SlCPK27 cross-links SlHY5 and SlPIF4 in brassinosteroid-dependent photo- and thermo-morphogenesis in tomato.

ELONGATED HYPOCOTOYL5 (HY5) and PHYTOCHROME INTERACTING FACTORs (PIFs) are two types of important light-related regulators of plant growth, however, their interplay remains elusive. Here, we report that the activated tomato (Solanum lycopersicum) HY5 (SlHY5) triggers the transcription of a Calcium-dependent Protein Kinase SlCPK27. SlCPK27 interacts with and phosphorylates SlPIF4 at Ser-252 and Ser-308 phosphosites to promote its degradation. SlPIF4 promotes hypocotyl elongation mainly by activating the transcription of SlDWF, a key gene in brassinosteroid (BR) biosynthesis. Such a SlHY5-SlCPK27-SlPIF4-BR cascade not only plays a crucial role in photomorphogenesis but also regulates thermomorphogenesis. Our results uncover a previously unidentified mechanism that integrates Ca2+ signaling with the light signaling pathways to regulate plant growth by modulating BR biosynthesis in response to changes in ambient light and temperature.

HY5 functions as a systemic signal by integrating BRC1-dependent hormone signaling in tomato bud outgrowth.

Light plays an important role in determining plant architecture, which greatly influences crop yield. However, the precise mechanisms by which light signaling regulates bud outgrowth remain to be identified. Here, we show that light regulates bud outgrowth via both HY5 and brassinosteroid (BR)-dependent pathways in tomato. Inactivation of the red-light photoreceptor PHYB, or deficiencies in PHYB or the blue-light photoreceptor CRY1a, inhibits bud outgrowth and leads to decreased accumulation of HY5 protein and increased transcript level of BRANCHED1 (BRC1), a central integrator of branching signals. HY5, functioning as a mobile systemic signal from leaves, promotes bud outgrowth by directly suppressing BRC1 transcript and activating the transcript of BR biosynthesis genes within the lateral buds in tomato. Furthermore, BRC1 prevents the accumulation of cytokinin (CK) and gibberellin (GA) by directly inhibiting the transcript of CK synthesis gene LOG4, while increasing the transcript levels of CK and GA degradation genes (CKX7, GA2ox4, and GA2ox5), leading to an arrest of bud outgrowth. Moreover, bud outgrowth occurs predominantly in the day due to the suppression of BRC1 transcript by HY5. These findings demonstrate that light-inducible HY5 acts as a systemic signaling factor in fine-tuning the bud outgrowth of tomato.

The RALF2-FERONIA-MYB63 module orchestrates growth and defense in tomato roots.

Plant secreted peptides RAPID ALKALINISATION FACTORs (RALFs), which act through the receptor FERONIA (FER), play important roles in plant growth. However, it remains unclear whether and how RALF-FER contributes to the trade-off of plant growth-defense. Here, we used a variety of techniques such as CRISPR/Cas9, protein-protein interaction and transcriptional regulation methods to investigate the role of RALF2 and its receptor FER in regulating lignin deposition, root growth, and defense against Fusarium oxysporum f. sp. lycopersici (Fol) in tomato (Solanum lycopersicum). The ralf2 and fer mutants show reduced primary root length, elevated lignin accumulation, and enhanced resistance against Fol than the wild-type. FER interacts with and phosphorylates MYB63 to promote its degradation. MYB63 serves as an activator of lignin deposition by regulating the transcription of dirigent protein gene DIR19. Mutation of DIR19 suppresses lignin accumulation, and reverses the short root phenotype and Fol resistance in ralf2 or fer mutant. Collectively, our results demonstrate that the RALF2-FER-MYB63 module fine-tunes root growth and resistance against Fol through regulating the deposition of lignin in tomato roots. The study sheds new light on how plants maintain the growth-defense balance via RALF-FER.

A transcriptional regulation of ERF15 contributes to ABA-mediated cold tolerance in tomato.

Cold stress is a major meteorological threat to crop growth and yield. Abscisic acid (ABA) plays important roles in plant cold tolerance by activating the expression of cold-responsive genes; however, the underlying transcriptional regulatory module remains unknown. Here, we demonstrated that the cold- and ABA-responsive transcription factor ETHYLENE RESPONSE FACTOR 15 (ERF15) positively regulates ABA-mediated cold tolerance in tomato. Exogenous ABA treatment significantly enhanced cold tolerance in wild-type tomato plants but failed to rescue erf15 mutants from cold stress. Transcriptome analysis showed that ERF15 was associated with the expression of cold-responsive transcription factors such as CBF1 and WRKY6. Further RT-qPCR assays confirmed that the ABA-induced increased in CBF1 and WRKY6 transcripts was suppressed in erf15 mutants when the plants were subjected to cold treatment. Moreover, yeast one-hybrid assays, dual-luciferase assays and electrophoretic mobility shift assays demonstrated that ERF15 activated the transcription of CBF1 and WRKY6 by binding their promoters. Silencing CBF1 or WRKY6 significantly decreased cold tolerance. Overall, our study identified the role of ERF15 in conferring ABA-mediated cold tolerance in tomato plants by activating CBF1 and WRKY6 expression. This study not only broadens our knowledge of the mechanism of ABA-mediated cold tolerance in plants but also highlights ERF15 as an ideal target gene for cold-tolerant crop breeding.

The phyB-dependent induction of HY5 promotes iron uptake by systemically activating FER expression.

Iron (Fe) deficiency affects global crop productivity and human health. However, the role of light signaling in plant Fe uptake remains uncharacterized. Here, we find that light-induced Fe uptake in tomato (Solanum lycopersicum L.) is largely dependent on phytochrome B (phyB). Light induces the phyB-dependent accumulation of ELONGATED HYPOCOTYL 5 (HY5) protein both in the leaves and roots. HY5 movement from shoots to roots activates the expression of FER transcription factor, leading to the accumulation of transcripts involved in Fe uptake. Mutation in FER abolishes the light quality-induced changes in Fe uptake. The low Fe uptake observed in phyB, hy5, and fer mutants is accompanied by lower photosynthetic electron transport rates. Exposure to red light at night increases Fe accumulation in wild-type fruit but has little effects on fruit of phyB mutants. Taken together, these results demonstrate that Fe uptake is systemically regulated by light in a phyB-HY5-FER-dependent manner. These findings provide new insights how the manipulation of light quality could be used to improve Fe uptake and hence the nutritional quality of crops.

Palladium-Catalyzed Multicomponent Cascade Reaction of 3-Hydroxypropionitrile: Synthesis of Substituted Dihydrochalcones.

Direct access to substituted dihydrochalcones from the easily available starting materials 3-hydroxypropionitrile derivatives and arylboronic acids is described. The procedure involves a multicomponent aryl addition/hydroxyl elimination/reduction Heck approach in the presence of a Pd catalyst with excellent functional group tolerance and a wide range of substrates. In addition, mixed 1,3-diarylation of 3-hydroxypropanenitrile using two arylboronic acids with different electronic properties was also achieved.

Physiological and Metabolic Changes in Tamarillo (Solanum betaceum) during Fruit Ripening.

Physiological and metabolic profiles in tamarillo were investigated to reveal the molecular changes during fruit maturation. The firmness, ethylene production, soluble sugar contents, and metabolomic analysis were determined in tamarillo fruit at different maturity stages. The firmness of tamarillo fruit gradually decreased during fruit ripening with increasing fructose and glucose accumulation. The rapid increase in ethylene production was found in mature fruit. Based on the untargeted metabolomic analysis, we found that amino acids, phospholipids, monosaccharides, and vitamin-related metabolites were identified as being changed during ripening. The contents of malic acid and citric acid were significantly decreased in mature fruits. Metabolites involved in phenylpropanoid biosynthesis, phenylalanine metabolism, caffeine metabolism, monoterpenoid biosynthesis, and thiamine metabolism pathways showed high abundance in mature fruits. However, we also found that most of the mature-enhanced metabolites showed reduced abundance in over-mature fruits. These results reveal the molecular profiles during tamarillo fruit maturing and suggest tamarillos have potential benefits with high nutrition and health function.

Herbivore-induced Ca2+ signals trigger a jasmonate burst by activating ERF16-mediated expression in tomato.

Herbivory severely affects plant growth, posing a threat to crop production. Calcium ion (Ca2+ ) signaling and accumulation of jasmonates (JAs) are activated in plant response to herbivore attack, leading to the expression of defense pathways. However, little is known about how the Ca2+ signal modulates JA biosynthesis. We used diverse techniques, including CRISPR/Cas9, UPLC-MS/MS and molecular biology methods to explore the role of ETHYLENE RESPONSE FACTOR 16 in Ca2+ signal-triggered JA burst during herbivore defense in tomato. Here we show that simulated herbivory induces GLUTAMATE RECEPTOR LIKE3.3/3.5 (GLR3.3/3.5)-dependent increases in electrical activity, Ca2+ influx and increases the abundance of CALMODULIN2 (CaM2) and ERF16 transcripts in tomato. The interaction between CaM2 and ERF16 promotes JA biosynthesis by enhancing the transcriptional activity of ERF16, which increases the activation of ERF16 expression and causes expression of LIPOXYGENASE D (LOXD), AOC and 12-OXO-PHYTODIENOIC ACID REDUCTASE 3 (OPR3), the key genes in JA biosynthesis. Mutation of CaM2 results in decreased JA accumulation, together with the expression of JA biosynthesis-related genes, leading to reduced resistance to the cotton bollworm Helicoverpa armigera. These findings reveal a molecular mechanism underpinning the Ca2+ signal-initiated systemic JA burst and emphasize the pivotal role of Ca2+ signal/ERF16 crosstalk in herbivore defense.

Ethylene response factors 15 and 16 trigger jasmonate biosynthesis in tomato during herbivore resistance.

Jasmonates (JAs) are phytohormones with crucial roles in plant defense. Plants accumulate JAs in response to wounding or herbivore attack, but how JA biosynthesis is triggered remains poorly understood. Here we show that herbivory by cotton bollworm (Helicoverpa armigera) induced both ethylene (ET) and JA production in tomato (Solanum lycopersicum) leaves. Using RNA-seq, ET mutants, and inhibitors of ET signaling, we identified ET-induced ETHYLENE RESPONSE FACTOR 15 (ERF15) and ERF16 as critical regulators of JA biosynthesis in tomato plants. Transcripts of ERF15 and ERF16 were markedly upregulated and peaked at 60 and 15 min, respectively, after simulated herbivore attack. While mutation in ERF16 resulted in the attenuated expression of JA biosynthetic genes and decreased JA accumulation 15 min after the simulated herbivory treatment, these changes were not observed in erf15 mutants until 60 min after treatment. Electrophoretic mobility shift assays and dual-luciferase assays demonstrated that both ERFs15 and 16 are transcriptional activators of LIPOXYGENASE D, ALLENE OXIDE CYCLASE, and 12-OXO-PHYTODIENOIC ACID REDUCTASE 3, key genes in JA biosynthesis. Furthermore, JA-activated MYC2 and ERF16 also function as the transcriptional activators of ERF16, contributing to dramatic increases in ERF16 expression. Taken together, our results demonstrated that ET signaling is involved in the rapid induction of the JA burst. ET-induced ERF15 and ERF16 function as powerful transcriptional activators that trigger the JA burst in response to herbivore attack.

ELONGATED HYPOCOTYL 5 mediates blue light-induced starch degradation in tomato.

Starch is the major storage carbohydrate in plants, and its metabolism in chloroplasts depends mainly on light. However, the mechanism through which photoreceptors regulate starch metabolism in chloroplasts is unclear. In this study, we found that the cryptochrome 1a (CRY1a)-mediated blue light signal is critical for regulating starch accumulation by inducing starch degradation through the transcription factor HY5 in chloroplasts in tomato. cry1a mutants and HY5-RNAi plants accumulated more starch and presented lower transcript levels of starch degradation-related genes in their leaves than wild-type plants. Blue light significantly induced the transcription of starch degradation-related genes in wild-type and CRY1a- or HY5-overexpressing plants but had little effect in cry1a and HY5-RNAi plants. Dual-luciferase assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation-qPCR revealed that HY5 could activate the starch degradation-related genes PWD, BAM1, BAM3, BAM8, MEX1, and DPE1 by directly binding to their promoters. Silencing of HY5 and these starch degradation-related genes in CRY1a-overexpressing plants led to increased accumulation of starch and decreased accumulation of soluble sugars. The findings presented here not only deepen our understanding of how light controls starch degradation and sugar accumulation but also allow us to explore potential targets for improving crop quality.

The MYC2-PUB22-JAZ4 module plays a crucial role in jasmonate signaling in tomato.

Jasmonates (JAs), a class of lipid-derived stress hormones, play a crucial role across an array of plant physiological processes and stress responses. Although JA signaling is thought to rely predominantly on the degradation of specific JAZ proteins by SCFCOI1, it remains unclear whether other pathways are involved in the regulation of JAZ protein stability. Here, we report that PUB22, a plant U-box type E3 ubiquitin ligase, plays a critical role in the regulation of plant resistance against Helicoverpa armigera and other JA responses in tomato. Whereas COI1 physically interacts with JAZ1/2/5/7, PUB22 physically interacts with JAZ1/3/4/6. PUB22 ubiquitinates JAZ4 to promote its degradation via the 26S proteasome pathway. Importantly, we observed that pub22 mutants showreduced resistance to H. armigera, whereas jaz4 single mutants and jaz1 jaz3 jaz4 jaz6 quadruple mutants have enhanced resistance. The hypersensitivity of pub22 mutants to herbivores could be partially rescued by JAZ4 mutation. Moreover, we found that expression of PUB22 can be transcriptionally activated by MYC2, thus forming a positive feedback circuit in JA signaling. We noticed that the PUB22-JAZ4 module also regulates other JA responses, including defense against B. cinerea, inhibition of root elongation, and anthocyanin accumulation. Taken together, these results indicate that PUB22 plays a crucial role in plant growth and defense responses, together with COI1-regulated JA signaling, by targeting specific JAZs.

A copper complex that combats triple negative breast cancer by restraining angiogenesis.

Angiogenesis and metastasis are major factors affecting the growth and invasion of triple negative breast cancer (TNBC). A phenanthroline copper(II) complex CPT8 modified with an alkyl chain-linked triphenylphosphonium group showed potent antiproliferative activity against a series of cancer cells including TNBC MDA-MB-231 cells. CPT8 induced mitophagy through activation of PINK1/Parkin and BNIP3 pathways in cancer cells due to damage to mitochondria. More importantly, CPT8 reduced the tube formation ability of human umbilical vein endothelial cells (HUVEC) through downregulation of nuclear factor erythroid 2-related factor 2 (Nrf2). The anti-angiogenic potential of CPT8 was confirmed by decreased vascular endothelial growth factor (VEGF) and CD34 expression in HUVEC. Moreover, CPT8 suppressed the expression of vascular endothelial cadherin and matrix metalloproteinases MMP2 and MMP9, leading to the inhibition of vasculogenic mimicry formation. CPT8 also weakened the metastatic potential of MDA-MB-231 cells. Downregulation of Ki67 and CD34 expression indicates that CPT8 suppressed tumor proliferation and vascularization in vivo, thus providing a unique metal drug candidate for the treatment of TNBC.

Molecular Characteristics and Prognostic Role of MFAP2 in Stomach Adenocarcinoma.

Molecular characteristics and prognostic role of MFAP2 were by no means stated. The MFAP2 expression and prognostic prices in this study, with Cox analysis, was employed to develop a predictive fee for MFAP2. To know about coexpression and practical networks associated with MFAP2, LinkedOmics and GEPIA2 have been used. MFAP2 expression has been increased and verified in many unbiased coalitions in TCGA-STAD tumor tissues. In addition, in each TCGA and various cohorts, increased MFAP2 was linked with lower survival. Evaluation by Cox revealed the unbiased danger to average survival, disease-specific survival, and progression-free survival of STAD used to be due to the elevated expression of MFAP2. Active community assessed the MFAP2, through which more than a few cancer-associated kinases and E2F household pathways are regulated, which shows that MFAP2 affects RNA transportation, oocyte meiosis, spliceosome, and ribosome biogenesis. MFAP2 can predict and is linked to the prediction of STAD independently. The closure of the MFAP2 link to the macrophage marker genes is, in particular, the achievable core of immune response.

The complete mitochondrial genome sequence of Scolopendra mutilans L. Koch, 1878 (Scolopendromorpha, Scolopendridae), with a comparative analysis of other centipede genomes.

Scolopendra mutilans L. Koch, 1878 is an important Chinese animal with thousands of years of medicinal history. However, the genomic information of this species is limited, which hinders its further application. Here, the complete mitochondrial genome (mitogenome) of S. mutilans was sequenced and assembled by next-generation sequencing. The genome is 15,011 bp in length, consisting of 13 protein-coding genes (PCGs), 14 tRNA genes, and two rRNA genes. Most PCGs start with the ATN initiation codon, and all PCGs have the conventional stop codons TAA and TAG. The S. mutilans mitogenome revealed nine simple sequence repeats (SSRs), and an obviously lower GC content compared with other seven centipede mitogenomes previously sequenced. After analysis of homologous regions between the eight centipede mitogenomes, the S. mutilans mitogenome further showed clear genomic rearrangements. The phylogenetic analysis of eight centipedes using 13 conserved PCG genes was finally performed. The phylogenetic reconstructions showed Scutigeromorpha as a separate group, and Scolopendromorpha in a sister-group relationship with Lithobiomorpha and Geophilomorpha. Collectively, the S. mutilans mitogenome provided new genomic resources, which will improve its medicinal research and applications in the future.

Design of Virtual Guiding Tasks With Haptic Feedback for Assessing the Wrist Motor Function of Patients With Upper Motor Neuron Lesions.

Impaired motor function is a common consequence of upper motor neuron lesions (UMNLs). Fine motor skills involved in small movements occurring in the fingers, hand, and wrist are usually regained by patient self-training at home. Most studies focus on the rehabilitation of the fingers but ignore the recovery of wrist motor function. In this paper, three virtual guiding tasks were designed to assess wrist motor functions, including the basic motor flexibility, motion stability, and a range of active motion. A haptic device was used to provide haptic feedback to users who performed virtual tasks in a virtual reality (VR) environment. In total, 46 healthy subjects and 10 UMNL patients were included to test the effectiveness of the designed tasks on improving wrist motor assessments. Quantitative performances, including the completion time, contact force, and motion trajectory, were automatically acquired during the tasks. Measurements for 95% of control subjects were used to establish normative references. Patient deficiencies in the wrist motor function were identified when their quantitative performances were outside the normative control ranges. The results suggest that the designed virtual tasks are sensitive for patients in the later period of rehabilitation, making the assessment suitable for using at home.

Systemic Root-Shoot Signaling Drives Jasmonate-Based Root Defense against Nematodes.

Shoot-root communication is crucial for plant adaptation to environmental changes. However, the extensive crosstalk between shoots and roots that controls the synthesis of jasmonates (JAs), in order to enhance defense responses against rhizosphere herbivores, remains poorly understood. Here, we report that the root-knot nematode (RKN) Meloidogyne incognita induces the systemic transmission of electrical and reactive oxygen species (ROS) signals from attacked tomato roots to the leaves, leading to an increased accumulation of JAs in the leaves. Grafting of 1.0-cm stem sections from mutants lacking GLUTAMATE RECEPTOR-LIKE 3.5 or the mutants deficient in RESPIRATORY BURST OXIDASE HOMOLOG 1 abolished the RKN-induced electrical signals and associated ROS and JA accumulation in the upper stems and leaves with attenuated resistance to RKN. Furthermore, the absence of systemic transmission of electrical and ROS signals compromised the activation of mitogen-activated protein kinases (MPKs) 1/2 in leaves. Silencing MPK1 or MPK2 abolished RKN-induced accumulation of JAs and associated resistance. These findings reveal a systemic signaling loop that integrates electrical, ROS, and JA signals to enhance the resistance in distal organs via root-shoot-root communication.

Polyaniline/Carbon Nanotubes Composite Modified Anode via Graft Polymerization and Self-Assembling for Microbial Fuel Cells.

Microbial fuel cells (MFCs) are promising devices for sustainable energy production, wastewater treatment and biosensors. Anode materials directly interact with electricigens and accept electrons between cells, playing an important role in determining the performance of MFCs. In this study, a novel carbon nanotubes (CNTs) and polyaniline (PANI) nanocomposite film modified Indium-tin oxide (ITO) anode was fabricated through graft polymerization of PANI after the modification of γ-aminopropyltriethoxysilane (APTES) on ITO substrate, which was followed by layer-by-layer (LBL) self-assembling of CNTs and PANI alternatively on its surface. (CNTs/PANI)n/APTES/ITO electrode with low charge transfer resistance showed better electrochemical behavior compared to the bare ITO electrode. Twelve layers of CNTs/PANI decorated ITO electrode with an optimal nanoporous network exhibited superior biocatalytic properties with a maximal current density of 6.98 µA/cm², which is 26-fold higher than that of conventional ITO electrode in Shewanella loihica PV-4 bioelectrochemical system. MFCs with (CNTs/PANI)12/APTES/ITO as the anode harvested a maximum output power density of 34.51 mW/m², which is 7.5-fold higher than that of the unmodified ITO electrode. These results demonstrate that (CNTs/PANI)12/APTES/ITO electrode has superior electrochemical and electrocatalytic properties compared to the bare ITO electrode, while the cellular toxicity of CNTs has an effect on the performance of MFC with (CNTs/PANI)n/APTES/ITO electrode.