Reduced H2O2 Accumulation Research Articles

The transcription factor ZmNAC84 increases maize salt tolerance by regulating ZmCAT1 expression

Salt stress severely affects plant growth and yield. The transcription factor NAC plays a variety of important roles in plant abiotic stress, but we know relatively little about the specific molecular mechanisms of NAC in antioxidant defense. Here, our genetic studies reveal the positive regulation of salt tolerance in maize by the transcription factor ZmNAC84. Under salt stress, overexpression of ZmNAC84 in maize increased the expression of ZmCAT1, enhanced CAT activity, and consequently reduced H2O2 accumulation, thereby improving salt stress tolerance in maize. Whereas RNA interference-mediated knockdown of ZmNAC84 produced the opposite effect. Subsequently, we found that ZmNAC84 directly binds to and regulates the expression of the ZmCAT1 promoter, and the hybridized material also demonstrated that ZmCAT1 is a downstream target gene of ZmNAC84. In addition, phenotypic and biochemical analyses indicated that ZmCAT1 positively regulated salt tolerance by regulating H2O2 accumulation under salt stress. Taken together, these results reveal the function of ZmNAC84 in regulating ZmCAT1-mediated antioxidant defense in response to salt stress in plants.

Effector CLas0185 targets methionine sulphoxide reductase B1 of Citrus sinensis to promote multiplication of 'Candidatus Liberibacter asiaticus' via enhancing enzymatic activity of ascorbate peroxidase 1.

Citrus huanglongbing (HLB) has been causing enormous damage to the global citrus industry. As the main causal agent, 'Candidatus Liberibacter asiaticus' (CLas) delivers a set of effectors to modulate host responses, while the modes of action adopted remain largely unclear. Here, we demonstrated that CLIBASIA_00185 (CLas0185) could attenuate reactive oxygen species (ROS)-mediated cell death in Nicotiana benthamiana. Transgenic expression of CLas0185 in Citrus sinensis 'Wanjincheng' enhanced plant susceptibility to CLas. We found that methionine sulphoxide reductase B1 (CsMsrB1) was targeted by the effector, and its abundance was elevated in CLas0185-transgenic citrus plants. Their interaction promoted CLas proliferation. We then determined that CsMsrB1 sustained redox state and enzymatic activity of ascorbate peroxidase 1 (CsAPX1) under oxidative stress. The latter reduced H2O2 accumulation and was associated with host susceptibility to CLas infection. Consistently, citrus plants expressing CLas0185 and CsMsrB1 conferred enhanced APX activity and decreased H2O2 content. Taken together, these findings revealed how CLas0185 benefits CLas colonization by targeting CsMsrB1, which facilitated the antioxidant activity and depressed ROS during pathogen infection.

Metagenomic and transcriptome insights into the disease resistance mechanisms in fresh-cut apple under pyridoxine treatment

Microbial spoilage poses a significant challenge to fresh-cut apples during storage, impacting quality and consumer health. This study explores pyridoxine’s (vitamin B6; VB6) potential mode of action to mitigate microbial spoilage in fresh-cut apples through physiological, transcriptomic, and metagenomic analysis. VB6 notably reduces microbial spoilage at room temperature by suppressing bacterial genera (Pseudomonas, Curtobacterium, Frigoribacterium) and fungal genera (Alternaria, Penicillium, Aureobasidium, Pseudogymnoascus) proliferation. Spoilage-causing identified biomarkers include species related to Pseudomonas, Curtobacterium, Frigoribacterium, and Penicillium. KEGG analysis suggests correlations of induced disease resistance with tropane alkaloids biosynthesis, plant-pathogen interaction, and phenylpropanoid biosynthesis pathways in fresh-cut apples. Moreover, VB6 treatment enhanced the antioxidant system via maintaining higher total phenolic contents, total flavonoid contents, and ascorbic acid under elevated catalase, superoxide dismutase, and ascorbate peroxidase enzymes activities, induced disease resistance under higher enzymes activities including phenylalanine ammonialyase, chitinase, cinnamate 4-hydroxylase, β-1,3-glucanase, and 4-coumarate: co-enzyme A ligase enzymes activities, maintained elevated VB6 contents, and reduced H2O2 accumulation. Notably, 6 elevated transcription factors involving WRKY, NAC and MYB might regulate disease resistance pathways in fresh-cut apples under VB6 treatment. This research establishes a theoretical and practical foundation for preserving post-processing quality of fresh-cut apples by restraining microbial proliferation through the proposed VB6-induced disease resistance regulatory mechanism.

Elucidating the molecular mechanisms in phytohormones-induced alleviation of postharvest physiological deterioration in cassava tuberous roots

Cassava is a staple food crop in tropical regions, serving as a significant source of carbohydrates. However, rapid postharvest physiological deterioration (PPD) significantly limits the shelf life and utilization of cassava. Despite its importance, the overlapping molecular mechanisms underlying phytohormones-induced PPD alleviation in cassava tuberous roots remain unknown. In this study, the phytohormones ethephon (ET), gibberellic acid (GA), and methyl jasmonate (MeJA) were applied to cassava tuberous roots after harvest. These treatments led to significantly reduced H2O2 accumulation and alleviation of PPD. Transcriptomic analysis revealed shared functions, including transcriptional regulation and oxidoreductase activity, among differentially expressed genes (DEGs) affected by ET, GA, or MeJA. A total of 305 genes encoding antioxidant enzymes were systematically identified, with 151 responsive to phytohormone treatments across ten families. Notably, the expression of 10 Glutathione S-transferase (GST) genes was induced by all three phytohormone treatments, while their expression was suppressed in the absence of phytohormones during PPD. By using co-expression network analysis, DNA-binding motif analysis, and experimental validation using yeast one-hybrid and dual-luciferase assays, a regulatory network involving MeMYB102 and MeZAT11 transcription factors (TFs) that directly promote the expression of MeGST42, MeGST43, MeGST44, and MeGST46 was established. Overall, this study enhances the understanding of the overlapping molecular mechanisms underlying phytohormones-induced alleviation of PPD in cassava, thereby providing genetic resources for developing PPD-resistant cassava.

Expression of the alfalfa gene MsMDHAR in Arabidopsis thaliana increases water stress tolerance.

The ascorbate-glutathione pathway plays an essential role in the physiology of vascular plants, particularly in their response to environmental stresses. This pathway is responsible for regulating the cellular redox state, which is critical for maintaining cell function and survival under adverse conditions. To study the involvement of the alfalfa monodehydroascorbate reductase (MsMDHAR) in water stress processes, Arabidopsis thaliana plants constitutively expressing the sequence encoding MsMDHAR were developed. Transgenic events with low and high MsMDHAR expression and ascorbate levels were selected for further analysis of drought and waterlogging tolerance. Under water stress, Arabidopsis transgenic plants generated higher biomass, produced more seeds, and had larger roots than wild type ones. This higher tolerance was associated with increased production of waxes and chlorophyll a at the basal level, greater stomatal opening and stability in regulating the relative water content and reduced H2O2 accumulation under stress conditions in transgenic plants. Overall, these results show that MsMDHAR is involved in plant tolerance to abiotic stresses. The data presented here also emphasises the potential of the MsMDHAR enzyme as a plant breeding tool to improve water stress tolerance.

ABA Inhibits Rice Seed Aging by Reducing H2O2 Accumulation in the Radicle of Seeds

The seed, a critical organ in higher plants, serves as a primary determinant of agricultural productivity, with its quality directly influencing crop yield. Improper storage conditions can diminish seed vigor, adversely affecting seed germination and seedling establishment. Therefore, understanding the seed-aging process and exploring strategies to enhance seed-aging resistance are paramount. In this study, we observed that seed aging during storage leads to a decline in seed vigor and can coincide with the accumulation of hydrogen peroxide (H2O2) in the radicle, resulting in compromised or uneven germination and asynchronous seedling emergence. We identified the abscisic acid (ABA) catabolism gene, abscisic acid 8′-hydroxylase 2 (OsABA8ox2), as significantly induced by aging treatment. Interestingly, transgenic seeds overexpressing OsABA8ox2 exhibited reduced seed vigor, while gene knockout enhanced seed vigor, suggesting its role as a negative regulator. Similarly, seeds pretreated with ABA or diphenyleneiodonium chloride (DPI, an H2O2 inhibitor) showed increased resistance to aging, with more robust early seedling establishment. Both OsABA8ox2 mutant seeds and seeds pretreated with ABA or DPI displayed lower H2O2 content during aging treatment. Overall, our findings indicate that ABA mitigates rice seed aging by reducing H2O2 accumulation in the radicle. This study offers valuable germplasm resources and presents a novel approach to enhancing seed resistance against aging.

MiR396b/GRF6 module contributes to salt tolerance in rice.

Salinity, as one of the most challenging environmental factors restraining crop growth and yield, poses a severe threat to global food security. To address the rising food demand, it is urgent to develop crop varieties with enhanced yield and greater salt tolerance by delving into genes associated with salt tolerance and high-yield traits. MiR396b/GRF6 module has previously been demonstrated to increase rice yield by shaping the inflorescence architecture. In this study, we revealed that miR396b/GRF6 module can significantly improve salt tolerance of rice. In comparison with the wild type, the survival rate of MIM396 and OE-GRF6 transgenic lines increased by 48.0% and 74.4%, respectively. Concurrent with the increased salt tolerance, the transgenic plants exhibited reduced H2O2 accumulation and elevated activities of ROS-scavenging enzymes (CAT, SOD and POD). Furthermore, we identified ZNF9, a negative regulator of rice salt tolerance, as directly binding to the promoter of miR396b to modulate the expression of miR396b/GRF6. Combined transcriptome and ChIP-seq analysis showed that MYB3R serves as the downstream target of miR396b/GRF6 in response to salt tolerance, and overexpression of MYB3R significantly enhanced salt tolerance. In conclusion, this study elucidated the potential mechanism underlying the response of the miR396b/GRF6 network to salt stress in rice. These findings offer a valuable genetic resource for the molecular breeding of high-yield rice varieties endowed with stronger salt tolerance.

Binucleate Rhizoctonia induced tomato resistance against Rhizoctonia solani via affecting antioxidants and cell wall reinforcement

Isolates of Rhizoctonia solani (AG-3 PT, AG-4 HG-I, AG-4 HG-II) and one binucleate Rhizoctonia sp. (BNR) belonging to AG-Bb were investigated for pathogenicity on tomato cultivar Mobil. The BNR isolate revealed the lowest virulence and it was used as biocontrol agent against R. solani AG-4 HG-II, which showed the highest virulence on tomato. Inoculation of tomato plants with the hypovirulent BNR isolate reduced the disease symptoms of R. solani and induced resistance. Resistance induction was observed not only on the plants simultaneously inoculated with BNR and R. solani, but also when the plants were inoculated by the BNR and R. solani with time intervals. The peroxidase (POX), superoxide dismutase (SOD) and catalase (CAT) activities and expression levels of the corresponding genes in tomato plants increased after R. solani or BNR inoculation. The highest level of antioxidant activities and expression of their genes, lignin and callose formation were observed in the plants inoculated with the BNR and R. solani, simultaneously. The BNR inoculation reduced H2O2 accumulation. The highest level of priming was observed for the POX among other antioxidants tested via application of the BNR. Treatment with potassium cyanide (as a POX inhibitor) reduced basal resistance and BNR-induced resistance (BNR-IR) via reduction of lignification and callose deposition in tomato plants. These findings demonstrated the role of antioxidant enzymes, mainly the POX, in both basal resistance and BNR-IR. Therefore, redox state and antioxidants are involved in cell wall strengthening via lignin and callose formation, as important defense components which decrease the pathogen progress in plant tissues.

LysM protein BdLM1 of Botryosphaeria dothidea plays an important role in full virulence and inhibits plant immunity by binding chitin and protecting hyphae from hydrolysis.

Botryosphaeria dothidea infects hundreds of woody plants and causes a severe economic loss to apple production. In this study, we characterized BdLM1, a protein from B. dothidea that contains one LysM domain. BdLM1 expression was dramatically induced at 6 h post-inoculation in wounded apple fruit, strongly increased at 7 d post-inoculation (dpi), and peaked at 20 dpi in intact shoots. The knockout mutants of BdLM1 had significantly reduced virulence on intact apple shoots (20%), wounded apple shoots (40%), and wounded apple fruit (40%). BdLM1 suppressed programmed cell death caused by the mouse protein BAX through Agrobacterium-mediated transient expression in Nicotiana benthamiana, reduced H2O2 accumulation and callose deposition, downregulated resistance gene expression, and promoted Phytophthora nicotianae infection in N. benthamiana. Moreover, BdLM1 inhibited the active oxygen burst induced by chitin and flg22, bound chitin, and protected fungal hyphae against degradation by hydrolytic enzymes. Taken together, our results indicate that BdLM1 is an essential LysM effector required for the full virulence of B. dothidea and that it inhibits plant immunity. Moreover, BdLM1 could inhibit chitin-triggered plant immunity through a dual role, i.e., binding chitin and protecting fungal hyphae against chitinase hydrolysis.

A LlWRKY33-LlHSFA4-LlCAT2 module confers resistance to Botrytis cinerea in lily.

Gray mold caused by Botrytis cinerea is one of the major threats in lily production. However, limited information is available about the underlying defense mechanism against B. cinerea in lily. Here, we characterized a nuclear-localized class A heat stress transcription factor (HSF)-LlHSFA4 from lily (Lilium longiflorum), which positively regulated the response to B. cinerea infection. LlHSFA4 transcript and its promoter activity were increased by B. cinerea infection in lily, indicating its involvement in the response to B. cinerea. Virus-induced gene silencing (VIGS) of LlHSFA4 impaired the resistance of lily to B. cinerea. Consistent with its role in lily, overexpression of LlHSFA4 in Arabidopsis (Arabidopsis thaliana) enhanced the resistance of transgenic Arabidopsis to B. cinerea infection. Further analysis showed that LlWRKY33 directly activated LlHSFA4 expression. We also found that both LlHSFA4 and LlWRKY33 positively regulated plant response to B. cinerea through reducing cell death and H2O2 accumulation and activating the expression of the reactive oxygen species (ROS) scavenging enzyme gene LlCAT2 (Catalase 2) by binding its prompter, which might contribute to reducing H2O2 accumulation in the infected area. Taken together, our data suggested that there may be a LlWRKY33-LlHSFA4-LlCAT2 regulatory module which confers B. cinerea resistance via reducing cell death and the ROS accumulation.

Cover Image:

Southern corn leaf blight (SCLB), caused by Bipolaris maydis (B. maydis), is one of the most devastating diseases aff ecting maize production. Zhang et al. demonstrate that a cytosol‐localized ascorbate peroxidase 1 (ZmAPX1) confers SCLB resistance by reducing H2O2 accumulation and activating the JA‐mediated defense signaling pathway. The cover image shows that ZmAPX1 provides a barrier for maize against B. maydis infection (Page 1196–1211).

Piriformospora indica colonization promotes the root growth of Dimocarpus longan seedlings

As a mutualistic endophyte, the host range of Piriformospora indica is very broad and its promoting effects on host plant growth and development have been increasingly reported. In this study, we investigated the influence of P. indica colonization on the growth of longan (Dimocarpus longan Lour.) seedlings and found that the root length and root number, chlorophyll content in leaf, and flavonoid contents in both leaf and stem of P. indica colonized longan seedlings were significantly higher than those of the noncolonized controls. To reveal its promoting mechanism, activities of several antioxidant enzymes, contents of some antioxidant related molecules and phytohormones were measured. Results showed that the peroxidase (POD) activity (indicator of rooting ability) and indole-3-acetic acid (IAA, a root growth promoting phytohormone) content in P. indica colonized roots were both significantly higher than controls. Moreover, P. indica colonization reduced the accumulations of hydrogen peroxide (H2O2), malondialdehyde (MDA) and jasmonic acid (JA) in longan roots. Our results indicated that P. indica colonization improved the rooting ability of longan through inducing the IAA biosynthesis and POD activity, facilitated and accelerated the root growth by reducing H2O2 accumulation and suppressing the JA accumulation.

PeMPK7 is induced in an ROS-dependent manner and confers poplar para-hydroxybenzoic acid stress resistance through the removal of ROS

The continuous cropping of poplar plantations will accumulate allelochemicals such as para-hydroxybenzoic acid (pHBA) in the soil, which restricts the growth and biomass of poplar. Mitogen-activated protein kinase (MAPK) cascades are widely involved in environmental stress responses in plants. However, little is known about the role of MAPK cascade genes in the response to pHBA stress until now. Here, a novel group C MAPK gene, PeMPK7, from Populus × euramericana was characterized. The PeMPK7 transcript was induced by exogenous pHBA and H2O2, and its induced transcription by pHBA was blocked by the reactive oxygen species (ROS) manipulators. Overexpression of PeMPK7 in poplar alleviated the phenotypic injury induced by pHBA stress, reduced H2O2 accumulation and alleviated the oxidative injury in leaf cells. The expression of the encoding genes and enzyme activity of the antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) were upregulated in transgenic poplars under pHBA conditions, which will help to remove excessive ROS. In addition, the enhanced soluble sugar and free proline in transgenic poplar seedlings may contribute to maintaining osmotic homeostasis under pHBA stress. Furthermore, the interaction between PeMPKK3 and PeMPK7 was confirmed. A schematic model was proposed to show the regulatory mechanism of PeMPK7 in response to pHBA stress.

The Heat Stress Transcription Factor LlHsfA4 Enhanced Basic Thermotolerance through Regulating ROS Metabolism in Lilies (Lilium Longiflorum).

Heat stress severely affects the annual agricultural production. Heat stress transcription factors (HSFs) represent a critical regulatory juncture in the heat stress response (HSR) of plants. The HsfA1-dependent pathway has been explored well, but the regulatory mechanism of the HsfA1-independent pathway is still under-investigated. In the present research, HsfA4, an important gene of the HsfA1-independent pathway, was isolated from lilies (Lilium longiflorum) using the RACE method, which encodes 435 amino acids. LlHsfA4 contains a typical domain of HSFs and belongs to the HSF A4 family, according to homology comparisons and phylogenetic analysis. LlHsfA4 was mainly expressed in leaves and was induced by heat stress and H2O2 using qRT-PCR and GUS staining in transgenic Arabidopsis. LlHsfA4 had transactivation activity and was located in the nucleus and cytoplasm through a yeast one hybrid system and through transient expression in lily protoplasts. Over expressing LlHsfA4 in Arabidopsis enhanced its basic thermotolerance, but acquired thermotolerance was not achieved. Further research found that heat stress could increase H2O2 content in lily leaves and reduced H2O2 accumulation in transgenic plants, which was consistent with the up-regulation of HSR downstream genes such as Heat stress proteins (HSPs), Galactinol synthase1 (GolS1), WRKY DNA binding protein 30 (WRKY30), Zinc finger of Arabidopsis thaliana 6 (ZAT6) and the ROS-scavenging enzyme Ascorbate peroxidase 2 (APX2). In conclusion, these results indicate that LlHsfA4 plays important roles in heat stress response through regulating the ROS metabolism in lilies.

Citrus rootstocks modify scion antioxidant system under drought and heat stress combination

Key messageThe activation of the antioxidant system under stress combination is a transmissible trait from the rootstock to the scion. Therefore, rootstock selection is key to improve crop performance and a sustainable production under changing climate conditions.Climate change is altering weather conditions such as mean temperatures and precipitation patterns. Rising temperatures, especially in certain regions, accelerates soil water depletion and increases drought risk, which affects agriculture yield. Previously, our research demonstrated that the citrus rootstock Carrizo citrange (Citrus sinensis × Poncirus trifoliata) is more tolerant than Cleopatra mandarin (C. reshni) to drought and heat stress combination, in part, due to a higher activation of the antioxidant system that alleviated damage produced by oxidative stress. Here, by using reciprocal grafts of both genotypes, we studied the importance of the rootstock on scion performance and antioxidant response under this stress combination. Carrizo rootstock, under stress combination, positively influenced Cleopatra scion by reducing H2O2 accumulation, increasing superoxide dismutase (SOD) and ascorbate peroxidase (APX) enzymatic activities and inducing SOD1, APX2 and catalase (CAT) protein accumulations. On the contrary, Cleopatra rootstock induced decreases in APX2 expression, CAT activity and SOD1, APX2 and CAT contents on Carrizo scion. Taken together, our findings indicate that the activation of the antioxidant system under stress combination is a transmissible trait from the rootstock to the scion and highlight the importance of the rootstock selection to improve crop performance and maintain citrus yield under the current scenario of climate change.

Kinetin mitigates Cd-induced damagesto growth, photosynthesis and PS II photochemistry of Trigonella seedlings by up-regulating ascorbate-glutathione cycle.

Cytokinins (CKs) plays a key role in plant adaptation over a range of different stress conditions. Here, we analyze the effects of a cytokinin (i.e., kinetin, KN) on the growth, photosynthesis (rate of O2 evolution), PS II photochemistry and AsA–GSH cycle in Trigonella seedlings grown under cadmium (Cd) stress. Trigonella seeds were sown in soil amended with 0, 3 and 9 mg Cd kg-1 soil, and after 15 days resultant seedlings were sprayed with three doses of KN, i.e.,10 μM (low, KNL), 50 μM (medium, KNM) and 100 μM (high, KNH); subsequent experiments were performed after 15 days of KN application, i.e., 30 days after sowing. Cadmium toxicity induced oxidative damage as shown by decreased seedling growth and photosynthetic pigment production (Chl a, Chl b and Car), rates of O2-evolution, and photochemistry of PS II of Trigonella seedlings, all accompanied by an increase in H2O2 accumulation. Supplementation with doses of KN at KNL and KNM significantly improved the growth and photosynthetic activity by reducing H2O2 accumulation through the up-regulation AsA–GSH cycle. Notably, KNL and KNM doses stimulated the rate of enzyme activities of APX, GR and DHAR, involved in the AsA–GSH cycle thereby efficiently regulates the level of AsA and GSH in Trigonella grown under Cd stress. The study concludes that KN can mitigate the damaging effects of Cd stress on plant growth by maintaining the redox status (>ratios: AsA/DHA and GSH/GSSG) of cells through the regulation of AsA-GSH cycle at 10 and 50 μM KN under Cd stress conditions. At 100 μM KN, the down-regulation of AsA-GSH cycle did not support the growth and PS II activity of the test seedlings.

Silicon-induced mitigation of drought stress in peanut genotypes (Arachis hypogaea L.) through ion homeostasis, modulations of antioxidative defense system, and metabolic regulations

Drought stress considered as a major environmental constraint that frequently limits crop production globally. In the current investigation, drought stress-induced alterations in growth, ion homeostasis, photosynthetic pigments, organic osmolytes, reactive oxygen species (ROS) generation, antioxidative components, and metabolic profile were examined in order to assess the role of silicon (Si) in mitigation of drought effects and to understand the drought adaptive mechanism in two contrasting peanut genotypes (GG7: fast growing and tall, TG26: slow growing and semi-dwarf). Si application significantly improved the leaf chlorophyll content, relative water content % (RWC %), growth and biomass in GG7 compared with TG26 genotype under water stress. Si supplementation considerably promotes the uptake and transport of mineral nutrients under drought condition in both the genotypes, which eventually promote plant growth. Exogenous application of Si protects the photosynthetic pigments from oxidative damage by reducing membrane lipid peroxidation and either maintained or reduced H2O2 accumulation in both the genotypes. The activity of enzymatic antioxidants like superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and glutathione reductase (GR) and non-enzymatic antioxidants like ascorbate (AsA) and glutathione (GSH) were either maintained or increased in both the genotypes in response to Si under drought as compared to those without Si. Silicon-induced higher accumulation of metabolites mainly sugars and sugar alcohols (talose, mannose, fructose, sucrose, cellobiose, trehalose, pinitol, and myo-inositol), amino acids (glutamic acid, serine, histidine, threonine, tyrosine, valine, isoleucine, and leucine) in GG7 genotype as compared to TG26, provides osmo-protection. Moreover, Si application increased phytohormones levels such as indole-3-acetic acid (IAA), gibberellic acid (GA3), jasmonic acid (JA), and zeatin in GG7 genotype under drought stress compared to non-Si treated seedlings suggesting its involvement in signaling pathways for drought adaptation and tolerance. Noteworthy increment in polyphenols (chlorogenic acid, caffeic acid, ellagic acid, rosmarinic acid, quercetin, coumarin, naringenin, and kaempferol) in the Si treated seedlings of GG7 genotype as compared to TG26 under drought stress suggests an efficient mechanism of ROS sequestration in GG7 genotype. Our findings provide comprehensive information on physiological, biochemical, and metabolic dynamics associated with Si-mediated water stress tolerance in peanut. This study indicates that the drought tolerance efficacy of peanut genotypes can be improved by Si application.

A Plasmopara viticola RXLR effector targets a chloroplast protein PsbP to inhibit ROS production in grapevine.

Pathogens secrete a large number of effectors that manipulate host processes to create an environment conducive to pathogen colonization. However, the underlying mechanisms by which Plasmopara viticola effectors manipulate host plant cells remain largely unclear. In this study, we reported that RXLR31154, a P. viticola RXLR effector, was highly expressed during the early stages of P. viticola infection. In our study, stable expression of RXLR31154 in grapevine (Vitis vinifera) and Nicotiana benthamiana promoted leaf colonization by P. viticola and Phytophthora capsici, respectively. By yeast two-hybrid screening, the 23-kDa oxygen-evolving enhancer 2 (VpOEE2 or VpPsbP), encoded by the PsbP gene, in Vitis piasezkii accession Liuba-8 was identified as a host target of RXLR31154. Overexpression of VpPsbP enhanced susceptibility to P. viticola in grapevine and P. capsici in N. benthamiana, and silencing of NbPsbPs, the homologs of PsbP in N. benthamiana, reduced P. capcisi colonization, indicating that PsbP is a susceptibility factor. RXLR31154 and VpPsbP protein were co-localized in the chloroplast. Moreover, VpPsbP reduced H2 O2 accumulation and activated the 1 O2 signaling pathway in grapevine. RXLR31154 could stabilize PsbP. Together, our data revealed that RXLR31154 reduces H2 O2 accumulation and activates the 1 O2 signaling pathway through stabilizing PsbP, thereby promoting disease.

Priming with γ-Aminobutyric Acid against Botrytis cinerea Reshuffles Metabolism and Reactive Oxygen Species: Dissecting Signalling and Metabolism.

The stress-inducible non-proteinogenic amino acid γ-aminobutyric acid (GABA) is known to alleviate several (a)biotic stresses in plants. GABA forms an important link between carbon and nitrogen metabolism and has been proposed as a signalling molecule in plants. Here, we set out to establish GABA as a priming compound against Botrytis cinerea in Arabidopsis thaliana and how metabolism and reactive oxygen species (ROS) are influenced after GABA treatment and infection. We show that GABA already primes disease resistance at low concentrations (100 µM), comparable to the well-characterized priming agent β-Aminobutyric acid (BABA). Treatment with GABA reduced ROS burst in response to flg22 (bacterial peptide derived from flagellum) and oligogalacturonides (OGs). Plants treated with GABA showed reduced H2O2 accumulation after infection due to increased activity of catalase and guaiacol peroxidase. Contrary to 100 µM GABA treatments, 1 mM exogenous GABA induced endogenous GABA before and after infection. Strikingly, 1 mM GABA promoted total and active nitrate reductase activity whereas 100 µM inhibited active nitrate reductase. Sucrose accumulated after GABA treatment, whereas glucose and fructose only accumulated in treated plants after infection. We propose that extracellular GABA signalling and endogenous metabolism can be separated at low exogenous concentrations.

HaASR1 gene cloned from a desert shrub, Haloxylon ammodendron, confers drought tolerance in transgenic Arabidopsis thaliana

Abiotic stresses pose adverse impact on plant growth and development and exploring the stress tolerance genes from dominant species is still urgent. The desert shrub, Haloxylon ammodendron, a C4 and succulent xero-halophyte species, has great drought and salt tolerance. Here, we reported the isolation and functions analysis of HaASR1 (abscisic acid, stress, and ripening) from H. ammodendron. HaASR1 interacts with HaPrxQ and HaBADH and may function in preventing PrxQ and BADH from denaturation in plants under drought stress. Overexpression of HaASR1 reduces the sensitivity of Arabidopsis plants to exogenous ABA and decreases the endogenous ABA level through down-regulating ABA biosynthesis genes under drought stress, which making the plants display less inhibition of shoot growth and enhanced drought tolerance. Overexpression of HaASR1 significantly enhances photosynthetic capacity of Arabidopsis plants under drought stress by reducing H2O2 accumulation through up-regulating AtCAT2, AtAPX1 and AtPrxQ and maintaining relatively integrity of chloroplasts. Overexpression of HaASR1 also significantly improves water-retaining ability. Taken together, we propose that HaASR1 plays critical roles in plant drought stress adaptation by making the plants partially defective in ABA signaling and participating in drought tolerance regulation through nonstomatal pathways, enhancing ROS-scavenging ability and water-retaining ability, which is different from the functions of previously reported ASR genes from other plant species. This research also reveals that HaASR1 gene from the desert shrub, Haloxylon ammodendron, has potential application values in improving abiotic stresses of other plant species by genetic engineering.