Genome-Wide Identification and Expression Analysis of SnRK2 Gene Family in Dormant Vegetative Buds of Liriodendron chinense in Response to Abscisic Acid, Chilling, and Photoperiod.

Protein kinases play an essential role in plants' responses to environmental stress signals. SnRK2 (sucrose non-fermenting 1-related protein kinase 2) is a plant-specific protein kinase that plays a crucial role in abscisic acid and abiotic stress responses in some model plant species. In apple, corn, rice, pepper, grapevine, Arabidopsis thaliana, potato, and tomato, a genome-wide study of the SnRK2 protein family was performed earlier. The genome-wide comprehensive investigation was first revealed to categorize the SnRK2 genes in the Liriodendron chinense (L. chinense). The five SnRK2 genes found in the L. chinense genome were highlighted in this study. The structural gene variants, 3D structure, chromosomal distributions, motif analysis, phylogeny, subcellular localization, cis-regulatory elements, expression profiles in dormant buds, and photoperiod and chilling responses were all investigated in this research. The five SnRK2 genes from L. chinense were grouped into groups (I-IV) based on phylogeny analysis, with three being closely related to other species. Five hormones-, six stress-, two growths and biological process-, and two metabolic-related responsive elements were discovered by studying the cis-elements in the promoters. According to the expression analyses, all five genes were up- and down-regulated in response to abscisic acid (ABA), photoperiod, chilling, and chilling, as well as photoperiod treatments. Our findings gave insight into the SnRK2 family genes in L. chinense and opened up new study options.

N-Methyltransferase CaASHH3 Acts as a Positive Regulator of Immunity against Bacterial Pathogens in Pepper.

Proteins with conserved SET domain play a critical role in plant immunity. However, the means of organization and functions of these proteins are unclear, particularly in non-model plants such as pepper (Capsicum annum L.). Herein, we functionally characterized CaASHH3, a member of class II (the ASH1 homologs H3K36) proteins in pepper immunity against Ralstonia solanacearum and Pseudomonas syringae pv tomato DC3000 (Pst DC3000). The CaASHH3 was localized in the nucleus, and its transcript levels were significantly enhanced by R. solanacearum inoculation (RSI) and exogenous application of salicylic acid (SA), methyl jasmonate (MeJA), ethephon (ETH), and abscisic acid (ABA). Knockdown of CaASHH3 by virus-induced gene silencing (VIGS) compromised peppers' resistance to RSI. Furthermore, silencing of CaASHH3 impaired hypersensitive-response (HR)-like cell death response due to RSI and downregulated defense-associated marker genes, including CaPR1, CaNPR1, and CaABR1. The CaASHH3 protein was revealed to affect the promoters of CaNPR1, CaPR1, and CaHSP24. Transiently over-expression of CaASHH3 in pepper leaves elicited HR-like cell death and upregulated immunity-related marker genes. To further study the role of CaASHH3 in plant defense in vivo, CaASHH3 transgenic plants were generated in Arabidopsis. Overexpression of CaASHH3 in transgenic Arabidopsis thaliana enhanced innate immunity against Pst DC3000. Furthermore, CaASHH3 over-expressing transgenic A. thaliana plants exhibited upregulated transcriptional levels of immunity-associated marker genes, such as AtNPR1, AtPR1, and AtPR2. These results collectively confirm the role of CaASHH3 as a positive regulator of plant cell death and pepper immunity against bacterial pathogens, which is regulated by signaling synergistically mediated by SA, JA, ET, and ABA.

Genome Engineering Technology for Durable Disease Resistance: Recent Progress and Future Outlooks for Sustainable Agriculture.

Crop production worldwide is under pressure from multiple factors, including reductions in available arable land and sources of water, along with the emergence of new pathogens and development of resistance in pre-existing pathogens. In addition, the ever-growing world population has increased the demand for food, which is predicted to increase by more than 100% by 2050. To meet these needs, different techniques have been deployed to produce new cultivars with novel heritable mutations. Although traditional breeding continues to play a vital role in crop improvement, it typically involves long and laborious artificial planting over multiple generations. Recently, the application of innovative genome engineering techniques, particularly CRISPR-Cas9-based systems, has opened up new avenues that offer the prospects of sustainable farming in the modern agricultural industry. In addition, the emergence of novel editing systems has enabled the development of transgene-free non-genetically modified plants, which represent a suitable option for improving desired traits in a range of crop plants. To date, a number of disease-resistant crops have been produced using gene-editing tools, which can make a significant contribution to overcoming disease-related problems. Not only does this directly minimize yield losses but also reduces the reliance on pesticide application, thereby enhancing crop productivity that can meet the globally increasing demand for food. In this review, we describe recent progress in genome engineering techniques, particularly CRISPR-Cas9 systems, in development of disease-resistant crop plants. In addition, we describe the role of CRISPR-Cas9-mediated genome editing in sustainable agriculture.

Entailing the Next-Generation Sequencing and Metabolome for Sustainable Agriculture by Improving Plant Tolerance.

Crop production is a serious challenge to provide food for the 10 billion individuals forecasted to live across the globe in 2050. The scientists' emphasize establishing an equilibrium among diversity and quality of crops by enhancing yield to fulfill the increasing demand for food supply sustainably. The exploitation of genetic resources using genomics and metabolomics strategies can help generate resilient plants against stressors in the future. The innovation of the next-generation sequencing (NGS) strategies laid the foundation to unveil various plants' genetic potential and help us to understand the domestication process to unmask the genetic potential among wild-type plants to utilize for crop improvement. Nowadays, NGS is generating massive genomic resources using wild-type and domesticated plants grown under normal and harsh environments to explore the stress regulatory factors and determine the key metabolites. Improved food nutritional value is also the key to eradicating malnutrition problems around the globe, which could be attained by employing the knowledge gained through NGS and metabolomics to achieve suitability in crop yield. Advanced technologies can further enhance our understanding in defining the strategy to obtain a specific phenotype of a crop. Integration among bioinformatic tools and molecular techniques, such as marker-assisted, QTLs mapping, creation of reference genome, de novo genome assembly, pan- and/or super-pan-genomes, etc., will boost breeding programs. The current article provides sequential progress in NGS technologies, a broad application of NGS, enhancement of genetic manipulation resources, and understanding the crop response to stress by producing plant metabolites. The NGS and metabolomics utilization in generating stress-tolerant plants/crops without deteriorating a natural ecosystem is considered a sustainable way to improve agriculture production. This highlighted knowledge also provides useful research that explores the suitable resources for agriculture sustainability.

Diversity and Taxonomic Distribution of Endophytic Bacterial Community in the Rice Plant and Its Prospective.

Endophytic bacterial communities are beneficial communities for host plants that exist inside the surfaces of plant tissues, and their application improves plant growth. They benefit directly from the host plant by enhancing the nutrient amount of the plant's intake and influencing the phytohormones, which are responsible for growth promotion and stress. Endophytic bacteria play an important role in plant-growth promotion (PGP) by regulating the indirect mechanism targeting pest and pathogens through hydrolytic enzymes, antibiotics, biocontrol potential, and nutrient restriction for pathogens. To attain these benefits, firstly bacterial communities must be colonized by plant tissues. The nature of colonization can be achieved by using a set of traits, including attachment behavior and motility speed, degradation of plant polymers, and plant defense evasion. The diversity of bacterial endophytes colonization depends on various factors, such as plants' relationship with environmental factors. Generally, each endophytic bacteria has a wide host range, and they are used as bio-inoculants in the form of synthetic applications for sustainable agriculture systems and to protect the environment from chemical hazards. This review discusses and explores the taxonomic distribution of endophytic bacteria associated with different genotypes of rice plants and their origin, movement, and mechanism of PGP. In addition, this review accentuates compressive meta data of endophytic bacteria communities associated with different genotypes of rice plants, retrieves their plant-growth-promoting properties and their antagonism against plant pathogens, and discusses the indication of endophytic bacterial flora in rice plant tissues using various methods. The future direction deepens the study of novel endophytic bacterial communities and their identification from rice plants through innovative techniques and their application for sustainable agriculture systems.

Molybdenum-Induced Regulation of Antioxidant Defense-Mitigated Cadmium Stress in Aromatic Rice and Improved Crop Growth, Yield, and Quality Traits.

Cadmium (Cd) stress causes serious disruptions in plant metabolism, physio-biochemical responses, crop yield, and grain quality characteristics. A pot experiment was conducted to investigate the role of molybdenum (Mo) in mitigating Cd-induced adversities on plant growth, yield attributes, and grain quality characteristics of a popular aromatic rice cultivar 'Xiangyaxiangzhan'. The Mo was applied at 0.15 mg kg-1 soil in both control (no Cd) and Cd-contaminated (100 mg kg-1) soils. A treatment with Mo-free (-Mo) soil was also maintained for comparison. The results showed that Cd toxicity significantly (p < 0.05) reduced plant dry biomass, grain yield, photosynthetic efficiency, and pigment contents, and impaired chloroplast ultra-structural configuration and simultaneously destabilized the plant metabolism owing to higher accumulation of hydrogen peroxide, electrolyte leakage, and malondialdehyde contents. However, Mo supply improved grain yield and 2-acetyl-1-pyrroline content by 64.75% and 77.09%, respectively, under Cd stress, suggesting that Mo supply mitigated Cd-provoked negative effects on yield attributes and grain quality of aromatic rice. Moreover, Mo supply enhanced photosynthesis, proline, and soluble protein content, and also strengthened plant metabolism and antioxidant defense through maintaining higher activities and transcript abundance of ROS-detoxifying enzymes at the vegetative, reproductive, and maturity stages of aromatic rice plants under Cd toxicity. Collectively, our findings indicated that Mo supply strengthened plant metabolism at prominent growth stages through an improved enzymatic and non-enzymatic antioxidant defense system, thereby increasing grain yield and quality characteristics of aromatic rice under Cd toxicity.

A basic helix-loop-helix transcription factor CabHLH113 positively regulate pepper immunity against Ralstonia solanacearum.

Pepper's (Capsicum annum) response to bacterial pathogen Ralstonia solanacearm inoculation (RSI) and abiotic stresses is known to be synchronized by transcriptional network; however, related molecular mechanisms need extensive experimentation. We identified and characterized functions of CabHLH113 -a basic helix-loop-helix transcription factor-in pepper immunity to R. solanacearum infection. The RSI and foliar spray of phytohormones, including salicylic acid (SA), methyl jasmonate (MeJA), ethylene (ETH), and absicic acid (ABA) induced transcription of CabHLH113 in pepper. Loss of function of CabHLH113 by virus-induced-gene-silencing (VIGS) compromised defense of pepper plants against RSI and suppressed relative expression levels of immunity-associated marker genes, i.e., CaPR1, CaNPR1, CaDEF1, CaHIR1 and CaABR1. Pathogen growth was significantly increased after loss of function of CabHLH113 compared with un-silenced plants with remarkable increase in pepper susceptibility. Besides, transiently over-expression of CabHLH113 induced HR-like cell death, H2O2 accumulation and up-regulation of defense-associated marker genes, e.g. CaPR1, CaNPR1, CaDEF1, CaHIR1 and CaABR1. Additionally, transient over-expression of CabHLH113 enhanced the transcriptional levels of CaWRKY6, CaWRKY27 and CaWRKY40. Conversely, transient over-expression of CaWRKY6, CaWRKY27 and CaWRKY40 enhanced the transcriptional levels of CabHLH113. Collectively, our results indicate that newly characterized CabHLH113 has novel defense functions in pepper immunity against RSI via triggering HR-like cell death and cellular levels of defense linked genes.

Metabolic Profiling and Transcriptome Analysis Reveal the Key Role of Flavonoids in Internode Coloration of Phyllostachys violascens cv. Viridisulcata.

Bamboo, being an ornamental plant, has myriad aesthetic and economic significance. Particularly, Phyllostachys violascens cv. Viridisulcata contains an internode color phenotype in variation in green and yellow color between the sulcus and culm, respectively. This color variation is unique, but the underlying regulatory mechanism is still unknown. In this study, we used metabolomic and transcriptomic strategies to reveal the underlying mechanism of variation in internode color. A total of 81 metabolites were identified, and among those, prunin as a flavanone and rhoifolin as a flavone were discovered at a high level in the culm. We also found 424 differentially expressed genes and investigated three genes (PvGL, PvUF7GT, and PvC12RT1) that might be involved in prunin or rhoifolin biosynthesis. Their validation by qRT-PCR confirmed high transcript levels in the culm. The results revealed that PvGL, PvUF7GT, and PvC12RT1 might promote the accumulation of prunin and rhoifolin which were responsible for the variation in internode color of P. violascens. Our study also provides a glimpse into phenotypic coloration and is also a valuable resource for future studies.

The structural characteristics and the substrate recognition properties of RNase ZS1.

TMS5 encodes an RNase ZS1 protein that can process ubiquitin-60S ribosomal protein L40 family (UbL40) mRNAs to regulate thermo-sensitive genic male sterility in rice. Despite the importance of this protein, the structural characteristics and substrate recognition properties of RNase ZS1 remain unclear. Here, we found that the variations in several conservative amino acids alter the activation of RNase ZS1, and its recognition of RNA substrates depends on the structure of RNA. RNase ZS1 acts as a homodimer. The conserved amino acids in or adjacent to enzyme center play a critical role in the enzyme activity of RNase ZS1 and the conserved amino acids that far from active center have little impact on its enzyme activity. The cleavage efficiency of RNase ZS1 for pre-tRNA-MetCAU35 and UbL401 mRNA with cloverleaf-like structure was higher than that of pre-tRNA-AspAUC9 and UbL404 mRNA with imperfect cloverleaf-like structure. This difference implies that the enzyme activity of RNase ZS1 depends on the cloverleaf-like structure of the RNA. Furthermore, the RNase ZS1 activity was not inhibited by the 5' leader sequence and 3' CCA motif of pre-tRNA. These findings provide new insights for studying the cleavage characteristics and substrate recognition properties of RNase ZS.

Maize endophytic microbial-communities revealed by removing PCR and 16S rRNA sequencing and their synthetic applications to suppress maize banded leaf and sheath blight.

Endophytic microbial-communities have specific beneficial functions and are considered key drivers for host plant health. The removing-PCR (R-PCR) is a simple culture-independent cost-effective method to identify endophytic microbial-communities. Microbial communities from maize plant grown in different soil types were identified and characterized via the R-PCR and 16S rRNA sequencing. Culture-dependent microbial community identified through 16S rRNA gene sequencing, further these bacterial communities screened for antagonistic assay against Rhizoctonia solani WH1, in vitro compatibility tests, plant-growth-promoting traits and BIOLOG identification. After that, synthetic-communities (SycomA and SycomB) were prepared by mixing different compatible bacterial-strains to use as an inoculant to suppress pathogens of maize. We identified 167 bacterial operational taxonomic units (OTUs) and unexpected 8 fungal OTUs through the R-PCR, whereas, 95 bacterial OTUs via 16S rRNA sequencing from maize leaves and roots. SycomA and SycomB treatments suppressed the disease level and promoted growth attributes more effectively as compare to the single bacterial-strain and control treatments. This study establishes an efficient approach to isolate, identify and characterize diverse endophytic microbial-community assembly in maize leaves and roots, to successfully apply particular microbes to improve crop growth in soils affected by soil-borne-pathogens.

Molecular Control and Application of Male Fertility for Two-Line Hybrid Rice Breeding.

The significance of the climate change may involve enhancement of plant growth as well as utilization of the environmental alterations in male fertility (MF) regulation via male sterility (MS) systems. We described that MS systems provide a fundamental platform for improvement in agriculture production and have been explicated for creating bulk germplasm of the two-line hybrids (EGMS) in rice as compared to the three-line, to gain production sustainability and exploit its immense potential. Environmental alterations such as photoperiod and/or temperature and humidity regulate MS in EGMS lines via genetic and epigenetic changes, regulation of the noncoding RNAs, and RNA-metabolism including the transcriptional factors (TFs) implication. Herein, this article enlightens a deep understanding of the molecular control of MF in EGMS lines and exploring the regulatory driving forces that function efficiently during plant adaption under a changing environment. We highlighted a possible solution in obtaining more stable hybrids through apomixis (single-line system) for seed production.

Local management and landscape structure determine the assemblage patterns of spiders in vegetable fields.

Both field- and landscape-scale factors can influence the predator communities of agricultural pests, but the relative importance and interactions between these scales are poorly understood. Focusing on spiders, an important taxon for providing biological control, we tested the influence of field- and landscape-scale factors on structuring the spider communities in a highly dynamic brassica agroecosystem. We found that local factors (pesticide-use and crop type) and forested landscape significantly influenced the abundance and species richness of spiders, whilst grassland patches significantly affected the spider species richness. Correlation results demonstrated that assemblage patterns of most spider families positively responded to the interplay between local factors and forest patches in the landscape. The spiders abundance was greatest in cauliflower crops surrounded with forest and grassland patches in landscape. Similarly, ordination analyses revealed that organic fields of cauliflower in forested landscapes had a strong positive association with the abundance and species richness of spiders. In contrast, insecticide and synthetic fertilizer-treated fields of Chinese cabbage in landscapes with little non-crop habitat reduced the abundance and species richness of spiders. Our results highlight the extent of interaction between local- and landscape-scale factors, help explain recently reported inconsistent effects of landscape factors on conservation biological control.

Q-SNARE protein FgSyn8 plays important role in growth, DON production and pathogenicity of Fusarium graminearum.

SNAREs (Soluble N-ethylmaleimide-sensitive factor attachment protein receptors) help intracellular vesicle trafficking and membrane fusion among eukaryotes. They are vital for growth and development of phyto-pathogenic fungi such as Fusarium graminearum which causes Fusarium Head Blight (FHB) of wheat and barley. The SNARE protein Syn8 and its homologues play many roles among different organisms. Here, we have characterized FgSyn8 in F. graminearum as a homologue of Syn8. We have integrated biochemical, microbiological and molecular genetic approaches to investigate the roles of this protein. Our results reveal that FgSyn8 is indispensable for normal vegetative growth, conidiation, conidial morphology and pathogenicity of F. graminearum. Deoxynivalenol (DON) biochemical assay reveals active participation of this protein in DON production of F. graminearum. This has further been confirmed by the production of bulbous structures among the intercalary hyphae. FgSyn8 mutant strain produced defects in perithecia formation which portrays its role in sexual reproduction. In summary, our results support that the SNARE protein FgSyn8 is required for vegetative growth, sexual reproduction, DON production and pathogenicity of F. graminearum.

A novel MYB transcription factor CaPHL8 provide clues about evolution of pepper immunity againstsoil borne pathogen.

MYB TFs in plants are of crucial importance not only for growth and development but also for plant defense against pathogens. CaPHL8, an MYB TF, was identified as a positive regulator of pepper defense against Ralstonia solanacerum inoculation (RSI). Phylogenetic evaluation and functional characterization of CaPHL8 revealed its role in pepper defense evolution. Analysis of the amino acid sequence of PHL8 demonstrates its maximum similarity with the MYB family transcription factor in other plants. Up-regulation of CaPHL8 was observed in pepper plants facing Ralstonia attack.. Consistently the GUS activity of pCaPHL8 showed significantly high activity under RSI as compared to mock-treated plants. The loss of function studies of CaPHL8 conducted through VIGS (virus-induced gene silencing) confirmed the reduced pepper immunity to R. solanacearum and impaired plant growth accompanied by high pathogen growth. Compromised pepper immunity in silenced plants was coupled with a reduction in transcription of defense linked marker genes. On the other hand, transiently overexpressing CaPHL8 (35S::CaPHL8-HA) in pepper caused a hypersensitive response, elevated H2O2 production and high expression of immunity associated marker genes. Stable expression of CaPHL8-HA protein was confirmed by Western blot. Additionally, unlike many other TFs, CaPHL8 is not involved in high-temperature stress tolerance as evident by phenotype and non-significant transcription of high temperature-tolerance related marker genes in pepper. So, all these findings confirm that CaPHL8 is induced by RSI, not by high temperature and high humidity (HTHH). It provides adaptive plasticity to pepper by activating defense to RSI by direct or indirect regulation of different immunity -associated genes.

Molecular regulation of pepper innate immunity and stress tolerance: An overview of WRKY TFs.

The WRKY transcription factors (TFs) family constitutes a major group of TFs in spermatophytes. Different studies have endorsed the considerable biological roles performed by WRKY TFs in plant growth, biotic and abiotic stress responses. Genomic and transcriptomic profiling facilitate us in understanding the WRKY genes in various plants and reveal how WRKY TFs perform their action in response to different plant stresses. WRKY TFs actively take part in metabolism including carbohydrate synthesis, senescence, and secondary metabolites production. Molecular organization of WRKY TFs in plants highlight most predicted outcome of multiple responses simultaneously. Repression and activation related to W-box and other such elements is controlled at transcriptional, translational and domain level. WRKY TFs are becoming more important in crop improvement because of their binding with downstream elements. Additionally, WRKY proteins intermingle with various other TFs for modulating plant immunity. However, WRKY TFs self-regulation and crosstalk between different signaling pathways using WRKY TFs still need extensive investigations. In this review, we focused characteristics of WRKY TFs in Capsicum annum and related research advancement on their functional involvement in plant responses to the challenges of high temperature stress and pathogens infection. We summarized information about Capsicum annum WRKY TFs on the basis of their functions, their target genes and signaling pathways. Moreover, the mechanisms for synergistic responses to various biotic and abiotic stresses, WRKY target genes and other TFs as well will be of more interest with increments in existing information.

Capsicum annuum HsfB2a Positively Regulates the Response to Ralstonia solanacearum Infection or High Temperature and High Humidity Forming Transcriptional Cascade with CaWRKY6 and CaWRKY40.

The responses of pepper (Capsicum annuum) plants to inoculation with the pathogenic bacterium Ralstonia solanacearum and to high-temperature-high-humidity (HTHH) conditions were previously found to be coordinated by the transcription factors CaWRKY6 and CaWRKY40; however, the underlying molecular mechanism was unclear. Herein, we identified and functionally characterized CaHsfB2a, a nuclear-localized heat shock factor involved in pepper immunity to R. solanacearum inoculation (RSI) and tolerance to HTHH. CaHsfB2a is transcriptionally induced in pepper plants by RSI or HTHH and by exogenous application of salicylic acid (SA), methyl jasmonate (MeJA), ethylene (ETH), or abscisic acid (ABA). Virus-induced gene silencing (VIGS) of CaHsfB2a significantly impaired pepper immunity to RSI, hampered HTHH tolerance, and curtailed expression of immunity- and thermotolerance-associated marker genes such as CaHIR1, CaNPR1, CaABR1, and CaHSP24. Likewise, transient overexpression of CaHsfB2a in pepper leaves induced hypersensitive response (HR)-like cell death and H2O2 accumulation and upregulated the above-mentioned marker genes as well as CaWRKY6 and CaWRKY40. Chromatin immunoprecipitation (ChIP) and microscale thermophoresis (MST) analysis revealed that CaHsfB2a bound the promoters of both CaWRKY6 and CaWRKY40. In a parallel experiment, we determined by ChIP-PCR and MST that CaHsfB2a was regulated directly by CaWRKY40 but indirectly by CaWRKY6. Cumulatively, our results suggest that CaHsfB2a positively regulates plant immunity against RSI and tolerance to HTHH, via transcriptional cascades and positive feedback loops involving CaWRKY6 and CaWRKY40.

CaWRKY22 Acts as a Positive Regulator in Pepper Response to RalstoniaSolanacearum by Constituting Networks with CaWRKY6, CaWRKY27, CaWRKY40, and CaWRKY58.

The WRKY web, which is comprised of a subset of WRKY transcription factors (TFs), plays a crucial role in the regulation of plant immunity, however, the mode of organization and operation of this network remains obscure, especially in non-model plants such as pepper (Capsicum annuum). Herein, CaWRKY22, a member of a subgroup of IIe WRKY proteins from pepper, was functionally characterized in pepper immunity against Ralstonia Solanacearum. CaWRKY22 was found to target the nuclei, and its transcript level was significantly upregulated by Ralstonia Solanacearum inoculation (RSI) and exogenously applied salicylic acid (SA), Methyl jasmonate (MeJA), or ethephon (ETH). Loss-of-function CaWRKY22, caused by virus-induced gene silencing (VIGS), enhanced pepper’s susceptibility to RSI. In addition, the silencing of CaWRKY22 perturbed the hypersensitive response (HR)-like cell death elicited by RSI and downregulated defense-related genes including CaPO2, CaPR4, CaACC, CaBPR1, CaDEF1, CaHIR1, and CaWRKY40. CaWRKY22 was found to directly bind to the promoters of CaPR1, CaDEF1, and CaWRKY40 by chromatin immuno-precipitation (ChIP) analysis. Contrastingly, transient overexpression of CaWRKY22 in pepper leaves triggered significant HR-like cell death and upregulated the tested immunity associated maker genes. Moreover, the transient overexpression of CaWRKY22 upregulated the expression of CaWRKY6 and CaWRKY27 while it downregulated of the expression of CaWRKY58. Conversely, the transient overexpression of CaWRKY6, CaWRKY27, and CaWRKY40 upregulated the expression of CaWRKY22, while transient overexpression of CaWRKY58 downregulated the transcript levels of CaWRKY22. These data collectively recommend the role of CaWRKY22 as a positive regulator of pepper immunity against R. Solanacearum, which is regulated by signaling synergistically mediated by SA, jasmonic acid (JA), and ethylene (ET), integrating into WRKY networks with WRKY TFs including CaWRKY6, CaWRKY27, CaWRKY40, and CaWRKY58.

Expression and functional evaluation of CaZNF830 during pepper response to Ralstonia solanacearum or high temperature and humidity.

Extensive transcriptional reprogramming after pathogen attack determines immunity to these invaders and plant development. Zinc finger (ZNF) transcription factors regulate important processes in plants such as development, vegetative activities and plant immunity. Despite their immense significance, majority of ZNF transcription factors (TF) involved in pepper immunity and resistance to heat stress have not been focused much. Herein, we identified and functionally characterized CaZNF830 in pepper defense against Ralstonia solanacearum inoculation (RSI) and tolerance to high temperature and high humidity (HTHH). Transient expression analysis of CaZNF830-GFP fusion protein in tobacco leaves revealed its localization to the nucleus. Transcription of CaZNF830 is induced in pepper plants upon RSI or HTHH. Consistent with this, fluorometric GUS enzymatic assay driven by pCaZNF830 presented significantly enhanced activity under RSI and HTHH in comparison with the control plants. The silencing of CaZNF830 by virus induced gene silencing (VIGS) significantly compromised pepper immunity against RSI with enhanced growth of Ralstonia solanacearum in pepper plants. Silencing of CaZNF830 also impaired tolerance to HTHH coupled with decreased expression levels of immunity and thermo-tolerance associated marker genes including CaHIR1, CaNPR1, CaPR1, CaABR1 and CaHSP24. By contrast, the transient over-expression of CaZNF830 in pepper leaves by infiltration of GV3101 cells containing 35S::CaZNF830-HA induced HR mimic cell death, H2O2 accumulation and activated the transcriptions of the tested defense-relative or thermo-tolerance associated marker genes. RT-PCR and immune-blotting assay confirmed the stable expression of HA-tagged CaZNF830 mRNA and protein in pepper. All these results suggest that CaZNF830 acts as a positive regulator of plant immunity against RSI or tolerance to HTHH, it is induced by RSI or HTHH and consequently activate pepper immunity against RSI or tolerance to HTHH by directly or indirectly transcriptional modulation of many defense-linked genes.

Basic leucine zipper domain transcription factors: the vanguards in plant immunity.

Regulation of spatio-temporal expression patterns of stress tolerance associated plant genes is an essential component of the stress responses. Eukaryotes assign a large amount of their genome to transcription with multiple transcription factors (TFs). Often, these transcription factors fit into outsized gene groups which, in several cases, exclusively belong to plants. Basic leucine zipper domain (bZIP) transcription factors regulate vital processes in plants and animals. In plants, bZIPs are implicated in numerous fundamental processes like seed development, energy balance, and responses to abiotic or biotic stresses. Systematic analysis of the information obtained over the last two decades disclosed a constitutive role of bZIPs against biotic stress. bZIP TFs are vital players in plant innate immunity due to their ability to regulate genes associated with PAMP-triggered immunity, effector-triggered immunity, and hormonal signaling networks. Expression analysis of studied bZIP genes suggests that exploration and functional characterization of novel bZIP TFs in planta is helpful in improving crop resistance against pathogens and environmental stresses. Our review focuses on major advancements in bZIP TFs and plant responses against different pathogens. The integration of genomics information with the functional studies provides new insights into the regulation of plant defense mechanisms and engineering crops with improved resistance to invading pathogens. Conclusively, succinct functions of bZIPs as positive or negative regulator mediate resistance to the plant pathogens and lay a foundation for understanding associated genes and TFs regulating different pathways. Moreover, bZIP TFs may offer a comprehensive transgenic gizmo for engineering disease resistance in plant breeding programs.