FaSnRK1a mediates salicylic acid pathways to enhance strawberry resistance to Botrytis cinerea

Strawberry is a major fruit crop worldwide because its nutritional and health benefits to human health,but its productivity is limited by Botrytis cinerea.Sucrose nonfermentation 1-related protein kinase 1(SnRK1)has a defense function against pathogens,but the function of SnRK1 in the defense response to B.cinerea in plants is still unclear.In this study,FaSnRK1a-OE and RNAi fruits were constructed and then inoculated with B.cinerea.The result reveals a positive role of Fa SnRK1a in the regulation of resistance to gray mold.FaSnRK1a affects SA content by regulating FaPAL1 and FaPAL2 expressions.The genes related to the SA signaling pathway(FaTGA1 and FaTGA2.1)were significantly increased/decreased in FaSnRK1a-OE or FaSnRK1a-RNAi fruit,respectively.FaSnRK1a interacted with the FaWRKY33.2 protein and negatively regulated FaWRKY33.2 expression,and FaWRKY33.2 acts as a repressor of disease resistance to B.cinerea.Finally,FaSnRK1a regulates the expression of six PR genes and the activities of antioxidant enzymes to boost defense response after B.cinerea inoculation.Our findings showed that FaSnRK1a increases the resistance of strawberry fruit to B.cinerea via SA signaling pathway and interaction with the FaWRKY33.2 transcription factor.

Salicylic acid positively regulates maize defenses against lepidopteran insects

In response to insect attack,plants use intricate signaling pathways,including phytohormones,such as jasmonate(JA),ethylene(ET),and salicylic acid(SA),to activate defenses.Maize(Zea mays)is one of the most important staple food crops around the world.Previous studies have shown that the JA and ET signaling play important roles in maize defense against insects,but little is known about whether and how SA regulates maize resistance to insect herbivores.In this study,we ectopically expressed the NahG(salicylate hydroxylase)gene in maize plants(NahG maize)to block the accumulation of SA.It was found that compared with the wild-type(WT)maize,the NahG-maize exhibited decreased resistance to the generalist insects Spodoptera litura and Spodoptera frugiperda and the specialist Mythimna separata,and the compromised resistance in the NahG maize was associated with decreased levels of defensive metabolites benzoxazinoids(Bxs)and chlorogenic acid(CA).Quantification of simulated S.litura feedinginduced JA,JA-isoleucine conjugate(JA-Ile),and ET in the WT and NahG maize indicated that SA does not regulate JA or JA-Ile,but positively controls ET.We provide evidence suggesting that the SA pathway does not crosstalk with the JA or the ET signaling in regulating the accumulation of Bxs and CA.Transcriptome analysis revealed that the bHLH,ERF,and WRKY transcription factors might be involved in SAregulated defenses.This study uncovers a novel and important phytohormone pathway in maize defense against lepidopterous larvae.

Investigating the mechanisms of isochorismate synthase:An approach to improve salicylic acid synthesis and increase resistance to Fusarium head blight in wheat

Salicylic acid(SA),a vital endogenous hormone,plays a crucial role in plant growth and the response to abiotic and biotic stress.Isochorismate synthase(ICS)and phenylalanine ammonia lyase(PAL)are critical rate-limiting enzymes for SA synthesis.Fusarium head blight(FHB)seriously threatens the safety of wheat production,but increasing the content of SA can enhance FHB resistance.However,the pathway of SA synthesis and regulation in wheat remains unknown.In this study,three wheat ICS(TaICSA,TaICSB,and TaICSD)were identified,and their functions were validated in vitro for isomerizing chorismate to isochorismate.The mutation of one or two homoeoalleles of TaICSA,TaICSB,and TaICSD in the wheat variety‘Cadenza’reduced SA levels under ultraviolet treatment and Fusarium graminearum infection,further enhancing sensitivity to FHB.Overexpression of TaICSA can significantly enhance SA levels and resistance to FHB.To further study SA synthesis pathways in wheat and avoid interference with pathogenicity related genes,the leaves of wild-type Cadenza and different TaICS mutant lines were subjected to ultraviolet treatment for transcriptomic analysis.The results showed that 37 PALs might be involved in endogenous SA synthesis,and 82 WRKY and MYB family transcription factors may regulate the expression of ICS and PAL.These results were further confirmed by RT-PCR.In conclusion,this study expands our knowledge of SA biosynthesis and identifies TaICSA,as well as several additional candidate genes that encode transcription factors for regulating endogenous SA levels,as part of an efficient strategy for enhancing FHB resistance in wheat.

OsbZIP53 Negatively Regulates Immunity Response by Involving in Reactive Oxygen Species and Salicylic Acid Metabolism in Rice

The basic region/leucine zipper(bZIP)transcription factors play important roles in plant development and responses to abiotic and biotic stresses.OsbZIP53 regulates resistance to Magnaporthe oryzae in rice by analyzing APIP5-RNAi transgenic plants.To further investigate the biological functions of OsbZIP53,we generated osbzip53 mutants using CRISPR/Cas9 editing and also constructed OsbZIP53 over-expression transgenic plants.Comprehensive analysis of phenotypical,physiological,and transcriptional data showed that knocking-out OsbZIP53 not only improved disease resistance by inducing a hypersensitivity response in plants,but also regulated the immune response through the salicylic acid pathway.Specifically,disrupting OsbZIP53 increased H2O2 accumulation by promoting reactive oxygen species generation through up-regulation of several respiratory burst oxidase homologs(Osrboh genes)and weakened H2O2 degradation by directly targeting OsMYBS1.In addition,the growth of osbzip53 mutants was seriously impaired,while OsbZIP53 over-expression lines displayed a similar phenotype to the wild type,suggesting that OsbZIP53 has a balancing effect on rice immune response and growth.

Effects of Chitosan and Salicylic Acid on Cold Resistance of Litchi under Low Temperature

This paper focused on the effect of spraying chitosan and salicylic acid to Litchi under low temperature stress conditions.The physiology and biochemistry of litchi were studied as well.Results showed that the appropriate concentration of chitosan and salicylic acid treatment could effectively reduce injury caused by low temperature to litchi,compared with water control,chlorophyll,proline,soluble protein content of litchi after treatment and the activity of protective enzyme increasing significantly.However,the accumulation of resistance could significantly be improved.Furthermore,when 1 000 mg/L chitosan combined with 50 mg/L salicylic acid,the litchi acquired the best cold resistance capability.

Effects of Acetylsalicylic Acid (ASA) and Ethylene Treatments onRipening and Softening of Postharvest Kiwifruit

Kiwifruit (Actinidia deliciosa (A. Chev.) C. F. Liang et A. R. Ferguson cv. Bruno) was used toinvestigate the effects of acetylsalicylic acid (ASA, 1.0 mmol/L, pH 3.5) and ethylene (100 mL/L) treat-ments on changes at endogenous salicylic acid (SA) levels and other senescence-related factors duringfruit ripening and softening at 20 ℃. The level of endogenous SA in ripening fruits declined and a closerelationship was observed between the change at endogenous SA level and the rate of fruit ripening andsoftening. ASA treatment elevated SA level in the fruit, slowed down the increases in lipoxygenase (LOX)and allene oxide synthase (AOS) activities, decreased the O22-. production in the preclimacteric phase andthe early phase of ethylene climacteric rise, maintained the stability of cell membrane, inhibited ethylenebiosynthesis, postponed the onset of the ethylene climacteric, and delayed the process of fruit ripeningand softening. On the contrary, application of ethylene to ripening kiwifruit resulted at a lower SA level, anaccelerated increases in the activities of LOX and AOS and the rate of O22-. production, an elevated relativeelectric conductivity and an advanced onset of ethylene climacteric, and a quicker fruit ripening andsoftening. It is suggested that the effects of ASA on ripening kiwifruit can be attributed to its ability toscavenge O22-. and/or to maintain stability of cell membrane.

Increase of β -1, 3-Glucanase and Chitinase Activities in Cotton Callus Cells Treated by Salicylic Acid and Toxin of Verticillium dahliae

The different resistance of cotton (Gossypium hirsutum L.) cultivars to crude toxin of Verticillium dah/iae(VD) was correlated with the activities of chitinase and β-1, 3-glucanase in callus cells. The activities of chitinase and β-1, 3-glucanase in the callus cells treated with the VD-toxin were increased to the higher level at earlier time point in resistant cultivars than these in the susceptible cultivars. Exogenous salicylic acid (SA) induced the accumulation of chitinase and β -1,3-glucanase, which resulted in the resistance of callus cells to the VD. toxin. Western blot using a polyclonal antibody against β -1,3-glucanase identified 28 kD protein that was induced by VD-toxin, SA, or VD-toxin plus SA.

Participation of H_2O_2 in Enhancement of Cold Chilling by Salicylic Acid in Banana Seedlings

The possible physiological mechanism of enhancement of cold tolerance by salicylic acid (SA) in banana seedlings ( Musa acuminata cv. Williams 8188) was explored. Measurements of leakage electrolyte after 2 d of recovery at 30/22 ℃ (day/night) following 3 d of cold stress at 7 ℃ showed that pretreatment with hydroponic solution containing SA 0.3-0.9 mmol/L as foliar spray under normal growth conditions (30/22 ℃) could significantly enhance cold tolerance of banana plants. The highest enhancing effect of SA occurred at 0.5 mmol/L and it showed the lowest leakage rate of electrolyte or smaller leaf wilting area after 2 d of recovery at normal temperature from 3 d of 7 ℃ or 5 ℃ cold stress. Higher concentrations (≥2.5 mmol/L) of SA, however, caused more electrolyte leakage, indicating that they aggravated chilling damage. Enhanced cold tolerance by SA could be related to H 2O 2 metabolism. Compared with water_treated seedlings (control), SA 0.5 mmol/L treatment inhibited activities of catalase (CAT) and ascorbate peroxidase (APX), increased peroxidase (POX) activity, but did not affect the activity of superoxide dismutase (SOD) under normal growth conditions, and these changes might lead to an accumulation of H 2O 2, whereas SA pretreatment enhanced the activities of CAT and APX, and reduced the increase in productions of H 2O 2 and thiobarbituric acid_reaction substances (TBARS) during subsequent 7 ℃ cold stress and recovery periods. Exogenous H 2O 2 treatments (1.5 -2.5 mmol/L) also increased cold tolerance of banana seedlings. Furthermore, pretreatment of banana seedlings with dimethylthiourea (a trap for H 2O 2) significantly inhibited cold tolerance induced by SA. These results suggested that endogenous H 2O 2 may be required for SA_enhanced cold tolerance. The significance of the interaction of SA, H 2O 2 and H 2O 2_metabolizing enzymes during cold stress has been discussed.

Effects of Salicylic Acid on Photosynthesis and Chlorophyll Fluorescence Characteristics of Fluecured Tobacco Leaf in Subdued Light

[Objective] This study aimed to test whether salicylic acid （SA） can im-prove the physiological functions of flue-cured tobacco under subdued light condition, and to determine the mechanism of its action. [Method] The tobacco plants under subdued light were foliar-sprayed with 100 mg/L of SA. Then, the physiological in-dices such as plant fresh weight and dry weight, chlorophyl content, photosynthetic parameters and chlorophyl fluorescence parameters were measured. SPSS17.0 and Excellwere adopted for variance analysis and significance test. [Result] The leaf photosynthetic rate （Pn）, stomatal conductance （Gs） and transpiration rate （Tr） of tobacco plants in subdued light were al decreased while the intercellular CO2 con-centration （Ci） was increased, suggesting that non-stomatal limitation led to the de-crease of Pn under weak light intensity stress. SA released the inhibition of tobacco plant growth in weak light, as it elevated the leaf photosynthetic rate, the maximum photochemical efficiency of PSⅡ, potential activity of PSⅡ, effective photochemical efficiency of PSⅡ and photochemical quenching coefficient in weak light significant-ly, and reduced the non-photochemical quenching coefficient. [Conclusion] SA has significant effects on the photosynthetic characteristics of flue-cured tobacco in weak light, and it can improve the synthesis or distribution of photosynthesis product, and the efficiency of light energy, conducive to plant growth and development.

Preparation and Application of Monoclonal Antibodies Specific for Salicylic Acid

Salicylic acid (SA) is widely distributed in many monocots and dicots and has many physiological effects. It can induce heat production in the thermogenic inflorescences of Arum lily([1]), block the biosynthesis of ethylene, and more attractively, it seems to be an important natural signal molecule in the induction of systemic acquired resistance (SAR) in tobacco, cucumber and other plants([2,3]). Studies in recent years showed that SA was also intimately related to the resistance of plants to aboitic stress, for example, SA increased chilling resistance of maize seedlings. Hence, SA has been accepted as a kind of new plant hormones. Up to date, the quantification of SA usually has been performed by HPLC[4,5], which often needs a large quantity of sample and a verbose pretreatment. Compared to HPLC, immunoassays, including radio-immunoassays (RIA) and enzyme-immunoassays (EIA), are easy to perform and have been widely used in the quantification of other plant hormones, such as IAA([6]), ABA([7,8]), GAs([9]), cytokinins et al([10]), and jasmonic acid (JA)([11]), and other low-molecular-weight, none-immunogenic compounds in plants([12]). Till now, only an indirect enzyme-linked immunosorbent assay (ELISA) for SA based on polyclonal antibodies (PAbs) has been developed by our group([13]), although Bennett et al([14]) had prepared SA PAbs using 4-aminosalicylic acid linked to KLH as immunogen in goat. However, the sensitivity of the ELISA we established formerly was relatively low, and also relatively larger quantity of sample is needed than other ELISAs for plant hormones. In this paper, an ELISA for SA based on monoclonal antibody raised against SA-NH-CH2-NH-KLH was introduced, and the fluctuation of SA content in cucumber leaves after inoculated with Pseudomonas syringae pv. syringae was determined.

Science Letters:Anodic oxidation of salicylic acid at Ta/BDD electrode

Boron-doped diamond （BDD） film electrodes using Ta as substrates were employed for anodic oxidation of salicylic acid （SA）. The effects of operational variables including initial concentration, current density, temperature and pH were examined. The results showed that BDD films deposited on the Ta substrates had high electrocatalytic activity for SA degradation. There was little effect ofpH on SA degradation. The current efficiency （CE） ,aas fbund to be dependent mainly on the initial SA concentration, current density and temperature. Chemical oxygen demand （COD） was reduced from 830 mg/L to 42 mg/L under a current density of200A/m^2 at 30℃.

Comparison of the Effects of H_2O_2 and Salicylic Acid on Alternative Respiratory Pathway in Aged Potato Tuber Slices

The influences of exogenous H 2O 2 and salicylic acid (SA) treatments on the alternative respiratory pathway (ARP) in aged potato (Solanum tuberosum L.) tuber slices were compared. The results showed that both H 2O 2(5.0 mmol/L) and SA (0.1 mmol/L) treatments had a significant inducing effect on ARP capacity (V alt ) and its ratio to total respiration (V alt /V t) in potato slices aged for 24 h. With a monoclonal antibody against the alternative oxidase (AOX), Western blotting results showed that both H 2O 2 and SA treatments increased the AOX expression levels in aged potato tuber slices. However, the results of oxygen isotope discrimination experiments showed that H 2O 2 had no influence on the in vivo ARP activity (ρV alt ) and its contribution to V t(expressed as ρV alt /V t) in potato slices aged for 24 h, but SA had a significant influence on the ρV alt and ρV alt /V t values of the aged potato tuber slices. These results indicate that there are differences between the effects of H 2O 2 and SA on ARP in plant tissues. Both of them possess the ability to induce ARP capacity through inducing AOX expression. However, SA can simultaneously stimulate the operation of ARP, but H 2O 2 can not.

Effects of Salicylic Acid on the Growth and Some Physiological Characters in Salt Stressed Tomato Plants （ Solanum L ycopersicum）

Excessive soil salinity is an important constraint limiting the distribution of plants in natural habitats, and is an increasingly severe agricultural problem in arid and semi-arid regions. Higher salinity levels caused significant reduction in growth parameters like leaf area, leaf length and root and shoot dry weight. Salicylic acid （SA）, a plant phenolic is now considered as a hormone-like endogenous regulator, and its role in the defence mechanisms against biotic stressors has been well documented. In recent years its role has been widely investigated in abiotic stress （salinity, drought, water deficit and so on）. The aim of the present work was to study the effects of salicylic acid on growth and some physiological characters of salt stressed tomato plants. The presence of salicylic acid at low concentration （0.01 mM） in culture medium riched with NaCl 100 mM （6 g·L^-1） improves the tolerance of tomato cv. Golden Sunrise to salinity. This amelioration results in stimulation of growth and development of plants. The applied of SA in saline medium induce： （i） an increase in chlorophyll content; （ii） a better supply of essential elements in plant growth, such as K＋; （iii） a decrease in toxic ions such Na＋ and CI in aerial organs; and （iv） an additional synthesis of organic solutes and osmoprotectors like proline and proteins. All these results suggest that salicylic acid could be successfully used in alleviating depressive effects of salt on the productivity of the cultivated tomato.

合成乙酰水杨酸(acetyl salicylic acid)实验教学研究

乙酰水杨酸的合成是大学生在生物与化工类有机合成中的一个经典性实验。目前各高校常采用王清廉的方法进行实验教学,笔者经多年的实验教学观察,发现学生按照该方法进行该实验时,常出现油状物而没有真正产物生成,或产物为假性产物(经性质检验为水杨酸本身)。另外,该实验即使在具有较好通风设备条件下进行,也会产生较大刺激气味且反应残余物对环境污染较大。为此,笔者从反应条件、器皿等方面着手进行该实验教学研究,并在保证教学双方的成功率、保护环境、节约药品和严格控制教学时间等方面已取得较理想的效果。

Effects of Exogenous Salicylic Acid Derivative on the Resistance to TMV and Activity of Defense Enzymes of Tobacco

[Objective] This study aimed to evaluate the effects of exogenous salicylic acid derivatives on tobacco resistance to TMV and activity of defense enzymes. [Method] The tobboco leaves were treated by exogenous salicylic acid derivatives. Then, the disease occurrence was observed, and the activity of phenylalanin ammo- nia lyase （PAL） and peroxidase （POX） were measured. [Result] Exogenous salicylic acid derivative increased the activities of PAL and POX, while did not influence the resistance to TMV. [Conclusion] The result provides a theoretical basis for the study of plant disease resistance mechanisms.

Application of advanced oxidation processes for removing salicylic acid from synthetic wastewaters

In this study,advanced oxidation processes(AOPs) such as anodic oxidation(AO),UV/H_2O_2 and Fenton processes(FP) were investigated for the degradation of salicylic acid(SA) in lab-scale experiments.Boron-doped diamond(BDD) film electrodes using Ta as substrates were employed for AO of SA.In the case of FP and UV/H_2O_2,most favorable experimental conditions were determined for each process and these were used for comparing with AO process.The study showed that the FP was the most effective process under aci...

Effects of exogenous salicylic acid on growth and H_2O_2-metabolizing enzymes in rice seedlings under lead stress

Salicylic acid （SA） was an essential component of the plant resistance to pathogens and also plays an important role in mediating plant response to some abiotic stress. The possible effects of SA on the growth and H2O2-metabolizing enzymes in rice seedlings under lead stress were studied. When rice seedlings grown in nutrient solution containing Pb^2＋ （0, 0.05, 0.15, 0.25 mmol/L） for 18 d, the plant biomass as well as the chlorophyll content of leaves decreased with increasing Pb concentration. The pre-treatment with SA （treated with 0.1 mmol/L SA for 48 h before Pb stress） partially protected seedlings from Pb toxicity. The chlorophyll contents were significant higher in leaves of Pb-exposed with SA pre-treatment seedlings than in Pb-exposed plants at the same Pb intensity. SA pre-treated alone could significantly increase the length of shoot and root of seedlings but the vigour difference was not marked under long-term exposure to Pb toxicity. SA pre-treated influence the H2O2 level in leaves of seedlings by up-regulating the activity of superoxide dismutase （SOD）, repressing the activity of catalase （CAT） and ascorbate peroxidase （APX） depending on the concentrations of Pb^2＋ in the growth medium. The results supported the conclusion that SA played a positive role in rice seedlings against Pb toxicity.

Physiological Mechanism of Salicylic Acid for Alleviation of Salt Stress in Rice

Soil salinity is one of the most important problems of crop production in estuarine and coastal zones. Improvement in salt tolerance of major food crops is an important way for the economic utilization of coastal zones. This study proved that the application of salicylic acid(SA) improved the growth and yield under salt stress conditions and investigated its physiological mechanisms for salt tolerance. The investigation on the effect of SA for salt tolerance during germination showed that the decreased rates of germination and growth(in terms of shoot and root lengths) by the salt stress were significantly increased by the SA application(SA + NaCl). The treatment of SA to the high and low saline soils enhanced the growth, yield and nutrient values of rice. The effects of SA on Na^+, K^+ and Cl~– ionic accumulation were traced under salt stress condition by inductively coupled plasma optical emission spectrometry and ion chromatography. It was revealed that the increased accumulation of Na^+ and Clˉ ions by the salt stress were reduced by SA application. An increased concentration of endogenous SA level was detected from the SA-treated rice varieties(ASD16 and BR26) by liquid chromatography electrospray Ionization-tandem mass spectrometry. The activities of antioxidant enzymes such as superoxide dismutase, catalase and peroxidase were increased by salt stress whereas decreased by the SA application. The study proved that the application of SA could alleviate the adverse effects of salt stress by the regulation of physiological mechanism in rice plants. In spite of salt stress, it can be applied to the coastal and estuarine regions to increase the rice production.

Salicylic acid alleviates postharvest chilling injury of sponge gourd(Luffa cylindrica)

Effect of salicylic acid（SA）on chilling injury（CI）of sponge gourd during storage（9 days,9°C）plus shelf life（2 days,20°C）was evaluated in this study.SA treatment at the concentration of 1.5 mmol L^-1 significantly reduced postharvest CI of sponge gourds.Besides,the application of SA could effectively decrease the electrolyte leakage,reduce the accumulation of malondialdehyde（MDA）and total phenolics,enhance the activities of antioxidant enzymes such as superoxide dismutase（SOD）,catalase（CAT）and ascorbate peroxidase（APX）,and inhibit the activities of phenylalanine ammonia-lyase（PAL）and polyphenol oxidase（PPO）.The beneficial effects of SA could be attributed to preserved membrane integrity,inhibited membrane peroxidation,enhanced antioxidant system and suppressed activities of browning related enzymes.In a sense,SA as a postharvest tool may be commercially used in alleviating CI of sponge gourd.

Response of Endogenous Salicylic Acid and Jasmonates to Mechanical Wounding in Pea Leaves

The roles of on endogenous jasmonates （JAs） and salicylic acid （SA） in wounding response were investigated. Pea （Pisum sativum L.） seedlings were treated with three different methods including mechanical wounding, JAs application, and SA application. The contents of endogenous JAs and SA, as well as the activities of the related enzymes were detected by enzyme-linked immunosorbent assay （ELISA）, high performance liquid chromatography （HPLC）, and spectrophotometer, respectively. The results showed that endogenous JA rapidly accumulated within 30 min after wounding. The increase in the activities of both lipoxygenase （LOX） and allene oxide synthase （AOS） lagged behind JAs burst. A second slight increase in JAs level was observed at 24 h after wounding treatment, and at the same time point, higher activities of LOX and AOS were also detected. Endogenous free SA content decreased accompanied with JAs burst. Effects of exogenous JA application were similar to those of wounding treatment on endogenous SA level and phenylalanine ammonia lyase （PAL） activity, whereas exogenous SA application led to the significant inhibition of LOX and AOS activities and the decrease of endogenous JAs level at the early stage of treatment. It is thus suggested that JAs burst and SA decrease in early response to wounding may constitute an important mechanism by which plant starts the related defense reaction and adapts to wounding stress.