Plant Physiology最新文献

Atypical basic helix-loop-helix (bHLH) transcription factors, which lack the basic region for DNA binding, are important elements of brassinosteroid (BR) signaling. Recently, our systematic characterization of the rice (Oryza sativa) INCREASED LEAF INCLINATION (ILI) subfamily of atypical bHLHs revealed their indispensable roles in BR-mediated growth and development. Here, we reported the isolation of two additional rice ILI-interacting atypical bHLHs, ATBS1-INTERACTING FACTOR 1 (OsAIF1)/OsbHLH176 and OsAIF2/OsbHLH178. Genetic and cytological analyses of the OsAIFs knockout mutants and overexpression lines revealed that OsAIF1 and OsAIF2 negatively regulate rice leaf inclination and grain size in a synergistic and redundant manner. Compared to the wild-type, osaif knockout mutants exhibited hypersensitivity to BR, while OsAIF1 and OsAIF2 overexpression lines showed greatly reduced sensitivity or complete insensitivity to BR, indicating that these two OsAIFs act as major negative regulators of rice BR signaling. As ILI-interacting negative atypical HLHs, OsAIF1 and OsAIF2 genetically counteracted the positive ILI subfamily of atypical HLHs. Moreover, OsAIF1 and OsAIF2 physically interacted with and antagonized OsbHLH92, a positive regulator of BR signaling, thereby modulating rice development and gene transcription. These findings suggested that the atypical HLHs (ILIs and OsAIF1/OsAIF2) and the bHLH (OsbHLH92) transcription factors form a triantagonistic cascade in rice, counteracting each other to fine-tune leaf angle and grain size through BR signaling. Our results provide insights into the mechanisms balancing BR signaling and growth in rice.

Transcription factors belonging to the large ethylene response factor (ERF) family are involved in plant responses to biotic and abiotic stresses. Among the ERFs, OCTADECANOID-RESPONSIVE ARABIDOPSIS 59 (ORA59) integrates ethylene and jasmonic acid signaling to regulate resistance to necrotrophic pathogens. The ERF group ERFVII encodes oxygen-labile proteins that are required for oxygen sensing and are stabilized by hypoxia established at the site of Botrytis (Botrytis cinerea) infection. Here, we show that ORA59 represses ERFVII protein activity to induce the expression of hypoxia-responsive genes in Arabidopsis (Arabidopsis thaliana). Moreover, inhibition of ethanol fermentation enhances plant tolerance to Botrytis, indicating a trade-off between the hypoxia and defense responses. In addition, ERFVII members and ORA59 are both involved in the downregulation of hypoxia-responsive genes during reoxygenation. Taken together, our results reveal that the ERFVII transcription factor-ORA59 module ensures that the multiple roles of ERFVII proteins are correctly balanced to favor plant tolerance to biotic or abiotic stresses.

Lignin is a critical component of the closing layer of the potato (Solanum tuberosum L.) tuber during healing; however, the molecular mechanism of its formation remains poorly understood. To elucidate the molecular mechanism of tuber healing, we screened the genes encoding transcription factors that regulate lignin synthesis(StMYB24/49/105/144/168, StWRKY19/20/22/23/34) and the key genes involved in lignin monomer synthesis (PHENYLALANINE AMMONIA LYASE 5 (StPAL5) and CINNAMYL ALCOHOL DEHYDROGENASE 14 (StCAD14)) for induced expression after wounding using transcriptome data. Dual-luciferase assay, yeast one-hybrid, electrophoretic mobility shift assay, and chromatin immunoprecipitation-qPCR assays revealed that StMYB168 could bind directly to the StPAL5 and StCAD14 promoters to activate their expression and that StWRKY20 enhanced this regulation with a synergistic effect. Y2H, bimolecular fluorophore complementation, and coimmunoprecipitation assays showed that StMYB168 interacted with StWRKY20 to form a MYB-WRKY complex. Furthermore, transient overexpression (OE) of StMYB168 and StWRKY20 in Nicotiana benthamiana leaves upregulated the expression of NbPAL and NbCAD10 and promoted lignin accumulation in the leaves. In addition, OE of StWRKY20 and StMYB168 together resulted in higher expression levels of NbPAL and NbCAD10 and higher levels of lignin monomer and total lignin. In contrast, silencing of StMYB168 and StWRKY20 in potato significantly reduced the lignin content of wounded tubers. In conclusion, StMYB168 and StWRKY20 are important regulators of lignin biosynthesis in potato tubers during healing and can positively regulate lignin biosynthesis by forming a complex. The elucidation of this regulatory module provides information on the regulatory mechanism of lignin monomer synthesis in wounded tubers at the transcriptional level.

CELL DIVISION CYCLE 5 (CDC5) is a R2R3-type MYB transcription factor, serving as a key component of modifier of snc1, 4-associated complex/NineTeen complex, which is associated with plant immunity, RNA splicing, and miRNA biogenesis. In this study, we demonstrate that mutation of CDC5 accelerates flowering in Arabidopsis (Arabidopsis thaliana). CDC5 activates the expression of FLOWERING LOCUS C (FLC) by binding to and affecting the enrichment of RNA polymerase II on FLC chromatin. Moreover, genetic analysis confirmed that CDC5 regulates flowering in an FLC-dependent manner. Furthermore, we characterized the interaction of CDC5 with the RNA polymerase-associated factor 1 (Paf1) complex and confirmed that CDC5, as part of the spliceosome, mediates genome-wide alternative splicing, as revealed by RNA-seq. CDC5 affected the splicing of flowering-associated genes such as FLC, SEF, and MAFs. Additionally, we also demonstrated that CDC5 contributes to the regulation of histone modification of FLC chromatin, which further promotes FLC expression. In summary, our results establish CDC5 as a key factor regulating flowering. This provides valuable insight for future research into plant flowering.

Nucleotide-binding leucine-rich repeat (NLR) proteins are intracellular immune receptors that activate innate immune responses upon sensing pathogen attack. However, the molecular mechanisms by which NLR proteins initiate downstream signal transduction pathways to counteract pathogen invasion remain poorly understood. In this study, we identified the wheat (Triticum aestivum) NLR protein Resistance Gene Analogs3 (TaRGA3), which was significantly upregulated during Puccinia striiformis f. sp. tritici (Pst) infection. TaRGA3 and its coiled-coil (CC) domain, localized to the cytoplasm and nucleus, can induce cell death in Nicotiana benthamiana. Virus-induced gene silencing and overexpression suggested that TaRGA3 contributed to wheat resistance to stripe rust by facilitating reactive oxygen species (ROS) accumulation. Yeast 2-hybrid, luciferase complementation imaging, and co-immunoprecipitation assays revealed that TaRGA3 interacted with wheat protein Ascorbate Peroxidase 6 (TaAPX6). Further analysis showed that TaAPX6 specifically targeted the CC domain of TaRGA3. The TaRGA3-TaAPX6 interplay led to reduced enzyme activity of TaAPX6. Notably, TaAPX6 negatively regulated wheat resistance to Pst by removing excessive ROS accompanying Pst-induced hypersensitive responses. Our findings reveal that TaRGA3 responding to Pst infection confers enhanced wheat resistance to stripe rust, possibly by suppressing TaAPX6-modulated ROS scavenging, and demonstrate that TaRGA3 can be used to engineer stripe rust resistance in wheat.

Low oxygen availability within plant cells arises during plant development but is exacerbated under environmental stress conditions. The group VII ETHYLENE RESPONSE FACTOR (ERFVII) transcription factors have been identified as pivotal regulators in the hypoxia response to abiotic stress. However, their roles in transcriptional regulation during biotic stresses remain less defined. In this study, we investigated the biological function and regulatory mechanism of soybean (Glycine max) ERFVII transcription factors during soybean cyst nematode (Heterodera glycines Ichinohe) infection. We provide evidence that soybean cyst nematode infection induces responses at the infection sites similar to those induced by hypoxia, characterized by the stabilization of ERFVII proteins and increased expression of hypoxia-responsive genes. Hypoxia pretreatment of soybeans enhances their resistance to nematode infection. We demonstrate that ERFVII members GmRAP2.12 and GmRAP2.3 act as transcriptional activators to drive the expression of GmPR10-09g, a member of the PR10 gene family highly induced by soybean cyst nematode and positively impacting nematode resistance. Transgenic hairy root analysis of nematode infection for either GmRAP2.12 or N-end rule pathway components (GmATE or GmPRT6) indicates a positive role of ERFVIIs in soybean defense responses against cyst nematode. The results of our study emphasize the important functions of GmERFVIIs in strengthening soybean's immune responses against cyst nematode by transcriptional activation of GmPR10.

Cotton (Gossypium hirsutum) fiber is a highly elongated single cell with a thickened cell wall. MYB transcription factors are important regulators of plant cell elongation; however, the molecular mechanism involved in regulating fiber elongation remains to be explored. Here, we present evidence that the R2R3-MYB transcription factor GhMYB4 negatively regulates cotton fiber cell elongation by suppressing the expression of 2 crucial genes previously reported to affect fiber development: lipid transfer protein 4 (GhLTP4) and sucrose transporter 12 (GhSWEET12). GhMYB4 is preferentially expressed in elongating fiber cells. Knockdown of GhMYB4 in cotton results in longer fiber cells, whereas overexpression of GhMYB4 in Arabidopsis leads to reduced plant height and root length. Transcriptomic and lipidomic analyses revealed that GhMYB4 is involved in coordinating 3 interconnected biological processes, namely lipid content regulation, auxin signaling, and sugar metabolism. Additionally, we showed that GhMYB4 inhibits the expression of GhLTP4 and GhSWEET12 by binding to the MYB cis-element (TTTAGTG) in their respective promoters. Interestingly, basic helix-loop-helix transcription factor 105 (GhbHLH105) and MYB transcription factor 212 (GhMYB212) counteract the inhibitory effects of GhMYB4 on the expression of GhLTP4 and GhSWEET12, respectively. These findings provide insights into the complex molecular mechanisms regulating plant cell elongation.

Inositol pyrophosphates are eukaryotic signaling molecules that have been recently identified as key regulators of plant phosphate sensing and homeostasis. Given the importance of phosphate to current and future agronomic practices, we sought to design plants, which could be used to sequester phosphate, as a step in a phytoremediation strategy. To achieve this, we expressed diadenosine and diphosphoinositol polyphosphate phosphohydrolase (DDP1), a yeast (Saccharomyces cerevisiae) enzyme demonstrated to hydrolyze inositol pyrophosphates, in Arabidopsis thaliana and pennycress (Thlaspi arvense), a spring annual cover crop with emerging importance as a biofuel crop. DDP1 expression in Arabidopsis decreased inositol pyrophosphates, activated phosphate starvation response marker genes, and increased phosphate accumulation. These changes corresponded with alterations in plant growth and sensitivity to exogenously applied phosphate. Pennycress plants expressing DDP1 displayed increases in phosphate accumulation, suggesting that these plants could potentially serve to reclaim phosphate from phosphate-polluted soils. We also identified a native Arabidopsis gene, Nucleoside diphosphate-linked moiety X 13 (NUDIX13), which we show encodes an enzyme homologous to DDP1 with similar substrate specificity. Arabidopsis transgenics overexpressing NUDIX13 had lower inositol pyrophosphate levels and displayed phenotypes similar to DDP1-overexpressing transgenics, while nudix13-1 mutants had increased levels of inositol pyrophosphates. Taken together, our data demonstrate that DDP1 and NUDIX13 can be used in strategies to regulate plant inositol pyrophosphates and could serve as potential targets for engineering plants to reclaim phosphate from polluted environments.

Jasmonates are a family of oxylipin phytohormones regulating plant development and growth and mediating "defense versus growth" responses. The upstream JA biosynthetic precursor cis-(+)-12-oxo-phytodienoic acid (cis-OPDA) acts independently of CORONATIVE INSENSITIVE 1-mediated JA signaling in several stress-induced and developmental processes. However, its perception and metabolism are only partially understood. An isoleucine analog of the biologically active JA-Ile, OPDA-Ile, was detected years ago in wounded leaves of flowering plants, opening up the possibility that conjugation of cis-OPDA to amino acids might be a relevant mechanism for cis-OPDA regulation. Here, we extended the analysis of amino acid conjugates of cis-OPDA and identified naturally occurring OPDA-Val, OPDA-Phe, OPDA-Ala, OPDA-Glu, and OPDA-Asp accumulating in response to biotic and abiotic stress in Arabidopsis (Arabidopsis thaliana). The OPDA amino acid conjugates displayed cis-OPDA-related plant responses in a JA-Ile-dependent manner. We also showed that the synthesis and hydrolysis of cis-OPDA amino acid conjugates are mediated by members of the amidosynthetase GRETCHEN HAGEN 3 and the amidohydrolase INDOLE-3-ACETYL-LEUCINE RESISTANT 1/ILR1-like families. Thus, OPDA amino acid conjugates function in the catabolism or temporary storage of cis-OPDA in stress responses instead of acting as chemical signals per se.