Differential coupling of gibberellin responses by Rht-B1c suppressor alleles and Rht-B1b in wheat highlights a unique role for the DELLA N-terminus in dormancy

During the Green Revolution, substantial increases in wheat (Triticum aestivum) yields were realized, at least in part, through the introduction of the Reduced height (Rht)-B1b and Rht-D1b semi-dwarfing alleles. In contrast to Rht-B1b and Rht-D1b, the Rht-B1c allele is characterized by extreme dwarfism and exceptionally strong dormancy. Recently, 35 intragenic Rht-B1c suppressor alleles were created in the spring wheat cultivar Maringa, and termed overgrowth (ovg) alleles. Here, 14 ovg alleles with agronomically relevant plant heights were reproducibly classified into nine tall and five semi-dwarf alleles. These alleles differentially affected grain dormancy, internode elongation rate, and coleoptile and leaf lengths. The stability of these ovg effects was demonstrated for three ovg alleles in different genetic backgrounds and environments. Importantly, two semi-dwarf ovg alleles increased dormancy, which correlated with improved pre-harvest sprouting (PHS) resistance. Since no negative effects on grain yield or quality were observed, these semi-dwarf ovg alleles are valuable for breeding to achieve adequate height reduction and protection of grain quality in regions prone to PHS. Furthermore, this research highlights a unique role for the first 70 amino acids of the DELLA protein, encoded by the Rht-1 genes, in grain dormancy

LARP7 suppresses P-TEFb activity to inhibit breast cancer progression and metastasis.

Transcriptional elongation by RNA polymerase (Pol) II is essential for gene expression during cell growth and differentiation. The positive transcription elongation factor b (P-TEFb) stimulates transcriptional elongation by phosphorylating Pol II and antagonizing negative elongation factors. A reservoir of P-TEFb is sequestered in the inactive 7SK snRNP where 7SK snRNA and the La-related protein LARP7 are required for the integrity of this complex. Here, we show that P-TEFb activity is important for the epithelial-mesenchymal transition (EMT) and breast cancer progression. Decreased levels of LARP7 and 7SK snRNA redistribute P-TEFb to the transcriptionally active super elongation complex, resulting in P-TEFb activation and increased transcription of EMT transcription factors, including Slug, FOXC2, ZEB2, and Twist1, to promote breast cancer EMT, invasion, and metastasis. Our data provide the first demonstration that the transcription elongation machinery plays a key role in promoting breast cancer progression by directly controlling the expression of upstream EMT regulators

Spt5 Cooperates with Human Immunodeficiency Virus Type 1 Tat by Preventing Premature RNA Release at Terminator Sequences

The human immunodeficiency virus type 1 (HIV-1) Tat protein activates transcription elongation by stimulating the Tat-activated kinase (TAK/p-TEFb), a protein kinase composed of CDK9 and its cyclin partner, cyclin T1. CDK9 is able to hyperphosphorylate the carboxyl-terminal domain (CTD) of the large subunit of RNA polymerase during elongation. In addition to TAK, the transcription elongation factor Spt5 is required for the efficient activation of transcriptional elongation by Tat. To study the role of Spt5 in HIV transcription in more detail, we have developed a three-stage Tat-dependent transcription assay that permits the isolation of active preinitiation complexes, early-stage elongation complexes, and Tat-activated elongation complexes. Spt5 is recruited in the transcription complex shortly after initiation. After recruitment of Tat during elongation through the transactivation response element RNA, CDK9 is activated and induces hyperphosphorylation of Spt5 in parallel to the hyperphosphorylation of the CTD of RNA polymerase II. However, immunodepletion experiments demonstrate that Spt5 is not required for Tat-dependent activation of the kinase. Chase experiments using the Spt5-depleted extracts demonstrate that Spt5 is not required for early elongation. However, Spt5 plays an important role in late elongation by preventing the premature dissociation of RNA from the transcription complex at terminator sequences and reducing the amount of polymerase pausing at arrest sites, including bent DNA sequences. This novel biochemical function of Spt5 is analogous to the function of NusG, an elongation factor found in Escherichia coli that enhances RNA polymerase stability on templates and shows sequence similarity to Spt5

Promiscuous, non-catalytic, tandem carbohydrate-binding modules modulate the cell-wall structure and development of transgenic tobacco (Nicotiana tabacum) plants

We have compared heterologous expression of two types of carbohydrate binding module (CBM) in tobacco cell walls. These are the promiscuous CBM29 modules (a tandem CBM29-1-2 and its single derivative CBM29-2), derived from a non-catalytic protein1, NCP1, of the Piromyces equi cellulase/hemicellulase complex, and the less promiscuous tandem CBM2b-1-2 from the Cellulomonas fimi xylanase 11A. CBM-labelling studies revealed that CBM29-1-2 binds indiscriminately to every tissue of the wild-type tobacco stem whereas binding of CBM2b-1-2 was restricted to vascular tissue. The promiscuous CBM29-1-2 had much more pronounced effects on transgenic tobacco plants than the less promiscuous CBM2b-1-2. Reduced stem elongation and prolonged juvenility, resulting in delayed flower development, were observed in transformants expressing CBM29-1-2 whereas such growth phenotypes were not observed for CBM2b-1-2 plants. Histological examination and electron microscopy revealed layers of collapsed cortical cells in the stems of CBM29-1-2 plants whereas cellular deformation in the stem cortical cells of CBM2b-1-2 transformants was less severe. Altered cell expansion was also observed in most parts of the CBM29-1-2 stem whereas for the CBM2b-1-2 stem this was observed in the xylem cells only. The cellulose content of the transgenic plants was not altered. These results support the hypothesis that CBMs can modify cell wall structure leading to modulation of wall loosening and plant growth

Elongation, rooting and acclimatization of micropropagated shoots from mature material of hybrid larch

Factors were defined for elongation, rooting and acclimatization of micropropagated shoots of Larix x eurolepis Henry initiated from short shoot buds of plagiotropic stecklings serially propagated for 9 years from an 8-year-old tree. Initiation and multiplication were on Schenk and Hildebrandt (SH) medium supplemented with 5 μM 6-benzyladenine (BA) and 1 μM indole-butyric acid (IBA). Stem elongation was obtained in 36% of the shoots on SH medium containing 0.5 μM BA and 63% of the remaining non-elongated shoots initiated stem elongation after transfer on SH medium devoid of growth regulators. Rooting involved 2 steps: root induction on Campbell and Durzan mineral salts and Murashige and Skoog organic elements, both half-strength (CD-MS/2), supplemented with 1 μM of both naphthaleneacetic acid (NAA) and IBA, and root elongation following transfer to CD-MS/2 medium devoid of growth regulators. Repeating this 2-step sequence yielded up to 67% rooted shoots. Acclimatization of plantlets ranged from 83% to 100%. Over 300 plants were transferred to the greenhouse; some showed plagiotropic growth

Cell aging preserves cellular immortality in the presence of lethal levels of damage.

Cellular aging, a progressive functional decline driven by damage accumulation, often culminates in the mortality of a cell lineage. Certain lineages, however, are able to sustain long-lasting immortality, as prominently exemplified by stem cells. Here, we show that Escherichia coli cell lineages exhibit comparable patterns of mortality and immortality. Through single-cell microscopy and microfluidic techniques, we find that these patterns are explained by the dynamics of damage accumulation and asymmetric partitioning between daughter cells. At low damage accumulation rates, both aging and rejuvenating lineages retain immortality by reaching their respective states of physiological equilibrium. We show that both asymmetry and equilibrium are present in repair mutants lacking certain repair chaperones, suggesting that intact repair capacity is not essential for immortal proliferation. We show that this growth equilibrium, however, is displaced by extrinsic damage in a dosage-dependent response. Moreover, we demonstrate that aging lineages become mortal when damage accumulation rates surpass a threshold, whereas rejuvenating lineages within the same population remain immortal. Thus, the processes of damage accumulation and partitioning through asymmetric cell division are essential in the determination of proliferative mortality and immortality in bacterial populations. This study provides further evidence for the characterization of cellular aging as a general process, affecting prokaryotes and eukaryotes alike and according to similar evolutionary constraints

Comparison of Pruning Regimes for Stone Pine (Pinus pinea L.) Using a Functional- Structural Plant Model

Functional-Structural Plant Models (FSPM) are becoming important tools for modeling the structure and growth of plants, including complex organisms like trees. These models combine the advantages of empirical, mechanistic, and structural models to simulate the growth of individual plant structures (branches, buds, leaves, etc.). This approach enables realistic evaluation of the plant’s response– including changes in structure and growth to different environmental conditions. We demonstrate the potential use of these models to evaluate individual tree growth under different management regimes (pruning). The data used in this study was obtained from 3-D measurements taken with a FASTRAK Polhemus digitizer, with specific attention given to bud creation and branching. Each branch segment was analyzed to estimate its age, enabling us to document annual structural changes. We use the XL programming language and a GroIMP environment to simulate and compare different pruning scenarios

Laboratory Evaluation of Mefluidide Effects on Elongation of Hydrilla and Eurasian Watermilfoil

The potential of mefluidide (N-(2,4-dimethyl-5[[trifluromethyl) sulfonyl] amino] phenol) acetamide) to act as a submersed aquatic plant growth regulator was evaluated using a laboratory bioassay system. Main stem elongation of hydrilla (Hydrilla verticillata (L.f.) Royle) and Eurasian watermilfoil (Myriophyllum spicatum L.) was effectively reduced by mefluidide at low concentrations. The lowest effective concentration of mefluidide that reduced stem length in Eurasian watermilfoil (100 yg a.i./L) was 5 times lower than that for hydrilla (500 yg a.i./L). Short-term net photosynthetic rates of these plants were not affected by mefluidide at concentrations as high as 1000 yg a.i./L. The minimum exposure time required to maintain an inhibitory effect for at least 28 days at a concentration of 500 yg ai.i./L was 3 to 7 days for Eurasian watermilfoil and 7 to 14 days for hydrilla. The results suggest that mefluidide is a more effective growth regulator for Eurasian watermilfoil than hydrilla. Exogenously applied gibberellic acid (GA) did not completely overcome the inhibitory effect of mefluidide even when GA was added at a high concentration (10-5 M). In addition, the internodal lengths of stems treated with mefluidide were not reduced as they were when treated with gibberellin synthesis inhibitors. The reduction of main stem elongation by mefluidide appeared to be due to the inhibition of new cell and tissue development at the stem tip rather than from inhibition of GA biosynthesis

Plant response to solar ultraviolet-B radiation in a southern South American Sphagnum peatland

1. Plant growth and pigmentation of the moss Sphagnum magellanicum and the vascular plants Empetrum rubrum, Nothofagus antarctica and Tetroncium magellanicum were measured under near-ambient (90% of ambient) and reduced (20%) ultraviolet-B (UV-B) radiation for three growing seasons in a Sphagnum peatland in Tierra del Fuego, Argentina (55°S). 2. Reduction of solar UV-B increased height growth but decreased volumetric density in S. magellanicum so that biomass production was not influenced during the 3 years. The morphology of vascular plants tended not to respond to UV-B reduction. 3. A 10-20% decrease in UV-B-absorbing compounds occurred in T. magellanicum under solar UV-B reduction. No effects were seen on chlorophyll or carotenoids in S. magellanicum, although, for UV-B-absorbing compounds, a significant interaction between UV-B and year suggests some response to solar UV-B reduction. 4. The climate-related growth of the dwarf shrub E. rubrum was assessed retrospectively by correlating an 8-year record of annual stem elongation with macroclimatic factors including solar UV-B and visible radiation, precipitation and temperature. 5. No significant negative correlations were found between annual E. rubrum stem elongation and ambient solar UV-B, the ratio of UV-B: visible radiation, or the 305-nm: 340-nm irradiance ratio for an 8-year record (1990-91 to 1997-98), nor was stem elongation affected by solar UV-B reduction in our experimental field plots after 3 years. 6. The role of solar UV-B radiation on plant growth in Sphagnum peatlands in Tierra del Fuego, Argentina, is likely to depend on the severity of stratospheric ozone depletion over the next several decades. The increases in ambient solar UV-B associated with ozone depletion over the last 20 years are less than the difference between our radiation treatments. Therefore, providing that the ozone layer substantially recovers by the middle of this century, only modest effects of increased solar UV-B on plant growth may be expected.Fil: Searles, Peter Stoughton. State University of Utah; Estados UnidosFil: Flint, Stephan D.. State University of Utah; Estados UnidosFil: Diaz, Susana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Rousseaux, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Ballare, Carlos Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Caldwell, Martyn M.. State University of Utah; Estados Unido

Identification of the prebiotic translation apparatus within the contemporary ribosome

A structural element that could have existed independently in the prebiotic era was identified at the active site of the contemporary ribosome. It is suggested to have functioned as a proto-ribosome catalyzing peptide bond formation and non-coded elongation in the same manner that contemporary ribosomes exert positional catalysis, namely by accommodating the reactants in stereochemistry favourable for inline nucleophilic attack. This simple apparatus is a dimer of self-folding RNA units that could have assembled spontaneously into a symmetrical pocket-like structure, sufficiently efficient to be preserved throughout evolution as the active site of modern ribosomes, thus presenting a conceivable starting point for translation.Here we discuss the proto-ribosome emergence hypothesis and show that the tendency for dimerization, a prerequisite for obtaining the catalytic centre, is linked to the fold of its two components, indicating functional selection at the molecular level in the prebiotic era and supporting the existence of dimeric proto-ribosome