Identification and Functional Analysis of CAP Genes from the Wheat Stripe Rust Fungus Puccinia striiformis f. sp. tritici.

Cysteine-rich secretory proteins (C), antigen 5 (A), and pathogenesis-related 1 proteins (P) comprise widespread CAP superfamily proteins, which have been proven to be novel virulence factors of mammalian pathogenic fungi and some plant pathogens. Despite this, the identification and function of CAP proteins in more species of plant pathogens still need to be studied. This work presents the identification and functional analysis of CAP superfamily proteins from Puccinia striiformis f. sp. tritici (Pst), an important fungal pathogen that causes wheat stripe rust on wheat worldwide. A total of six CAP genes were identified in the Pst genome, designated as PsCAP1-PsCAP6. Five PsCAP proteins, including PsCAP1, PsCAP2, PsCAP3, PsCAP4, and PsCAP5, have N-terminal signal peptides secreted with the yeast signal sequence trap assay. Single-nucleotide polymorphism (SNP) analysis indicated that they showed a low level of intraspecies polymorphism. The expression abundance of PsCAP genes at different Pst infection stages was detected by RT-qPCR, and most of them were highly expressed during Pst infection on wheat and also Pst sexual reproduction on barberry (Berberis shensiana). Noticeably, the silencing of these six PsCAP genes by BSMV-mediated HIGS indicated that PsCAP1, PsCAP4, and PsCAP5 contribute significantly to Pst infection in wheat. These results indicate that PsCAP proteins may act as virulence factors during Pst infection, which also provides insights into Pst pathogenicity.

Research on Electric Field-Induced Catalysis Using Single-Molecule Electrical Measurement.

The role of catalysis in controlling chemical reactions is crucial. As an important external stimulus regulatory tool, electric field (EF) catalysis enables further possibilities for chemical reaction regulation. To date, the regulation mechanism of electric fields and electrons on chemical reactions has been modeled. The electric field at the single-molecule electronic scale provides a powerful theoretical weapon to explore the dynamics of individual chemical reactions. The combination of electric fields and single-molecule electronic techniques not only uncovers new principles but also results in the regulation of chemical reactions at the single-molecule scale. This perspective focuses on the recent electric field-catalyzed, single-molecule chemical reactions and assembly, and highlights promising outlooks for future work in single-molecule catalysis.

The overlooked role of UV185 induced high-energy excited states in the dephosphorization of organophosphorus pesticide by VUV/persulfate.

Vacuum ultraviolet (VUV) based advanced oxidation processes (AOPs) recently attracted widespread interests. However, the role of UV185 in VUV is only considered to be generating a series of active species, while the effect of photoexcitation has long been overlooked. In this work, the role of UV185 induced high-energy excited state for the dephosphorization of organophosphorus pesticides was studied using malathion as a model. Results showed malathion degradation was highly related to radical yield, while its dephosphorization was not. It was UV185 rather than UV254 or radical yield that was responsible for malathion dephosphorization by VUV/persulfate. DFT calculation results demonstrated that the polarity of P-S bond was further increased during UV185 excitation, favoring dephosphorization while UV254 did not. The conclusion was further supported by degradation path identification. Moreover, despite the fact that anions (Cl-, SO42- and NO3-) considerably affected radical yield, only Cl- and NO3- with high molar extinction coefficient at 185 nm significantly affected dephosphorization. This study shed light on the crucial role of excited states in VUV based AOPs and provided a new idea for the development of mineralization technology of organophosphorus pesticides.

CaREM1.4 interacts with CaRIN4 to regulate Ralstonia solanacearum tolerance by triggering cell death in pepper.

Remorins, plant-specific proteins, have a significant role in conferring on plants the ability to adapt to adverse environments. However, the precise function of remorins in resistance to biological stress remains largely unknown. Eighteen CaREM genes were identified in pepper genome sequences based on the C-terminal conserved domain that is specific to remorin proteins in this research. Phylogenetic relations, chromosomal localization, motif, gene structures, and promoter regions of these remorins were analyzed and a remorin gene, CaREM1.4, was cloned for further study. The transcription of CaREM1.4 in pepper was induced by infection with Ralstonia solanacearum. Knocking down CaREM1.4 in pepper using virus-induced gene silencing (VIGS) technologies reduced the resistance of pepper plants to R. solanacearum and downregulated the expression of immunity-associated genes. Conversely, transient overexpression of CaREM1.4 in pepper and Nicotiana benthamiana plants triggered hypersensitive response-mediated cell death and upregulated expression of defense-related genes. In addition, CaRIN4-12, which interacted with CaREM1.4 at the plasma membrane and cell nucleus, was knocked down with VIGS, decreasing the susceptibility of Capsicum annuum to R. solanacearum. Furthermore, CaREM1.4 reduced ROS production by interacting with CaRIN4-12 upon co-injection in pepper. Taken together, our findings suggest that CaREM1.4 may function as a positive regulator of the hypersensitive response, and it interacts with CaRIN4-12, which negatively regulates plant immune responses of pepper to R. solanacearum. Our study provides new evidence for comprehending the molecular regulatory network of plant cell death.

Non-radical degradation of organic pharmaceuticals by g-C3N4 under visible light irradiation: The overlooked role of excitonic energy transfer.

In this work, an excitonic energy transfer (EET) based non-radical mechanism was proposed for the degradation of organic pharmaceuticals by graphitic carbon nitride (g-C3N4) under visible light irradiation. Using diclofenac (DCF) as a model molecule, the competition between single electron transfer (SET) and EET was studied through modulating the exciton binding energy of g-C3N4. The different mechanisms of SET and EET for DCF degradation were predicted by DFT calculation, and further confirmed by their different degradation pathways. When EET played an important role, the rationality of some very popular radical scavengers, such as p-BQ, TEMPOL and furfuryl alcohol must be reconsidered. In addition, humic acid (HA) had a distinct effect on EET and SET. Specifically, HA enhanced the EET process through photosensitization, but suppressed SET through radical quenching effect. The effect of HA on DCF degradation depended on the contribution ratio of SET and ET.

The degradation of municipal solid waste incineration leachate by UV/persulfate and UV/H2O2 processes: The different selectivity of SO4•- and •OH.

In this work, the different selectivity of SO4•- and •OH towards municipal solid waste incineration leachates (MSWILs) was studied by a comparative study of UV/persulfate (PS) and UV/H2O2. Results showed SO4•- preferentially mineralized carbon atoms of higher average oxidation state, while •OH showed a two-stage mechanism of partial oxidation and mineralization successively. Electron spin resonance (ESR) analysis showed SO4•- had superior selectivity towards MSWILs than •OH, and Fe(II) would significantly affect the selectivity via forming Fe-MSWILs complex. As the consequence, Fe(II) showed slightly negative effect on UV/PS, but greatly enhanced the performance of UV/H2O2/Fe(II). High concentration of Cl- affected the degradation of non-fluorescent substances by UV/PS, while SO42- and NO3- showed no effect. In contrast, anions showed no effect on UV/H2O2. In addition, •OH preferentially attacked large molecules, but SO4•- showed no selectivity. This study further revealed the selectivity of SO4•- and •OH in the treatment of hypersaline wastewater, and provided theoretical support for the development of targeted technology.

Comparative evaluation of MSW incineration leachate treatment by heterogeneous catalytic O3 and UV/O3: The unexpected contribution of high salinity and overlooked role of excited state.

The efficiency and mechanism of heterogeneous catalytic O3 and UV/O3 for municipal solid waste (MSW) incineration leachate advanced treatment was systematically compared. Prior to comparison, catalyst used in heterogenous catalytic O3 and operation parameters for each technology were optimized. The COD removal of CuO@Al2O3/O3 under its optimal parameters was 57.2%, which failed to meet the standard (≥75%). In contrast, the COD removal by UV/O3 could be 82.3%. The superior efficiency of UV/O3 over CuO@Al2O3/O3 could be summarized into three aspects: (I) Cu bounded ·OH (≡Cu-O·) preferentially attacked hydrophilic groups, while free hydroxyl radical (·OH) was non-selective, thus UV/O3 exhibited a unique three-stage mechanism; (II) The oxidation potential of ≡Cu-O· was higher than that of ·OH, therefore was more vulnerable to the negative effect of radical self-quenching; (III) The existence of UV-induced excited states made organics in UV/O3 more active than in CuO@Al2O3/O3 system, thus high concentration of anions enhanced COD removal in UV/O3 but affected that in CuO@Al2O3/O3. The study further revealed the characteristics of heterogeneous catalytic O3 and UV/O3, and UV induced excited state should be considered in UV-based advanced oxidation processes (AOPs).

Inactivation of a wheat protein kinase gene confers broad-spectrum resistance to rust fungi.

Wheat crops are frequently devastated by pandemic stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). Here, we identify and characterize a wheat receptor-like cytoplasmic kinase gene, TaPsIPK1, that confers susceptibility to this pathogen. PsSpg1, a secreted fungal effector vital for Pst virulence, can bind TaPsIPK1, enhance its kinase activity, and promote its nuclear localization, where it phosphorylates the transcription factor TaCBF1d for gene regulation. The phosphorylation of TaCBF1d switches its transcriptional activity on the downstream genes. CRISPR-Cas9 inactivation of TaPsIPK1 in wheat confers broad-spectrum resistance against Pst without impacting important agronomic traits in two years of field tests. The disruption of TaPsIPK1 leads to immune priming without constitutive activation of defense responses. Taken together, TaPsIPK1 is a susceptibility gene known to be targeted by rust effectors, and it has great potential for developing durable resistance against rust by genetic modifications.

Transcriptional repression of TaNOX10 by TaWRKY19 compromises ROS generation and enhances wheat susceptibility to stripe rust.

Reactive oxygen species (ROS) are vital for plant immunity and regulation of their production is crucial for plant health. While the mechanisms that elicit ROS production have been relatively well studied, those that repress ROS generation are less well understood. Here, via screening Brachypodium distachyon RNA interference mutants, we identified BdWRKY19 as a negative regulator of ROS generation whose knockdown confers elevated resistance to the rust fungus Puccinia brachypodii. The three wheat paralogous genes TaWRKY19 are induced during infection by virulent P. striiformis f. sp. tritici (Pst) and have partially redundant roles in resistance. The stable overexpression of TaWRKY19 in wheat increased susceptibility to an avirulent Pst race, while mutations in all three TaWRKY19 copies conferred strong resistance to Pst by enhancing host plant ROS accumulation. We show that TaWRKY19 is a transcriptional repressor that binds to a W-box element in the promoter of TaNOX10, which encodes an NADPH oxidase and is required for ROS generation and host resistance to Pst. Collectively, our findings reveal that TaWRKY19 compromises wheat resistance to the fungal pathogen and suggest TaWRKY19 as a potential target to improve wheat resistance to the commercially important wheat stripe rust fungus.

Insight into the role of Fe in the synergetic effect of persulfate/sulfite and Fe2O3@g-C3N4 for carbamazepine degradation.

In this work, the role of Fe in the synergetic effect of persulfate/sulfite and Fe2O3@g-C3N4 (FCN) for carbamazepine (CBZ) degradation was studied. Unexpectedly, Fe2O3 in FCN plays very different roles for sulfite [S(IV)] and persulfate (PS) activation. Specifically, since photo-generated holes (h+) can transform S(IV) into SO4-, and photo-generated electrons (e-) can accelerate Fe(III) reduction which promotes transition metal based S(IV) activation, a synergetic effect of photocatalysis and Fe is observed in FCN/S(IV)/vis system. In contrast, in FCN/PS/vis system, both Fe(III)/Fe(II) cycle and PS activation compete for e-. Since PS is a stronger electron acceptor, Fe(III) reduction by e- is limited. Therefore, the contribution of Fe2O3 in FCN/S(IV)/vis system is 3 times higher than that in FCN/PS/vis system. Initial pH affects CBZ removal by changing surface charge of catalysts and oxidants species, while the effect varies for different catalysts and oxidants. This study provides new insight into the synergetic effect of photocatalysis and transition metal for SO4- generation, which contributes to catalyst design for environmental application.

Insight into the synergetic effect of photocatalysis and transition metal on sulfite activation: Different mechanisms for carbamazepine and diclofenac degradation.

Sulfite [S(IV)] is a promising alternative for sulfate radical-based advanced oxidation processes (SR-AOPs). Transition metal and photocatalysis are generally considered to have a synergetic effect for S(IV) activation. However, the study shows that the synergetic effect is target specific. Herein, an ultra-small Fe2O3 clusters deposited graphitic carbon nitride is synthesized and used for S(IV) activation. For carbamazepine (CBZ) degradation, photogenerated holes can transform S(IV) into sulfate radical and photogenerated electrons can accelerate Fe(II)/Fe(III) cycle, which account for the synergetic effect. In contrast, the degradation of diclofenac (DCF) depends on the excitation of DCF rather than photocatalyst. Instead of radical precursor, S(IV) acts as the electron transfer bridge between excited DCF and photocatalyst. Thus, the deposition of Fe2O3 negatively affects DCF degradation. Density Functional Theory calculation shows that the first excited state rather than the ground state of diclofenac is more suitable for reactive site prediction, which confirms the photosensitization-like degradation mechanism. Moreover, the effects of pH and coexisted anions varies for CBZ and DCF. The study shed light on the synergetic effect of transition metal and photocatalysis for S(IV) activation, and also open an avenue for the study of target specific mechanisms for AOPs.

Comparative in Silico Analysis of Ferric Reduction Oxidase (FRO) Genes Expression Patterns in Response to Abiotic Stresses, Metal and Hormone Applications.

The ferric reduction oxidase (FRO) gene family is involved in various biological processes widely found in plants and may play an essential role in metal homeostasis, tolerance and intricate signaling networks in response to a number of abiotic stresses. Our study describes the identification, characterization and evolutionary relationships of FRO genes families. Here, total 50 FRO genes in Plantae and 15 'FRO like' genes in non-Plantae were retrieved from 16 different species. The entire FRO genes have been divided into seven clades according to close similarity in biological and functional behavior. Three conserved domains were common in FRO genes while in two FROs sub genome have an extra NADPH-Ox domain, separating the function of plant FROs. OsFRO1 and OsFRO7 genes were expressed constitutively in rice plant. Real-time RT-PCR analysis demonstrated that the expression of OsFRO1 was high in flag leaf, and OsFRO7 gene expression was maximum in leaf blade and flag leaf. Both genes showed vigorous expressions level in response to different abiotic and hormones treatments. Moreover, the expression of both genes was also substantial under heavy metal stresses. OsFRO1 gene expression was triggered following 6 h under Zn, Pb, Co and Ni treatments, whereas OsFRO7 gene expression under Fe, Pb and Ni after 12 h, Zn and Cr after 6 h, and Mn and Co after 3 h treatments. These findings suggest the possible involvement of both the genes under abiotic and metal stress and the regulation of phytohormones. Therefore, our current work may provide the foundation for further functional characterization of rice FRO genes family.

CLD1/SRL1 modulates leaf rolling by affecting cell wall formation, epidermis integrity and water homeostasis in rice.

Leaf rolling is considered as one of the most important agronomic traits in rice breeding. It has been previously reported that SEMI-ROLLED LEAF 1 (SRL1) modulates leaf rolling by regulating the formation of bulliform cells in rice (Oryza sativa); however, the regulatory mechanism underlying SRL1 has yet to be further elucidated. Here, we report the functional characterization of a novel leaf-rolling mutant, curled leaf and dwarf 1 (cld1), with multiple morphological defects. Map-based cloning revealed that CLD1 is allelic with SRL1, and loses function in cld1 through DNA methylation. CLD1/SRL1 encodes a glycophosphatidylinositol (GPI)-anchored membrane protein that modulates leaf rolling and other aspects of rice growth and development. The cld1 mutant exhibits significant decreases in cellulose and lignin contents in secondary cell walls of leaves, indicating that the loss of function of CLD1/SRL1 affects cell wall formation. Furthermore, the loss of CLD1/SRL1 function leads to defective leaf epidermis such as bulliform-like epidermal cells. The defects in leaf epidermis decrease the water-retaining capacity and lead to water deficits in cld1 leaves, which contribute to the main cause of leaf rolling. As a result of the more rapid water loss and lower water content in leaves, cld1 exhibits reduced drought tolerance. Accordingly, the loss of CLD1/SRL1 function causes abnormal expression of genes and proteins associated with cell wall formation, cuticle development and water stress. Taken together, these findings suggest that the functional roles of CLD1/SRL1 in leaf-rolling regulation are closely related to the maintenance of cell wall formation, epidermal integrity and water homeostasis.

REL2, A Gene Encoding An Unknown Function Protein which Contains DUF630 and DUF632 Domains Controls Leaf Rolling in Rice.

BACKGROUND: Rice leaves are important energy source for the whole plant. An optimal structure will be beneficial for rice leaves to capture light energy and exchange gas, thus increasing the yield of rice. Moderate leaf rolling and relatively erect plant architecture may contribute to high yield of rice, but the relevant molecular mechanism remains unclear. RESULTS: In this study, we identified and characterized a rolling and erect leaf mutant in rice and named it as rel2. Histological analysis showed that the rel2 mutant has increased number of bulliform cells and reduced size of middle bulliform cells. We firstly mapped REL2 to a 35-kb physical region of chromosome 10 by map-based cloning strategy. Further analysis revealed that REL2 encodes a protein containing DUF630 and DUF632 domains. In rel2 mutant, the mutation of two nucleotide substitutions in DUF630 domain led to the loss-of-function of REL2 locus and the function of REL2 could be confirmed by complementary expression of REL2 in rel2 mutant. Further studies showed that REL2 protein is mainly distributed along the plasma membrane of cells and the REL2 gene is relatively higher expressed in younger leaves of rice. The results from quantitative RT-PCR analysis indicated that REL2 functioning in the leaf shape formation might have functional linkage with many genes associated with the bulliform cells development, auxin synthesis and transport, etc. CONCLUSIONS: REL2 is the DUF domains contained protein which involves in the control of leaf rolling in rice. It is the plasma membrane localization and its functions in the control of leaf morphology might involve in multiple biological processes such as bulliform cell development and auxin synthesis and transport.