Dual-Cation Electrolytes for High-Power and High-Energy LTO//AC Hybrid Capacitors

Dual-cation electrolyte systems, which contain two cations [Li+ and spiro-1,1′-bipyrrolidinium (SBP+), are proposed to enhance the power capability of hybrid capacitors composed of thick Li4Ti5O12 (LTO) negative (200 μm) and activated carbon (AC) positive electrodes (400 μm), which thus reduces the resistive overvoltage in the system. Detailed studies of the mass transport properties based on the combination of spectroscopy and electrochemical analysis have shown that the presence of SBP+, despite slower Li+ transport in the electrolyte bulk, further reduces overvoltage associated with migration limitation in the thick LTO electrode macropores. This study on the dual-cation electrolyte quantifies the influence of the addition of a supporting electrolyte and shows interest in SBPBF4 addition for increasing the output power density of hybrid capacitors with a thick electrode configuration

Coevolution and hierarchical interactions of Tomato mosaic virus and the resistance gene Tm-1.

During antagonistic coevolution between viruses and their hosts, viruses have a major advantage by evolving more rapidly. Nevertheless, viruses and their hosts coexist and have coevolved, although the processes remain largely unknown. We previously identified Tm-1 that confers resistance to Tomato mosaic virus (ToMV), and revealed that it encodes a protein that binds ToMV replication proteins and inhibits RNA replication. Tm-1 was introgressed from a wild tomato species Solanum habrochaites into the cultivated tomato species Solanum lycopersicum. In this study, we analyzed Tm-1 alleles in S. habrochaites. Although most part of this gene was under purifying selection, a cluster of nonsynonymous substitutions in a small region important for inhibitory activity was identified, suggesting that the region is under positive selection. We then examined the resistance of S. habrochaites plants to ToMV. Approximately 60% of 149 individuals from 24 accessions were resistant to ToMV, while the others accumulated detectable levels of coat protein after inoculation. Unexpectedly, many S. habrochaites plants were observed in which even multiplication of the Tm-1-resistance-breaking ToMV mutant LT1 was inhibited. An amino acid change in the positively selected region of the Tm-1 protein was responsible for the inhibition of LT1 multiplication. This amino acid change allowed Tm-1 to bind LT1 replication proteins without losing the ability to bind replication proteins of wild-type ToMV. The antiviral spectra and biochemical properties suggest that Tm-1 has evolved by changing the strengths of its inhibitory activity rather than diversifying the recognition spectra. In the LT1-resistant S. habrochaites plants inoculated with LT1, mutant viruses emerged whose multiplication was not inhibited by the Tm-1 allele that confers resistance to LT1. However, the resistance-breaking mutants were less competitive than the parental strains in the absence of Tm-1. Based on these results, we discuss possible coevolutionary processes of ToMV and Tm-1

Coevolution and Hierarchical Interactions of <em>Tomato mosaic virus</em> and the Resistance Gene <em>Tm-1</em>

<div><p>During antagonistic coevolution between viruses and their hosts, viruses have a major advantage by evolving more rapidly. Nevertheless, viruses and their hosts coexist and have coevolved, although the processes remain largely unknown. We previously identified <em>Tm-1</em> that confers resistance to <em>Tomato mosaic virus</em> (ToMV), and revealed that it encodes a protein that binds ToMV replication proteins and inhibits RNA replication. <em>Tm-1</em> was introgressed from a wild tomato species <em>Solanum habrochaites</em> into the cultivated tomato species <em>Solanum lycopersicum</em>. In this study, we analyzed <em>Tm-1</em> alleles in <em>S. habrochaites</em>. Although most part of this gene was under purifying selection, a cluster of nonsynonymous substitutions in a small region important for inhibitory activity was identified, suggesting that the region is under positive selection. We then examined the resistance of <em>S. habrochaites</em> plants to ToMV. Approximately 60% of 149 individuals from 24 accessions were resistant to ToMV, while the others accumulated detectable levels of coat protein after inoculation. Unexpectedly, many <em>S. habrochaites</em> plants were observed in which even multiplication of the <em>Tm-1</em>-resistance-breaking ToMV mutant LT1 was inhibited. An amino acid change in the positively selected region of the Tm-1 protein was responsible for the inhibition of LT1 multiplication. This amino acid change allowed Tm-1 to bind LT1 replication proteins without losing the ability to bind replication proteins of wild-type ToMV. The antiviral spectra and biochemical properties suggest that <em>Tm-1</em> has evolved by changing the strengths of its inhibitory activity rather than diversifying the recognition spectra. In the LT1-resistant <em>S. habrochaites</em> plants inoculated with LT1, mutant viruses emerged whose multiplication was not inhibited by the <em>Tm-1</em> allele that confers resistance to LT1. However, the resistance-breaking mutants were less competitive than the parental strains in the absence of <em>Tm-1</em>. Based on these results, we discuss possible coevolutionary processes of ToMV and <em>Tm-1</em>.</p> </div

Inhibition of <i>in vitro</i> RNA replication of ToMV mutants by Tm-1 variants.

<p>The genomic RNA of TLIle, ToMV-L, LT1, T21, LT1^E979K, or LT1^D1097Y and the mRNA for tm-1^GCR26, Tm-1^GCR237, or Tm-1^I91T proteins were translated in mdBYL. The translation mixtures of the Tm-1 variants were mixed with the viral RNA-translated mixtures, followed by RNA replication reaction as described in the <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#s4" target="_blank">Materials and Methods</a> section. The amount of added Tm-1 mRNA were approximately 9 (lanes 2, 5, 8), 42 (lanes 3, 6, 9), or 126 (lanes 4, 7, 10) times as much as viral RNA on a molar basis. Mock-translated mixture was added as a control (lane 1). The positions of the genomic RNA (G) and the replicative form RNA (RF) are indicated. Asterisks represent the background signals.</p

ToMV-L-resistant and -susceptible <i>S. habrochaites</i> have distinct amino acid sequences in the positively selected region of Tm-1.

<p>Deduced amino acid sequences of the Tm-1 protein of five ToMV-L-resistant and -susceptible <i>S. habrochaites</i> plants from the indicated accessions were aligned. The positively selected region (79–112) is indicated. Identical amino acid residues to those of Tm-1^GCR237 are indicated by dots.</p

A small region of the <i>Tm-1</i> gene is under positive selection in <i>S. habrochaites</i>.

<p>(A) Predicted domain structure of the Tm-1 protein by the NCBI Conserved Domain Database. A region encoded by the alternative exon (46–263) is underlined. (B) Detection of natural selection in the <i>Tm-1</i> alleles from <i>S. habrochaites</i>. The ratio of nonsynonymous/synonymous substitutions (ω) in each codon was inferred by omegaMap <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Wilson1" target="_blank">[33]</a>. ω>1, ω = 1, and ω<1 suggest positive selection, neutral evolution, and negative selection, respectively. The region where posterior probability of positive selection (ω>1) exceeds 95% is indicated (from 79^th to 112^th codon). (C) Sliding window analysis of Tajima's <i>D</i> of the <i>Tm-1</i> alleles from <i>S. habrochaites</i>. The confidence limits of <i>D</i> for neutral evolution <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Tajima1" target="_blank">[35]</a> are shown as dashed lines.</p

Accumulation of ToMV-L CP in <i>S. habrochaites</i> accessions.

a<p>high level accumulation,</p>b<p>low level accumulation,</p>c<p>not detectable.</p><p>In underlined accessions, more than 70% of plant individuals did not accumulate detectable amounts of ToMV-L CP.</p

Tm-1^I91T binds LT1 replication proteins, but not LT1^E979K or LT1^D1097Y.

<p>The genomic RNA of TLIle, ToMV-L, LT1, T21, LT1^E979K, or LT1^D1097Y were translated in mdBYL; mixed with mdBYL in which tm-1^GCR26-FLAG, Tm-1^GCR237-FLAG, or Tm-1^I91T-FLAG mRNA were translated; and immunoprecipitated using anti-FLAG antibody-conjugated agarose. Mock-translation was performed as controls and indicated as no viral RNA or no FLAG RNA. Protein samples before (Input) or after (IP: anti-FLAG) FLAG purification were analyzed by Western blotting using anti-130K protein or anti-FLAG antibodies. Positions of the replication proteins (130K and 180K proteins) and FLAG-tagged tm-1^GCR26, Tm-1^GCR237, or Tm-1^I91T proteins are indicated.</p

Schematic representation of ToMV mutant genomes with different sensitivities to <i>Tm-1</i> alleles.

<p>Positions of amino acid residue changes in <i>Tm-1</i>-resistance-breaking mutants are shown. Amino acid residues identical to ToMV-L are indicated by dots. LT1 and T21 are <i>Tm-1^GCR237</i>-breaking mutants <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Meshi2" target="_blank">[20]</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Strasser1" target="_blank">[28]</a> and TLIle is a <i>tm-1^GCR26</i>-sensitive mutant <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Ishibashi3" target="_blank">[30]</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002975#ppat.1002975-Hamamoto1" target="_blank">[50]</a>. LT1^E979K and LT1^D1097Y were characterized in this study. MT: methyltransferase domain, Hel: helicase domain, Pol: RNA-dependent RNA polymerase domain, CP: coat protein.</p