Search Results (114)

Moist chilling is widely used to overcome seed dormancy in zoysiagrass (Zoysia japonica Steud.), but the coordinated physiological and molecular basis remains unclear. Here, freshly matured seeds were subjected to moist chilling at 4 °C in darkness for 0 (Control), 1 (CS1), 2 (CS2), 3 (CS3), or 4 weeks (CS4) and then transferred to germination conditions (30/20 °C, day/night). Prolonged moist chilling progressively improved dormancy release: final germination percentage increased from 40.5% (Control) to 73.5% (CS4), accompanied by a higher germination index and earlier, faster cumulative germination dynamics. Moist chilling also enhanced early seedling vigor, with stronger treatment differentiation in root elongation than in shoot growth. Physiologically, abscisic acid (ABA) content declined while gibberellic acid (GA) content increased, resulting in an elevated GA/ABA ratio with prolonged chilling. Metabolic activation was evidenced by increased α-amylase activity, greater soluble sugar and soluble protein accumulation, and stimulated oxygen uptake. In addition, CAT, SOD, and POD activities were enhanced under prolonged moist chilling, whereas H₂O₂ levels remained relatively stable, suggesting that redox adjustment during dormancy release was characterized by strengthened antioxidant buffering rather than pronounced oxidative accumulation. qRT-PCR supported a mechanistic transition from dormancy maintenance to germination execution, showing moist chilling-associated regulation of ABA/GA metabolism and signaling genes (e.g., NCED, CYP707A, ABI3/ABI5, and GA20ox) and downstream metabolic modules (e.g., GAMYB, AMY, ISA, INV, and HXK1), together with concurrent modulation of respiration- and ROS-related markers (e.g., AOX1a, RBOH, and CAT). Correlation analysis linked germination performance most strongly with α-amylase activity, oxygen uptake, and the GA/ABA ratio. Collectively, our data support a working model in which moist chilling rebalances the ABA–GA gate and activates downstream metabolic and redox adjustment modules to promote dormancy release and improve germination performance in zoysiagrass, providing practical markers for optimizing seed establishment through moist chilling treatment. Full article

Eucalyptus is a highly diverse genus of the Myrtaceae family that is planted worldwide. Many changes occur during callus development, an important process during in vitro plant regeneration. In this study, we conducted methylome and transcriptome analyses to reveal such changes. The results showed that differentially expressed genes between E. camaldulensis (voucher ID: c0009; high embryogenic potential) and E. grandis × urophylla (voucher ID: j0017; low embryogenic potential) during callus development were enriched in plant hormone signal transduction and MAPK (Mitogen-activated protein kinase) signaling pathways. qRT-PCR analysis showed AHP, BAK1, BSK, CRE1, GID1, MKS1, PR-1, PYL, RbohD, and TCH4 could be involved in the callus development and plantlet regeneration capacity. The differences observed in regenerative potential during callus maturation between the two species under study provide a reliable molecular basis for the study of Eucalyptus regeneration mechanisms. Full article

Drought stress severely limits maize growth, and enhancing drought resistance remains a central objective of crop improvement. Plant-specific NAC (NAM/ATAF/CUC) transcription factors are critical regulators of abiotic stress responses. Previously, we identified ZmNAC20—a gene rapidly induced by both drought and abscisic acid (ABA)—as a positive regulator of stomatal closure and drought survival when over-expressed; however, its direct target genes and downstream regulatory network remained elusive. Here, we used DAP-seq to identify 3537 ZmNAC20 binding peaks genome-wide, revealing four enriched cis-motifs and 1439 target genes. GO and KEGG analyses showed pronounced enrichment in intracellular signal transduction. The intersection of RNA-seq of drought-stressed ZmNAC20-over-expressing and wild-type B104 seedlings with DAP-seq outputs defined 80 potential direct targets (48 up-, 32 down-regulated). Among the activated genes, ZmNAC20 binds and up-regulates genes of the ROS-producing enzyme ZmRBOH8, BURP domain-containing protein ZmBURP4, gibberellin (GA) catabolic enzyme ZmGA2OX6, and HD-Zip transcription factor ZmHB56. These results unveil a multi-layered drought-responsive network through which ZmNAC20 integrates hormone and ROS signaling, providing a molecular blueprint for breeding drought-resilient maize. Full article

Background: Black spot disease severely constrains Chinese cabbage production. Methods: To elucidate the defence mechanisms underlying this response, transcriptomic and metabolomic profiles were analysed in leaves of the Chinese cabbage line 904B at 24 h post-inoculation (hpi) with Alternaria brassicicola. In parallel, gene silencing and overexpression were conducted for BraPBL, an RLCK family member in Chinese cabbage. Results: The Chinese cabbage line 904B exhibited marked suppression of cytokinin and auxin signalling, coupled with enhanced expression of genes involved in ethylene and jasmonic acid signalling. Multiple secondary metabolites exhibited differential changes, specifically the sterol compound 4,4-dimethyl-5alpha-cholest-7-en-3beta-ol was significantly upregulated in the treatment group. These metabolites were primarily enriched in the indole alkaloid metabolism and glycerolipid metabolism pathways. Concurrently, BraPBL exhibits increasing expression with prolonged infection. BraPBL overexpression enhances resistance to black spot disease, whereas silencing reduces resistance. Subcellular localization confirmed BraPBL at the plasma membrane. Overexpression of BraPBL upregulates the reactive oxygen species-related gene RBOH and the signal transduction-related gene MEKK1, whilst simultaneously activating the JA pathway. Conclusions: Overall, 904B activates defence-related hormones while suppressing growth and development-related hormones during early infection. Secondary metabolites, particularly the sterol compound 4,4-dimethyl-5alpha-cholest-7-en-3beta-ol, play key roles in defence, and BraPBL functions as a black spot disease–related defence gene in Chinese cabbage. Full article

Candidatus Liberibacter spp. can infect most citrus plants and rely entirely on phloem sieve tube cells of the host plant for survival. Candidatus Liberibacter primarily contains Ca. L. asiaticus (CLas), Ca. L. africanus (CLaf), and Ca. L. americanus (CLam). Among these, CLas is the most harmful and widely distributed and is the primary pathogen of the devastating citrus disease Huanglongbing (HLB). Effectors are among the core weapons secreted by pathogens into plant cells to attack the plant immune system. In this study, we focused on CLas-specific effectors and those that are highly expressed during the infection stage to identify essential virulence effectors. Using secretion signal peptide prediction analysis, 40 candidate effectors with potential secretory capabilities were identified. Transient expression of these candidate effectors in Nicotiana benthamiana revealed their impact on pattern-triggered immunity, including INF-induced cell death and microbial pattern-induced reactive oxygen species (ROS) bursts, and the resistance of N. benthamiana to the bacterial pathogen Pst DC3000. 10 candidate effectors capable of suppressing plant immunity were identified. The stable expression of these candidate effectors in Arabidopsis showed that several candidate effectors enhanced plant susceptibility to Pst DC3000 and inhibited flg22-induced ROS production and MAPK activation. Among the three candidate effectors that significantly suppressed ROS burst, one effector, E3 (CLIBASIA_03085), interacts with the plant NADPH oxidase RbohD, a key enzyme responsible for ROS production. This suggests that E3 likely inhibits ROS accumulation by directly targeting RbohD. Here, we identified multiple candidate effectors capable of suppressing microbial pattern-triggered immunity that may be essential virulence factors for CLas infection, enhancing our understanding of CLas pathogenesis. Full article

This study explored the regulatory effects of exogenous glucose (Glu) and sucrose (Suc) on the growth performance and physiological mechanisms of cucumber seedlings under salt stress. Using two cucumber cultivars as experimental materials, pot experiments demonstrated that salt stress significantly suppressed seedling growth, decreased chlorophyll content, and triggered oxidative damage. However, pretreatment with exogenous sugars effectively mitigated these adverse effects by maintaining photosynthetic efficiency, enhancing the activities of key antioxidant enzymes—superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)—and reducing the accumulation of reactive oxygen species (ROS) and membrane lipid peroxidation. Transcriptomic analysis revealed that the two sugars differentially modulated antioxidant pathways and transcription factor networks to synergistically enhance salt tolerance. Specifically, sucrose preferentially activated POD, whereas glucose specifically induced APX and RbohD. Furthermore, glucose upregulated NAC and ERF family genes, while sucrose suppressed certain WRKY members. Both sugars contributed to the restoration of auxin and abscisic acid (ABA) signaling pathways. This study provides a theoretical foundation for the role of sugar signaling in enhancing crop resistance to abiotic stress. Full article

Plant respiratory burst oxidase homolog (Rboh) genes are integral to the production of reactive oxygen species (ROS) and the regulation of stress responses. Here, bioinformatic techniques were employed to identify eight PgRboh genes (PgRbohA–H) in the genome of pomegranate (Punica granatum L.) and conduct a systematic analysis of this family. The findings showed that all PgRbohs proteins possess characteristic NADPH oxidase domains and are predicted to be localized on the cell membrane. Experimental verification confirmed the membrane localization of PgRbohD and PgRbohE proteins. Phylogenetic analysis categorized the PgRbohs proteins into six distinct groups, suggesting potential functional divergence among these groups. Promoter analysis revealed a significant presence of cis-acting elements responsive to low-temperature and methyl jasmonate (MeJA). The expression of PgRboh genes was found to be tissue-specific. Additionally, real-time PCR (RT-qPCR) was used to analyze expression patterns in response to low-temperature stress that involves multiple PgRboh genes in the cold response process. Overall, our results lay an important foundation for subsequent studies on the cold resistance function of pomegranate Rboh genes and provides new ideas for the breeding of new cold-resistant pomegranate varieties. Full article

Gray mold disease, caused by Botrytis cinerea, severely impacts grape production worldwide. Although proanthocyanidins (PAs) contribute to fungal pathogen resistance, their role in grape defense against B. cinerea remains unclear. Here, we demonstrate that VvMYBPA1, a key transcriptional regulator of PA biosynthesis, negatively modulates B. cinerea resistance in grape berries. While infection suppressed endogenous VvMYBPA1, its agroinfiltration-mediated transient overexpression in berries elevated susceptibility, paralleling reduced β-1,3-glucanase (BGL) and polyphenol oxidase (PPO) activities. Additionally, VvMYBPA1 overexpression elevated VvRBOHs’ expression and reduced peroxidase (POD) activity, resulting in excessive hydrogen peroxide (H₂O₂) accumulation and more cell death. Our results reveal that VvMYBPA1 negatively regulates B. cinerea resistance by disrupting antioxidant enzyme activity and ROS homeostasis, providing new insights into the interplay between PA biosynthesis and fungal defense mechanisms. Full article

Huanglongbing (HLB), caused by Candidatus Liberibacter asiaticus (CLas), is the most devastating disease threatening global citrus production. Although no commercial citrus varieties exhibit complete HLB resistance, genotype-specific tolerance variations remain underexplored. This study conducted a comparative transcriptomic profiling of six commercially citrus cultivars in South China, four susceptible cultivars (C. reticulata cv. Tankan, Gongkan, Shatangju, and C. sinensis Osbeck cv. Newhall), and two tolerant cultivars (C. limon cv. Eureka; C. maxima cv Guanxi Yu) to dissect molecular mechanisms underlying HLB responses. Comparative transcriptomic analyses revealed extensive transcriptional reprogramming, with tolerant cultivars exhibiting fewer differentially expressed genes (DEGs) and targeted defense activation compared to susceptible genotypes. The key findings highlighted the genotype-specific regulation of starch metabolism, where β-amylase 3 (BAM3) was uniquely upregulated in tolerant varieties, potentially mitigating starch accumulation. Immune signaling diverged significantly: tolerant cultivars activated pattern-triggered immunity (PTI) via receptor-like kinases (FLS2) and suppressed ROS-associated RBOH genes, while susceptible genotypes showed the hyperactivation of ethylene signaling and oxidative stress pathways. Cell wall remodeling in susceptible cultivars involved upregulated xyloglucan endotransglucosylases (XTH), contrasting with pectin methylesterase induction in tolerant Eureka lemon for structural reinforcement. Phytohormonal dynamics revealed SA-mediated defense and NPR3/4 suppression in Eureka lemon, whereas susceptible cultivars prioritized ethylene/JA pathways. These findings delineate genotype-specific strategies in citrus–CLas interactions, identifying BAM3, FLS2, and cell wall modifiers as critical targets for breeding HLB-resistant cultivars through molecular-assisted selection. This study provides a foundational framework for understanding host–pathogen dynamics and advancing citrus immunity engineering. Full article

Seed germination and early seedling growth are pivotal stages that define crop establishment and yield potential. Conventional agrochemicals used to improve these processes often raise environmental concerns, highlighting the need for sustainable alternatives. In this study, we demonstrated that water treated with cylindrical dielectric barrier discharge (c-DBD) plasma, enriched with nitric oxide (NO) and reactive nitrogen species (RNS), markedly enhanced onion (Allium cepa) seed germination and seedling vigor. The plasma-treated water (PTW) promoted rapid imbibition, broke dormancy, and accelerated germination rates beyond 98%. Seedlings irrigated with PTW exhibited significantly increased biomass, root and shoot length, chlorophyll content, and antioxidant enzyme activities, accompanied by reduced lipid peroxidation. Transcriptomic profiling revealed that PTW orchestrated a multifaceted regulatory network by upregulating gibberellin biosynthesis genes (GA3OX1/2), suppressing abscisic acid signaling components (ABI5), and activating phenylpropanoid metabolic pathways (PAL, 4CL) and antioxidant defense genes (RBOH1, SOD). These molecular changes coincided with elevated NO₂⁻ and NO₃⁻ levels and finely tuned hydrogen peroxide dynamics, underpinning redox signaling crucial for seed activation and stress resilience. Our findings establish plasma-generated NO-enriched water as an innovative, eco-friendly technology that leverages redox and hormone crosstalk to stimulate germination and early growth, offering promising applications in sustainable agriculture. Full article

The application of an environmentally friendly plant growth regulator to regulate plant growth and development represents a promising strategy for sustainable agriculture. Thymol is a kind of plant-derived natural compound. We have found that thymol is a potential biostimulant with the capability to trigger plant defense against abiotic stresses. Little is known about whether and how thymol modulates plant root system architecture. In this study, physiological, histochemical, and molecular approaches were applied to identify the role of thymol in promoting lateral root development in watermelon seedlings. Thymol significantly promoted LRP (lateral root primordia) initiation and lateral root formation. Rboh (respiratory burst oxidase homolog)-dependent reactive oxygen species (ROS) generation was involved in thymol-promoted lateral root development from LRP. Then, the Rboh gene family with nine members (ClRboh1–ClRboh9) was identified from watermelon genome. Thymol significantly induced the expression of a set of ClRbohs in roots. These results suggested that thymol was able to stimulate lateral root formation by triggering Rboh-dependent ROS production. These findings may help understand the biological function of thymol as an elicitor of lateral root in both applied and fundamental study. Full article

The plant respiratory burst oxidase homologs (RBOHs) are crucial enzymes responsible for the production of reactive oxygen species (ROS) in plants, playing a pivotal role in regulating various aspects of plant growth, development, and stress responses. While RBOH family members have been identified across a wide range of plant species, the functions and characteristics of the RBOH gene family in oats remain poorly understood. In this study, 35 members of the RBOH gene family in the oat genome were identified using bioinformatics approaches. Conserved motif and gene structure analyses revealed that most AsRBOH genes contain Motif4 and Motif5. Phylogenetic tree analysis demonstrated that the AsRBOHs can be classified into five distinct subfamilies. Synteny analysis indicated that AsRBOHs share the highest number of syntenic gene pairs with wheat. Additionally, cis-regulatory element analysis identified several elements associated with drought and hypoxia-specific responses in AsRBOHs. Expression analysis using qRT-PCR showed that 28 AsRBOH genes were upregulated under drought stress, while 18 were downregulated under salt stress. Notably, the genes 7DG1382190 and 7AG1225850 were found to be involved in both drought and salt stress responses. In conclusion, these findings provide a valuable foundation for future functional studies of the AsRBOH gene family in oats, offering insights that could contribute to the improvement and innovation of oat varieties and germplasm. Full article

Living organisms are constantly exposed to various DNA damaging agents. While the mechanisms of DNA damage and DNA repair are well understood, the impact of these agents on RNA secondary structure and subsequent function remains elusive. In this study, we explore the effects of DNA damaging reagent methyl methanesulfonate (MMS) on arabidopsis gene expression and RNA secondary structure using the dimethyl sulfate (DMS) mutational profiling with sequencing (DMS-MaPseq) method. Our analyses reveal that changes in transcriptional levels and mRNA structure are key factors in response to DNA damaging agents. MMS treatment leads to the up-regulation of arabidopsis RBOHs (respiratory burst oxidase homologues) and alteration in the RNA secondary structure of GSTF9 and GSTF10, thereby enhancing mRNA translation efficiency. Redox homeostasis manipulated by RBOHs and GSTFs plays a crucial role in MMS-induced primary root growth inhibition. In conclusion, our findings shed light on the effects of DNA damaging agents on RNA structure and potential mRNA translation, which provide a new insight to understand the mechanism of DNA damage. Full article

Low temperature stress represents a significant abiotic stress factor affecting rice yields. While the structure and some of the functions of cell cycle protein-dependent protein kinase inhibitor (CKI) family proteins have been the subject of study, their relevance to cold tolerance in rice has been less investigated. In this study, we cloned OsEL2 (LOC_Os03g01740) and constructed anti-expression lines of this gene. The resulting lines exhibited significant cold sensitivity and displayed greater oxidative damage than wild type Nippobare (Nip). However, the activities of antioxidant enzymes, such as catalase (CAT), were significantly elevated in OsEL2-AX plants in comparison to Nip following exposure to 4 °C stress. RNA sequencing revealed the presence of 18,822 differential genes, with the majority of them being expressed with temporal specificity. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that a considerable number of differentially expressed genes (DEGs) are involved in the metabolism of amino acids, lipids, and terpenoids. Weighted gene co-expression network analysis (WGCNA) revealed a close relationship between the genes in the turquoise and light green modules and rice cold tolerance traits. These genes were predominantly enriched in terpene metabolism and the metabolism of various plant secondary metabolites, suggesting that OsEL2 influences rice cold tolerance through the metabolism of these two classes of substances. An analysis of the genes within these two modules using transcription factor (TF) enrichment and KEGG enrichment revealed that they are predominantly regulated by mitogen-activated protein kinase (MAPK) and ethylene signaling pathways. Furthermore, we found that tryptophan metabolism, phenylalanine metabolism, and monoterpene synthesis were enriched in down-regulated pathway enrichment analysis. In addition, we also found that the MAPK signaling pathway was enriched in the KEGG enrichment analysis of AX2 with Nip. The results demonstrate that anti-expression of OsEL2 is associated with a notable decline in rice tolerance to cold stress. Full article

B-box (BBX) transcription factors play crucial roles in plant growth, development, and defense responses to biotic and abiotic stresses. In this study, we cloned a BBX transcription factor gene, CsCOL9I, from cucumber and analyzed its role in the plant’s defense against the feeding of Bemisia tabaci. CsCOL9 is expressed throughout all developmental stages in cucumber, with the highest expression in the leaves. CsCOL9 is induced by B. tabaci feeding, salicylic acid (SA), methyl jasmonate (MeJA), and hydrogen peroxide (H₂O₂). Cucumber plants with CsCOL9 silence (TRV2-CsCOL9) and overexpression (1301-CsCOL9) were obtained and analyzed. After CsCOL9 silencing, survival rates and host selectivity for B. tabaci increased; however, the expression levels of genes encoding enzymes (CsSOD, CsRBOH, CsPOD), activities of superoxide dismutase (SOD) and peroxidase (POD), and content of H₂O₂ in plants were all reduced. CsCOL9 overexpression led to decreased survival rates and host selectivity for B. tabaci. Conversely, the expression levels of genes (CsSOD, CsRBOH and CsPOD), activities of SOD and POD, and content of H₂O₂ increased after CsCOL9 overexpression in plants. Collectively, our results demonstrate CsCOL9 positively regulates cucumber resistance to B. tabaci by activating reactive oxygen species bursts. This study lays a theoretical foundation for the application of CsCOL9 in cucumber resistance breeding and green pest control of B. tabaci. Full article