Phenylalanine Ammonia-lyase Activity Research Articles

Responses of plant immune system and rhizosphere soil microbiome to the elicitor BAR11 in Arabidopsis thaliana

Microbial elicitors have been shown to boost plant immunity by inducing defense responses to reduce plant disease. However, little is known about the changes in plant microbiome and metabolism in the process of enhancing plant immunity with elicitors. The protein elicitor BAR11, from Saccharothrix yanglingensis Hhs.015, induces defense responses in Arabidopsis thaliana that enhances resistance to pathogens. In this study, bar11 was inserted into Col-0 A. thaliana to obtain BAR11-Trans plant by Agrobacterium-mediated immersion transformation. BAR11-Trans exhibited an elevated defense level against Pseudomonas syringae pv. tomato DC3000 while experiencing a decline in biomass production of above-ground parts. In the process, BAR11-Trans increased the activity of phenylalanine ammonia lyase (PAL) and catalase (CAT), and up-regulated genes related to plant defense pathways. Furthermore, BAR11-Trans decreased root tip reactive oxygen species (ROS) levels while increasing ROS burst in the leaves. Soil transplantation experiments showed that soil planted with BAR11-Trans could enhance the resistance of Col-0 A. thaliana to DC3000. Analysis of A. thaliana rhizosphere soil through 16S rRNA amplified sequencing revealed that BAR11-Trans increased the relative abundance and diversity of the rhizosphere microbial community, leading to the recruitment of more plant probiotics. Additionally, the accumulation of kaempferitrin and robinin in BAR11-Trans influenced the physicochemical properties of rhizosphere soil and the composition of the bacterial community. In summary, BAR11-Trans exhibited heightened defense levels compared to Col-0, leading to increased secretion of secondary metabolites and the recruitment of a greater number of microorganisms to adapt to the environment. These findings offer novel insights for the potential application of elicitors in agricultural disease control.

Production of Phenylpropanoids, Naphthodianthrones and Antioxidant Status of Hypericum perforatum L. Transgenic Shoots

Fifteen transgenic shoot clones (TS A-TS O) regenerated from Hypericum perforatum hairy roots were evaluated for growth characteristics, phenylpropanoid, and naphthodianthrone production, as well as non-enzymatic and enzymatic antioxidant status. Transgenic shoots displayed higher biomass production and non-altered phenotypes compared to control shoots. Phenylpropanoid production in TS was not related to phenylalanine ammonia lyase activity indicating that alternative enzymes could be involved in the regulation of phenylpropanoid/flavonoid biosynthesis. The positive correlation between phenylpropanoids (phenolic acids, flavonoids, flavan-3-ols, tannins) and non-enzymatic antioxidant assays suggested that these compounds are the main contributors to the redox state and radical scavenging properties of TS clones. More importantly, enhanced naphthodianthrone production (hypericin, pseudohypericin and protopseudohypericin) in TS was associated with the number of leaf dark glands. Concerning the enzymatic antioxidant status of TS, guaiacol peroxidase was shown as the main enzyme that turns down H2O2 levels, while the upregulation of superoxide dismutase coincided with decreased O2•− production. The efficiency of antioxidant enzymes for O2•− and H2O2 neutralization decreased the level of MDA and consequently delayed lipid peroxidation in TS clones. Altogether, H. perforatum transgenic shoots could be proposed as a prospective biotechnological system for scale-up production of naphthodianthrones and phenylpropanoids with antioxidant properties.

Evaluating the imazethapyr herbicide mediated regulation of phenol and glutathione metabolism and antioxidant activity in lentil seedlings.

The imidazolinone group of herbicides generally work for controlling weeds by limiting the synthesis of the aceto-hydroxy-acid enzyme, which is linked to the biosynthesis of branched-chain amino acids in plant cells. The herbicide imazethapyr is from the class and the active ingredient of this herbicide is the same as other herbicides Contour, Hammer, Overtop, Passport, Pivot, Pursuit, Pursuit Plus, and Resolve. It is commonly used for controlling weeds in soybeans, alfalfa hay, corn, rice, peanuts, etc. Generally, the herbicide imazethapyr is safe and non-toxic for target crops and environmentally friendly when it is used at low concentration levels. Even though crops are extremely susceptible to herbicide treatment at the seedling stage, there have been no observations of its higher dose on lentils (Lens culinaris Medik.) at that stage. The current study reports the consequence of imazethapyr treatment on phenolic acid and flavonoid contents along with the antioxidant activity of the phenolic extract. Imazethapyr treatment significantly increased the activities of several antioxidant enzymes, including phenylalanine ammonia lyase (PAL), phenol oxidase (POD), glutathione reductase (GR), and glutathione-s-transferase (GST), in lentil seedlings at doses of 0 RFD, 0.5 RFD, 1 RFD, 1.25 RFD, 1.5 RFD, and 2 RFD. Application of imazethapyr resulted in the 3.2 to 26.31 and 4.57-27.85% increase in mean phenolic acid and flavonoid content, respectively, over control. However, the consequent fold increase in mean antioxidant activity under 2, 2- diphenylpicrylhdrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assay system was in the range of 1.17-1.85 and 1.47-2.03%. Mean PAL and POD activities increased by 1.63 to 3.66 and 1.71 to 3.35-fold, respectively, in agreement with the rise in phenolic compounds, indicating that these enzyme's activities were modulated in response to herbicide treatment. Following herbicide treatments, the mean thiol content also increased significantly in corroboration with the enhancement in GR activity in a dose-dependent approach. A similar increase in GST activity was also observed with increasing herbicide dose.

Exploring the biochemical dynamics in faba bean (Vicia faba L. minor) in response to Orobanche foetida Poir. parasitism under inoculation with different rhizobia strains.

In Tunisia, Orobanche foetida Poir. is considered an important agricultural biotic constraint on faba bean (Vicia faba L.) production. An innovative control method for managing this weed in faba bean is induced resistance through inoculation by rhizobia strains. In this study, we explored the biochemical dynamics in V. faba L. minor inoculated by rhizobia in response to O. foetida parasitism. A systemic induced resistant reaction was evaluated through an assay of peroxidase (POX), polyphenol oxidase (PPO) and phenyl alanine ammonialyase (PAL) activity and phenolic compound and hydrogen peroxide (H2O2) accumulation in faba bean plants infested with O. foetida and inoculated with rhizobia. Two rhizobia strains (Mat, Bj1) and a susceptible variety of cultivar Badi were used in a co-culture Petri dish experiment. We found that Mat inoculation significantly decreased O. foetida germination and the number of tubercles on the faba bean roots by 87% and 88%, respectively. Following Bj1 inoculation, significant decreases were only observed in O. foetida germination (62%). In addition, Mat and Bj1 inoculation induced a delay in tubercle formation (two weeks) and necrosis in the attached tubercles (12.50% and 4.16%, respectively) compared to the infested control. The resistance of V. faba to O. foetida following Mat strain inoculation was mainly associated with a relatively more efficient enzymatic antioxidative response. The antioxidant enzyme activity was enhanced following Mat inoculation of the infected faba bean plant. Indeed, increases of 45%, 67% and 86% were recorded in the POX, PPO and PAL activity, respectively. Improvements of 56% and 12% were also observed in the soluble phenolic and H2O2 contents. Regarding inoculation with the Bj1 strain, significant increases were only observed in soluble phenolic and H2O2 contents and PPO activity (especially at 45 days after inoculation) compared to the infested control. These results imply that inoculation with the rhizobia strains (especially Mat) induced resistance and could bio-protect V. faba against O. foetida parasitism by inducing systemic resistance, although complete protectionwas not achieved by rhizobia inoculation. The Mat strain could be used as a potential candidate for the development of an integrated method for controlling O. foetida parasitism in faba bean.

Effect of total phenolic content (TPC) and activity of phenylalanine ammonia-lyase (PAL) on spot blotch of wheat incited by Bipolaris sorokiniana

Effect of total phenolic content (TPC) and activity of phenylalanine ammonia-lyase (PAL) on spot blotch of wheat incited by Bipolaris sorokiniana

Response of cassava (Manihot esculenta Crantz) genotypes to natural infestation by scale insect pest Stictococcus vayssierei Richard (Hemiptera: Stictococcidae)

Cassava is mostly grown for its starchy roots, which ensure food security. However, it is heavily attacked by the African root and tuber scale (ARTS) Stictococcus vayssierei in Central Africa. This pest is a severe constraint to the production of cassava, food and income security for smallholder farmers. Crop resistance development through the selection of varieties with resistant traits against targeted pests is a promising approach to pest control. This study investigated cassava genotypes' response to natural infestation and determined their resistance levels against S. vayssierei. Six cassava genotypes (two local and four improved) were planted in a completely randomized block design with four replicates. Agronomic parameters and ARTS density were evaluated at 3, 6, 9 and 12 months after planting (MAP). Biochemical content was determined on the pith and cortex of 12 MAP aged tuberous roots. As a result, the improved Excel variety recorded the highest scale density per plant with 102.83 ± 4.14 ARTS/P at 9 MAP. At 12 MAP, high activity of total cyanide (69.18 ± 0.88 and 69.16 ± 1.44 mg/kg) and phenylalanine ammonia-lyase (0.142 ± 0.020 and 0.145 ± 0.010 ΔA/min/mg) were observed in the cortex of the tuberous roots of the improved varieties TMS 96/0023 and TMS 92/0057 which were colonized by the lowest ARTS density. The local variety (Douma) had a high content of total phenols (44.87 ± 1.15 µg/g) in the pith. It also produced the highest yield (23.8 ± 2.9 t ha-1). Varieties TMS 96/0023, TMS 92/0057 and Douma may be the most suitable varieties for the control of ARTS stress.

Influence of different pre-cooling methods on the postharvest storage of ‘Kyoho’ grapes (Vitis labrusca × vinifera ‘Kyoho’)

Abstract Various pre-cooling methods for ‘Kyoho’ grapes were selected, such as forced-air pre-cooling (FAPC), vacuum pre-cooling, natural convection pre-cooling (NCPC), electrolyzed water pre-cooling, cold water pre-cooling, and fluid ice pre-cooling. The aim of this study was to determine the most suitable pre-cooling method for grapes. The storage temperature of grapes was (4±0.5) °C with a relative humidity of 85%, and physiological parameters were measured every 5 d. All pre-cooling methods, except for NCPC, were able to complete the pre-cooling process within 20 min. Interestingly, water and ice cooling methods were not suitable for pre-cooling grapes, probably due to microbial growth and damage to cellular tissue. In contrast, three groups of air pre-cooling methods were more effective in preserving freshness, the best of which was FAPC. It was found that FAPC inhibited grape weight loss, moisture loss, respiration rate, total viable count, malondialdehyde, and relative electrolyte leakage, while stimulating the activities of superoxide dismutase and phenylalanine ammonia-lyase. It also suppressed the activity of polyphenol oxidase. Additionally, in the FAPC group, optimal hardness, titratable acidity, and vitamin C content were observed. Among all the tested methods, FAPC demonstrated better efficacy in delaying the softening and senescence of ‘Kyoho’ grapes, thereby preserving fruit quality and nutritional content.

The phenylalanine ammonia-lyase inhibitor AIP induces rice defence against the root-knot nematode Meloidogyne graminicola.

The phenylalanine ammonia-lyase (PAL) enzyme catalyses the conversion of l-phenylalanine to trans-cinnamic acid. This conversion is the first step in phenylpropanoid biosynthesis in plants. The phenylpropanoid pathway produces diverse plant metabolites that play essential roles in various processes, including structural support and defence. Previous studies have shown that mutation of the PAL genes enhances disease susceptibility. Here, we investigated the functions of the rice PAL genes using 2-aminoindan-2-phosphonic acid (AIP), a strong competitive inhibitor of PAL enzymes. We show that the application of AIP can significantly reduce the PAL activity of rice crude protein extracts invitro. However, when AIP was applied to intact rice plants, it reduced infection of the root-knot nematode Meloidogyne graminicola. RNA-seq showed that AIP treatment resulted in a rapid but transient upregulation of defence-related genes in roots. Moreover, targeted metabolomics demonstrated higher levels of jasmonates and antimicrobial flavonoids and diterpenoids accumulating after AIP treatment. Furthermore, chemical inhibition of the jasmonate pathway abolished the effect of AIP on nematode infection. Our results show that disturbance of the phenylpropanoid pathway by the PAL inhibitor AIP induces defence in rice against M. graminicola by activating jasmonate-mediated defence.

Quality evaluation of fresh pistachios (Pistacia vera L.) cultivars coated with chitosan/TiO2 nanocomposite

Fresh pistachios are rich in dietary fiber, minerals and unsaturated fatty acids, but they have a short shelf life. This investigation examined the effect of pre-harvest foliar application with chitosan (500 and 1000 mg. L−1), nano-chitosan (250 and 500 mg. L−1), and chitosan/TiO2 nanocomposite (250 and 500 mg. L−1) coating films on the postharvest physiology and storage of fresh pistachios (Pistacia vera cvs. Akbari and Ahmad Aghaei) cultivar during storage at 4 ± 0.5 °C. It was found that, fresh pistachios' shelf life could by increased by up to 30 days by the use of chitosan/TiO2 nanocomposite coating for foliar application. The decay index of the composite coated fruits was 4–6 % lower than that of the control group, and after 50–60 days the bacterial contamination appeared in cultivars; respectively. The nanocomposite treatments reduced the fruits weight between 30 and 40 %, which was 15 % higher that of than uncoated fruits. The pre-harvest application of chitosan/TiO2 coating reduced microbial contamination, weight loss, phenylalanine ammonialyase (PAL) activity and saturated fatty acids, and increased unsaturated fatty acids, antioxidant properties, sensory properties, essential minerals, superoxide dismutase (SOD), quality indicators and shelf life. These results demonstrated that the chitosan/TiO2 (250 and 500 mg. L−1) coating film effectively preserved the nutrient composition, sensory quality, nutritional value, antioxidant capacity and shelf life of fresh pistachio.

Agricultural wastes: a new promising source for phenylalanine ammonia-lyase as anticancer agent.

The present study aims to investigate the physicochemical characteristics of phenylalanine ammonia-lyase (PAL) extracted from agricultural waste and its potential use as an anticancer agent in comparison to microbial PAL. We extracted and partially purified PAL from agricultural waste sources. We assessed the temperature and pH range of PAL and determined enzyme kinetics parameters including Michaelis constants (Km), maximum velocity (Vmax), and specificity constant values (Vmax/Km). Additionally, we examined the effects of different storage temperatures on PAL activity. In our analysis, we compared the efficacy of agricultural waste-derived PAL with PAL from Rhodotorula glutinis. The results demonstrated that PAL extracted from agricultural waste exhibited significantly higher specific activity (Vmax/Km) compared to its microbial counterpart. The agricultural waste-derived PAL displayed a stronger affinity for phenylalanine, as indicated by a lower Km value than the microbial PAL did. Furthermore, PAL from agricultural waste maintained activity across a broader temperature and pH range (15-75°C, pH 5-11), in contrast to microbial PAL (20-60°C, pH 5.5-10). Importantly, the PAL derived from agricultural waste exhibited superior stability, retaining over 90% of its activity after 6months of storage at room temperature (25°C), whereas microbial PAL lost more than 70% of its activity under similar storage conditions. In anticancer experiments against various cancer cell lines, agricultural waste-derived PAL demonstrated greater anticancer activity compared to microbial PAL. These findings suggest that PAL sourced from agricultural waste has the potential to be a safe and effective natural anticancer agent.

Investigation of Melatonin Incorporated CMC-Gelatin Based Edible Coating on the Alleviation of Chilling Injury Induced Pericarp Browning in Longkong.

Longkong (Aglaia dookkoo Griff.) fruit is prone to rapid pericarp browning and shortened shelf life (<7 days) under prolonged low-temperature storage. This study investigates the effect of an edible coating, comprising carboxymethyl cellulose (CMC) and gelatin in a fixed 3:1 ratio, integrated with various concentrations of melatonin (MT) (0.4, 0.8, and 1.2 mM/L) to mitigate chilling injury in longkong fruit. Coated longkong fruits were stored at 13 °C with 90% relative humidity for 18 days and underwent physicochemical evaluations every three days. Samples coated with CMC-Gel without MT and uncoated fruits were served as controls. The findings indicated that the CMC-Gel-MT coating significantly mitigated pericarp browning, chilling injury, weight loss, and respiration rate increase under extended cold storage conditions. High concentrations of MT (≥0.8 mM/L) in the coating notably inhibited the activities of cellular degrading enzymes such as lipoxygenase and phospholipase D. This inhibition contributed to reduced membrane permeability, lower reactive oxygen species accumulation (H2O2, OH-, O2-), and decreased malondialdehyde levels in the longkong pericarp. Furthermore, the CMC-Gel-MT coating increased the activity of phenylalanine ammonia lyase, leading to an enhancement in phenolic content. Consequently, it improved the fruit's ability to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,20-azino-di-3-ethylbenzthiazoline sulfonic acid) radicals. Control samples exhibited high levels of pericarp browning-related enzymes (polyphenol oxidase, peroxidase), whereas CMC-Gel-MT-coated fruits, particularly at higher MT concentrations, showed significant reductions in those enzyme activities. In conclusion, incorporating high concentrations of MT in a CMC-Gel-based edible coating is a promising alternative for mitigating chilling injury in longkong fruit.

Role of Acetic Acid and Nitric Oxide against Salinity and Lithium Stress in Canola (Brassica napus L.).

In this study, canola (Brassica napus L.) seedlings were treated with individual and combined salinity and lithium (Li) stress, with and without acetic acid (AA) or nitric acid (NO), to investigate their possible roles against these stresses. Salinity intensified Li-induced damage, and the principal component analysis revealed that this was primarily driven by increased oxidative stress, deregulation of sodium and potassium accumulation, and an imbalance in tissue water content. However, pretreatment with AA and NO prompted growth, re-established sodium and potassium homeostasis, and enhanced the defense system against oxidative and nitrosative damage by triggering the antioxidant capacity. Combined stress negatively impacted phenylalanine ammonia lyase activity, affecting flavonoids, carotenoids, and anthocyanin levels, which were then restored in canola plants primed with AA and NO. Additionally, AA and NO helped to maintain osmotic balance by increasing trehalose and proline levels and upregulating signaling molecules such as hydrogen sulfide, γ-aminobutyric acid, and salicylic acid. Both AA and NO improved Li detoxification by increasing phytochelatins and metallothioneins, and reducing glutathione contents. Comparatively, AA exerted more effective protection against the detrimental effects of combined stress than NO. Our findings offer novel perspectives on the impacts of combining salt and Li stress.

Estimation of phenyl alanine ammonialyase (PAL) and some phytoalexins in Vicia faba plants infected with spot disease caused by Alternaria alternata

Biological agents are an important approach to controlling many fungal phytopathogens; plants produce phytoalexins such as phenol, alkaloids and tannins as a response to the biotic stresses. The results showed that the addition of the biological control of Trichoderma harzianum to Alternaria alternata caused an increase in the activity of the enzyme phenylalanine ammonialyase (PAL) after 20 days from the start of the greenhouse experiment, which was 6.21 mmol/min in the Spinal variety, compared with the enzyme activity in the treatment with A. alternata alone, which was 5.51 mmol/min. The variety Netherland mulch recorded a lower phenolic content in all treatments, and the treatment with biological control T. harzianum in addition to A. alternata had the highest content as it reached 4.50 mg/g on day 20, Netherland mulch variety showed the most amount of alkaloids and tannins than others during different periods. The plants treated with the biological control of T. harzianum contained the most alkaloids on day 20, which was 5.15% It decreased to 3.65% on day 30. The most tannins in Netherland mulch variety were in the plants treated with biological control T. harzianum alone, which was 3.93% on day 20, compared to the plants treated with A. alternata alone, which was 0.71%. The addition of biological control of T. harzianum to A. alternata increased the plant amount of tannins, which was 2.01%. The study would help to understand the role of T. harzianum as a biological control to the response of the varieties to the formation of plant defenses, including phytoalexins.

Shading and Planting Material Type Effects on Some Physiological and Biochemical Compounds Synthesis in Desmodium adscendens Plants Leaves in the Domestication Phase, Daloa, Côte d'Ivoire

Desmodium adscendens (Sw.) is a plant of the Fabaceae Family, a wild medicinal plant frequently found in industrial plantations, where it is exposed to pesticide residues. This makes species unsuitable for use as a raw material in pharmaceutical industry. To this end, domestication is one of best ways of cleaning up the species. During this process, the evaluation of a few physiological and biochemical compounds would be an indicator for the choice of planting material and the ecology species. Two types of plant material (seeds and stem) grown under three types of shade (accentuated, moderate, no shade) were tested. The leaves of various 90-day-old plants were used to determine physiological and biochemical parameters various. Results showed that chlorophyll a, carotenoids, polyphenols, total sugars, protein, proline and the activities of tyrosine ammonia-lyase, polyphenol oxidase and catalase were higher in leaves with moderate and no shading. However, under heavy shading, chlorophyll b content, phenylalanine ammonia lyase (PAL) and peroxidase enzyme activities were higher. Whatever shading type, the chl (a) content remained lower than that of chl (b). Also, total sugars, proline, ascorbate peroxidase and catalase were more abundant in the leaves of plants grown from stem. In contrast, phenolics, proteins and PPO are more abundant in leaves of seedlings.

Calcium chloride treatment maintains quality and enhances antioxidant capacity of fresh-cut carrots through modulating DcCMLs-DcCAMTA3-mediated phenylpropanoid pathway

Whitening and microbial infection are two major reasons to induce quality deterioration in fresh-cut carrots. Calcium chloride (CaCl2) could enhance the quality attributes in fresh-cut carrots, but the molecular mechanism is still not illuminated. In this study, CaCl2 treatment suppressed the increases of whiteness index and total bacterial count as well as enhanced the antioxidant capacity of fresh-cut carrots. In addition, CaCl2 treatment promoted phenolics accumulation by upregulating the transcripts and enzyme activity of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H) and 4-coumarate coenzyme A ligase (4CL) in phenylpropanoid pathway. However, this treatment inhibited the increase of enzyme activity and gene expression of cinnamyl alcohol dehydrogenase (CAD), thus suppressing lignin synthesis and whitening process in fresh-cut carrots. Moreover, a calmodulin-binding transcription activator (CAMTA) transcription factor DcCAMTA3, was induced in response to CaCl2 treatment. DcCAMTA3 activated the transcription of DcPAL1, DcC4H and Dc4CL1 genes and repressed the expression of DcCAD1 gene by directly targeting the CG-box. DcCML3/11, a calmodulin-like protein, interacted with DcCAMTA3, and this interaction enhanced DcCAMTA3-mediated transcriptional activation of DcPAL1, DcC4H and Dc4CL1 and repression of DcCAD1. Therefore, CaCl2 treatment is effective in maintaining quality and enhancing antioxidant capacity of fresh-cut carrots by the cooperative action of DcCMLs and DcCAMTA3 in positively regulating phenylpropanoid pathway.

Acetic acid delays fresh-cut cassava browning through fine-tunning redox homeostasis

Cassava is one of the important commercial crops in tropical and subtropical regions. However, cassava browning is an important factor restricting its postharvest storage, shelf life and quality. Therefore, suitable approaches are needed to control fresh-cut cassava browning. Acetic acid alleviates the effects of different abiotic stresses and prolongs the shelf life of food. The study aimed to study the effects of exogenous acetic acid on postharvest browning in fresh-cut cassava. The results showed that acetic acid treatment strongly delayed the discoloration during storage through reducing the activities of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), and inhibiting the generation of reactive oxygen species (ROS) and malondialdehyde (MDA), as well as accelerating loss of total phenolic and flavonoid. Notably, acetic acid treatment markedly promoted the accumulation of acetic acid, delayed loss of total starch and had slight impact on jasmonic acid (JA). In addition, JA treatment also delayed discoloration during cassava storage. These findings suggested that the delay of cassava browning via acetic acid treatment could be attributed to maintenance of redox homeostasis by modulating repair of oxidatively damaged proteins, providing a new way to prevent the browning of fresh-cut cassava and elucidates the anti-browning mechanism of acetic acid treatment.

Seed Soaking Using Methanol Kalanchoe pinnata Leaf Extracts Induces Rice Resistance against Bacterial Leaf Blight

Bacterial leaf blight (Xanthomonas oryzae pv. oryzae) is a significant rice disease. Aqueous crude extracts of Kalanchoe pinnata were shown to induce rice resistance against the disease. This study aims at testing the disease-reducing effects of K. pinnata leaf extracts using the liquid–liquid extraction method with three different solvents (dichloromethane, methanol, and water). This serves as a basis to select appropriate extracts for effective disease control. Three concentrations (1, 1.5, and 2%) of each extract were tested using seed soaking. The extracts did not show adverse effects on seed germination and seedling growth. Methanol extracts showed significantly different effects compared to those of the untreated control. The involvement of induced resistance in the disease reduction was shown through activities of the four defense-related and antioxidant enzymes, i.e., peroxidase (POX), catalase (CAT), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL). Using 1% methanol extract, activities of POX and CAT involved in hydrogen peroxide production in rice tissues increased 1–4 days after pathogen inoculation (DAI) and remained at high levels until 6 DAI. Activities of PPO and PAL involved in resistance signaling pathways significantly increased after pathogen inoculation. Activities of the four enzymes generally increased after pathogen inoculation and reached higher levels with extract applications.

Methyl jasmonate conferred Arsenic tolerance in Thymus kotschyanus by DNA hypomethylation, stimulating terpenoid metabolism, and upregulating two cytochrome P450 monooxygenases

Arsenic (As) is a highly cytotoxic element impairing normal cellular functions, and its bioremediation has become one of the environmental concerns. This study explored the molecular and physiological responses of thyme (Thymus kotschyanus) seedlings to incorporating As (0 and 10 mgl-1) and methyl jasmonate (MJ; 0 and 10 µM) into the culture medium. The MJ treatment reinforced root system and mitigated the As cytotoxicity risk. MJ contributed to hypomethylation, a potential adaptation mechanism for conferring the As tolerance. Two cytochrome P450 monooxygenases, including CYP71D178 and CYP71D180 genes, were upregulated in response to As and MJ. The MJ treatment contributed to up-regulation in the γ-terpinene synthase (TPS) gene, a marker gene in the terpenoid metabolism. The As presence reduced photosynthetic pigments (chlorophylls and carotenoids), while the MJ utilization alleviated the As toxicity. The MJ supplementation increased proline accumulation and soluble phenols. The application of MJ declined the toxicity sign of As on the concentration of proteins. The activities of peroxidase, catalase, and phenylalanine ammonia-lyase (PAL) enzymes displayed an upward trend in response to As and MJ treatments. Taken collective, MJ can confer the As tolerance by triggering DNA hypomethylation, regulating CYPs, and stimulating primary and secondary metabolism, especially terpenoid.

Induction of Resistance in Mandarin Fruits Against Penicillium digitatum, the Causative Agent of Green Mold Infection

This study was conducted at the College of Agricultural Engineering Sciences / University of Baghdad during 2021-2022. This work aimed to evaluate the efficiency of Sodium Bicarbonate(SB) and Phylex to protect and induce mandarin fruits’ resistance against the green mold pathogen, Penicillium digitatum at 4ºCstorage conditions. The molecular diagnosis by PCR technology using the primers ITS1 and ITS4, confirmed the identification of P. digitatum. The genetic tree showed that the two Iraqi isolates(K1andK2) were belonging to one group, matched by 98% with the universal isolates registered in the GenBank of China (MZ8819 and MT448740.1) and Portugal(OK0949001). The best treatment method was when the wounded fruits were immersed in Phylex at 4ml/ liter for an hour, then inoculated with a drop of fungal spore’s suspension (103 spores.ml-1). In this treatment, no fungal infection was observed on the fruits after 42days compared to the Inoculation with P.digitatum(15.27%) under refrigerated condition(4ºC). The highest rate of peroxidase (POX) and Phenylalanine ammonia-lyase (PAL) activity was reported 194.22, unite (estimated based on the changes of spectrophotometer OD / minute / gram fresh weight) and 26.33 µg cinnamic acid / hour / gram fresh weight respectively. The above results indicated that the Phylex was more efficient to induce fruit’s resistance against P.

Biocontrol potential of lipopeptides produced by the novel Bacillus altitudinis strain TM22A against postharvest Alternaria rot of tomato

Bacillus genus exhibits antagonistic potential against a broad range of pathogenic fungi. This study presents the antifungal potential of lipopeptides (LPs) produced by a novel strain Bacillus altitudinis TM22A to suppress the infection of Alternaria rot on tomato. We investigated the antifungal effect of ten Bacillus species against Alternaria alternata, where four Bacillus strains TM22A, MCM61, S2, and CPCF54 exhibited antifungal activity. The LPs extracted from TM22A, MCM61, S2, and CPCF54 restricted mycelial growth and conidial germination of A. alternata in vitro with the highest inhibition produced by TM22A. Liquid chromatography and mass spectrometry (LC-MS) analysis revealed the presence of surfactin, fengycin, iturin, bacillibactin, and bacilysin in strain TM22A at 1036.69, 1435.76, 1085.58, 883.26, and 271.13 m/z respectively. In an In planta assay LPs from TM22A caused the lowest lesion diameter and disease incidence on tomato fruit. Furthermore, the activity of defense enzymes viz, catalase (CAT), polyphenol oxidase (PPO), peroxidase (POD), superoxide dismutase (SOD), and phenylalanine ammonia-lyase (PAL) was enhanced in Bacillus LPs treated tomato fruit with the more pronounced impact by TM22A. Postharvest quality analysis revealed little impact on ascorbic acid, fruit pH, total soluble solids (TSS), titratable acidity (TA), weight loss, and fruit firmness compared to healthy control. This is the first comprehensive study to reveal the antifungal effect of Bacillus-derived LPs against postharvest Alternaria rot of tomato and its role in defense elicitation to protect the fruit from pathogen invasion.