Jasmonate activates secondary cell wall biosynthesis through MYC2-MYB46 module.

Formation of secondary cell wall (SCW) is tightly regulated spatiotemporally by various developmental and environmental signals. Successful fine-tuning of the trade-off between SCW biosynthesis and stress responses requires a better understanding of how plant growth is regulated under environmental stress conditions. However, the current understanding of the interplay between environmental signaling and SCW formation is limited. The lipid-derived plant hormone jasmonate (JA) and its derivatives are important signaling components involved in various physiological processes including plant growth, development, and abiotic/biotic stress responses. Recent studies suggest that JA is involved in SCW formation but the signaling pathway has not been studied for how JA regulates SCW formation. We tested this hypothesis using the transcription factor MYB46, a master switch for SCW biosynthesis, and JA treatments. Both the transcript and protein levels of MYB46, a master switch for SCW formation, were significantly increased by JA treatment, resulting in the upregulation of SCW biosynthesis. We then show that this JA-induced upregulation of MYB46 is mediated by MYC2, a central regulator of JA signaling, which binds to the promoter of MYB46. We conclude that this MYC2-MYB46 module is a key component of the plant response to JA in SCW formation.

OsWRKY7 contributes to pattern-triggered immunity against Xanthomonas oryzae pv. oryzae.

Rice is a staple crop continually threatened by bacterial and fungal pathogens. OsWRKY transcription factors are involved in various disease responses. However, the functions of many OsWRKYs are still elusive. In this study, we demonstrated that OsWRKY7 enhances rice immunity against Xanthomonas oryzae pv. oryzae (Xoo). OsWRKY7 localized in the nucleus, and gene expression of OsWRKY7 was induced by Xoo inoculation. The OsWRKY7-overexpressing lines showed enhanced resistant phenotype against Xoo, and gene expressions of OsPR1a, OsPR1b, and OsPR10a were significantly increased in the transgenic lines after Xoo inoculation. Moreover, OsWRKY7 activated the OsPR promoters, and the promoter activities were synergistically upregulated by flg22. Genetic- and cell-based analysis showed OsWRKY7 is involved in pattern-triggered immunity against Xoo. These results suggest that OsWRKY7 plays a role as a positive regulator of disease resistance to Xoo through pattern-triggered immunity.

Map-Based Cloning and Characterization of a Major QTL Gene, FfR1, Which Confers Resistance to Rice Bakanae Disease.

Bakanae disease (BD), caused by the fungal pathogen Fusarium fujikuroi, is a serious threat to rice production worldwide. Breeding elite rice varieties resistant to BD requires the identification of resistance genes. Previously, we discovered a resistant quantitative trait locus (QTL), qFfR1, in a Korean japonica rice variety, Nampyeong. In this study, we fine-mapped qFfR1 with a Junam*4/Nampyeong BC3F3 population and delimited its location to a 37.1 kb region on chromosome 1. Complementation experiments with seven candidate genes in this region revealed that OsI_02728 is the gene for qFfR1. This gene encodes a protein with a typical leucine-rich repeat (LRR) receptor-like protein structure. RNA-sequencing-based transcriptomic analysis revealed that FfR1 induces the transcription of defense genes, including lignin and terpenoid biosynthesis genes, pathogenesis-related genes, and thionin genes. These results may facilitate investigations into the molecular mechanisms underlying BD resistance, including molecular patterns of Fusarium fujikuroi interacting with FfR1 and players working in signal transduction pathways downstream of FfR1, and the breeding of new BD-resistant varieties by providing a BD resistance gene with its precise selection marker. This will contribute to efficient control of BD, which is becoming more prevalent according to temperature rises due to climate change.

SNF1-related protein kinase 1 represses Arabidopsis growth through post-translational modification of E2Fa in response to energy stress.

Cellular sugar starvation and/or energy deprivation serves as an important signaling cue for the live cells to trigger the necessary stress adaptation response. When exposed to cellular energy stress (ES) conditions, the plants reconfigure metabolic pathways and rebalance energy status while restricting vegetative organ growth. Despite the vital importance of this ES-induced growth restriction, the regulatory mechanism underlying the response remains largely elusive in plants. Using plant cell- and whole plant-based functional analyses coupled with extended genetic validation, we show that cellular ES-activated SNF1-related protein kinase 1 (SnRK1.1) directly interacts with and phosphorylates E2Fa transcription factor, a critical cell cycle regulator. Phosphorylation of E2Fa by SnRK1.1 leads to its proteasome-mediated protein degradation, resulting in S-phase repression and organ growth restriction. Our findings show that ES-dependently activated SnRK1.1 adjusts cell proliferation and vegetative growth for plants to cope with constantly fluctuating environments.

OsWRKY114 Is a Player in Rice Immunity against Fusarium fujikuroi.

Every year, invasive pathogens cause significant damage to crops. Thus, identifying genes conferring broad-spectrum resistance to invading pathogens is critical for plant breeding. We previously demonstrated that OsWRKY114 contributes to rice (Oryza sativa L.) immunity against the bacterial pathovar Xanthomonas oryzae pv. oryzae (Xoo). However, it is not known whether OsWRKY114 is involved in defense responses to other pathogens. In this study, we revealed that OsWRKY114 enhances innate immunity in rice against the fungal pathogen Fusarium fujikuroi, which is the causal agent of bakanae disease. Transcript levels of various gibberellin-related genes that are required for plant susceptibility to F. fujikuroi were reduced in rice plants overexpressing OsWRKY114. Analysis of disease symptoms revealed increased innate immunity against F. fujikuroi in OsWRKY114-overexpressing rice plants. Moreover, the expression levels of OsJAZ genes, which encode negative regulators of jasmonic acid signaling that confer immunity against F. fujikuroi, were reduced in OsWRKY114-overexpressing rice plants. These results indicate that OsWRKY114 confers broad-spectrum resistance not only to Xoo but also to F. fujikuroi. Our findings provide a basis for developing strategies to mitigate pathogen attack and improve crop resilience to biotic stress.

OsWRKY114 Inhibits ABA-Induced Susceptibility to Xanthomonas oryzae pv. oryzae in Rice.

The phytohormone abscisic acid (ABA) regulates various aspects of plant growth, development, and stress responses. ABA suppresses innate immunity to Xanthomonas oryzae pv. oryzae (Xoo) in rice (Oryza sativa), but the identity of the underlying regulator is unknown. In this study, we revealed that OsWRKY114 is involved in the ABA response during Xoo infection. ABA-induced susceptibility to Xoo was reduced in OsWRKY114-overexpressing rice plants. OsWRKY114 attenuated the negative effect of ABA on salicylic acid-dependent immunity. Furthermore, OsWRKY114 decreased the transcript levels of ABA-associated genes involved in ABA response and biosynthesis. Moreover, the endogenous ABA level was lower in OsWRKY114-overexpressing plants than in the wild-type plants after Xoo inoculation. Taken together, our results suggest that OsWRKY114 is a negative regulator of ABA that confers susceptibility to Xoo in rice.

Identification of a novel NPR1 homolog gene, OsNH5N16, which contributes to broad-spectrum resistance in rice.

Over half of the earth's population consumes rice as the primary food crop for dietary calories. However, severe loss of rice yield occurs due to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) and bakanae disease caused by Fusarium fujikuroi (F. fujikuroi). Therefore, broad-spectrum resistance (BSR) to these pathogens is essential for rice cultivation. The Nonexpressor of Pathogenesis-Related Genes1 (NPR1), which is related to the signal molecule salicylic acid (SA) and the expression of pathogenesis-related (PR) genes, is a key regulator of systemic acquired resistance (SAR). Although five NPR1 homologs (NHs) have been identified in rice thus far, their cellular and biological functions remain largely unexplored. In this study, we identified a novel rice NH gene from Oryza sativa L. cv. Dongjin. The genetic variation of single nucleotide polymorphisms in OsNH5 caused a single amino acid substitution of asparagine for serine at residue 16. OsNH5N16 was mainly located in the nucleus, and its transcription was induced by Xoo. We generated transgenic rice lines constitutively expressing OsNH5N16 to investigate its function. Plants that overexpressed OsNH5N16 displayed enhanced BSR to Xoo and F. fujikuroi compared with wild varieties, and the transcription of PR genes such as OsPR1, GLUC, and CHIT2 was considerably upregulated. Moreover, we revealed that SA increases the transcription of OsNH5N16 and the promoter activity of OsPR1 regulated by OsNH5N16. These results showed that OsNH5N16 enhances BSR by regulating the expression of PR genes related to SAR and it is controlled by SA at the transcriptional and post-translational levels. This is the first report on the innate immune response conferring BSR associated with NH5.

The Capsicum baccatum-Specific Truncated NLR Protein CbCN Enhances the Innate Immunity against Colletotrichum acutatum.

Chili pepper (Capsicumannuum) is an important fruit and spice used globally, but its yield is seriously threatened by anthracnose. Capsicum baccatum is particularly valuable as it carries advantageous disease resistance genes. However, most of the genes remain to be identified. In this study, we identified the C. baccatum-specific gene CbCN, which encodes a truncated nucleotide-binding and leucine-rich repeat protein in the anthracnose resistant chili pepper variety PBC80. The transcription of CbCN was greater in PBC80 than it was in the susceptible variety An-S after Colletotrichum acutatum inoculation. In order to investigate the biological function of CbCN, we generated transgenic tobacco lines constitutively expressing CbCN. Notably, CbCN-overexpressing transgenic plants exhibited enhanced resistance to C. acutatum compared to wild-type plants. Moreover, the expression of pathogenesis-related (PR) genes was remarkably increased in a CbCN-overexpressing tobacco plants. In order to confirm these results in chili pepper, we silenced the CbCN gene using the virus-induced gene silencing system. The anthracnose resistance and expressions of PR1, PR2, and NPR1 were significantly reduced in CbCN-silenced chili peppers after C. acutatum inoculations. These results indicate that CbCN enhances the innate immunity against anthracnose caused by C. acutatum by regulating defense response genes.

Identification of the Capsicum baccatum NLR Protein CbAR9 Conferring Disease Resistance to Anthracnose.

Anthracnose is caused by Colletotrichum species and is one of the most virulent fungal diseases affecting chili pepper (Capsicum) yield globally. However, the noble genes conferring resistance to Colletotrichum species remain largely elusive. In this study, we identified CbAR9 as the causal locus underlying the large effect quantitative trait locus CcR9 from the anthracnose-resistant chili pepper variety PBC80. CbAR9 encodes a nucleotide-binding and leucine-rich repeat (NLR) protein related to defense-associated NLRs in several other plant species. CbAR9 transcript levels were induced dramatically after Colletotrichum capsici infection. To explore the biological function, we generated transgenic Nicotiana benthamiana lines overexpressing CbAR9, which showed enhanced resistance to C. capsici relative to wild-type plants. Transcript levels of pathogenesis-related (PR) genes increased markedly in CbAR9-overexpressing N. benthamiana plants. Moreover, resistance to anthracnose and transcript levels of PR1 and PR2 were markedly reduced in CbAR9-silenced chili pepper fruits after C. capsici infection. Our results revealed that CbAR9 contributes to innate immunity against C. capsici.

Rice glutaredoxin GRXS15 confers broad-spectrum resistance to Xanthomonas oryzae pv. oryzae and Fusarium fujikuroi.

Rice is a particularly widely consumed food crop globally, but its yield is seriously damaged by bacterial blight due to Xanthomonas oryzae pv. oryzae (Xoo) and bakanae disease due to Fusarium fujikuroi (F. fujikuroi). However, broad-spectrum resistance (BSR) to both Xoo and F. fujikuroi remains largely elusive. In this study, we showed that rice monothiol glutaredoxin GRXS15 localizes in mitochondria and the nucleus, and its transcription is induced by Xoo. Transgenic rice lines constitutively expressing OsGRXS15 showed enhanced disease resistance to Xoo and F. fujikuroi, while CRISPR/Cas9-based knockout mutants showed reduced resistance compared with the wild-type plants. The transcription of pathogenesis-related (PR) genes was significantly induced in OsGRXS15-expressing plants. The rice transcription factor OsWRKY65 was identified as a binding partner, and it directly interacted with OsGRXS15 in the nucleus. Moreover, we revealed that the interaction of OsGRXS15 and OsWRKY65 results in the upregulation of OsPR1. These results suggested that OsGRXS15 interacts with transcription factors, and it confers BSR through regulating the expression of genes related to pathogen response. This is the first report on the nuclear function associated with the monothiol glutaredoxin GRXS15.

Rice transcription factor WRKY114 directly regulates the expression of OsPR1a and Chitinase to enhance resistance against Xanthomonas oryzae pv. oryzae.

Rice (Oryza sativa L.) is a global staple crop, but its yield is severely threatened by bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo). The OsWRKY transcription factor family play a central role in innate plant immunity against Xoo, but the various biological functions of a large number of OsWRKYs remain to be understood. We characterized the role of OsWRKY114 against Xoo. OsWRKY114 has transcriptional activity in yeast and localizes in the nucleus. When OsWRKY114 is overexpressed in transgenic plants they show enhanced disease resistance against Xoo compared with wild types. By using genetic- and cell-based functional analyses, we showed OsWRKY114 directly associates with the promoters of OsPR1a and Chitinase and increases the promoter activities. These results suggest that OsWRKY114 enhances the innate immunity of Asian rice against Xoo through direct activation of defense genes that include OsPR1a and chtinase. This is the first report to functionally characterize OsWRKY114 in Xoo infection.

Regulatory role of BOTRYTIS INDUCED KINASE1 in ETHYLENE INSENSITIVE3-dependent gene expression in Arabidopsis.

KEY MESSAGE: Arabidopsis BIK1 negatively regulates EIN3-depedent gene expression as an immediate cellular response. BIK1 localizes to the plasma membrane and its autophosphorylation and kinase activity involves in EIN3 repression. BOTRYTIS INDUCED KINASE1 (BIK1) is a multifunctional receptor-like kinase that involves in ethylene-mediated plant defense signaling. The loss of function BIK1 becomes insensitive to ethylene, but it still accumulates a higher level of ETHYLENE INSENSITIVE3 (EIN3) that serves as the key transcription activator in ethylene signaling. To unequivocally elucidate BIK1 function on EIN3 regulation in ethylene signaling, we took a combined approach of transient expression assay and stable expression analysis of BIK1. In our cell-based functional assay BIK1 destabilized EIN3 and down-regulated EIN3-dependent transcription. Membrane localization and autophosphorylation of BIK1 were required for full repression of EIN3 function, but its kinase activity potential compromised such regulatory action. Consistently, the analysis of transgenic plants verified BIK1 function on EIN3 repression. Our findings have clarified that autophosphorylated BIK1 in the plasma membrane negatively regulates EIN3-dependent gene expression. Thus, ethylene insensitivity in bik1 appears to be an indirect or a feedback long-term response.

Plant translational reprogramming for stress resilience.

Organisms regulate gene expression to produce essential proteins for numerous biological processes, from growth and development to stress responses. Transcription and translation are the major processes of gene expression. Plants evolved various transcription factors and transcriptome reprogramming mechanisms to dramatically modulate transcription in response to environmental cues. However, even the genome-wide modulation of a gene's transcripts will not have a meaningful effect if the transcripts are not properly biosynthesized into proteins. Therefore, protein translation must also be carefully controlled. Biotic and abiotic stresses threaten global crop production, and these stresses are seriously deteriorating due to climate change. Several studies have demonstrated improved plant resistance to various stresses through modulation of protein translation regulation, which requires a deep understanding of translational control in response to environmental stresses. Here, we highlight the translation mechanisms modulated by biotic, hypoxia, heat, and drought stresses, which are becoming more serious due to climate change. This review provides a strategy to improve stress tolerance in crops by modulating translational regulation.

The rice SnRK family: biological roles and cell signaling modules.

Stimulus-activated signaling pathways orchestrate cellular responses to control plant growth and development and mitigate the effects of adverse environmental conditions. During this process, signaling components are modulated by central regulators of various signal transduction pathways. Protein phosphorylation by kinases is one of the most important events transmitting signals downstream, via the posttranslational modification of signaling components. The plant serine and threonine kinase SNF1-related protein kinase (SnRK) family, which is classified into three subgroups, is highly conserved in plants. SnRKs participate in a wide range of signaling pathways and control cellular processes including plant growth and development and responses to abiotic and biotic stress. Recent notable discoveries have increased our understanding of how SnRKs control these various processes in rice (Oryza sativa). In this review, we summarize current knowledge of the roles of OsSnRK signaling pathways in plant growth, development, and stress responses and discuss recent insights. This review lays the foundation for further studies on SnRK signal transduction and for developing strategies to enhance stress tolerance in plants.

Heat shock factor binding protein BrHSBP1 regulates seed and pod development in Brassica rapa.

Plant heat shock factor binding proteins (HSBPs) are well known for their implication in the negative regulation of heat stress response (HSR) pathways. Herein, we report on the hitherto unknown functions of HSBP1 in Brassica rapa (BrHSBP1). BrHBSP1 was found to be predominant in flower buds and young leaves, while its segmental duplicate, BrHSBP1-like, was abundant in green siliques. Exposure to abiotic stress conditions, such as heat, drought, cold, and H2O2, and to phytohormones was found to differentially regulate BrHSBP1. The activity of BrHSBP1-GFP fusion proteins revealed their cellular localization in nuclei and cytosols. Transgenic overexpression of BrHSBP1 (BrHSBP1OX) improved pod and seed sizes, while CRISPR-Cas BrHSBP1 knock-out mutants (Brhsbp1_KO) were associated with aborted seed and pod development. The transcriptomic signatures of BrHSBP1OX and Brhsbp1_KO lines revealed that 360 and 2381 genes, respectively, were differentially expressed (Log2FC≥2, padj<0.05) expressed relative to control lines. In particular, developmental processes, including plant reproductive structure development (RSD)-related genes, were relatively downregulated in Brhsbp1_KO. Furthermore, yeast two-hybrid assays confirmed that BrHSBP1 can physically bind to RSD and other genes. Taking the findings together, it is clear that BrHSBP1 is involved in seed development via the modulation of RSD genes. Our findings represent the addition of a new regulatory player in seed and pod development in B. rapa.

GmMPK6 Positively Regulates Salt Tolerance through Induction of GmRbohI1 in Soybean.

Salt stress is a critical environmental stress that impairs plant growth and development, especially in crop productivity; therefore, understanding the salt response in plants is the basis for their development of salt tolerance. Under salinity, soybean mitogen-activated protein kinase 6 (GmMPK6) is activated and positively regulates reactive oxygen species (ROS) generation. However, it is not yet elucidated how GmMPK6 regulates ROS generation and its role in salt tolerance. Here, we show that GmMPK6, solely activated in NaCl treatment, and gene expression of GmRbohI1 was not only reduced by MPK inhibitor SB202190 in NaCl treatment, but also increased in a GMKK1-expressing protoplast. Furthermore, SB202190 and the NADPH-oxidase inhibitor, diphenyleneiodonium chloride, increased susceptibility to salt stress. The expression of GmRD19A was induced by NaCl treatment, but this expression was compromised by SB202190. Consequently, we revealed that GmMPK6 induces ROS generation through the transcriptional regulation of GmRbohI1 and increases salt tolerance in soybean.

Challenges Facing CRISPR/Cas9-Based Genome Editing in Plants.

The development of plant varieties with desired traits is imperative to ensure future food security. The revolution of genome editing technologies based on the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has ushered in a new era in plant breeding. Cas9 and the single-guide RNA (sgRNA) form an effective targeting complex on a locus or loci of interest, enabling genome editing in all plants with high accuracy and efficiency. Therefore, CRISPR/Cas9 can save both time and labor relative to what is typically associated with traditional breeding methods. However, despite improvements in gene editing, several challenges remain that limit the application of CRISPR/Cas9-based genome editing in plants. Here, we focus on four issues relevant to plant genome editing: (1) plant organelle genome editing; (2) transgene-free genome editing; (3) virus-induced genome editing; and (4) editing of recalcitrant elite crop inbred lines. This review provides an up-to-date summary on the state of CRISPR/Cas9-mediated genome editing in plants that will push this technique forward.

Climate change impedes plant immunity mechanisms.

Rapid climate change caused by human activity is threatening global crop production and food security worldwide. In particular, the emergence of new infectious plant pathogens and the geographical expansion of plant disease incidence result in serious yield losses of major crops annually. Since climate change has accelerated recently and is expected to worsen in the future, we have reached an inflection point where comprehensive preparations to cope with the upcoming crisis can no longer be delayed. Development of new plant breeding technologies including site-directed nucleases offers the opportunity to mitigate the effects of the changing climate. Therefore, understanding the effects of climate change on plant innate immunity and identification of elite genes conferring disease resistance are crucial for the engineering of new crop cultivars and plant improvement strategies. Here, we summarize and discuss the effects of major environmental factors such as temperature, humidity, and carbon dioxide concentration on plant immunity systems. This review provides a strategy for securing crop-based nutrition against severe pathogen attacks in the era of climate change.

SNF1-Related Protein Kinase 1 Activity Represses the Canonical Translational Machinery.

Protein biosynthesis is achieved through translation, which consumes enormous energy. Therefore, under conditions of limited energy supply, translation progress should be strictly coordinated. Sucrose non-fermenting kinase1 (SNF1)-related protein kinase 1 (SnRK1) is an evolutionarily conserved master regulator of cellular energy stress signaling in plants. Rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana) SnRK1 enhance hypoxia tolerance and induce the expression of stress-related genes. However, whether SnRK1 modulates protein synthesis in plants is unknown. In this study, using translational reporter constructs transfected in Arabidopsis protoplasts we showed that the expression of OsSnRK1A and AtSnRK1.1 decreases the abundance of canonical proteins without affecting their encoding transcript levels and protein stability. Moreover, the loading of total mRNAs and GFP mRNAs into the heavy polysome fraction which is normally translated was attenuated in transgenic Arabidopsis lines constitutively expressing OsSnRK1A or AtSnRK1.1. Taken together, these results suggest that OsSnRK1A and AtSnRK1.1 suppress protein translation to maintain energy homeostasis.