Relativistic effects and two-body currents in (H)((-->)e(')p)n using out-of-plane detection.

Measurements of the (2)H((-->)e,e(')p)n reaction were performed with the out-of-plane magnetic spectrometers (OOPS) at the MIT-Bates Linear Accelerator. The longitudinal-transverse, f(LT) and f(')(LT), and the transverse-transverse, f(TT), interference responses at a missing momentum of 210 MeV/c were simultaneously extracted in the dip region at Q2 = 0.15 (GeV/c)(2). In comparison to models of deuteron electrodisintegration, the data clearly reveal strong effects of relativity and final-state interactions and the importance of two-body meson-exchange currents and isobar configurations. We demonstrate that such effects can be disentangled by extracting these responses using the novel out-of-plane technique.

Lilliputian mutant of maize lacks cell elongation and shows defects in organization of actin cytoskeleton.

The maize mutant lilliputian is characterized by miniature seedling stature, reduced cell elongation, and aberrant root anatomy. Here, we document that root cells of this mutant show several defects in the organization of actin filaments (AFs). Specifically, cells within the meristem lack dense perinuclear AF baskets and fail to redistribute AFs during mitosis. In contrast, mitotic cells of wild-type roots accumulate AFs at plasma membrane-associated domains that face the mitotic spindle poles. Both mitotic and early postmitotic mutant cells fail to assemble transverse arrays of cortical AFs, which are characteristic for wild-type root cells. In addition, early postmitotic cells show aberrant distribution of endoplasmic AF bundles that are normally organized through anchorage sites at cross-walls and nuclear surfaces. In wild-type root apices, these latter AF bundles are organized in the form of symmetrically arranged conical arrays and appear to be essential for the onset of rapid cell elongation. Exposure of wild-type and cv. Alarik maize root apices to the F-actin drugs cytochalasin D and latrunculin B mimics the phenotype of lilliputian root apices. In contrast to AFs, microtubules are more or less normally organized in root cells of lilliputian mutant. Collectively, these data suggest that the LILLIPUTIAN protein, the nature of which is still unknown, impinges on plant development via its action on the actin cytoskeleton.

Search for quadrupole strength in the electroexcitation of the delta+(1232).

High-precision 1H(e,e'p)pi(0) measurements at Q2 = 0.126 (GeV/c)2 are reported, which allow the determination of quadrupole amplitudes in the gamma*N-->Delta transition; they simultaneously test the reliability of electroproduction models. The derived quadrupole-to-dipole ( I = 3/2) amplitude ratios, R(SM) = (-6.5+/-0.2(stat+sys)+/-2.5(mod))% and R(EM) = (-2.1+/-0.2(stat+sys)+/-2.0(mod))%, are dominated by model error. Previous R(SM) and R(EM) results should be reconsidered after the model uncertainties associated with the method of their extraction are taken into account.

Dap (Defective aleurone pigmentation) mutations affect maize aleurone development.

Dap (Defective aleurone pigmentation) is the designation for mutations in maize that give rise to a characteristic dappled endosperm phenotype, consisting of patches of purple tissue, of variable size and shape, on a yellow background. Features shared by all Dap mutants are: dominant expression when they are maternally derived, lack of expression or transmission when they originate from pollen, failure to recover homozygous Dap genotypes, reduced frequency of Dap seeds in the progeny of outcrosses of Dap/+ females, association of the dappled phenotype with reduction in seed size. The mutants so far tested, six in all, can be grouped into two classes, one including male-transmissible (MT) isolates not expressed in the endosperm if their contribution is paternal, and a second class of isolates (NMT) that are permanently lost following paternal transmission. We suggest that the NMT mutations are on a chromosome that carries an intercalary deletion. Assuming linkage between the mutant and the deletion, selection against the deficient chromosome during male gametogenesis would account for the failure to recover Dap seeds in the progeny of Dap/+ male parents. We have obtained genetic evidence supporting this hypothesis. This interpretation, however, does not apply to MT alleles. For these, other mechanisms, such as imprinting and/or dosage effects may be proposed. The mutable pattern in the endosperm to which all Dap mutants give rise is an intriguing phenotype which remains to be clarified. An unexpected finding is that aleuronic and subaleuronic cells corresponding to the colourless areas are abnormal in shape and anthocyanin biosynthesis is blocked in these cells. This finding calls for further investigation in light of a possible connection between flavonoid precursors and cell shape.

Light-Dependent Spatial and Temporal Expression of Pigment Regulatory Genes in Developing Maize Seeds.

Both light and developmental stimuli are directly involved in the regulation of plant gene expression. In maize, activation of the anthocyanin pathway represents an excellent model system for studying the interactions between an external factor, such as light, and internal factors that regulate plant and seed development. By analyzing in detail the aleurone and pericarp seed layers, different developmental windows for light have been found in the two tissues[mdash]the former in the advanced stages of development and the latter in the early stages of seed development. Transcriptional control of the structural genes involved in anthocyanin deposition within the pericarp is known to be exerted by the Sn and pl genes, whereas the aleurone is controlled by the R and C1 regulatory genes. By using in situ hybridization analysis, we detected tissue-specific expression of Sn and R in the seed layers, revealing a correlation between structural gene activation and anthocyanin accumulation. In addition, RNA gel blot analysis revealed that Sn expression is enhanced by light, whereas the R gene expression is not. However, the light-induced expression of the myb-type genes C1 and pl, detected by reverse transcriptase-polymerase chain reaction, was found to be the limiting factor for conferring the developmental competence of the pericarp and the aleurone layers to light responsiveness.

Antiparallel expression of the sense and antisense transcripts of maize alpha-tubulin genes.

In all eukaryotes alpha- and beta-tubulins are encoded by small families of closely related genes and are highly conserved. In Zea mays, at least six different alpha-tubulin coding sequences are known. We describe the isolation from scutellar nodes of the maize inbred line W22 of a clone (CTM5) coding for an alpha-tubulin. On the basis of the 3' end nucleotide sequence, this clone can be assigned to the already reported tua4 gene. Northern analysis demonstrates that CTM5 encodes a 1.5 kb transcript, which is expressed in different tissues of the seed and of the seedling. In order to define the spatial and temporal expression of alpha-tubulin genes, in situ hybridization experiments were performed on these tissues. Unexpectedly, a specific signal was detected with both antisense and sense RNA strands. Temporal and spatial distribution of the two RNAs, however, shows that high levels of the two transcripts are always discordant. In tissues where sense transcripts are highly abundant (embryos at various developmental stages, root tips, pollen grains), the antisense transcripts are expressed in relatively small amounts, while in pericarp, coleoptile, leaves, and scutellar node, where antisense transcripts accumulate, the sense transcript only reaches a very low level. Northern analysis using single-stranded DNA probes confirmed the presence of an antisense transcript of 1.5 kb, prompting speculation about the role of this transcript in the regulation of the expression of alpha-tubulin genes.

Genetic and molecular analysis of Sn, a light-inducible, tissue specific regulatory gene in maize.

The Sn locus of maize is functionally similar to the R and B loci, in that Sn differentially controls the tissue-specific deposition of anthocyanin pigments in certain seedling and plant cells. We show that Sn shows molecular similarity to the R gene and have used R DNA probes to characterize several Sn alleles. Northern analysis demonstrates that all Sn alleles encode a 2.5 kb transcript, which is expressed in a tissue-specific fashion consistent with the distribution of anthocyanins. Expression of the Sn gene is light-regulated. However, the Sn: bol3 allele allows Sn mRNA transcription to occur in the dark, leading to pigmentation in dark-grown seedlings and cob integuments. We report the isolation of genomic and cDNA clones of the light-independent Sn: bol3 allele. Using Sn cDNA as a probe, the spatial and temporal expression of Sn has been examined. The cell-specific localization of Sn mRNA has been confirmed by in situ hybridization using labelled antisense RNA probes. According to its proposed regulatory role, expression of Sn precedes and, in turn, causes a coordinate and tissue-specific accumulation of mRNA of structural genes for pigment synthesis and deposition, such as A1 and C2. The functional and structural relationship between R, B, Lc and Sn is discussed in terms of an evolutionary derivation from a single ancestral gene which gave rise this diverse gene family by successive duplication events.

Heterochromatin heterogeneity in Drosophila chromosomes detected by AluI-banding.

In situ selective digestion of mitotic chromosomes from three Drosophila species (virilis, hydei, and funebris), having a chromosome number 2n = 12, was achieved with AluI restriction endonuclease. The distribution of AluI-bands, revealing a heterogeneity within heterochromatin, was compared with that of quinacrine-bands observed in standard chromosomes. The results confirmed a species-specific AluI-banding pattern, heterochromatin being selectively digested in D. virilis and D. funebris, but unaffected by the enzyme in D. hydei. The overlapping of AluI-bands and Q-bands in D. virilis and in D. funebris is discussed, since these data seem to favour the hypothesis that the induction of AluI-bands does not depend only on the presence of enzyme targets, but also on a suitable conformation of the chromatin.

High resolution of heterochromatin of Drosophila melanogaster by distamycin A.

A DNA-binding AT-specific antibiotic, distamycin A, was used as inhibitor of the condensation process of the heterochromatic regions in Drosophila melanogaster embryonic cells. By this treatment the structural organization of heterochromatin at interphase is preserved until metaphase. The different patterns observed are interpreted as chronological steps in the condensation process.

Relationship between sister chromatid exchanges and DNA replication in somatic cells of Drosophila melanogaster.

In Drosophila melanogaster cell lines and larval neuroblast cells, two aspects of the phenomenon of sister chromatid exchanges were analyzed: (1) the frequency of SCEs in relation to the ploidy level (comparing diploid and tetraploid cells) and in relation to the cell type (comparing embryonic and larval cells) (2) the localization of the sites of exchange with reference to eu- and heterochromatin. A good correlation between SCE frequency and genome size in the same cell type (in distant species also), but a significant difference in the SCE rate between different cell types within the same species, were found. The results confirmed also the non-random distribution of SCEs in the different portions of the genome since a preferential localization in the euchromatin was clearly demonstrated. Moreover, a direct proportionality between SCE frequency and the length of the S phase was supposed, favouring the hypothesis of a relationship between the phenomenon of sister chromatid exchanges and DNA replication.

Karyotype evolution in cell lines of Drosophila melanogaster.

The chromosomal changes occurred in two independent cell lines (GM2 and GM3) of Drosophila melanogaster maintained in medium supplemented with serum and in serum-free medium were compared. In both culture conditions and in both lines a chromosomal evolution was revealed. Structural and numerical variations were analysed. The breaks giving rise to rearrangements were at heterochromatic level. Moreover, a tetraploidisation followed by loss of chromosomes or of portions of chromosomes recalls an analogous cycle observed in human cells.