Biosynthesis of L-ascorbic acid in plants: new pathways for an old antioxidant

The biosynthetic pathway of L-ascorbic acid (vitamin C) in plants has been established for several years. However, recent reports describe alternative pathways, revealing a more complex picture of L-ascorbic acid biosynthesis than had been expected. GDP-L-gulose and myo-inositol are proposed as new intermediates in L-ascorbic acid biosynthesis, indicating that part of the animal pathway might also be operating in plants. Enzymatic studies on the GDP-mannose-3 0 ,5 0 -epimerase and L-galactono-1,4-lactone dehydrogenase suggest that they are important regulatory steps for L-ascorbic acid biosynthesis

The Arabidopsis Synaptotagmin1 is enriched in endoplasmic reticulum-plasma membrane contact sites and confers cellular resistance to mechanical stresses

Eukaryotic endoplasmic reticulum (ER)-plasma membrane (PM) contact sites are evolutionarily conserved microdomains that have important roles in specialized metabolic functions such as ER-PM communication, lipid homeostasis, and Ca2+ influx. Despite recent advances in knowledge about ER-PM contact site components and functions in yeast (Saccharomyces cerevisiae) and mammals, relatively little is known about the functional significance of these structures in plants. In this report, we characterize the Arabidopsis (Arabidopsis thaliana) phospholipid binding Synaptotagmin1 (SYT1) as a plant ortholog of the mammal extended synaptotagmins and yeast tricalbins families of ER-PM anchors. We propose that SYT1 functions at ER-PM contact sites because it displays a dual ER-PM localization, it is enriched in microtubule-depleted regions at the cell cortex, and it colocalizes with Vesicle-Associated Protein27-1, a known ER-PM marker. Furthermore, biochemical and physiological analyses indicate that SYT1 might function as an electrostatic phospholipid anchor conferring mechanical stability in plant cells. Together, the subcellular localization and functional characterization of SYT1 highlights a putative role of plant ER-PM contact site components in the cellular adaptation to environmental stresses

Small violations of unitarity, the phase in Bs mixing and visible t->cZ decays at the LHC

We show that it is possible to accommodate the observed size of the phase in $B^0_s$--$\bar B^0_s$, mixing in the framework of a model with violation of $3\times 3$ unitarity. This violation is associated to the presence of a new $Q=2/3$ isosinglet quark $T$, which mixes both with $t$ and $c$ and has a mass not exceeding 500 GeV. The crucial point is the fact that this framework allows for $\chi\equiv\arg(-V_{ts}V_{cb}V_{tb}^*V_{cs}^*)$ of order $\lambda$, to be contrasted with the situation in the Standard Model, where $\chi$ is constrained to be of order $\lambda^2$. We point out that this scenario implies rare top decays $t\to cZ$ at a rate observable at the LHC and $|V_{tb}|$ significantly different from unity. In this framework, one may also account for the observed size of $D^0$--$\bar D^0$ mixing without having to invoke long distance contributions. It is also shown that in the present scenario, the observed size of $D^0$--$\bar D^0$ mixing constrains $\chi^\prime\equiv\arg(-V_{cd}V_{us}V_{cs}^*V_{ud}^*)$ to be of order $\lambda^4$, which is significantly smaller than what is allowed in generic models with violations of $3\times 3$ unitarity.Comment: 20 pages. References added. Additional observables considered, updated numerical examples, conclusions unchange

Vitamin C Content in Fruits: Biosynthesis and Regulation

Throughout evolution, a number of animals including humans have lost the ability to synthesize ascorbic acid (ascorbate, vitamin C), an essential molecule in the physiology of animals and plants. In addition to its main role as an antioxidant and cofactor in redox reactions, recent reports have shown an important role of ascorbate in the activation of epigenetic mechanisms controlling cell differentiation, dysregulation of which can lead to the development of certain types of cancer. Although fruits and vegetables constitute the main source of ascorbate in the human diet, rising its content has not been a major breeding goal, despite the large inter- and intraspecific variation in ascorbate content in fruit crops. Nowadays, there is an increasing interest to boost ascorbate content, not only to improve fruit quality but also to generate crops with elevated stress tolerance. Several attempts to increase ascorbate in fruits have achieved fairly good results but, in some cases, detrimental effects in fruit development also occur, likely due to the interaction between the biosynthesis of ascorbate and components of the cell wall. Plants synthesize ascorbate de novo mainly through the Smirnoff-Wheeler pathway, the dominant pathway in photosynthetic tissues. Two intermediates of the Smirnoff-Wheeler pathway, GDP-D-mannose and GDP-L-galactose, are also precursors of the non-cellulosic components of the plant cell wall. Therefore, a better understanding of ascorbate biosynthesis and regulation is essential for generation of improved fruits without developmental side effects. This is likely to involve a yet unknown tight regulation enabling plant growth and development, without impairing the cell redox state modulated by ascorbate pool. In certain fruits and developmental conditions, an alternative pathway from D-galacturonate might be also relevant. We here review the regulation of ascorbate synthesis, its close connection with the cell wall, as well as different strategies to increase its content in plants, with a special focus on fruits

Searching for New Physics in Leptonic Decays of Bottomonium

New Physics can show up in various well-known processes already studied in the Standard Model, in particular by modifying decay rates to some extent. In this work, I examine leptonic decays of $\Upsilon$ vector resonances of bottomonium below $B\bar{B}$ production, subsequent to a magnetic dipole radiative structural transition of the vector resonance yielding a pseudoscalar continuum state, searching for the existence of a light Higgs-like neutral boson that would imply a slight but experimentally measurable breaking of lepton universality.Comment: LaTeX, 12 pages, 1 EPS figur

Phosphorylation regulates cellulose biosynthesis regulation.

Plants comprise over 82% of all biomass on Earth, a third of which is cellulose, making it the most abundant organic compound¹. Cellulose is also essential for plant development and defense against multiple stresses. But, despite its relevance, there remains much to be discovered about its biosynthetic regulation to improve crop’s tolerance to biotic and abiotic stresses. We have described the Tetratricopeptide Thioredoxin‐Like (TTL² proteins as regulators of the cellulose synthase complex (CSC) under cellulose‐deficient conditions³. We found that TTLs are required to maintain cellulose synthesis under salt stress by relocalising from the cytosol to the CSCs, promoting the polymerization of microtubules to form a stress‐resilient cortical microtubule array, and interacting with the CSCs to stabilize them at the plasma membrane. We are currently investigating how TTLs are targeted to the CSCs. We have found that TTL3 is a substrate for a kinase that when mutated show cellulose‐defective phenotypes under stresses that affect cell wall integrity. We are currently analysing how the CSCs and microtubules behave under abiotic stress in mutants for this kinase. This will provide new insights into how changes in phosphorylation status regulate the activity and dynamic localization of these proteins.This work was funded by the Spanish Ministry for Science and Innovation (PID2020-114419RB-I00MCIN/AEI/10.13039/501100011033 ) to MAB. The Andalusian Research Plan co-financed by the European Union (PAIDI 2020- PY20_00084 and UMA20-FEDERJA-023) to MAB. FP was supported by FPU19/02219 fellowships and EMBO Scientific Exchange Grant 10026 and VAS was supported by an Emerging Investigator research project (UMA20-FEDERJA -007) and co-financed by the “Programa Operativo FEDER 2014-2020” and by the “Consejería de Economía y Conocimiento de la Junta de Andalucía”. Funding from the University of Zürich, and the Swiss National Science Foundation grants no. 31003A_182625 and no. 310030_212382 (to CZ). Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Golgi Apparatus-Localized Synaptotagmin 2 Is Required for Unconventional Secretion in Arabidopsis

BACKGROUND: Most secretory proteins contain signal peptides that direct their sorting to the ER and secreted via the conventional ER/Golgi transport pathway, while some signal-peptide-lacking proteins have been shown to export through ER/Golgi independent secretory pathways. Hygromycin B is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus that is active against both prokaryotic and eukaryotic cells. The hygromycin phosphotransferase (HYG(R)) can phosphorylate and inactivate the hygromycin B, and has been widely used as a positive selective marker in the construction of transgenic plants. However, the localization and trafficking of HYG(R) in plant cells remain unknown. Synaptotagmins (SYTs) are involved in controlling vesicle endocytosis and exocytosis as calcium sensors in animal cells, while their functions in plant cells are largely unclear. METHODOLOGY/PRINCIPAL FINDINGS: We found Arabidopsis synaptotagmin SYT2 was localized on the Golgi apparatus by immunofluorescence and immunogold labeling. Surprisingly, co-expression of SYT2 and HYG(R) caused hypersensitivity of the transgenic Arabidopsis plants to hygromycin B. HYG(R), which lacks a signal sequence, was present in the cytoplasm as well as in the extracellular space in HYG(R)-GFP transgenic Arabidopsis plants and its secretion is not sensitive to brefeldin A treatment, suggesting it is not secreted via the conventional secretory pathway. Furthermore, we found that HYG(R)-GFP was truncated at carboxyl terminus of HYG(R) shortly after its synthesis, and the cells deficient SYT2 failed to efficiently truncate HYG(R)-GFP,resulting in HYG(R)-GFP accumulated in prevacuoles/vacuoles, indicating that SYT2 was involved in HYG(R)-GFP trafficking and secretion. CONCLUSION/SIGNIFICANCE: These findings reveal for the first time that SYT2 is localized on the Golgi apparatus and regulates HYG(R)-GFP secretion via the unconventional protein transport from the cytosol to the extracelluar matrix in plant cells

Genetic diversity and population structure of Pepino mosaic virus in tomato crops of Spain and Morocco

Pepino mosaic virus (PepMV, genus Potexvirus) is an emergent and highly infectious pathogen responsible for economically important diseases in tomato crops. An extensive survey of tomato plants showing PepMV-like symptoms was carried out in 2017 to study the PepMV genetic diversity and populations structure in different tomato-producing areas of Spain and Morocco. Molecular dot-blot hybridization analysis showed that virus populations from Spain and Morocco were mainly composed of isolates belonging to the Chilean 2 (CH2) strain, although isolates of the European (EU) strain were detected in significant proportions in Spanish populations, mainly in mixed infections. A few isolates of the American (US1) strain were also detected in Tenerife (Canary Islands, Spain) crops. Eighty-five isolates were randomly selected and sequenced in the genomic region that encodes the triple gene block and capsid protein genes. Our phylogenetic and population genetics analyses confirmed the presence of the CH2, EU and US1 PepMV strains. Despite the high genetic similarity observed within populations, variants were maintained at low frequency under purifying selection, and differentiation among more geographically distant locations was identified, with potential gene flow contributing to the shaping of the PepMV populations structur

Differential gene expression profile in omental adipose tissue in women with polycystic ovary syndrome

10 pages, 2 figures, 5 tables.CONTEXT: The polycystic ovary syndrome (PCOS) is frequently associated with visceral obesity, suggesting that omental adipose tissue might play an important role in the pathogenesis of the syndrome. OBJECTIVE: The objective was to study the expression profiles of omental fat biopsy samples obtained from morbidly obese women with or without PCOS at the time of bariatric surgery. DESIGN: This was a case-control study. SETTINGS: We conducted the study in an academic hospital. PATIENTS: Eight PCOS patients and seven nonhyperandrogenic women submitted to bariatric surgery because of morbid obesity. INTERVENTIONS: Biopsy samples of omental fat were obtained during bariatric surgery. MAIN OUTCOME MEASURE: The main outcome measure was high-density oligonucleotide arrays. RESULTS: After statistical analysis, we identified changes in the expression patterns of 63 genes between PCOS and control samples. Gene classification was assessed through data mining of Gene Ontology annotations and cluster analysis of dysregulated genes between both groups. These methods highlighted abnormal expression of genes encoding certain components of several biological pathways related to insulin signaling and Wnt signaling, oxidative stress, inflammation, immune function, and lipid metabolism, as well as other genes previously related to PCOS or to the metabolic syndrome. CONCLUSION: The differences in the gene expression profiles in visceral adipose tissue of PCOS patients compared with nonhyperandrogenic women involve multiple genes related to several biological pathways, suggesting that the involvement of abdominal obesity in the pathogenesis of PCOS is more ample than previously thought and is not restricted to the induction of insulin resistance.This work was supported by PI020578, PI020741, PI050341, PI050551, RCMN C03/08, and RGDM 03/212 from Fondo de Investigación Sanitaria, Instituto de Salud Carlos III, and Grants 08.6/0021/2003 and GR/SAL/0137/2004 from the Consejería de Educación y Cultura, Comunidad de Madrid, Spain.Peer reviewe