Tandem photoaffinity labeling of a target protein using a linker with biotin, alkyne and benzophenone groups and a bioactive small molecule with an azide group

<div><p>A novel linker containing biotin, alkyne and benzophenone groups (<b>1</b>) was synthesized to identify target proteins using a small molecule probe. This small molecule probe contains an azide group (azide probe) that reacts with an alkyne in <b>1</b> via an azide–alkyne Huisgen cycloaddition. Cross-linking of benzophenone to the target protein formed a covalently bound complex consisting of the azide probe and the target protein via <b>1</b>. The biotin was utilized via biotin–avidin binding to identify the cross-linked complex. To evaluate the effectiveness of <b>1</b>, it was applied in a model system using an allene oxide synthase (AOS) from the model moss <i>Physcomitrella patens</i> (PpAOS1) and an AOS inhibitor that contained azide group (<b>3</b>)<b>.</b> The cross-linked complex consisting of PpAOS1, <b>1</b> and <b>3</b> was resolved via SDS–PAGE and visualized using a chemiluminescent system. The method that was developed in this study enables the effective identification of target proteins.</p></div

Induction of plant disease resistance upon treatment with yeast cell wall extract

<p>It has been reported that treatment with yeast cell wall extract (YCWE) induces <i>PDF1</i> and <i>PR</i>-<i>1</i> gene expression; these transcripts are important markers of plant disease resistance, though the detailed signaling mechanisms that induce these defense responses are still unknown. In this report, we found that YCWE treatment triggered rice cell suspension cultures to accumulate phenylalanine (Phe), <i>cis</i>-12-oxo-phytodienoic acid (OPDA), 12-hydroxyjasmonoyle isoleucine (12OHJA-Ile), and azelaic acid (AzA). YCWE treatment also reduced endogenous triacylglycerol (TG) content. The addition of ¹³C-uniform-labeled oleic, linoleic and linolenic acids to the rice cell suspension cultures gave rise to ¹³C-uniform-labeled AzA. It was also found that YCWE treatment for <i>Arabidopsis thaliana</i> resulted in accumulations of OPDA, AzA, Phe, and camalexin together with enhanced resistance against <i>Botrytis cinerea</i> infection. This suggested that YCWE treatment upon plants may activate JA and AzA signaling systems to induce plant disease resistance.</p> <p>Yeast cell wall extract (YCWE) treatment induces plant defense response.</p

<i>N</i>¹, <i>N</i>¹⁴-diferuloylspermine as an antioxidative phytochemical contained in leaves of <i>Cardamine fauriei</i>

<p>Most Brassicaceae vegetables are ideal dietary sources of antioxidants beneficial for human health. <i>Cardamine fauriei</i> (Ezo-wasabi in Japanese) is a wild, edible Brassicaceae herb native to Hokkaido, Japan. To clarify the main antioxidative phytochemical, an 80% methanol extraction from the leaves was fractionated with Diaion® HP-20, Sephadex® LH-20, and Sep-Pak® C18 cartridges, and the fraction with strong antioxidant activity depending on DPPH method was purified by HPLC. Based on the analyses using HRESIMS and MS/MS, the compound might be <i>N</i>¹, <i>N</i>¹⁴-diferuloylspermine. This rare phenol compound was chemically synthesized, whose data on HPLC, MS and ¹H NMR were compared with those of naturally derived compound from <i>C. fauriei</i>. All results indicated they were the same compound. The radical-scavenging properties of diferuloylspermine were evaluated by ORAC and ESR spin trapping methods, with the diferuloylspermine showing high scavenging activities of the ROO^·, O₂^·−, and HO^· radicals as was those of conventional antioxidants.</p

Additional file 1: Table S1. of RNA-seq-based evaluation of bicolor tepal pigmentation in Asiatic hybrid lilies (Lilium spp.)

Summary of assembled sequences from the tepal parts of the Asiatic hybrid lily Lollypop. Table S3. Primers used for quantitative RT-PCR (qRT-PCR) analysis. Figure S1. Phenylpropanoid, anthocyanin, and cinnamic acid derivative biosynthesis pathways in lily tepals. Enzymes whose genes are up-regulated in upper tepals (estimated by qRT-PCR) are shown in blue. 3GT, anthocyanidin 3-O-glucosyltransferase; 3RT, anthocyanidin-3-glucoside rhamnosyltransferase; 4CL, 4-coumaroyl: CoA-ligase; 7GT, anthocyanidin-3-rutinoside 7-glucosyltransferase; ANS, anthocyanidin synthase; CHI, chalcone isomerase; CHS, chalcone synthase; C3H, p-coumarate 3-hydroxylase; C4H, cinnamate 4-hydroxylase; DFR, dihydroflavonol 4-reductase; F3H, flavanone 3-hydroxylase; F3′H, flavonoid 3′-hydroxylase; FLS, flavonol synthase; GST, glutathione S-transferase; HCT, shikimate O-hydroxycinnamoyl transferase; MATE, multidrug and toxic compound extrusion transporter; PAL, phenylalanine ammonia-lyase. Figure S2. HPLC analysis of anthocyanins and CADs in upper tepals (upper) and tepal bases (basal) of lily cultivars. A: Absorbance at 525 nm (anthocyanins) of the tepal extracts in Lollypop, and cyanidin 3-O-glycoside (Cy3G) and cyanidin 3-O-rutinoside (Cy3R) standards. B: Absorbance at 340 nm (CADs) of the tepal extracts in six cultivars. Figure S3. Alignment of predicted amino acid sequences of isoforms annotated as LhPAL1, LhPAL2, and LhPAL3 (A), and LhCHSa and LhCHSb (B). Letters on black and grey backgrounds indicate identical and similar amino acids, respectively. Asterisks indicate stop codons. Figure S4. Relative expression levels of c30288_g1 (HCT), c10735_g1 (MYB3), c25442_g1 (MYB8), c25442_g2, c24227_g1 (R3-MYB), c24227_g2 (R3-MYB), c18278_g2 (R3-MYB), c36339_g1 (SPL9), and c16635-g1 (RCP1) in upper tepals and tepal bases of Lollypop during floral development (St 1–5). ACTIN was used to normalize the expression of target genes. Values and vertical bars indicate the mean ± standard error (n = 3). The same letters above the columns indicate that the values are not statistically significant (p <0.05) by Tukey’s HSD. Figure S5. Relative expression levels of LhMYB12 in Sugar Love and WD40 in Sugar Love and Montreux in upper tepals and tepal bases during floral development (St 1–5, A) and flowers of the cultivars Montreux and Sugar Love (B). ACTIN was used to normalize the expression of target genes. Values and vertical bars indicate the means ± standard error (n = 3). The same letters above the columns indicate that the values are not statistically significant (p <0.05) by Tukey’s HSD. Figure S6. Putative miR828 and pri-miR828 sequences in Lollypop. A: Putative miR828 and its target site appeared in c22900_g1 (MYB12). B: Sequence alignment of c13793_g1 and pri-miR828 in Glycine max [GmPri-miR828a (NR_126648) and GmPri-miR828b (NR_126651)], Vitis vinifera [VvPri-miR828a (NR_127861) and VvPri-miR828b (LM611741)], and Malus domestica [MdPri-miR828b (NR_120979) and MdPri-miR828a (NR_120978)]. (PDF 1261 kb

Additional file 2: Table S2. of RNA-seq-based evaluation of bicolor tepal pigmentation in Asiatic hybrid lilies (Lilium spp.)

Annotation and expression summary of differentially expressed genes. (XLSX 216 kb

Isolation, Structural Elucidation, and Biological Evaluation of a 5‑Hydroxymethyl-2-furfural Derivative, Asfural, from Enzyme-Treated Asparagus Extract

A novel 5-hydroxymethyl-2-furfural (HMF; <b>1</b>) derivative, which is named asfural (compound <b>2</b>), was isolated from enzyme-treated asparagus extract (ETAS) along with HMF (<b>1</b>) as a heat shock protein 70 (HSP70) inducible compound. The structure of compound <b>2</b> was elucidated on the basis of its spectroscopic data from HREIMS and NMR, whereas the absolute configuration was determined using chiral HPLC analysis, compared to two synthesized compounds, (<i>S</i>)- and (<i>R</i>)-asfural. As a result, compound <b>2</b> derived from ETAS was assigned as (<i>S</i>)-(2-formylfuran-5-yl)­methyl 5-oxopyrrolidine-2-carboxylate. When compound <b>2</b>, synthesized (<i>S</i>)- and (<i>R</i>)-asfural, and HMF (<b>1</b>) were evaluated in terms of HSP70 mRNA expression-enhancing activity in HL-60 cells, compound <b>2</b> and (<i>S</i>)-asfural significantly increased the expression level in a concentration-dependent manner. HMF (<b>1</b>) also showed significant activity at 0.25 mg/mL

GTR1 is a jasmonic acid and jasmonoyl-l-isoleucine transporter in <i>Arabidopsis thaliana</i>

<p>Jasmonates are major plant hormones involved in wounding responses. Systemic wounding responses are induced by an electrical signal derived from damaged leaves. After the signaling, jasmonic acid (JA) and jasmonoyl-l-isoleucine (JA-Ile) are translocated from wounded to undamaged leaves, but the molecular mechanism of the transport remains unclear. Here, we found that a JA-Ile transporter, GTR1, contributed to these translocations in <i>Arabidopsis thaliana</i>. <i>GTR1</i> was expressed in and surrounding the leaf veins both of wounded and undamaged leaves. Less accumulations and translocation of JA and JA-Ile were observed in undamaged leaves of <i>gtr1</i> at 30 min after wounding. Expressions of some genes related to wound responses were induced systemically in undamaged leaves of <i>gtr1</i>. These results suggested that GTR1 would be involved in the translocation of JA and JA-Ile in plant and may be contributed to correct positioning of JA and JA-Ile to attenuate an excessive wound response in undamaged leaves.</p> <p>JA and JA-Ile translocations by GTR1 would be contributed to their correct positioning to attenuate an excessive wound response in undamaged leaves.</p