Genome-Wide Identification, Phylogeny and Expression Analysis of Subtilisin (SBT) Gene Family under Wheat Biotic and Abiotic Stress.

The subtilisin-like protease (SBT) family is widely known for its role in stress resistance to a number of stressors in different plant species, but is rarely studied in wheat. Subtilisin-like serine proteases (SBTs) are serine proteolytic enzymes that hydrolyze proteins into small peptides, which bind to receptors as signal molecules or ligands and participate in signal transduction. In this study, we identified 255 putative SBT genes from the wheat reference genome and then divided these into seven clades. Subsequently, we performed syntenic relation analysis, exon-intron organization, motif composition, and cis-element analysis. Further, expression analysis based on RNA-seq and tissue-specific expression patterns revealed that TaSBT gene family expression has multiple intrinsic functions during various abiotic and biotic stresses. Analysis of RNA-seq expression assays and further validation through qRT PCR suggested that some of the TaSBT genes have significant changes in expression levels during Pst interaction. TaSBT7, TaSBT26, TaSBT102, and TaSBT193 genes showed increasing expression levels during compatible and non-compatible interactions, while the expression levels of TaSBT111 and TaSBT213 showed a decreasing trend, indicating that these members of the wheat SBT gene family may have a role in wheat's defense against pathogens. In conclusion, these results expand our understanding of the SBT gene family, and provide a valuable reference for future research on the stress resistance function and comprehensive data of wheat SBT members.

High-energy-density polymer dielectrics via compositional and structural tailoring for electrical energy storage.

Dielectric capacitors with higher working voltage and power density are favorable candidates for renewable energy systems and pulsed power applications. A polymer with high breakdown strength, low dielectric loss, great scalability, and reliability is a preferred dielectric material for dielectric capacitors. However, their low dielectric constant limits the polymer to achieve satisfying energy density. Therefore, great efforts have been made to get high-energy-density polymer dielectrics. By compositional and structural tailoring, the synergic integrations of the multiple components and optimized structural design effectively improved the energy storage properties. This review presents an overview of recent advancements in the field of high-energy-density polymer dielectrics via compositional and structural tailoring. The surface/interfacial engineering conducted on both microscale and macroscale for polymer dielectrics is the focus of this review. Challenges and the promising opportunities for the development of polymer dielectrics for capacitive energy storage applications are presented at the end of this review.

Genome-wide association study revealed TaHXK3-2A as a candidate gene controlling stomatal index in wheat seedlings.

Stomata are important channels for the control of gas exchange between plants and the atmosphere. To examine the genetic architecture of wheat stomatal index, we performed a genome-wide association study (GWAS) using a panel of 539 wheat accessions and 450 678 polymorphic single nucleotide polymorphisms (SNPs) that were detected using wheat-specific 660K SNP array. A total of 130 SNPs were detected to be significantly associated with stomatal index in both leaf surfaces of wheat seedlings. These significant SNPs were distributed across 16 chromosomes and involved 2625 candidate genes which participate in stress response, metabolism and cell/organ development. Subsequent bulk segregant analysis (BSA), combined with GWAS identified one major haplotype on chromosome 2A, that is responsible for stomatal index on the abaxial leaf surface. Candidate gene association analysis revealed that genetic variation in the promoter region of the hexokinase gene TaHXK3-2A was significantly associated with the stomatal index. Moreover, transgenic analysis confirmed that TaHXK3-2A overexpression in wheat decreased the size of leaf pavement cells but increased stomatal density through the glucose metabolic pathway, resulting in drought sensitivity among TaHXK3-2A transgenic lines due to an increased transpiration rate. Taken together, these results provide valuable insights into the genetic control of the stomatal index in wheat seedlings.

The LONELY GUY gene family: from mosses to wheat, the key to the formation of active cytokinins in plants.

LONELY GUY (LOG) was first identified in a screen of rice mutants with defects in meristem maintenance. In plants, LOG codes for cytokinin riboside 5'-monophosphate phosphoribohydrolase, which converts inactive cytokinin nucleotides directly to the active free bases. Many enzymes with the PGGxGTxxE motif have been misannotated as lysine decarboxylases; conversely not all enzymes containing this motif are cytokinin-specific LOGs. As LOG mutants clearly impact yield in rice, we investigated the LOG gene family in bread wheat. By interrogating the wheat (Triticum aestivum) genome database, we show that wheat has multiple LOGs. The close alignment of TaLOG1, TaLOG2 and TaLOG6 with the X-ray structures of two functional Arabidopsis thaliana LOGs allows us to infer that the wheat LOGs 1-11 are functional LOGs. Using RNA-seq data sets, we assessed TaLOG expression across 70 tissue types, their responses to various stressors, the pattern of cis-regulatory elements (CREs) and intron/exon patterns. TaLOG gene family members are expressed variously across tissue types. When the TaLOG CREs are compared with those of the cytokinin dehydrogenases (CKX) and glucosyltransferases (CGT), there is close alignment of CREs between TaLOGs and TaCKXs reflecting the key role of CKX in maintaining cytokinin homeostasis. However, we suggest that the main homeostatic mechanism controlling cytokinin levels in response to biotic and abiotic challenge resides in the CGTs, rather than LOG or CKX. However, LOG transgenics and identified mutants in rice variously impact yield, providing interesting avenues for investigation in wheat.

Genetically Dissecting the Novel Powdery Mildew Resistance Gene in Wheat Breeding Line PBDH1607.

Powdery mildew is one of the most destructive diseases in wheat production. Identifying novel resistance genes and deploying them in new cultivars is the most effective approach to minimize wheat losses caused by powdery mildew. In this study, wheat breeding line PBDH1607 showed high resistance to powdery mildew at both the seedling and adult plant stages. Genetic analysis of the seedling data demonstrated that the resistance was controlled by a single dominant gene, tentatively designated PmPBDH. The ΔSNP index based on bulked segregant RNA sequencing indicated that PmPBDH was associated with an interval of about 30.8 Mb (713.5 to 744.3 Mb) on chromosome arm 4AL. Using newly developed markers, we mapped PmPBDH to a 3.2-cM interval covering 7.1 Mb (719,055,516 to 726,215,121 bp). This interval differed from those of Pm61 (717,963,176 to 719,260,469 bp), MlIW30 (732,769,506 to 732,790,522 bp), and MlNSF10 (729,275,816 to 731,365,462 bp) reported on the same chromosome arm. PmPBDH also differed from Pm61, MlIW30, and MlNSF10 by its response spectrum, origin, or inheritance mode, suggesting that PmPBDH should be a new Pm gene. In the candidate interval, five genes were found to be associated with PmPBDH via time course gene expression analysis, and thus they are candidate genes of PmPBDH. Six closely linked markers, including two kompetitive allele-specific PCR markers, were confirmed to be applicable for tracking PmPBDH in marker-assisted breeding.

E-ffective: A Visual Analytic System for Exploring the Emotion and Effectiveness of Inspirational Speeches.

What makes speeches effective has long been a subject for debate, and until today there is broad controversy among public speaking experts about what factors make a speech effective as well as the roles of these factors in speeches. Moreover, there is a lack of quantitative analysis methods to help understand effective speaking strategies. In this paper, we propose E-ffective, a visual analytic system allowing speaking experts and novices to analyze both the role of speech factors and their contribution in effective speeches. From interviews with domain experts and investigating existing literature, we identified important factors to consider in inspirational speeches. We obtained the generated factors from multi-modal data that were then related to effectiveness data. Our system supports rapid understanding of critical factors in inspirational speeches, including the influence of emotions by means of novel visualization methods and interaction. Two novel visualizations include E-spiral (that shows the emotional shifts in speeches in a visually compact way) and E-script (that connects speech content with key speech delivery information). In our evaluation we studied the influence of our system on experts' domain knowledge about speech factors. We further studied the usability of the system by speaking novices and experts on assisting analysis of inspirational speech effectiveness.

Plant Growth Regulators INCYDE and TD-K Underperform in Cereal Field Trials.

Using plant growth regulators to alter cytokinin homeostasis with the aim of enhancing endogenous cytokinin levels has been proposed as a strategy to increase yields in wheat and barley. The plant growth regulators INCYDE and CPPU inhibit the cytokinin degrading enzyme cytokinin oxidase/dehydrogenase (CKX), while TD-K inhibits the process of senescence. We report that the application of these plant growth regulators in wheat and barley field trials failed to enhance yields, or change the components of yields. Analyses of the endogenous cytokinin content showed a high concentration of trans-zeatin (tZ) in both wheat and barley grains at four days after anthesis, and statistically significant, but probably biologically insignificant, increases in cisZ-O-glucoside, along with small decreases in cZ riboside (cZR), dihydro Z (DHZ), and DHZR and DHZOG cytokinins, following INCYDE application to barley at anthesis. We discuss possible reasons for the lack of efficacy of the three plant growth regulators under field conditions and comment on future approaches to manipulating yield in the light of the strong homeostatic mechanisms controlling endogenous cytokinin levels.

Concurrent overexpression of amino acid permease AAP1(3a) and SUT1 sucrose transporter in pea resulted in increased seed number and changed cytokinin and protein levels.

Using pea as our model crop, we sought to understand the regulatory control over the import of sugars and amino acids into the developing seeds and its importance for seed yield and quality. Transgenic peas simultaneously overexpressing a sucrose transporter and an amino acid transporter were developed. Pod walls, seed coats, and cotyledons were analysed separately, as well as leaves subtending developing pods. Sucrose, starch, protein, free amino acids, and endogenous cytokinins were measured during development. Temporal gene expression analyses (RT-qPCR) of amino acid (AAP), sucrose (SUT), and SWEET transporter family members, and those from cell wall invertase, cytokinin biosynthetic (IPT) and degradation (CKX) gene families indicated a strong effect of the transgenes on gene expression. In seed coats of the double transgenics, increased content and prolonged presence of cytokinin was particularly noticeable. The transgenes effectively promoted transition of young sink leaves into source leaves. We suggest the increased flux of sucrose and amino acids from source to sink, along with increased interaction between cytokinin and cell wall invertase in developing seed coats led to enhanced sink activity, resulting in higher cotyledon sucrose at process pea harvest, and increased seed number and protein content at maturity.

Characterization of the Powdery Mildew Resistance Gene in Wheat Breeding Line KN0816 and Its Evaluation in Marker-Assisted Selection.

Wheat powdery mildew, caused by Blumeria graminis (DC.) Speer f. sp. tritici is a destructive disease seriously threatening yield and quality of common wheat (Triticum aestivum L., 2n=6x=42, AABBDD). Characterization of resistance genes against powdery mildew is useful in parental selection and for developing disease-resistant cultivars. Chinese wheat breeding line KN0816 has superior agronomic performance and resistance to powdery mildew at all growth stages. Genetic analysis using populations of KN0816 crossed with different susceptible parents indicated that a single dominant gene, tentatively designated PmKN0816, conferred seedling resistance to different B. graminis f. sp. tritici isolates. Using a bulked segregant analysis, PmKN0816 was mapped to the Pm6 interval on chromosome arm 2BL using polymorphic markers linked to the cataloged genes Pm6, Pm52, and Pm64, and flanked by the markers CISSR02g-6 and CIT02g-2, both with genetic distances of 0.7 cM. Analysis of closely linked molecular markers indicated that the marker alleles of PmKN0816 differed from those of other powdery mildew resistance genes on 2BL, including Pm6, Pm33, Pm51, Pm64, and PmQ. Based on the genetic and physical locations and response pattern to different B. graminis f. sp. tritici isolates, PmKN0816 is most likely a new powdery mildew resistance gene and possesses effective resistance to all the 14 tested B. graminis f. sp. tritici isolates. In view of the elite agronomic performance of KN0816 combined with the resistance, PmKN0816 is expected to become a valuable resistance gene in wheat breeding. To transfer PmKN0816 to different genetic backgrounds using marker-assisted selection (MAS), closely linked markers of PmKN0816 were evaluated, and four of them (CIT02g-2, CISSR02g-6, CIT02g-10, and CIT02g-17) were confirmed to be applicable for MAS in different genetic backgrounds.

Diagnostic Kompetitive Allele-Specific PCR Markers of Wheat Broad-Spectrum Powdery Mildew Resistance Genes Pm21, PmV, and Pm12 Developed for High-Throughput Marker-Assisted Selection.

Wheat powdery mildew is a devastating disease that seriously threatens yield worldwide. Utilization of host resistance is considered an effective strategy to minimize powdery mildew damage. Pm21, PmV, and Pm12 confer broad-spectrum resistance to wheat powdery mildew in China, of which Pm21 and PmV are allelic genes derived from the 6VS chromosome of Dasypyrum villosum, and Pm12 is derived from the 6SS chromosome of Aegilops speltoides and most likely orthologous to the former two genes. To accurately and efficiently transfer and pyramid these genes using marker-assisted selection (MAS), distinctive single-nucleotide polymorphisms (SNPs) among the exon sequences of Pm21, PmV, and Pm12 and their homologous sequences in the common wheat genome were identified and then used for developing diagnostic Kompetitive Allele-Specific PCR (KASP) markers. The markers were validated in different genotypes including transgenic vectors, transgenic lines, translocation lines, resistance stocks with documented Pm genes, and in multiple susceptible cultivars without Pm genes. As a result, we initially developed a KASP marker that can simultaneously diagnose Pm21, Pm12, and PmV. Subsequently, we obtained a highly diagnostic KASP marker for each of the three genes that could distinguish among the three genes and also accurately distinguish them from other resistant stocks with documented Pm genes and from multiple susceptible genotypes. Compared with previously reported markers, the highly diagnostic KASP markers developed in this study have the advantages of low cost, easy assay, accuracy, and potentially high throughput for MAS.

Cytokinin glucosyl transferases, key regulators of cytokinin homeostasis, have potential value for wheat improvement.

The cytokinins, which are N6 -substituted adenine derivatives, control key aspects of crop productivity. Cytokinin levels are controlled via biosynthesis by isopentenyl transferase (IPT), destruction by cytokinin oxidase/dehydrogenase (CKX), and inactivation via glucosylation by cytokinin glucosyl transferases (CGTs). While both yield components and tolerance to drought and related abiotic stressors have been positively addressed via manipulation of IPT and/or CKX expression, much less attention has been paid to the CGTs. As naming of the CGTs has been unclear, we suggest COGT, CNGT, CONGT and CNOGT to describe the O-, N- and dual function CGTs. As specific CGT mutants of both rice and arabidopsis showed impacts on yield components, we interrogated the wheat genome database, IWGSC RefSeq v1.0 & v2.0, to investigate wheat CGTs. Besides providing unambiguous names for the 53 wheat CGTs, we show their expression patterns in 70 developmental tissues and their response characteristics to various stress conditions by reviewing more than 1000 RNA-seq data sets. These revealed various patterns of responses and showed expression generally being more limited in reproductive tissues than in vegetative tissues. Multiple cis-regulatory elements are present in the 3 kb upstream of the start codons of the 53 CGTs. Elements associated with abscisic acid, light and methyl jasmonate are particularly over-represented, indicative of the responsiveness of CGTs to the environment. These data sets indicate that CGTs have potential value for wheat improvement and that these could be targeted in TILLING or gene editing wheat breeding programmes.

Knocking down of Polo-like kinase 2 inhibits cell proliferation and induced cell apoptosis in human glioma cells.

AIMS: Polo-like kinase 2 (PLK2) belongs to a family of serine/threonine kinases, and it is involved in tumorigenesis. The present study aimed to explore the potential clinical significance of PLK2 in the development of gliomas. MAIN METHODS: Immunohistochemistry (IHC) was performed to detect the expression of PLK2 in glioma tissues. Cell proliferation and apoptosis were determined by Cell Counting Kit 8 (CCK8) and flow cytometry analysis, respectively. KEY FINDINGS: PLK2 expression gradually increased with the degree of glioma malignancy. High PLK2 expression was associated with a poor prognosis in glioma. Short hairpin RNAs targeting PLK2 (shPLK2) inhibited the viability and induced apoptosis of glioma cells, both in vitro and in vivo. Ring finger protein 180 (RNF180), an E3 ubiquitin ligase, interacted with PLK2 and induced the ubiquitination of PLK2. Overexpression of PLK2 in glioma cells significantly inhibited RNF180 upregulation-induced cell apoptosis. The expression level of RNF180 gradually decreased with the degree of glioma malignancy. SIGNIFICANCE: Knocking down of PLK2 may suppress the glioma development through cancer cell proliferation inhibition and cell apoptosis promotion. Furthermore, RNF180 may mediate the ubiquitination of PLK2. The present findings may help improve the clinical management of glioma in the future.

Targeting Cytokinin Homeostasis in Rapid Cycling Brassica rapa with Plant Growth Regulators INCYDE and TD-K.

Modifying the cytokinin content in plants is a means of improving plant productivity. Here, we report the development and biological activity of compound TD-K (1-(furan-2-ylmethyl)-3-(1,2,3-thiadiazol-5-yl)urea)which is related to thidiazuron. TD-K-which exhibited extremely high antisenescence activity in the wheat leaf bioassay-and INCYDE (2-chloro-6-(3-methoxyphenyl)aminopurine)-a plant growth regulator reported to inhibit cytokinin oxidase/dehydrogenase (CKX), an enzyme involved in the degradation of the plant hormone cytokinin-were selected for investigation of their effects on the model plant Rapid Cycling Brassica rapa (RCBr). We monitored the expression of BrCKX and isopentenyl transferase (BrIPT), which codes for the key cytokinin biosynthesis enzyme, in developing leaves following INCYDE and TD-K application. Growth room experiments revealed that INCYDE increased RCBr seed yield per plant, but only when applied multiple times and when grown in 5 mM KNO3. Expression in control leaves showed transient, high levels of expression of BrCKX and BrIPT at true leaf appearance. Following INCYDE application, there was a rapid and strong upregulation of BrCKX3, and a transient downregulation of BrIPT1 and BrIPT3. Interestingly, the upregulation of BrCKX3 persisted in a milder form throughout the course of the experiment (16 days). TD-K also upregulated BrCKX3. However, in contrast to INCYDE, this effect disappeared after two days. These results suggest that both compounds (CKX inhibitor and cytokinin TD-K) influenced cytokinin homeostasis in RCBr leaves, but with different mechanisms.

Corrigendum: Immune and Clinical Features of CD96 Expression in Glioma by in silico Analysis.

[This corrects the article DOI: 10.3389/fbioe.2020.00592.].

Immune and Clinical Features of CD96 Expression in Glioma by in silico Analysis.

Background: Immune checkpoints target regulatory pathways in T cells that enhance antitumor immune responses and elicit durable clinical responses. As a novel immune checkpoint, CD96 is an attractive key target for cancer immunotherapy. However, there has been no integrative investigation of CD96 in glioma. Our study explored the relationship between CD96 expression and clinical prognosis in glioma. Methods: RNA and clinical data for a total of 1,001 samples were included in this study, including 325 samples from the Chinese Glioma Genome Atlas (CGGA) database and 676 samples from The Cancer Genome Atlas (TCGA) dataset. The R programming language was employed to perform statistical analysis and draw figures. Results: CD96 had a consistently positive relationship with glioblastoma and was highly enriched in IDH-wildtype and mesenchymal subtype glioma. Gene ontology enrichment and gene set variation analysis analyses suggested that CD96 was mostly involved in immune functions and was especially related to T cell-mediated immune response in glioma. Subsequent immune infiltration analysis showed that CD96 was positively correlated with infiltrating levels of CD4 + T and CD8 + T cells, macrophages, neutrophils, and DCs in glioblastoma multiforme and low-grade glioma. Additionally, CD96 was tightly associated with other immune checkpoints, including PD-1, CTLA-4, TIGIT, and TIM-3. Univariate and multivariate Cox analysis demonstrated that CD96 acts as an independent indicator of poor prognosis in glioma. Conclusion: CD96 expression was increased in malignant phenotype and negatively associated with overall survival in glioma. CD96 also showed a positive correlation with other immune checkpoints, immune response, and inflammatory activity. Our findings indicate that CD96 is a promising clinical target for further immunotherapeutic use in glioma patients.

Bulked Segregant RNA-Seq Reveals Distinct Expression Profiling in Chinese Wheat Cultivar Jimai 23 Responding to Powdery Mildew.

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most destructive fungal diseases threatening global wheat production. Host resistance is well known to be the most efficient method to control this disease. However, the molecular mechanism of wheat powdery mildew resistance (Pm) is still unclear. To analyze the molecular mechanism of Pm, we used the resistant wheat cultivar Jimai 23 to investigate its potential resistance components and profiled its expression in response to powdery mildew infection using bulked segregant RNA-Seq (BSR-Seq). We showed that the Pm of Jimai 23 was provided by a single dominant gene, tentatively designated PmJM23, and assigned it to the documented Pm2 region of chromosome 5DS. 3,816 consistently different SNPs were called between resistant and susceptible parents and the bulked pools derived from the combinations between the resistant parent Jimai23 and the susceptible parent Tainong18. 58 of the SNPs were assigned to the candidate region of PmJM23. Subsequently, 3,803 differentially expressed genes (DEGs) between parents and bulks were analyzed by GO, COG and KEGG pathway enrichment. The temporal expression patterns of associated genes following Bgt inoculation were further determined by RT-qPCR. Expression of six disease-related genes was investigated during Bgt infection and might serve as valuable genetic resources for the improvement of durable resistance to Bgt.

Characterization of the Powdery Mildew Resistance Gene in the Elite Wheat Cultivar Jimai 23 and Its Application in Marker-Assisted Selection.

Powdery mildew infection of wheat (Triticum aestivum L.), caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive disease that threatens yield and quality worldwide. The most effective and preferred means for the control of the disease is to identify broad-spectrum resistance genes for breeding, especially the genes derived from elite cultivars that exhibit desirable agronomic traits. Jimai 23 is a Chinese wheat cultivar with superior agronomic performance, high-quality characteristics, and effective resistance to powdery mildew at all growth stages. Genetic analysis indicated that powdery mildew resistance in Jimai 23 was mediated by a single dominant gene, tentatively designated PmJM23. Using bulked segregant RNA-Seq (BSR-Seq), a series of markers was developed and used to map PmJM23. PmJM23 was then located at the Pm2 locus on the short arm of chromosome 5D (5DS). Resistance spectrum analysis demonstrated that PmJM23 provided a broad resistance spectrum different from that of the documented Pm2 alleles, indicating that PmJM23 is most likely a new allele of Pm2. In view of these combined agronomic, quality, and resistance findings, PmJM23 is expected to be a valuable resistance gene in wheat breeding. To efficiently use PmJM23 in breeding, the closely linked markers of PmJM23 were evaluated and confirmed to be applicable for marker-assisted selection (MAS). Using these markers, a series of resistant breeding lines with high resistance and desirable agronomic performance was selected from the crosses involving PmJM23, resulting in improved powdery mildew resistance of these lines.

Integration of Transcriptome Resequencing and Quantitative Proteomics Analyses of Collagenase VII-Induced Intracerebral Hemorrhage in Mice.

OBJECTIVE: Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and morbidity rates. Our aim was to comprehensively analyze transcriptome and proteome in an experimental ICH model. METHODS: All mice were divided into ICH model (n = 3) and sham groups (n = 3). ICH was induced by collagenase VII. The ipsilateral hemisphere was used for whole transcriptome and proteomics resequencing. After preprocessing, differentially expressed lncRNAs (DElncRNAs), mRNAs (DEmRNAs), miRNAs (DEmiRNAs), and DEproteins between ICH and sham groups were identified. Functional enrichment analysis was performed using the clusterProfiler package, followed by protein-protein interaction (PPI) analysis. After that, the Pearson correlation coefficient between DEmRNAs and DElncRNAs or between DEmRNAs and DEproteins was calculated. DElncRNAs with similar functions were analyzed by the GOSemSim package. After prediction of DEmiRNA-DEmRNA and DElncRNA-DEmiRNA relationships, a competing endogenous RNA (ceRNA) network was constructed. Several DEmRNAs and DElncRNAs were validated in ipsilateral hemisphere tissues of the ICH model and control groups using RT-qPCR and western blot. RESULTS: Between the ICH and sham groups, 31 DElncRNAs, 367 DEmRNAs, 35 DEmiRNAs, and 96 DEproteins were identified. DEmRNAs were mainly enriched in inflammation, such as cytokine-cytokine receptor interaction, IL-17, and TNF signaling pathways. A PPI network of DEmRNAs was constructed and hub genes were identified, such as IL6 (degree = 59), TNF (degree = 44), and CXCR2 (degree = 39). 24 DElncRNAs with similar functions were identified, including 15 up- and 9 down-regulated lncRNAs. After integration of DEmiRNA-DEmRNA and DElncRNA-DEmiRNA relationships, we constructed a ceRNA network, composed of 71 DEmRNAs, 17 DEmiRNAs, and 12 DElncRNAs. RT-qPCR and western blot results confirmed that C3, Fga, and Slc4a1 proteins were more lowly expressed and Penk was more highly expressed in ICH than control groups, which could become potential markers for ICH. CONCLUSION: Our findings identified ICH-related DE-RNAs and proteins and potential molecular mechanisms of ICH by transcriptome resequencing and quantitative proteomic analyses.

Cytokinin dehydrogenase: a genetic target for yield improvement in wheat.

The plant hormone group, the cytokinins, is implicated in both qualitative and quantitative components of yield. Cytokinins have opposing actions in shoot and root growth-actions shown to involve cytokinin dehydrogenase (CKX), the enzyme that inactivates cytokinin. We revise and provide unambiguous names for the CKX gene family members in wheat, based on the most recently released wheat genome database, IWGSC RefSeq v1.0 & v2.0. We review expression data of CKX gene family members in wheat, revealing tissue-specific gene family member expression as well as sub-genome-specific expression. Manipulation of CKX in cereals shows clear impacts on yield, root growth and orientation, and Zn nutrition, but this also emphasizes the necessity to unlink promotive effects on grain yield from negative effects of cytokinin on root growth and uptake of mineral nutrients, particularly Zn and Fe. Wheat is the most widely grown cereal crop globally, yet is under-research compared with rice and maize. We highlight gaps in our knowledge of the involvement of CKX for wheat. We also highlight the necessity for accurate analysis of endogenous cytokinins, acknowledging why this is challenging, and provide examples where inadequate analyses of endogenous cytokinins have led to unjustified conclusions. We acknowledge that the allohexaploid nature of bread wheat poses challenges in terms of uncovering useful mutations. However, we predict TILLING followed by whole-exome sequencing will uncover informative mutations and we indicate the potential for stacking mutations within the three genomes to modify yield components. We model a wheat ideotype based on CKX manipulation.