of a murine B cell lymphoma. Isolation and characterization of idiotypic variants

mAb directed toward the idiotype of the 38C13 murine B cell lymphoma can be used to treat and cure a high percentage of mice challenged previously with an otherwise lethal dose of tumor cells. Tumors developing in animals despite antibody therapy were examined by immunofluorescence and found to demonstrate either loss of surface Ig, or expression of an altered idiotype that no longer bound the antibody used for treatment. Further immunofluorescence analysis of the variant tumors revealed individual patterns of cross-reactivity with anti-38C13 idiotype mAb other than that used for therapy. The variant tumor cells were fused to myeloma cells and hybrids were isolated which secreted large quantities of the altered idiotype proteins. Polyclonal antibodies and mAb prepared against the mutant proteins demonstrated cross-reactivity with the original 38C13 protein and its other variants. But the variants and wild type cells could be distinguished from each other by their patterns of reactivity with the panels of anti-idiotype antibodies. Differences in apparent m.w. were demonstrated in the L chains of each of the mutant proteins. Southern blot analysis of the H chain locus of these mutants established that they were all clonally related; however, the L chain loci were grossly different. Thus, rare cells with alteration in their Ig L chain genes and expressed proteins can give rise to idiotype variants in this B cell tumor.     

Inhibition of growth hormone-stimulated lipolysis by somatostatin, insulin, and insulin-like growth factors (somatomedins) in vitro

The effects of somatostatin, insulin, insulin-like growth factor I (IGF-I), and insulin-like growth factor II (IGF-II)/MSA on growth hormone (GH) (1 microgram/ml)-induced lipolysis were examined employing chicken adipose tissue in vitro. Basal and GH-stimulated glycerol release were inhibited by somatostatin (1 ng/ml) and by IGF-II/MSA (10 and 100 ng/ml). Insulin and IGF-I (10 and 100 ng/ml) completely inhibited the lipolytic response to GH without affecting basal glycerol release. Insulin and IGF-I were equipotent in inhibiting GH-induced lipolysis while IGF-II is only 16% as potent as insulin.