Phenylalanine Ammonia-lyase Enzyme Research Articles

Effectiveness of Plant-Induced Resistance Against Root-Knot Nematode Depends on the Policy of Using Inducer on the Host Plant.

This research was conducted to determine the relationship between plant defense responses and the extent of treatment applied to either the aerial parts or roots of the plant. The experimental treatments included different methods of application (spraying versus soil drenching), varying treatment areas (one-sixth, one-third, half, or all of the plant's aerial parts and roots) with SA, and infecting the plants with root-knot nematodes. Evaluation of plant growth and nematode pathogenicity indices in the greenhouse section, H2O2 accumulation rate, and phenylalanine ammonia lyase enzyme activity (in aerial parts and roots) were carried out in biochemical experiments. The results showed that treating less than one-third of the aerial parts with salicylic acid (SA) did not significantly impact plant growth or nematode pathogenicity indices. However, it did lead to a notable increase in hydrogen peroxide (H2O2) accumulation, while phenylalanine ammonia lyase (PAL) enzyme activity remained unchanged. In contrast, treating more than one-third of the aerial parts resulted in decreased nematode pathogenicity and enhanced production of defense compounds. Notably, treatments targeting the roots consistently demonstrated a more pronounced effect on nematode suppression and increased defense compound levels, emphasizing the importance of root treatment, as this is where nematodes are primarily present. Overall, the study highlights the differential impact of treatment location and extent on plant defense mechanisms and suggests that strategic targeting of either aerial or root tissues can optimize plant responses against nematode attacks.

Foliar application of sodium nitroprusside alters the physicochemical properties, antioxidant capacities, and enzymatic activities of strawberry cv. Camarosa

Strawberry (Fragaria × ananassa) is a horticultural crop known for its sensitivity to mechanical damage and susceptibility to postharvest decay. In recent years, various strategies have been implemented to enhance both the yield and quality of strawberries, among which the application of nitric oxide-producing compounds has garnered special attention. The present study aimed to investigate the effects of varying concentrations of sodium nitroprusside (SNP), specifically 0, 200, 400, and 600 μM, on strawberries (cv. Camarosa) cultivated in a soilless culture system. It was attempted to identify optimal treatment concentrations that would improve the quality and yield of the strawberries. The analysis of variance revealed significant differences (p ≤ 0.01) in all morphological and phytochemical properties, as well as antioxidant and enzymatic activities, between the treated samples and the control group. Notably, the highest concentrations of total phenolics, phenylalanine ammonia-lyase (PAL) enzyme activity, guaiacol peroxidase enzyme activity, and potassium content in the fruit were recorded at the 400 μM SNP treatment. Specifically, these values were 6.67 mg GAE 100 g⁻1 FW, 57.42 nmol g⁻1 FW min⁻1, 0.183 μmol H2O2 min-1 100 ml-1 extract, and 5.9% DW, respectively. Furthermore, the 200 μM SNP treatment yielded the highest ascorbic acid content (0.587 mg AA 100 g-1 FW) and the lowest 50% inhibitory concentration for free radicals at 44.18 μl. In contrast, the 600 μM treatment resulted in the highest total flavonoid content (0.529 mg QE 100 g⁻1 FW). In conclusion, the findings indicated that SNP treatment can effectively enhance the yield and improve the quality and marketability of the strawberry fruit.

Exogenous phenylalanine application effects on phytochemicals, antioxidant activity, HPLC profiling, and PAL and CHS genes expression in table grapes (Vitis vinifera cv. ‘Qzl Ouzum’)

BackgroundGrape (Vitis vinifera L.) is one of the most important fruit products globally and has a high nutritional value with potent antioxidant and anti-cancer activities. In current years, phenylalanine application has been particularly noticed for enhancing the nutritional quality of horticultural crops. With the aim of quality improvement, the effects of foliar application of phenylalanine at 5 concentrations (0, 100, 500, 1000, and 2000 μM) on Vitis vinifera cv. ‘Qzl Ouzum’ berry compositions were studied. The studied parameters included antioxidant activity, phenolics, flavonoids, anthocyanin, catalase and phenylalanine ammonia-lyase enzyme activity, and phenolic compounds content based on HPLC analyses (myricetin, quercetin, kaempferol, syringetin, catechin, gallic acid, caffeic acid, p-coumaric acid, resveratrol).ResultsPhenylalanine at 1000 μM increased total phenols, flavonoids, anthocyanins, and PAL enzyme activity. In addition, HPLC analysis revealed a significant accumulation of individual phenolic compounds by phenylalanine treatment. The highest values were recorded in the treatments with 100 and 500 μM phenylalanine for most of the phenolic components. Real-time quantitative RT-PCR results showed that the expression of PAL and CHS genes was induced by phenylalanine. The highest PAL and CHS gene expression was observed at 500 μM and then at 1000 μM phenylalanine treatment.ConclusionsWith the use of phenylalanine, the activity of the PAL and CHS enzyme and PAL and CHS gene expression were significantly increased and led to a greater accumulation of phenolic compounds and antioxidant activiety. This study suggests that the use of phenylalanine as a main precursor for the synthesis of phenolic compounds, can improve the phenolic composition of grapes and could be a practical approach to advance fruit quality in the production of grape cv. ‘Qzl Ouzum’.

Antifungal Properties of Bioactive Compounds Isolated from Fucus vesiculosus Supercritical Carbon Dioxide Extract.

The exploration of natural antifungal substances from algal origins is significant due to the increasing resistance of pathogens to conventional antifungal agents and the growing consumer demand for natural products. This manuscript represents the inaugural investigation into the antifungal attributes of bioactive compounds extracted from Fucus vesiculosus via supercritical carbon dioxide (scCO2) extraction utilizing contemporary countercurrent chromatography (CCC). In aligning with the prospective utilization of this extract within the agricultural sector, this study also serves as the preliminary report demonstrating the capability of Fucus vesiculosus scCO2 extract to enhance the activity of plant resistance enzymes. The fractions obtained through CCC were subjected to evaluation for their efficacy in inhibiting the macrospores of Fusarium culmorum. The CCC methodology facilitated the successful separation of fatty acids (reaching up to 82.0 wt.% in a given fraction) and fucosterol (attaining up to 79.4 wt.% in another fraction). All CCC fractions at the concentration of 1.0% were found to inhibit 100% of Fusarium culmorum growth. Moreover, Fucus vesiculosus scCO2 extract was able to activate plant resistance enzymes (Catalase, Ascorbic Peroxidase, Guaiacol Peroxidase, Phenylalanine Ammonia-Lyase, and Phenylalanine Ammonia-Lyase Activity).

Stereoselective Synthesis of Aliphatic α‐Amino Acids by Chemo‐Enzymatic Hydroamination‐Hydrodesulfurisation

Phenylalanine ammonia lyase (PAL) enzymes have been extensively exploited to produce differently substituted arylalanine analogues, but their use in the stereoselective synthesis of aliphatic fatty amino acids has never been addressed. In this work, a two‐step hydroamination‐hydrodesulfurisation process has been investigated for the production of aliphatic L‐α‐amino acids from thienylacrylic acids, easily accessible by condensation from the corresponding arylaldehyde. Wild‐type PALs from Planctomyces brasiliensis and Streptomyces rimosus were selected as the most promising candidates and improved by site‐directed mutagenesis, thus unlocking the hydroamination of a broad panel of 10 different thienylacrylic acids bearing aliphatic and aromatic substituents. The subsequent hydrodesulfurisation step, accomplished by means of inexpensive Raney nickel in mild conditions and aqueous medium, afforded the corresponding aliphatic α‐amino acids in 10‐63% overall isolated yield and perfect enantiopurity. This chemo‐enzymatic process represents the first example of aliphatic amino acid production using PALs, circumventing a major limitation of those biocatalysts.

Manipulating flavonoid biosynthesis in Trigonella persica through controlled spectral lighting

Trigonella persica Boiss., is renowned for its rich phytochemical profile, particularly the presence of the flavonol quercetin. This study explored the effects of various light treatments, including blue, red, blue-red (1:1) radiation (BRR), and pink fluorescent light (PFL), on the biochemical and molecular mechanisms governing quercetin and flavonoid biosynthesis in T. persica at different growth stages. Our results showed that light treatments significantly influenced the activity of key enzymes phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) during germination and vegetative growth, with blue light inducing higher PAL and TAL activities compared to control conditions. High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses revealed that 48-h-old sprouts grown under red light exhibited the highest levels of flavonoid components and phenolic acids, with catechin as the predominant flavonoid. Notably, BRR treatment led to elevated concentrations of the bioavailable quercetin-3-rhamnoside in 48-h-old sprouts, while 15-day-old plants grown under PFL conditions showed a significant accumulation of the sulfated quercetin-3-sulfate. Real-time PCR analysis demonstrated that BRR upregulated the expression of flavonoid biosynthesis genes PAL, chalcone synthase (CHS), and chalcone isomerase (CHI) in sprouts, whereas PFL treatment induced higher expression of these genes, as well as cinnamate 4-hydroxylase (C4H), in aerial parts. These findings suggest that targeted light treatments, particularly blue and red LED light, can enhance the accumulation of bioavailable quercetin-3-rhamnoside during T. persica germination and sprouts exhibit higher levels of flavonoids and phenolic acids than aerial parts during different vegetative growth stages.

Changes in Postharvest Quality and Physiological Attributes of Strawberry Fruits Influenced by l-Phenylalanine.

Strawberry (Fragaria × ananassa) is a popular fruit with rich nutrients and a delicious taste. But this fruit is very vulnerable to diseases and decay. Therefore, l-phenylalanine (Phe) (0, 4, 8 mM) was considered to improve biochemical characteristics and activity of antioxidant enzymes in strawberry fruit cv. Sabrina under cold storage (5, 10, 15 days). After treatment and storage, traits including weight loss, total phenol (TP), antioxidant capacity, ascorbic acid, total anthocyanin (TA), total flavonoid (TF), malondialdehyde (MDA), soluble protein content and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and guaiacol peroxidase (POD) and phenylalanine ammonialyase (PAL) were evaluated at 5-day intervals. Our findings showed that the treatment of l-phenylalanine in different concentrations prevented the weight loss of the fruit compared to the control and maintained and increased TP, antioxidant capacity, ascorbic acid, TA, TF, soluble protein and SOD, CAT, APX, POD, and PAL enzymes activity. Also, Phe decreased the MDA content and peroxidation of lipid. The results showed that 4 mM Phe is the best treatment for improving phytochemical characteristics and maintaining fruit quality. The findings indicated that Phe treatment may be useful to improve quality and increase postharvest shelf life in strawberry fruits.

Hexanal-containing dip treatment enhances kiwifruit resistance to Botrytis cinerea

This study evaluated the effectiveness of a hexanal-based enhanced freshness formulation (EFF), containing antioxidants such as geraniol, α-tocopherol and ascorbic acid, to inhibit the fungal decay caused by Botrytis cinerea in ‘Y368’ kiwifruit. In vitro and in vivo experiments using three treatments: control (untreated), 0.03 and 0.06 % (v/v) EFF were set up. In the in vitro experiment, mycelial growth and spore germination of Botrytis cinerea exposed to EFF were examined. In the in vivo experiment, disease incidence, lesion diameter, activity of defense-related enzymes (Phenylalanine Ammonia Lyase, Guaiacol Peroxidase, β-1,3-glucanase and Chitinase), total phenolics and flavonoids were evaluated. The results demonstrated that 0.03 and 0.06 % EFF treatments reduced mycelial growth of Botrytis cinerea by 41 and 89 %, respectively. The lesion diameter of the 0.03 and 0.06 % EFF-treated fruit was 28 and 76 % smaller than the control. Furthermore, EFF-treated fruit exhibited significantly higher activities of defense-related enzymes compared to the control, with total phenolic content increasing by 1.2 and 1.3-fold for the 0.03 and 0.06 % treatments, respectively. These findings provide compelling evidence regarding the efficacy of EFF in suppressing Botrytis cinerea and highlight the role of defense-related enzymes in enhancing the fruit's resistance to decay. Overall, the study underscores the potential of EFF as a valuable and practical method for controlling fungal decay in kiwifruit during postharvest storage.

Nutrition in phenylketonuria

Phenylketonuria (PKU) is a genetic metabolic disease resulting from a deficiency in the enzyme phenylalanine hydroxylase. This defect prevents the conversion of phenylalanine to tyrosine, and as a result, the level of phenylalanine in the body increases abnormally. The accumulation of this amino acid may cause brain damage and grows development in these patients.The primary approach to treating the disease involves dietary management that restricts protein and phenylalanine intake, often alongside Tetrahydrobiopterin (BH4) supplementation. Medical foods specifically formulated for these patients are utilized to supply the necessary energy, protein, and tyrosine. Other treatment options, including the use of large neutral amino acids (LNAAs) and the enzyme Phenylalanine ammonia lyase (PAL), can be utilized to manage symptoms in adult patients. Advancements in gene therapy aimed at correcting defective genes in patients may help restore phenylalanine metabolism, though further research in this area is necessary. Effective personal management of phenylketonuria necessitates enhanced knowledge and collaboration among healthcare professionals to achieve optimal treatment outcomes for patients. This article emphasis the nutritional management of individuals with phenylketonuria across various stages of their lives.

Synergistic Effect of Arbuscular Mycorrhizal Fungi and Germanium on the Growth, Nutritional Quality, and Health-Promoting Activities of Spinacia oleracea L.

Arbuscular mycorrhizal fungi (AMF) and the antioxidant germanium (Ge) are promising tools for boosting bioactive compound synthesis and producing healthier foods. However, their combined effect remains unexplored. This study demonstrates the synergistic impact of AMF and Ge on the growth, metabolite accumulation, biological activities, and nutritional qualities of Spinacia oleracea L. (spinach), a globally significant leafy vegetable. Individually, Ge and AMF increased biomass by 68.1% and 22.7%, respectively, while their combined effect led to an 86.3% increase. AMF and Ge also improved proximate composition, with AMF-Ge interaction enhancing crude fiber and mineral content (p < 0.05). Interestingly, AMF enhanced photosynthesis-related parameters (e.g., total chlorophyll) in Ge treated plants, which in turn increased carbohydrate accumulation. This accumulation could provide a route for the biosynthesis of amino acids, organic acids, and fatty acids, as evidenced by increased essential amino acid and organic acid levels. Consistently, the activity of key enzymes involved in amino acids biosynthesis (e.g., glutamine synthase (GS), methionine biosynthase (MS), lysine biosynthase (LS)) showed significant increments. Furthermore, AMF improved fatty acid levels, particularly unsaturated fatty acids in Ge-treated plants compared to the control. In addition, increased phenylalanine provided a precursor for the production of antioxidants (e.g., phenols and flavonoids), through the action of the enzyme phenylalanine ammonia-lyase (PAL), resulting in improved antioxidant activity gains as indicated by FRAP, ABTS, and DPPH assays. This study is the first to show that Ge enhances the beneficial effect of AMF on spinach, improving growth and nutritional quality, with promising implications for agricultural practices.

Prolonging Cyclamen Flower Vase Life via 8-HQS and AgNO3 Treatments in a Controlled Release System

The current study applied electrospun fibers containing 8-hydroxyquinoline-5-sulfonic acid (8-HQS) (100, 150, and 200 mg L−1) and silver nitrate (AgNO3) (15 and 20 mg L−1) to enhance the longevity and qualitative parameters of two cyclamen cultivars. The results indicated that the vase life of the flowers treated with 20 mg L−1 AgNO3 and 200 mg L−1 8-HQS was higher than that of the other treatments (16 days). Cyclamens treated with 8-HQS (100 mg L−1) + AgNO3 (15 mg L−1) and 8-HQS (100 mg L−1) + AgNO3 (20 mg L−1) had lower polyphenol oxidase (PPO) activity than the control group. Also, it was observed that the higher phenylalanine ammonia-lyase (PAL) enzyme activity of cyclamens in the 8-HQS (150 mg L−1) + AgNO3 (15 mg L−1) treatment was longer than that of the other treatments and control groups. Treatment with 100 mg L−1 8-HQS and 20 mg L−1 AgNO3 showed higher peroxidase (POD) activity than the different treatments. This research indicated that adding AgNO3 and 8-HQS to electrospun fibers is a promising method for enhancing the longevity and maintaining the quality of these cut flowers.

Inoculation with Jeotgalicoccus sp. improves nutritional quality and biological value of Eruca sativa by enhancing amino acid and phenolic metabolism and increasing mineral uptake, unsaturated fatty acids, vitamins, and antioxidants.

Plant growth-promoting bacteria (PGPB) are considered a promising tool for triggering the synthesis of bioactive compounds in plants and to produce healthy foods. This study aimed to demonstrate the impact of PGPB on the growth, accumulation of primary and secondary metabolites, biological activities, and nutritional qualities of Eruca sativa (arugula), a key leafy vegetable worldwide. To this end, Jeotgalicoccus sp. (JW0823), was isolated and identified by using partial 16S rDNA-based identification and phylogenetic analysis. The findings revealed that JW0823 significantly boosted plant biomass production by about 45% (P<0.05) and enhanced pigment contents by 47.5% to 83.8%. JW0823-treated plants showed remarkable improvements in their proximate composition and vitamin contents, with vitamin E levels increasing by 161.5%. JW0823 induced the accumulation of bioactive metabolites including antioxidants, vitamins, unsaturated fatty acids, and essential amino acids, thereby improving the nutritional qualities of treated plants. An increase in the amounts of amino acids was recorded, with isoleucine showing the highest increase of 270.2%. This was accompanied by increased activity of the key enzymes involved in amino acid biosynthesis, including glutamine synthase, dihydrodipicolinate synthase, cystathionine γ-synthase, and phenylalanine ammonia-lyase enzymes. Consequently, the total antioxidant and antidiabetic activities of the inoculated plants were enhanced. Additionally, JW0823 improved antimicrobial activity against several pathogenic microorganisms. Overall, the JW0823 treatment is a highly promising method for enhancing the health-promoting properties and biological characteristics of E. sativa, making it a valuable tool for improving the quality of this important leafy vegetable.

Effect of Nitrogen Fertilizer on Capsaicinoids and Related Metabolic Substances of Dried Chili Pepper Fruit

Reducing fertilizer pollution is an important direction for modern ecological agriculture. Commonly, excessive nitrogen is applied to pepper. Capsaicin is one of the most important economic qualities of spicy peppers, but the effect of nitrogen on capsaicin is still inconsistent. This study aims to elucidate the impact of nitrogen fertilizer on capsaicin accumulation and to provide guidance on fertilizer application on highly spicy chili peppers. The experiment was conducted with five nitrogen fertilizer concentrations: N1 (urea 750 kg ha−1), N2 (urea 562.5 kg ha−1), N3 (urea 375 kg ha−1), N4 (urea 187.5 kg ha−1), and N0 (no nitrogen fertilizer). Nitrogen treatment was applied to two varieties with different spiciness levels from the seedling stage. The fruits in different layers of pepper plants were sampled on the 20th, 35th, and 50th day after anthesis, and the fruits’ size, content of capsaicin, capsaicin precursors, capsaicin competitors, as well as capsaicin-related enzyme activities and gene expression level were analyzed. The results indicate that, when applying N2 and N3, both chili pepper varieties exhibited higher fruit length, diameter, weight, and yield values. There were increased contents of total phenol, flavonoids, and tannins in both fruit varieties with N2 application. Moreover, the placenta weights of the bottom, middle, and top layers of the fruits on the 35th day were improved by 40.14%, 26.80%, and 55.91% for ‘Honglong 23’ and 55.10%, 37.04%, and 75.56% for ‘Hongxi’ compared with N0. At the same time, under N2 treatment, the phenylalanine ammonia-lyase (PAL) enzyme activity of capsaicin synthase significantly increased. In contrast, the capsaicin-degrading enzyme activities of peroxidase (POD) and polyphenol oxidase (PPO) decreased notably. The expression levels of capsaicin-synthetic genes such as phenylalanine cleavage enzyme gene (PAL), acyltransferase gene (AT3), 4-Coumaroyl coenzyme A ligase gene (4CL), cinnamate 4-hydroxylase gene (C4H), caffeoyl coenzyme A-3-oxo-methyltransferase gene (COMT), paminotransferase gene (PAMT), and hydroxycinnamyltransferase gene (HCT) were up-regulated in N2 fruits, which led to a significant increase in capsaicin content compared with the other four nitrogen amounts. A further reduction in nitrogen application to N3 and N4 resulted in a decrease in the precursor substance’s total phenol content and PAL activity and an increase in the competitive substance’s flavonoid, lignin, POD, and PPO enzyme activities. At the same time, the expression levels of capsaicinoid synthetic genes were downregulated in the N3 and N4 treatments, leading to a low content of total capsaicinoids. The capsaicinoid content showed a trend of 35th day &gt; 50th day &gt; 20th day for both varieties. Additionally, the contents of total capsaicinoids, total phenols, flavonoids, and lignins, as well as PAL enzyme activity, and the expression levels of PAL, AT3, 4CL, C4H, COMT, PAMT, and HCT exhibited characteristics of bottom layers &gt; middle layers &gt; top layers. The activities of POD and PPO gradually increased from the bottom to the top layers. The N2 (562.5 kg ha−1) treatment resulted in increases in placenta mass, maximum capsaicinoid precursor substance of total phenol content, and synthase enzyme activity, as well as decreases in capsaicinoid competing substances and degradative enzyme activity, so there were more substances available for capsaicin synthesis. Combined with the higher fruit weight and capsaicinoid content in the N2 treatment, N2 was considered a suitable nitrogen fertilizer dosage for highly spicy chili pepper cultivation.

Nano-enhanced defense: Titanium-enriched Alginate–Bentonite coating augments Bacillus amyloliquefaciens D203 efficacy against Magnaporthe oryzae in Kenyan rice cultivation

Rice blast disease, caused by Magnaporthe oryzae, poses a significant threat to global rice production, necessitating the development of effective and sustainable management strategies. Biological control using beneficial microbes like Bacillus amyloliquefaciens has emerged as a promising approach due to its ability to enhance plant resistance and reduce disease incidence. Nano-encapsulation of bacteria, which involves embedding beneficial microbes within nanomaterials, offers a novel method to improve the stability, survival, and efficacy of these biocontrol agents. This study evaluated the capacity of encapsulated Bacillus amyloliquefaciens D203, embedded within an alginate-bentonite coating infused with titanium nanoparticles (TNs), to stimulate defense responses in rice seedlings challenged by the Magnaporthe oryzae the causal agent of rice blast disease. Encapsulation was achieved using the extrusion technique, with some modifications. Using a completely randomized design, the experiment was conducted in a greenhouse, with four treatments replicated four times. The experiment used the popular Kenyan rice variety "BASMATI 370”. The study investigated the impact of strain D203 on the incidence, severity, and area under disease progress curves related to M. oryzae, as well as the expression of defense-related enzymes. The results demonstrated that rice plants derived from seeds coated with the D203 encapsulated B. amyloliquefaciens strain exhibited higher levels of defense-related enzyme expression, including peroxidase (POD), phenylalanine ammonia-lyase (PAL), superoxide dismutase (SOD) and catalase (CAT), compared to controls. In addition, the incidence and severity of the disease were markedly lower in plants treated with encapsulated B. amyloliquefaciens compared to controls, sometimes paralleling the efficacy of hexaconazole treatment. These findings suggest that the encapsulation of strain D203 has the potential to enhance resistance against rice blast disease by inducing systemic resistance through the production of antioxidant enzymes.

Host Plants and Fertilization Mediated Life History of American Serpentine Leaf Miner, Liriomyza trifolii (Burgess) (Diptera: Agromyzidae).

Herbivorous insects depend on the host plant to optimize their overall reproductive success, and balanced fertilization may alter the plant's quality against herbivory. Life history traits of the Liriomyza trifolii (Burgess) were determined under laboratory conditions using either unfertilized and fertilized plants of bean [Phaseolus vulgaris L. (Fabaceae)], chrysanthemum [Chrysanthemum × morifolium (Asteraceae)], potato [Solanum tuberosum (Solanaceae)], bell pepper [Capsicum annuum (Solanaceae)], and tomato [Solanum lycopersicum (Solanaceae)]. Results indicated that L. trifolii completed development on all studied unfertilized and fertilized plants. Nevertheless, a higher performance of the leaf miner was observed on bean and bell pepper plants compared to the other plants. Furthermore, there was an interaction of the host plant and fertilization with Calcium Aria or Sitam negatively affecting the fitness-related traits of the leaf miner. Application of these fertilizers resulted in delayed immature development of L. trifolii, decreased survival rate, and reduced adult longevity and fecundity. The activity of cinnamyl alcohol dehydrogenase (CAD), peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) enzymes, as well as phenolic, flavonoid, and lignin content were higher in Calcium Aria + Sitam fertilized plants, intermediate in Calcium Aria and Sitam treated plants, and the lower in unfertilized plants. The development and survival of L. trifolii on different host plants, considering fertilization options, become important for deploying cultural control practices against this important pest species.

Volatiles emitted by Pseudomonas aurantiaca ST-TJ4 trigger systemic plant resistance to Verticillium dahliae

Verticillium dahliae is among the most devastating fungal pathogens, causing significant economic harm to agriculture and forestry. To address this problem, researchers have focused on eliciting systemic resistance in host plants through utilizing volatile organic compounds (VOCs) produced by biological control agents. Herein, we meticulously measured the quantity of V. dahliae pathogens in plants via RTqPCR, as well as the levels of defensive enzymes and pathogenesis-related (PR) proteins within plants. Finally, the efficacy of VOCs in controlling Verticillium wilt in cotton was evaluated. Following treatment with Pseudomonas aurantiaca ST-TJ4, the expression of specific VdEF1-α genes in cotton decreased significantly. The incidence and disease indices also decreased following VOC treatment. In cotton, the salicylic acid (SA) signal was strongly activated 24 h posttreatment; then, hydrogen peroxide (H2O2) levels increased at 48 h, and peroxidase (POD) and catalase (CAT) activities increased to varying degrees at different time points. The malondialdehyde (MDA) content and electrolyte leakage in cotton treated with VOCs were lower than those in the control group, and the expression levels of chitinase (CHI) and PR genes (PR10 and PR17), increased at various time points under the ST-TJ4 treatment. The activity of phenylalanine ammonia lyase (PAL) enzymes in cotton treated with VOCs was approximately 1.26 times greater than that in control plants at 24 h，while the contents of phenols and flavonoids increased significantly in the later stage. Additionally, 2-undecanone and 1-nonanol can induce a response in plants that enhances disease resistance. Collectively, these findings strongly suggest that VOCs from ST-TJ4 act as elicitors of plant defence and are valuable natural products for controlling Verticillium wilt.

Improving blueberry cold storage quality: the effect of preharvest hexanal application on chilling injuries and antioxidant defense mechanisms.

Blueberries are vulnerable to chilling injury (CI). This can lead to limited longevity when they are subjected to cold storage conditions. This study investigated the effectiveness of a preharvest spray containing 0.02% hexanal in reducing CI and improving the postharvest storage quality of 'Star' and 'Biloxi' blueberries. The blueberries were stored for a period of 5 weeks at 2 °C and in 90% relative humidity (RH). The findings revealed that the preharvest hexanal spraying of both cultivars delayed senescence by mitigating CI, as evidenced by the bolstering of the antioxidant defense system through increased superoxide dismutase (SOD), ascorbate peroxidase (APX), peroxidase (POD), catalase (CAT), and phenylalanine ammonia lyase (PAL) enzyme activity. The treated fruit also maintained elevated levels of total phenol content (TPC), total flavonoids (TFC), and vitamin C, demonstrating enhanced free radical scavenging capacity (FRSC), while exhibiting reduced polyphenoloxidase (PPO) activity, and reduced malondialdehyde (MDA), and H2O2 content in comparison with the control group. The preharvest hexanal treatment also suppressed fruit softening by maintaining greater firmness and higher membrane stability index (MSI) scores, inhibiting the activity of polygalacturonase (PG), pectinmethylesterase (PME), xylanase, and α-amylase, and reducing microbial counts (MC) and incidence of decay (DI) in comparison with the control. Preharvest hexanal treatment also improved the overall storage quality by reducing weight loss, total soluble solids (TSS), pH, and the TSS/acid ratio, while increasing titratable acidity (TA) in comparison with the control during cold storage. The findings suggest that hexanal, as a preharvest application, delays senescence effectively and preserves overall quality by enhancing cold tolerance through antioxidant defense mechanisms in blueberry storage under cold conditions. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Humic acid and grafting as sustainable agronomic practices for increased growth and secondary metabolism in cucumber subjected to salt stress

Salinity stress poses a significant treat to crop yields and product quality worldwide. Application of a humic acid bio stimulant and grafting onto tolerant rootstocks can both be considered sustainable agronomic practices that can effectively ameliorate the negative effects of salinity stress. This study aimed to assess the above mentioned ameliorative effects of both practices on cucumber plants subjected to saline environments. To attain this goal a factorial experiment was carried out in the form of a completely randomized design with three replications. The three factors considered were (a) three different salinity levels (0, 5, and 10 dS m−1 of NaCl), (b) foliar application of humic acid at three levels (0, 100, and 200 mg L−1), and (c) both grafted and ungrafted plants. Vegetative traits including plant height, fresh and dry weight and number of leaf exhibited a significant decrease under increasing salinity stress. However, the application of humic acid at both levels mitigated these effects compared to control plants. The reduction in relative water content (RWC) of the leaf caused by salinity, was compensated by the application of humic acid and grafting. Thus, the highest RWC (86.65%) was observed in grafting plants with 0 dS m−1 of NaCl and 20 mg L−1 of humic acid. Electrolyte leakage (EL) increased under salinity stress, but the application of humic acid and grafting improved this trait and the lowest amount of EL (26.95%) was in grafting plants with 0 dS m−1 of NaCl and 20 mg L−1 of humic acid. The highest amount of catalase (0.53 mmol H2O2 g−1 fw min−1) and peroxidase (12.290 mmol H2O2 g−1 fw min−1) enzymes were observed in the treatment of 10 dS m−1 of NaCl and 200 mg L−1 humic acid. The highest amount of total phenol (1.99 mg g−1 FW), total flavonoid (0.486 mg g−1 FW), total soluble carbohydrate (30.80 mg g−1 FW), soluble protein (34.56 mg g−1 FW), proline (3.86 µg g−1 FW) was in grafting plants with 0 dS m−1 of NaCl and 200 mg L−1 of humic acid. Phenolic acids and phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO) enzymes increased with increasing salinity and humic acid levels. Contrary to humic acid, salt stress increased the sodium (Na+) and chlorine (Cl−) and decreased the amount of potassium (K+) and calcium (Ca2+) in the root and leaf of ungrafted cucumber. However, the application 200 mg L−1 humic acid appeared to mitigate these effects, thereby suggesting a potential role in moderating physiological processes and improving growth of cucumber plants subjected to salinity stress. According to the obtained results, spraying of humic acid (200 mg L−1) and the use of salt resistant rootstocks are recommended to increase tolerance to salt stress in cucumber. These results, for the first time, clearly demonstrated that fig leaf gourd a new highly salt-tolerant rootstock, enhances salt tolerance and improves yield and quality of grafted cucumber plants by reducing sodium transport to the shoot and increasing the amount of compatible osmolytes.

Genome-wide analysis of R2R3-MYB transcription factors in poplar and functional validation of PagMYB147 in defense against Melampsora magnusiana

Main conclusionTranscription of PagMYB147 was induced in poplar infected by Melampsora magnusiana, and a decline in its expression levels increases the host's susceptibility, whereas its overexpression promotes resistance to rust disease.Poplars are valuable tree species with diverse industrial and silvicultural applications. The R2R3-MYB subfamily of transcription factors plays a crucial role in response to biotic stresses. However, the functional studies on poplar R2R3-MYB genes in resistance to leaf rust disease are still insufficient. We identified 191 putative R2R3-MYB genes in the Populus trichocarpa genome. A phylogenetic analysis grouped poplar R2R3-MYBs and Arabidopsis R2R3-MYBs into 33 subgroups. We detected 12 tandem duplication events and 148 segmental duplication events, with the latter likely being the main contributor to the expansion of poplar R2R3-MYB genes. The promoter regions of these genes contained numerous cis-acting regulatory elements associated with response to stress and phytohormones. Analyses of RNA-Seq data identified a multiple R2R3-MYB genes response to Melampsora magnusiana (Mmag). Among them, PagMYB147 was significantly up-regulated under Mmag inoculation, salicylic acid (SA) and methyl jasmonate (MeJA) treatment, and its encoded product was primarily localized to the cell nucleus. Silencing of PagMYB147 exacerbated the severity of Mmag infection, likely because of decreased reactive oxygen species (ROS) production and phenylalanine ammonia-lyase (PAL) enzyme activity, and up-regulation of genes related to ROS scavenging and down-regulation of genes related to PAL, SA and JA signaling pathway. In contrast, plants overexpressing PagMYB147 showed the opposite ROS accumulation, PAL enzyme activity, SA and JA-related gene expressions, and improved Mmag resistance. Our findings suggest that PagMYB147 acts as a positive regulatory factor, affecting resistance in poplar to Mmag by its involvement in the regulation of ROS homeostasis, SA and JA signaling pathway.

Combined Ascorbic Acid and Mild Heat Treatment to Improve the Quality of Fresh-Cut Carrots.

Mild heat (MH) treatment and ascorbic acid (AsA) addition can improve the quality of fresh-cut produce when used individually; however, their combined effect remains unclear. Herein, fresh-cut carrots were used as models to explore the effects of MH (50 °C)-AsA (0.5%) on quality properties including reactive oxygen species (ROS) metabolism, antioxidants, lignin metabolism, naturally present microbes, and inoculated pathogens (Escherichia coli O157: H7 and Salmonella Typhimurium) during storage (0-5 d, 4 °C). The results indicate that the antioxidant properties in the MH-AsA group were consistent with those of single treatments, resulting in a consistent ROS-scavenging effect. From day 3-5, lignin synthesis was significantly inhibited by MH-AsA as compared with single treatments, probably because the two enzymes (phenylalanine ammonia-lyase and peroxidase) responsible for lignin synthesis exhibited lower expressions. Microbial analysis revealed that MH-AsA treatment led to the lowest counts of both pathogens and aerobic mesophilic bacteria at 0-5 d. Conversely, the inhibitory effect of MH-AsA treatment on mold and yeast was consistent with the single treatments. These results suggest that MH-AsA is a low-cost and safe approach to improve the physiological characteristics of fresh-cut produce while reducing microbial risk.