Salicylic Acid Synthesis Research Articles

Candidate genes for grape white rot resistance based on SMRT and Illumina sequencing

BackgroundWhite rot is one of the most dangerous fungal diseases and can considerably affect grape berry production and quality. However, few studies have focused on this disease, and thus, finding candidate white rot resistance genes is of great importance for breeding resistant grapevine cultivars. Based on field observations and indoor experiments, the cultivars “Victoria” and “Zhuosexiang” showed significant differences in white rot resistance. For understanding the molecular mechanisms behind it, different phenotypes of grapevine leaves were used for RNA sequencing via Illumina and single-molecule real-time (SMRT) sequencing technology.ResultsA transcript library containing 53,906 reads, including known and novel transcripts, was constructed following the full-length transcriptome sequencing of the two grapevine cultivars. Genes involved in salicylic acid (SA) and jasmonic acid (JA) synthesis pathways showed different expression levels. Furthermore, four key transcription factors (TFs), NPR1, TGA4, Pti6, and MYC2, all involved in the SA and JA signal pathways were identified, and the expression profile revealed the different regulation of the pathogenesis related protein1 (PR1) resistance gene, as mediated by the four TFs.ConclusionsFull-length transcript sequencing can substantially improve the accuracy and integrity of gene prediction and gene function research in grapevine. Our results contribute to identify candidate resistance genes and improve our understanding of the genes and regulatory mechanisms involved in grapevine resistance to white rot.

Polyamine-Induced Hormonal Changes in eds5 and sid2 Mutant Arabidopsis Plants.

Polyamines are multifaceted compounds which play a role in regulating plant growth and stress tolerance in interactions with plant hormones. The aim of the present study was to reveal how exogenous polyamines influence the synthesis of salicylic acid, with a special emphasis on the effect of salicylic acid deficiency on the polyamine metabolism and polyamine-induced changes in other plant hormone contents. Our hypothesis was that the individual polyamines induced different changes in the polyamine and salicylic acid metabolism of the wild type and salicylic acid-deficient Arabidopsis mutants, which in turn influenced other hormones. To our knowledge, such a side-by-side comparison of the influence of eds5-1 and sid2-2 mutations on polyamines has not been reported yet. To achieve our goals, wild and mutant genotypes were tested after putrescine, spermidine or spermine treatments. Polyamine and plant hormone metabolism was investigated at metabolite and gene expression levels. Individual polyamines induced different changes in the Arabidopsis plants, and the responses were also genotype-dependent. Polyamines upregulated the polyamine synthesis and catabolism, and remarkable changes in hormone synthesis were found especially after spermidine or spermine treatments. The sid2-2 mutant showed pronounced differences compared to Col-0. Interactions between plant hormones may also be responsible for the observed differences.

Role of Salicylic Acid and Components of the Phenylpropanoid Pathway in Basal and Cultivar-Related Resistance of Oilseed Rape (Brassica napus) to Verticillium longisporum.

Enhanced resistance is a key strategy of controlling ‘Verticillium stem striping’ in Brassica napus caused by the soil-borne vascular pathogen Verticillium longisporum. The present study analyses the role of a broad range of components in the phenylpropanoid and salicylic acid (SA) pathways in basal and cultivar-related resistance of B. napus towards V. longisporum. A remarkable increase of susceptibility to V. longisporum in SA-deficient transgenic NahG plants indicated an essential role of SA in basal resistance of B. napus to V. longisporum. Accordingly, elevated SA levels were also found in a resistant and not in a susceptible cultivar during early asymptomatic stages of infection (7 dpi), which was associated with increased expression of PR1 and PR2. In later symptomatic stages (14 or 21 dpi), SA responses did not differ anymore between cultivars varying in resistance. In parallel, starting at 7 dpi, an overall increase in phenylpropanoid syntheses developed in the resistant cultivar, including the activity of some key enzymes, phenylalanine ammonium lyase (PAL), cinnamyl alcohol dehydrogenase (CAD) and peroxidase (POX) and the expression of key genes, PAL4, CCoAMT, CCR, POX. As a consequence, a remarkable increase in the levels of phenolic acids (t-cinnamic acid, p-coumaric acid, caffeic acid, ferulic acid, sinapic acid) occurred associated with cultivar resistance. A principal component analysis including all 27 traits studied indicated that component 1 related to SA synthesis (PR1, PR2, POX, level of free SA) and component 2 related to lignin synthesis (level of free ferulic acid, free p-coumaric acid, conjugated t-cinnamic acid) were the strongest factors to determine cultivar-related resistance. This study provides evidence that both SA and phenolic acid synthesis are important in cultivar-related resistance, however, with differential roles during asymptomatic and symptomatic stages of infection.

Drought Stress–induced Physiological and Metabolic Changes in Leaves of Two Oil Tea Cultivars

Oil tea (Camellia oleifera) is an important edible oil tree. However, its growth and yield are strongly limited by drought. This study investigated the physiological and metabolic responses of two common oil tea cultivars, Huajin and Changlin53, to moderate and severe drought stress. Based on the photosynthetic and physiological indices, ‘Changlin53’ may be more tolerant to drought than ‘Huajin’. A total of 41 key metabolites induced by drought stress, including 12 amino acids, 12 organic acids, 10 carbohydrates, 3 fatty acids, and 4 phenols, have been identified by liquid chromatography-mass spectrometry. Under moderate drought stress, the contents of carbohydrates, amino acids, and some organic acids in ‘Changlin53’ were significantly increased; however, under severe drought stress, the contents of soluble sugars were decreased and the synthesis ability of amino acids and organic acids were enhanced. The glutamic acid–mediated proline biosynthesis pathway and salicylic acid synthesis were continuously upregulated in ‘Changlin53’ under moderate and severe drought stress, which could regulate osmotic pressure and maintain intracellular environmental stability. Under moderate drought stress, the contents of monosaccharides, amino acids, and organic acids increased in ‘Huajin’ leaves. Furthermore, the shikimic acid–mediated secondary metabolite synthesis pathway was weakened. More secondary metabolites were used to increase glycolysis and tricarboxylic acid cycle to accelerate energy production and to enhance the glutamic acid–mediated proline biosynthesis pathway, which are necessary to increase osmotic regulation. Under severe drought stress, the contents of carbohydrates, organic acids, and some amino acids were significantly decreased in ‘Huajin’ leaves, indicating serious damage. These results deepened our understanding of the mechanisms involved in oil tea drought tolerance, which will help improve water management of oil tea seedlings.

CsBZIP40, a BZIP transcription factor in sweet orange, plays a positive regulatory role in citrus bacterial canker response and tolerance.

Citrus bacterial canker (CBC) caused by Xanthomonas citri subsp. citri (Xcc) is a systemic bacterial disease that affects citrus plantations globally. Biotic stress in plants has been linked to a group of important transcription factors known as Basic Leucine Zippers (BZIPs). In this study, CsBZIP40 was functionally characterized by expression analysis, including induction by Xcc and hormones, subcellular localization, over-expression and RNAi silencing. CsBZIP40 belongs to group D of the CsBZIP family of transcription factors and localizes in the nucleus, potentially serving as a transcriptional regulator. In wild type (WT) plants CsBZIP40 can be induced by plant hormones in addition to infection by Xcc which has given insight into its involvement in CBC. In the present study, over-expression of CsBZIP40 conferred resistance to Xcc while its silencing led to Xcc susceptibility. Both over-expression and RNAi affected salicylic acid (SA) production and expression of the genes involved in the SA synthesis and signaling pathway, in addition to interaction of CsBZIP40 with CsNPR1, as detected by a GST pull-down assay. Taken together, the results of this study confirmed the important role of CsBZIP40 in improving resistance to citrus canker through the SA signaling pathway by the presence of NPR1 to activate PR genes. Our findings are of potential value in the breeding of tolerance to CBC in citrus fruits.

Calcium-dependent protein kinase 5 links calcium signaling with N-hydroxy-l-pipecolic acid- and SARD1-dependent immune memory in systemic acquired resistance.

Systemic acquired resistance (SAR) prepares infected plants for faster and stronger defense activation upon subsequent attacks. SAR requires an information relay from primary infection to distal tissue and the initiation and maintenance of a self-maintaining phytohormone salicylic acid (SA)-defense loop. In spatial and temporal resolution, we show that calcium-dependent protein kinase CPK5 contributes to immunity and SAR. In local basal resistance, CPK5 functions upstream of SA synthesis, perception, and signaling. In systemic tissue, CPK5 signaling leads to accumulation of SAR-inducing metabolite N-hydroxy-L-pipecolic acid (NHP) and SAR marker genes, including Systemic Acquired Resistance Deficient 1 (SARD1) Plants of increased CPK5, but not CPK6, signaling display an 'enhanced SAR' phenotype towards a secondary bacterial infection. In the sard1-1 background, CPK5-mediated basal resistance is still mounted, but NHP concentration is reduced and enhanced SAR is lost. The biochemical analysis estimated CPK5 half maximal kinase activity for calcium, K50 [Ca2+ ], to be c.100nM, close to the cytoplasmic resting level. This low threshold uniquely qualifies CPK5 to decode subtle changes in calcium, a prerequisite to signal relay and onset and maintenance of priming at later time points in distal tissue. Our data explain why CPK5 functions as a hub in basal and systemic plant immunity.

PUB4, a CERK1-Interacting Ubiquitin Ligase, Positively Regulates MAMP-Triggered Immunity in Arabidopsis.

Lysin motif (LysM) receptor-like kinase CERK1 is a co-receptor essential for plant immune responses against carbohydrate microbe-associated molecular patterns (MAMPs). Concerning the immediate downstream signaling components of CERK1, receptor-like cytoplasmic kinases such as PBL27 and other RLCK VII members have been reported to regulate immune responses positively. In this study, we report that a novel CERK1-interacting E3 ubiquitin ligase, PUB4, is also involved in the regulation of MAMP-triggered immune responses. Knockout of PUB4 resulted in the alteration of chitin-induced defense responses, indicating that PUB4 positively regulates reactive oxygen species generation and callose deposition but negatively regulates MAPK activation and defense gene expression. On the other hand, detailed analyses of a double knockout mutant of pub4 and sid2, a mutant of salicylic acid (SA) synthesis pathway, showed that the contradictory phenotype of the pub4 mutant was actually caused by abnormal accumulation of SA in this mutant and that PUB4 is a positive regulator of immune responses. The present and recent findings on the role of PUB4 indicate that PUB4 is a unique E3 ubiquitin ligase involved in the regulation of both plant immunity and growth/development.

Synthesis of Three Bioactive Aromatic Compounds by Introducing Polyketide Synthase Genes into Engineered Escherichia coli.

Intermediates in aromatic amino acid biosynthesis can serve as substrates for the synthesis of bioactive compounds. In this study we used two intermediates in the shikimate pathway of Escherichia coli, chorismate and anthranilate, to synthesize three bioactive compounds: 4-hydroxycoumarin (4-HC), 2,4-dihydroxyquinoline (DHQ), and 4-hydroxy-1-methyl-2(1H)-quinolone (NMQ). We introduced genes for the synthesis of salicylic acid from chorismate to supply the substrate for 4-HC and the gene encoding N-methyltransferase for the synthesis of N-methylanthranilate from anthranilate. Polyketide synthases and coenzyme (Co)A ligases were tested to determine the optimal combination of genes for the synthesis of each compound. We also tested several constructs and identified the best one for increasing levels of endogenous substrates for chorismate, anthranilate, and malonyl-CoA. With the use of these strategies, 255.4 mg/L 4-HC, 753.7 mg/L DHQ, and 17.5 mg/L NMQ were synthesized. This work provides a basis for the synthesis of diverse coumarin and quinoline derivatives with potential medical applications.

Changes in the composition and function of bacterial communities during vermicomposting may explain beneficial properties of vermicompost

Vermicomposting is the process by which organic waste is broken down through the synergistic actions of earthworms and microbial communities. Although vermicomposting has been shown to effectively reduce organic biomass and generate high-quality fertilizer for plants, little is known about the bacterial communities that are involved in this decomposition process. Since optimization of vermicomposting for commercial use necessitates additional knowledge of the underlying biological processes, this study sought to characterize the bacterial succession involved in the vermicomposting of Scotch broom (Cytisus scoparius), a leguminous shrub that has become invasive around the world with consequences for the dynamics and productivity of the ecosystems they occupy. Scotch broom was processed in a pilot-scale vermireactor for 91 days with the earthworm species Eisenia andrei. Samples were taken at the initiation of vermicomposting, and days 14, 42 and 91, representing both active and mature stages of vermicomposting. Significant changes (P < 0.0001) in the bacterial community composition (richness and evenness) were observed throughout the process. Increases in taxonomic diversity were accompanied by increases in functional diversity of the bacterial community, including metabolic capacity, streptomycin and salicylic acid synthesis, and nitrification. These results highlight the role of bacterial succession during the vermicomposting process and provide evidence of microbial functions that may explain the beneficial effects of vermicompost on soil and plants.

Identification of salicylic acid-independent responses in an Arabidopsis phosphatidylinositol 4-kinase beta double mutant.

We have recently shown that an Arabidopsis thaliana double mutant of type III phosphatidylinositol-4-kinases (PI4Ks), pi4kβ1β2, constitutively accumulated a high level of salicylic acid (SA). By crossing this pi4kβ1β2 double mutant with mutants impaired in SA synthesis (such as sid2 impaired in isochorismate synthase) or transduction, we demonstrated that the high SA level was responsible for the dwarfism phenotype of the double mutant. Here we aimed to distinguish between the SA-dependent and SA-independent effects triggered by the deficiency in PI4Kβ1 and PI4Kβ2. To achieve this we used the sid2pi4kβ1β2 triple mutant. High-throughput analyses of phytohormones were performed on this mutant together with pi4kβ1β2 and sid2 mutants and wild-type plants. Responses to pathogens, namely Hyaloperonospora arabidopsidis, Pseudomonas syringae and Botrytis cinerea, and also to the non-host fungus Blumeria graminis, were also determined. Callose accumulation was monitored in response to flagellin. We show here the prominent role of high SA levels in influencing the concentration of many other tested phytohormones, including abscisic acid and its derivatives, the aspartate-conjugated form of indole-3-acetic acid and some cytokinins such as cis-zeatin. We show that the increased resistance of pi4kβ1β2 plants to the host pathogens H. arabidopsidis, P. syringae pv. tomato DC3000 and Bothrytis cinerea is dependent on accumulation of high SA levels. In contrast, accumulation of callose in pi4kβ1β2 after flagellin treatment was independent of SA. Concerning the response to Blumeria graminis, both callose accumulation and fungal penetration were enhanced in the pi4kβ1β2 double mutant compared to wild-type plants. Both of these processes occurred in an SA-independent manner. Our data extensively illustrate the influence of SA on other phytohormone levels. The sid2pi4kβ1β2 triple mutant revealed the role of PI4Kβ1/β2 per se, thus showing the importance of these enzymes in plant defence responses.

BTB-BACK Domain E3 Ligase MdPOB1 Suppresses Plant Pathogen Defense against Botryosphaeria dothidea by Ubiquitinating and Degrading MdPUB29 Protein in Apple.

Apple ring rot is a severe disease that affects the yield and quality of apple fruits worldwide. However, the underlying molecular mechanism that involved in this process still remains largely unexplored. Here, we report that apple POZ/BTB CONTAINING-PROTEIN 1 (MdPOB1), a BTB-BACK domain E3 ligase protein, functions to suppress apple pathogen defense against Botryosphaeria dothidea (B. dothidea). Both in vitro and in vivo assays indicated that MdPOB1 interacted directly with and degraded apple U-box E3 ligase MdPUB29, a well-established positive regulator of plant innate immunity, through the ubiquitin/26S proteasome pathway. A series of transgenic analyses in apple fruits demonstrated that MdPOB1 affected apple pathogen defense against B. dothidea at least partially, if not completely, via regulating MdPUB29. Additionally, it was found that the apple pathogen defense against B. dothidea was correlated with the H2O2 contents and the relative expression of salicylic acid (SA) synthesis- and SA signaling-related genes, which might be regulated via degradation of MdPUB29 by MdPOB1. Overall, our findings provide new insights into the mechanism of the MdPOB1 modulation of apple ring rot resistance, which occur by directly regulating potential downstream target protein MdPUB29 for proteasomal degradation in apple.

Salicylic acid‐sensitised titanium dioxide for photocatalytic degradation of fast green FCF under visible light irradiation

Synthesis and modification of salicylic acid (SA) adsorbed titanium dioxide (SA-TiO2) was proposed through a sol–gel reaction following annealing treatment. The prepared SA-TiO2 exhibited visible light absorption because of the formation of an intramolecular ligand to metal charge transfer transition. The photocatalytic activity of SA-TiO2 for the degradation of fast green FCF under visible light irradiation was evaluated. The experimental results suggest that the enhanced photocatalytic activity of SA-TiO2 is attributed to the visible light absorption ability. Moreover, •OH and •O2 − radicals are major active species involved in dye degradation. The practicality of SA-TiO2 was validated through the degradation of various dyestuff in environmental water samples under visible light irradiation.

Suppression of tryptophan synthase activates cotton immunity by triggering cell death via promoting SA synthesis.

Primary metabolism plays an important role in plant growth and development, however the relationship between primary metabolism and the adaptive immune response is largely unknown. Here, we employed RNA interference (RNAi), virus-induced gene silencing (VIGS) technology, phytohormone profiling, genetic studies, and transcriptome and metabolome analysis to investigate the function of the tryptophan synthesis pathway in the resistance of cotton to V.dahliae. We found that knock-down of GbTSA1 (Tryptophan Synthase α) and GbTSB1 (tryptophan synthase β) induced a spontaneous cell death phenotype in a salicylic acid (SA)-dependent manner and enhanced resistance to V.dahliae in cotton plants. Metabolome analysis showed that indole and indolic metabolites were highly accumulated in GbTSA1- or GbTSB1-silenced plants. Transcriptomic analysis showed that exogenous indole promotes the expression levels of genes involved in SA synthesis and the defense response. Similarly, indole application strongly enhanced cotton resistance to V.dahliae. These results suggested that metabolic intermediates in the Trp synthesis pathway may be a signal to activate SA synthesis. These results also provided a strategy to elicit plant defense responses by the application of indole.

Transcriptome analysis reveals pathways facilitating the growth of tobacco powdery mildew in Arabidopsis

Powdery mildew (PM) fungi are biotrophic pathogens that rely on living hosts to survive and thrive. However, their colonization is restricted by host defenses at both the penetration and post-penetration stages. The tobacco PM strain Golovinomyces cichoracearum (Gc) SICAU1 has overcome penetration resistance of Arabidopsis but its growth is arrested by post-penetration resistance. While Gc SICAU1 only poorly grows in Arabidopsis Col-0 wild-type plants, it can sustainably grow for more than 20 days on the same infected leaves of the double mutant pad4–1 sid2–1 that is defective in both the synthesis and signaling of salicylic acid (SA). To understand the underlying molecular mechanisms, we conducted a comparative transcriptome analysis between Col-0 and pad4–1 sid2–1 in response to Gc SICAU1. We found that 4811 genes were differentially expressed more than four-fold between any two of the measured seven time points (0, 1, 3, 6, 8, 10 and 12 days post-inoculation). Gene expression pattern analysis suggests that differential expression of 348 genes and 190 genes may explain resistance in Col-0 and susceptibility in pad4–1 sid2–1, respectively. Gene Ontology (GO) analysis suggests that Gc SICAU1 might be arrested in Col-0 by both pattern-triggered immunity and SA-dependent defense. By contrast, its sustained growth in pad4–1 sid2–1 may be attributable to the activation of a detoxification pathway that is normally repressed by the SA-signaling pathway. Taken together, our results suggest that multiple distinct, yet interconnected pathways control the growth of tobacco powdery mildew in Arabidopsis.

The Salt-Stress Response of the Transgenic Plum Line J8-1 and Its Interaction with the Salicylic Acid Biosynthetic Pathway from Mandelonitrile.

Salinity is considered as one of the most important abiotic challenges that affect crop productivity. Plant hormones, including salicylic acid (SA), are key factors in the defence signalling output triggered during plant responses against environmental stresses. We have previously reported in peach a new SA biosynthetic pathway from mandelonitrile (MD), the molecule at the hub of the cyanogenic glucoside turnover in Prunus sp. In this work, we have studied whether this new SA biosynthetic pathway is also present in plum and the possible role this pathway plays in plant plasticity under salinity, focusing on the transgenic plum line J8-1, which displays stress tolerance via an enhanced antioxidant capacity. The SA biosynthesis from MD in non-transgenic and J8-1 micropropagated plum shoots was studied by metabolomics. Then the response of J8-1 to salt stress in presence of MD or Phe (MD precursor) was assayed by measuring: chlorophyll content and fluorescence parameters, stress related hormones, levels of non-enzymatic antioxidants, the expression of two genes coding redox-related proteins, and the content of soluble nutrients. The results from in vitro assays suggest that the SA synthesis from the MD pathway demonstrated in peach is not clearly present in plum, at least under the tested conditions. Nevertheless, in J8-1 NaCl-stressed seedlings, an increase in SA was recorded as a result of the MD treatment, suggesting that MD could be involved in the SA biosynthesis under NaCl stress conditions in plum plants. We have also shown that the plum line J8-1 was tolerant to NaCl under greenhouse conditions, and this response was quite similar in MD-treated plants. Nevertheless, the MD treatment produced an increase in SA, jasmonic acid (JA) and reduced ascorbate (ASC) contents, as well as in the coefficient of non-photochemical quenching (qN) and the gene expression of Non-Expressor of Pathogenesis-Related 1 (NPR1) and thioredoxin H (TrxH) under salinity conditions. This response suggested a crosstalk between different signalling pathways (NPR1/Trx and SA/JA) leading to salinity tolerance in the transgenic plum line J8-1.

Isoproturon-Induced Salicylic Acid Confers Arabidopsis Resistance to Isoproturon Phytotoxicity and Degradation in Plants.

This study identified the effect of salicylic acid on degradation of isoproturon in Arabidopsis. Three T-DNA insertion mutant lines pal1- 1, pal1- 2, and eps1- 1 defective in salicylic acid synthesis were tested, which showed higher isoproturon accumulation and a toxic symptom in the mutants. When treated with 5 mg/L salicylic acid, these lines displayed a lower level of isoproturon and showed an attenuated toxic symptom. An RNA-sequencing study identified 2651 (1421 up and 1230 down) differentially expressed genes (DEGs) in eps1- 1 and 2211 (1556 up and 655 down) in pal1- 2 mutant plants (>2.0 fold change, p < 0.05). Some of the DEGs covered Phase I-III reaction components, like glycosyltransferases (GTs) and ATP-binding cassette transporters (ABCs). Using ultra performance liquid chromatography-time-of-flight-tandem-mass spectrometer/mass spectrometer (UPLC/Q-TOF-MS/MS), 13 Phase I and four Phase II metabolites were characterized. Of these, two metabolites 1-OH-isopropyl-benzene-O-glucoside and 4-isopropylphenol-S-2-methylbutanoyl-serine, have been identified and reported for the first time.

Transcriptome approach to understand the potential mechanisms of resistant and susceptible alfalfa (Medicago sativa L.) cultivars in response to aphid feeding

Plant breeding for resistance to agricultural pests is an essential element in the development of integrated crop management systems, however, the molecular and genetic mechanisms underlying resistance are poorly understood. In this pilot study, we conducted a transcriptomic analysis of a resistant (R) and susceptible (S) variety of alfalfa, with (+A) or without (–A) aphids (totally four treatments). We used the resistant cultivar Zhongmu 1 and the susceptible cultivar Soca. A total of 3 549 mRNAs were differentially expressed, of which 1 738 up-regulated and 1 307 down-regulated genes were identified in S+A/S–A plants, while 543 up-regulated and 331 down-regulated genes were identified in the R+A/R–A plants. KEGG analysis mapped 112 and 546 differentially expressed genes to 8 and 17 substantially enriched pathways for Zhongmu 1 and Soca, respectively. Six shared pathways were linked to plant resistance traits, including phenylpropanoid biosynthesis associated with salicylic acid synthesis, and linoleic acid metabolism associated with both jasmonic acid and flavonoid biosynthesis. Ultimately, we proposed a preliminary regulatory mechanism of alfalfa cultivar resistance response to aphids feeding based on transcriptome analyses and published documents.

Enhanced tolerance to a combination of heat stress and drought in Arabidopsis plants deficient in ICS1 is associated with modulation of photosynthetic reaction center proteins.

Plants are exposed to multiple abiotic stresses that simultaneously occur under natural environmental conditions. Studies deciphering acclimation of plants to stress combinations are, however, still scarce. ISOCHORISMATE SYNTHASE 1 (ICS1) is known as a crucial enzyme required for synthesis of salicylic acid and phylloquinone, one of the components of the photosystem I complex. Although the significance of ICS1 in the regulation of abiotic stress response and pathogen defense in plants has been evidenced in previous studies, the role of this enzyme in the acclimation of plants to stress combinations is still largely unknown. In this study, we demonstrated the enhanced tolerance of Arabidopsis salicylic acid induction deficient 2-1(sid2-1) mutant deficient in ICS1 to a combination of heat stress and drought. H2 O2 -dependent stomatal closure and accumulation of total soluble sugars are associated with the enhanced tolerance of sid2-1 plants to this stress combination. In addition, sid2-1 plants showed higher accumulation of reaction center proteins (D1 and D2) in photosystem II accompanied by enhanced expression of transcripts involved in repair of these reaction center proteins. Furthermore, investigation of chlorophyll fluorescence indicated that mechanisms for dissipating the excess energy might be activated in sid2-1 plants specifically under a combination of heat stress and drought. Taken together, our findings suggest that maintenance of photosynthetic apparatus as well as prevention of excess water loss might enhance the tolerance of sid2-1 plants deficient in ICS1 to a combination of heat stress and drought.

Mechanisms of Cr(VI) resistance by endophytic Sphingomonas sp. LK11 and its Cr(VI) phytotoxic mitigating effects in soybean (Glycine max L.)

Chromium Cr(VI) is highly toxic and leads to impaired phenotypic plasticity of economically important crops. The current study assessed an endophytic-bacteria assisted metal bio-remediation strategy to understand stress-alleviating mechanisms in Glycine max L (soybean) plants inoculated with Sphingomonas sp. LK11 under severe Cr(VI) toxicity. The screening analysis showed that high Cr concentrations (5.0 mM) slightly suppressed LK11 growth and metal uptake by LK11 cells, while significantly enhancing indole-3-acetic acid (IAA) production. Endophytic LK11 significantly upregulated its antioxidant system compared to control by enhancing reduced glutathione (GSH), catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities to counteract Cr-induced oxidative stress. Cr toxicity induced cell morphological alteration, as shown by SEM-EDX analysis and triggered significant lipid peroxidation. The interaction between LK11 and soybean in Cr-contaminated soil significantly increased plant growth attributes and down-regulated the synthesis of endogenous defense-related phytohormones, salicylic acid and abscisic acid, by 20% and 37%, respectively, and reduced Cr translocation to the roots, shoot, and leaves. Additionally, Cr-induced oxidative stress was significantly reduced in LK11-inoculated soybean, regulating metal responsive reduced GSH and enzymatic antioxidant CAT. Current findings indicate that LK11 may be a suitable candidate for the bioremediation of Cr-contaminated soil and stimulation of host physiological homeostasis.

Ralstonia solanacearum Type III Effector RipAL Targets Chloroplasts and Induces Jasmonic Acid Production to Suppress Salicylic Acid-Mediated Defense Responses in Plants.

Ralstonia solanacearum is the causal agent of bacterial wilt disease of plants. This pathogen injects more than 70 type III effector proteins called Rips (Ralstonia-injected proteins) into plant cells to succeed in infection. One of the Rips, RipAL, contains a putative lipase domain that shared homology with Arabidopsis DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1). RipAL significantly suppressed pattern-triggered immunity in leaves of Nicotiana benthamiana. Subcellular localization analyses suggest that RipAL localizes to chloroplasts and targets chloroplast lipids in plant cells. Notably, the expression of RipAL markedly increased the jasmonic acid (JA) and JA-isoleucine levels, and induced the expressions of JA-signaling marker genes in plant leaves. Simultaneously, RipAL greatly reduced the salicylic acid (SA) level and decreased the expression levels of SA-signaling marker genes. Mutations in two putative catalytic residues in the DAD1-like lipase domain abolished the ability of RipAL to induce JA production and suppress SA signaling. Infection of R. solanacearum also induced JA production and simultaneously decreased the SA level in susceptible pepper leaves in a ripAL-dependent manner. The growth of R. solanacearum enhanced in plants with silenced CaICS1, which encodes the SA synthesis enzyme isochorismate synthase 1. These results indicate that SA signaling is involved in the defense response against R. solanacearum and that R. solanacearum uses RipAL to induce JA production and suppress SA signaling in plant cells.