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At least 181 records · Page 10

Selected humanization of yeast U1 snRNP leads to global suppression of pre-mRNA splicing and mitochondrial dysfunction in the budding yeast

The recognition of the 5′ splice site (5′ ss) is one of the earliest steps of pre-mRNA splicing. To better understand, the mechanism and regulation of 5′ ss recognition, we selectively humanized components of the yeast U1 (yU1) snRNP to reveal the function of these components in 5′ ss recognition and splicing. We targeted U1C and Luc7, two proteins that interact with and stabilize the yU1 snRNA and the 5′ ss RNA duplex. We replaced the zinc-finger (ZnF) domain of yeast U1C (yU1C) with its human counterpart, which resulted in a cold-sensitive growth phenotype and moderate splicing defects. We next added an auxin-inducible degron to yeast Luc7 (yLuc7) protein (to mimic the lack of Luc7Ls in human U1 snRNP). We found that Luc7-depleted yU1 snRNP resulted in the concomitant loss of Prp40 and Snu71 (two other essential yU1 snRNP proteins), and further biochemical analyses suggest a model of how these three proteins interact with each other in the U1 snRNP. The loss of these proteins resulted in a significant growth retardation accompanied by a global suppression of pre-mRNA splicing. The splicing suppression led to mitochondrial dysfunction as revealed by a release of Fe 2+ into the growth medium and an induction of mitochondrial reactive oxygen species. Together, these observations indicate that the human U1C ZnF can substitute that of yeast, Luc7 is essential for the incorporation of the Luc7–Prp40–Snu71 trimer into yU1 snRNP, and splicing plays a major role in the regulation of mitochondrial function in yeast.

Biochemistry & Molecular Biology↗

Solar Photovoltaic (PV) Manufacturing Expansions in the United States, 2017-2019: Motives, Challenges, Opportunities, and Policy Context

This report analyzes U.S. PV manufacturing announcements from 2017 through July-2019 and explores the potential impact of tariffs on the competitiveness of U.S. PV module manufacturing. First, the fraction of announcements that resulted in established facilities in the time frame specified is analyzed, as well as firm nationality and the existence of any other U.S. facilities for the firm. The prevalence of different PV product types among the announcements was also analyzed, as well as the type of facilities selected for development (greenfield, existing PV factories, or repurposed buildings). The cost to build these facilities was also analyzed in the context of their capacity, the value of incentives received by state and local governments, and the number of jobs created. Case studies of Auxin Solar, First Solar, Hanwha Q Cells, JinkoSolar, LG, Silfab, Solaria, and SunPower were conducted and highlight five factors influencing firms’ decisions to expand their U.S. PV production capacity. These included: 1) proximity to demand, including supply contracts and demand for domestic content 2) tariffs and other incentives. The Section 201 tariffs on PV module imports were often described helpful but tariffs on cell imports (Section 201) or materials (Section 301, Section 232) were often described as counterproductive since U.S. supply chains do not exist for many necessary goods. The corporate tax decrease and local incentives were often mentioned as helpful. 3) the ability to move quickly in order to maximize the benefit of the Section 201 tariffs. This affected decisions to select localities with faster permitting or moving into existing buildings. 4) competition with scale, where competing with Asian PV imports or avoiding imports of material from China was challenging due to the larger scale of Asian manufacturers. Additionally, smaller firms expressed concerns that larger firms may not need their U.S. facilities to profitable. 5) access to capital, which was a particular challenge for smaller firms. Several U.S. tariffs affect the economics of PV cells, modules, and balance-of-module (BOM) materials, some of which are applicable to a long list of countries and others applicable only to China or Taiwan. We analyze how the tariffs interact to affect the prices of c-Si modules sourced under various assumptions about domestic and imported components and assembly. We estimate module prices under a range of baseline scenarios featuring U.S.-assembled modules with Southeast Asian cells (not from China or Taiwan). When all BOM components are sourced from the United States, baseline module prices are $0.37/W (below the 2.5-GW Section 201 cell cap) to $0.42/W (above the 2.5-GW cell cap). If all BOM components are sourced from China, the module price in this baseline U.S.-assembly scenario can reach up to $0.60/W. Imported module prices are lower than or within the range of the U.S.-assembled baseline prices under some conditions. Importing bifacial modules from Southeast Asia is the lowest-priced alternative, at $0.35/W, because bifacial modules are excluded from the Section 201 tariffs. Importing Southeast Asia modules that contain U.S. cells results in a price of $0.38/W, also owing to a Section 201 exclusion. Importing Southeast Asian modules that have Southeast Asian cells yields a price of $0.44/W, and importing Chinese modules that have U.S. cells yields a price of $0.58/W. These results suggest that U.S. tariff policy in 2019 may encourage U.S. assembly of mono-facial c-Si modules if the 2.5-GW cell import cap is not reached and Chinese BOM imports are negligible. However, some imported module options may offer the lowest prices if the 2.5-GW cell import cap is reached and/or U.S. module manufacturers must rely on significant Chinese BOM content. The analysis shows the 2.5-GW cell cap may be exceeded in 2019 and following years. It is also likely that U.S. module manufacturers will need to use at least some Chinese BOM content, since U.S. module manufacturers have stated that U.S. supply chains do not exist (or do not exist at a sufficient scale) for most BOM components, which makes them at least partially reliant on Chinese BOM and thus subject to Section 301 tariffs.

14 SOLAR ENERGY↗

HSQC spectra of lignin isolated from poplar stems

Here we present a curated dataset of two-dimensional heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) spectra of lignin isolated from stems of genetically engineered poplar through auxin signaling gene modification. The plants were grown in greenhouse with temperatures between 21 and 23 °C. Plants were harvested and the aboveground stems were cut off an approximately five-inch-long segment from the bottom end of the plant stem, debarked and air-dried for three weeks. The dried stem samples were Wiley milled (mesh size 20), Soxhlet-extracted with toluene/ethanol for 24 h to remove extractives. The extracted biomass was ball-milled in a Retsch PM100 planetary ball mill using a porcelain jar with ceramic balls at 600 rpm for 2 h (in 5 min on and 5 min off cycles to avoid excessive sample heating). The ball-milled materials were then subjected to enzymatic hydrolysis for 48 h followed by centrifugation and washing with deionized water. The solid residue was freeze-dried to recover the lignin. The dry stem lignin samples were dissolved in deuterated dimethyl sulfoxide (d6) and transferred into a 5 mm tube. 13C–1H HSQC experiments were performed in a Bruker Avance III HD 500 MHz NMR spectrometer operating at a frequency of 125.12 MHz for the 13C nucleus using a standard Bruker pulse sequence on a Prodigy platform cryoprobe. The NMR spectra were acquired under the following acquisition conditions: 230 ppm spectral width in F1 (13C) dimension with 256 data points and 12 ppm spectral width in F2 (1H) dimension with 2048 data points, a 90° pulse, a one bond C–H coupling constant of 145 Hz, a 1.0 s pulse delay, and 64 scans. Spectra were processed using the Bruker TopSpin 3.6 software. Additional meta data is embedded in the raw spectra figures.

HSQC, lignin, poplar, stems, CBI↗

VPS45 is required for both diffuse and tip growth of Arabidopsis thaliana cells

Introduction VPS45 belongs to the Sec1/Munc18 family of proteins, which interact with and regulate Qa-SNARE function during membrane fusion. We have shown previously that Arabidopsis thaliana VPS45 interacts with the SYP61/SYP41/VTI12 SNARE complex, which locates on the trans -Golgi network (TGN). It is required for SYP41 stability, and it functions in cargo trafficking to the vacuole and in cell expansion. It is also required for correct auxin distribution during gravitropism and lateral root growth. Results As vps45 knockout mutation is lethal in Arabidopsis, we identified a mutant, vps45-3 , with a point mutation in the VPS45 gene causing a serine 284-to-phenylalanine substitution. The VPS45-3 protein is stable and maintains interaction with SYP61 and SYP41. However, vps45-3 plants display severe growth defects with significantly reduced organ and cell size, similar to vps45 RNAi transgenic lines that have reduced VPS45 protein levels. Root hair and pollen tube elongation, both processes of tip growth, are highly compromised in vps45-3 . Mutant root hairs are shorter and thicker than those of wild-type plants, and are wavy. These root hairs have vacuolar defects, containing many small vacuoles, compared with WT root hairs with a single large vacuole occupying much of the cell volume. Pollen tubes were also significantly shorter in vps45-3 compared to WT. Discussion We thus show that VPS45 is essential for proper tip growth and propose that the observed vacuolar defects lead to loss of the turgor pressure needed for tip growth.

59 BASIC BIOLOGICAL SCIENCES↗

Bioenergy sorghum nodal root bud development: morphometric, transcriptomic and gene regulatory network analysis

Bioenergy sorghum’s large and deep nodal root system and associated microbiome enables uptake of water and nutrients from and deposition of soil organic carbon into soil profiles, key contributors to the crop’s resilience and sustainability. The goal of this study was to increase our understanding of bioenergy sorghum nodal root bud development. Sorghum nodal root bud initiation was first observed on the stem node of the 7 th phytomer below the shoot apex. Buds were initiated near the upper end of the stem node pulvinus on the side of the stem opposite the tiller bud, then additional buds were added over the next 6-8 days forming a ring of 10-15 nascent nodal root buds around the stem. Later in plant development, a second ring of nodal root buds began forming on the 17 th stem node immediately above the first ring of buds. Overall, nodal root bud development can take ~40 days from initiation to onset of nodal root outgrowth. Nodal root buds were initiated in close association with vascular bundles in the rind of the pulvinus. Stem tissue forming nascent nodal root buds expressed sorghum homologs of genes associated with root initiation (WOX4), auxin transport (LAX2, PIN4), meristem activation (NGAL2), and genes involved in cell proliferation. Expression of WOX11 and WOX5, genes involved in root stem niche formation, increased early in nodal root bud development followed by genes encoding PLTs, LBDs (LBD29), LRP1, SMB, RGF1 and root cap LEAs later in development. A nodal root bud gene regulatory network module expressed during nodal root bud initiation predicted connections linking PFA5, SPL9 and WOX4 to genes involved in hormone signaling, meristem activation, and cell proliferation. A network module expressed later in development predicted connections among SOMBRERO, a gene involved in root cap formation, and GATA19, BBM, LBD29 and RITF1/RGF1 signaling. Overall, this study provides a detailed description of bioenergy sorghum nodal root bud development and transcriptome information useful for understanding the regulation of sorghum nodal root bud formation and development.

09 BIOMASS FUELS↗

Potential Biocontrol Activities of Populus Endophytes against Several Plant Pathogens Using Different Inhibitory Mechanisms

The plant microbiome can be used to bolster plant defense against abiotic and biotic stresses. Some strains of endophytes, the microorganisms within plants, can directly inhibit the growth of plant fungal pathogens. A previously isolated endophyte from wild Populus (poplar), WPB of the species Burkholderia vietnamiensis, had robust in vitro antifungal activity against pathogen strains that are highly virulent and of concern to Pacific Northwest agriculture: Rhizoctonia solani AG-8, Fusarium culmorum 70110023, and Gaemannomyces graminis var. tritici (Ggt) ARS-A1, as well as activity against the oomycete, Pythium ultimum 217. A direct screening method was developed for isolation of additional anti-fungal endophytes from wild poplar extracts. By challenging pathogens directly with dilute extracts, eleven isolates were found to be inhibitory to at least two plant pathogen strains and were therefore chosen for further characterization. Genomic analysis was conducted to determine if these endophyte strains harbored genes known to be involved in antimicrobial activities. The newly isolated Bacillus strains had gene clusters for production of bacillomycin, fengicyn, and bacillibactin, while the gene cluster for the synthesis of sessilin, viscosin and tolaasin were found in the Pseudomonas strains. The biosynthesis gene cluster for occidiofungin (ocf) was present in the Burkholderia vietnamiensis WPB genome, and an ocf deletion mutant lost inhibitory activity against 3 of the 4 pathogens. The new isolates lacked the gene cluster for occidiofungin implying they employ different modes of action. Other symbiotic traits including nitrogen fixation, phosphate solubilization, and the production of auxins and siderophores were investigated. Although it will be necessary to conduct in vivo tests of the candidates with pathogen-infected agricultural crops, the wild poplar tree microbiome may be a rich source of beneficial endophyte strains with potential for biocontrol applications against a variety of pathogens and utilizing varying modes of action.

59 BASIC BIOLOGICAL SCIENCES↗

Growth responses of plants to gravity

Gravity effects on auxin transport, growth and foliage spread of green plants for efficient radiation capture, using horizontal clinostat experiments

Lyon, C. J.↗

Pathogenetic validation of the use of biological protective agents and early treatment in cases of radiation injury simulating radiation effects under space flight conditions

In considering a radiation safety system for space flights, the various measures to protect man against radiation include drug prophylaxis. At the present time a great deal of experimental material has been accumulated on the prevention and treatment of radiation injuries. Antiradiation effectiveness has been established for sulfur- and nitrogen-containing substances, auxins, cyanides, polynucleotides, mucopolysaccharides, lipopolysaccharides, aminosaccharides, synthetic polymers, vitamins, hormones, amino acids and other compounds which can be divided into two basic groups - biological and chemical protective agents.

Rogozkin, V. D.↗

Development of guayule (Parthenium argentatum) research in cell culture

Utilizing the lateral buds of known high rubber producing plants as explants in culture medium specifically designed to engender shoot development and to prevent callus formation, unlimited numbers of replicate plants can be produced. Each has the same genotype as the parent. This procedure has long been used to rid plants of virus, the latter generally does not occur in the embryonic tissues of the bud; it also, by virtue of its axenic nature, eliminates all microorganisms characteristic of the parent plant. Auxins were found essential to callus formation, but since the latter is known to bring about chromosomal aberrations, it was avoided. The cytokinin benzylaminopurine strongly stimulated shoot growth, and the number of regenerated buds on the inoculum was proportional to its concentration. These buds produced shoots several centimeters in length which were caused to root on medium containing indolebutyric acid. Transferred to the septic condition of soil, the plantlets were gradually brought into full sunlight where they showed a brief vegetative growth with production of mature leaves, and flowered.

Ball, E. A.↗

Some results from studies on the effects of weightlessness on the growth of epiphytic orchids

Epidendrum orchids were placed in a Malakhit-2 micro-greenhouse aboard the Soyuz-36-Salyut-6 space station to test their growth under weightless conditions. Growth occurred but was less than in control plants left on Earth; cells were smaller and parenchymal development slowed in all tissues. Stems, roots, and leaves were smaller. The number of stomas on the leaves was about the same as in the controls, but, because of the smaller leaf size, there were more per unit area. A modeling experiment using a clinostat revealed a large decrease in gibberellin activity and auxin activity. It was assumed that weightlessness primarily affects gibberellin biosynthesis, inhibiting cell growth. Reestablishment of growth compound activity upon return of the plants to Earth was indicated by the fact that the orchids resumed growth thereafter.

Cherevchenko, T. M.↗

Protein synthesis in geostimulated root caps

A study is presented of the processes occurring in the root cap of corn which are requisite for the formation of root cap inhibitor and which can be triggered or modulated by both light and gravity. The results of this study indicate the importance of protein synthesis for light-induced gravitropic bending in roots. Root caps in which protein synthesis is prevented are unable to induce downward bending. This suggests that light acts by stimulating proteins which are necessary for the translation of the gravitropic stimulus into a growth response (downward bending). The turnover of protein with time was also examined in order to determine whether light acts by stimulating the synthesis of unique proteins required for downward growth. It is found that auxin in combination with light allows for the translation of the gravitropic stimulus into a growth response at least in part through the modification of protein synthesis. It is concluded that unique proteins are stimulated by light and are involved in promoting the downward growth in roots which are responding to gravity.

Feldman, L. J.↗

Mechanism of shoot gravitropism

A better understanding of the cellular basis of plant shoot gravitropism was sought. A critical evaluation of the role of auxin gravitropism was provided. An alternative hypothesis which links Ca(42) fluxes to the asymmetric growth that leads to gravicurvature was evaluated.

Rayle, D. L.↗

The role of gravity in leaf blade curvatures

In the past year we have gained useful information on several aspects of leaf blade growth. The most important observations are as follows: The C(14)-1AA moves preferentially in a gravipositive dorsiventral direction through the blade. This movement is inhibited by inversion of the blade. The responding cells in leaf blade hyponasty are in the lower epidermis and bundle sheath cells. Two additional responses in the leaf were characterized. In addition to blade curvature, the leaf shows petiole curvature and changes in the liminal angle subtended by the pulvinus. Ethylene production was studied under a number of conditions. The blade, rather than the petiole or pulvinus, is the principal site of auxin-promoted ethylene synthesis. The effects of a variety of agents on the blade, including gibberellic acid, abscisic acid, vanadate, low pH buffers, and blue light were reviewed.

Hayes, A. B.↗

Polyamines as Possible Modulators of Gravity-induced Calcium Transport in Plants

Data from various laboratories indicate a probable relationship between calcium movement and some aspects of graviperception and tropistic bending responses. The movement of calcium in response to gravistimulation appears to be rapid, polar and opposite in direction to polar auxin transport. What might be the cause of such rapid Ca(2+) movement? Data from studies on polyamine (PA) metabolism may furnish a clue. A transient increase in the activity of ornithine decarboxylase (ODC) and titers of various PAs occurs within 60 seconds after hormonal stimulation of animal cells, followed by Ca(2+) transport out of the cells. Through the use of specific inhibitors, it was shown that the enhanced PA synthesis from ODC was essential not only for Ca(2+) transport, but also for Ca(2+) transport-dependent endocytosis and the movement of hexoses and amino acids across the plasmalemma. In plants, rapid changes in arginine decarboxylase (ADC) activity occur in response to various plant stresses. Physical stresses associated with gravisensor displacement and reorientation of a plant in the gravitational field could similarly activate ADC and that resultant increases in PA levels might initiate transient perturbations in Ca(2+) homeostasis.

Galston, A. W.↗

Gravitropism in Leafy Dicot Stems

A polarizing research microscope with rotating stage and associated camera equipment were ordered, and techniques of fixation and preparation of specimens were perfected for studying possible changes in orientation of cellulose microfibrils in cell walls of gravistimulated dicot stems. Acid ethephon solutions or acid without ethephon caused elongation of stem tissues where they were applied; stems bent away from the side of application. Acid solutions applied to the bottom of horizontal stems greatly delayed bending. Research in tissue sensitivity changes during gravitropic bending of soybean hypocotyls while immersed in auxin and in castor bean stems is also reported.

Salisbury, F. B.↗

Modification of cytogenetic and physiological effects of space flight factors by biologically active compounds

Physiological and cytogenetic changes in the Welsh onion plants induced by a short (82 days) and long term (522 days) space flight are expressed in decrease of seed germination, inhibition of stem growth, depression of cell division in root meristem, and increase in the number of structural chromosome rearrangements. The treatment of such plants with solutions of a-tocopherol, auxin, and kinetin decreased the level of chromosome aberrations to the control one and normalized cell divisions and growth partly or completely.

Aliyev, A. A.↗