Phenylalanine Ammonia-lyase Activity Research Articles

Selenium nanoparticles conferred drought tolerance in tomato plants by altering the transcription pattern of microRNA-172 (miR-172), bZIP, and CRTISO genes, upregulating the antioxidant system, and stimulating secondary metabolism.

Drought stress is one of the major limiting factors for the production of tomato in Iran. In this study, the efficiency of selenate and Se nanoparticle (SeNP) foliar application on tomato plants was assessed to vestigate mitigating the risk associated with water-deficit conditions. Tomato plants were treated with SeNPs at the concentrations of 0 and 4mg L-1; after the third sprays, the plants were exposed to water-deficit conditions. The foliar spraying with SeNPs not only improved growth, yield, and developmental switch to the flowering phase but also noticeably mitigated the detrimental risk associated with the water-deficit conditions. Gene expression experiments showed a slight increase in expression of microRNA-172 (miR-172) in the SeNP-treated plants in normal irrigation, whereas miR-172 displayed a downregulation trend in response to drought stress. The bZIP transcription factor and CRTISO genes were upregulated following the SeNP and drought treatments. Drought stress significantly increased the H2O2 accumulation that is mitigated with SeNPs. The foliar spraying with Se or SeNPs shared a similar trend to alleviate the negative effect of drought stress on the membrane integrity. The applied supplements also conferred drought tolerance through noticeable improvements in the non-enzymatic (ascorbate and glutathione) and enzymatic (catalase and peroxidase) antioxidants. The SeNP-mediated improvement in drought stress tolerance correlated significantly with increases in the activity of phenylalanine ammonia-lyase, proline, non-protein thiols, and flavonoid concentrations. SeNPs also improved the fruit quality regarding K, Mg, Fe, and Se concentrations. It was concluded that foliar spraying with SeNPs could mitigate the detrimental risk associated with the water-deficit conditions.

Oligogalacturonides reduced postharvest losses by alleviating mechanical damage in Goji berry (Lycium barbarum Miller)

Goji berry (Lycium barbarum) is a typical berry with a tender and thin pericarp, high moisture content and hollow interior, making it highly susceptible to mechanical damage caused by harvest and transportation. As a consequence, this susceptibility results in considerable economic losses within the fresh produce industry due to the degradation of fruit quality and subsequent decay. Here, oligogalacturonides (OGs), a well-characterized plant damage-associated molecular pattern, were applied to reduce postharvest loss and extend the shelf life of berries of the goji cultivar NingQi1. The loss rate in the OGs treatment was 44.45 % lower than that in the 0.8 mm compression treatments on Day 6, and the OGs treatment also effectively inhibited softening and delayed weight loss. The activity of superoxide dismutase increased by 27.4 % on the 4th day after OGs treatment, accompanied by a decrease in malondialdehyde content ranging from 14.2 % to 24.8 % on days 2 to 4 compared to the control group. Furthermore, the phenylalanine ammonia-lyase activity of OGs-treated goji berries was 57.0 % higher than that of the control at 6th days, which slowed the reduction in secondary metabolites such as total flavonoids. The findings of this study indicated that OGs may reduce postharvest losses caused by mechanical damage, extend the shelf life of goji berry by reducing softening and weight loss, and reduce wounding injury by minimizing free radical form of reactive oxygen species.

Proper doses of brassinolide enhance somatic embryogenesis in different competent Korean pine cell lines during embryogenic callus differentiation.

Somatic embryogenesis of Korean pine (Pinus koraiensis Sieb. Et Zucc.), an ecologically and econimically very important conifer species, was hindered by the gradually weakens and fast runaway of the embryogenicity and embryo competence of the embryogenic callus. Brassinolide (BL) has shown the enhancing capability of somatic embryo regeneration. For checking the function of BL in this issue, we applied different concentrations of BL to Korean pine callus materials exhibiting different embryogenic capacities and subsequently monitored the physiological alterations and hormone dynamics of the embryogenic callus. Our study revealed that calli with different embryogenic strengths responded differently to different concentrations of BL, but the effect after the addition of BL was very uniform. The addition of BL during the proliferation phase of embryogenic callus may help to stimulate the biological activity of callus during the proliferation process and improve the level of cell metabolism, which is accompanied by a reduction in storage substances. BL could reduce the level of endogenous auxin IAA in embryogenic callus and increase the level of abscisic acid to regulate cell division and differentiation. In addition, the MDA content in the callus was significantly decreased and the activity of antioxidant enzymes was significantly increased after the addition of BL. During the proliferation of embryogenic callus, BL was added to participate in the metabolism of phenylpropane in the cells and to increase the activity of phenylalanine ammonia-lyase and the content of lignin in the cells. We deduced that the proper doses of BL for Korean pine embryogenic callus culture were as follow: calli with low, high and decreasing embryogenicity were subcultured after the addition of 0.75 mg/L, 0.35 mg/L, 2.00 mg/L BL, respectively, during proliferation culture stage.

Modulatory role of nitric oxide in wound healing of potato tubers

IntroductionWounding of potato (Solanum tuberosum L.) tubers affects the postharvest storage qualities and marketability. To mitigate wound-related losses, it is necessary to accelerate wound healing (WH) responses of potato tubers. While role of nitric oxide (NO) in healing of wounded biological tissues is well known, its impact on WH responses of potato tubers has not been examined thoroughly. The aim of this study was to elucidate the potential modulatory role of NO in WH response of potato tubers by determining the activity of key enzymes associated with NO biosynthesis.MethodsUsing a model wounding system, tuber tissue discs were extracted from mini tubers of cv. Russet Burbank to examine their WH responses. Tuber discs were treated with NO related chemicals, such as NO scavenger, nitric oxide synthase (NOS) inhibitor, and NO donors. The effect of these treatments on in vitro production of NO, activity of nitrate reductase, NOS, and phenylalanine ammonia-lyase were determined. The accumulation of suberin polyphenolics (SPP) in wounded parenchyma cells, an indicator of early suberization process, was analyzed histologically.ResultsTreatment of tuber tissues with the higher dose of an NO scavenger (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) at the time of wounding completely ceased SPP accumulation between 0 h to 3 d after wounding. Increased production of NO and enhanced activity of NOS were also observed immediately after wounding of the tuber tissues. The results suggest that NO is a key biological modulator in WH responses of potato tubers and could be optimized as a potential exogenous treatment to counter wound-related losses.

Endogenous compound chenodeoxycholic acid in potatoes can delay enzymatic browning by inducing antioxidant capacity

Enzymatic browning is a primary factor affecting the quality of fresh-cut potatoes. Our previous research demonstrated that warming the tubers after cold storage can alleviate the enzymatic browning of fresh-cut potatoes. In this research, we explored if some new endogenous compounds play roles in browning inhibitory effects. Utilizing a non-targeted metabolomics approach on tubers subjected to pre-cutting different temperature treatments, thirteen differential metabolites in potatoes were revealed, and five compounds with anti-browning effects were screened out. Among these five compounds, chenodeoxycholic acid (CDCA) exhibited the best inhibitory effect on browning and has not been reported in plants till now. The changes in CDCA content in tubers of pre-cutting different temperature treatments were further confirmed by LC-MS/MS. Subsequent experiments showed that the anti-browning effect of pre-cutting CDCA treatment was much better than post-cutting. Pre-cutting CDCA treatment only at a low concentration of 5 mg L−1 for a 10 min soaking and then standing for 6 h extended the shelf life of fresh potato slices to 5 d from less than 1 d of control. Not a big difference in polyphenol oxidase (PPO) activity and tyrosine contents were caused by pre-cutting CDCA treatment compared with control. However, pre-cutting CDCA treatment caused a notable reduction in the contents of malondialdehyde (MDA), superoxide anion (O2-), H2O2, and lipoxygenase (LOX) activity, and elevated the activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). Meanwhile, CDCA suppressed phenylalanine ammonia-lyase (PAL) activity and reduced the contents of both individual and total phenolic compounds. These results suggest that CDCA, an endogenous compound in potato tuber, can improve the anti-browning ability of fresh-cut potatoes mainly by inducing the enhancement of antioxidant capacity.

Zinc and copper toxicity in basil (Ocimum basilicum L.) seedlings: Role of melatonin in mitigating stress

The effects of melatonin (MT) on the growth and antioxidant responses of basil (Ocimum basilicum L.) exposed to copper and zinc excess stress are investigated. For this purpose, seedlings were exposed to different concentrations of copper oxide (0, 50 and 150 µM), zinc oxide (0, 50 and 100 µM) and melatonin (0 and 100 µM) for 14 days. Growth and biochemical parameters, the activity of some antioxidant enzymes, and phenylalanine ammonia lyase (PAL) and some of the main nutritional elements in the shoot and root of seedlings were measured. The results of this study showed that the excess of copper (Cu 50 and 150 µM) and zinc (Zn 50 and 100 µM) causes a decrease in growth, and a higher oxidative stress indicator such as hydrogen peroxide and malondialdehyde (MDA) content. The homeostasis balance of mineral nutrition of basil seedlings was also disrupted under copper and zinc stress. Melatonin treatment in Cu, Zn stressed seedlings improved growth parameters by reducing the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) and increasing the activity of PAL and antioxidant enzymes. Melatonin also regulated the distribution of minerals in basil seedlings under Cu and Zn stress conditions. The results of this research showed that MT can improve the tolerance of basil seedlings against copper and zinc stress by modulate growth and antioxidant responses and regulating mineral nutrition.

Influence of myoinositol on post-ripening and softening of Prunus salicina ‘Wushan plum’

To investigate the impact of myoinositol (MI) immersion treatment on the storage characteristics of Prunus salicina ‘Wushan Plum’, we assessed myoinositol content, ascorbic acid content, weight loss, decay rate, alteration in color, titratable acidity (TA), soluble solids (TSS), firmness, cell wall metabolism-related compounds, enzyme activities, and levels of gene expression throughout a 15-day storage period at 25 °C. The results revealed that MI treatment could increase myoinositol and ascorbic acid contents of fruit during the pre-storage period, effectively delay the color transformation and decay progression of P. salicina ‘Wushan Plum’ while maintaining elevated levels of TA and TSS. Furthermore, it suppressed the increase in weight loss and the decline in firmness. Notably, the delay in softening was more pronounced in the group treated with 100 mg L−1 MI compared to other groups. MI treatment regulated the expression levels of various genes associated with cell wall metabolism, which resulted in reduced activities of polygalacturonase (PG), pectate lyase (PL), pectin methylesterase (PME), cellulase (Cx), and β-glucosidase (β-Glu), and increased activities of phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD). Furthermore, MI treatment maintained higher levels of covalently bound pectin (CSP), cellulose, and lignin, while decreasing the levels of water-soluble pectin (WSP) in the fruit. As a result, MI treatment could delay the depolymerization of cell wall polysaccharides and further delay the quality deterioration of P. salicina ‘Wushan Plum’ fruits under ambient storage environment and prolong the storage period.

Methyl jasmonate improves resistance in scab-susceptible Red Delicious apple by altering ROS homeostasis and enhancing phenylpropanoid biosynthesis

Apple (Malus domestica) is an economically important rosaceous fruit crop grown at temperate climate zones. Nevertheless, its production is severely affected by scab disease caused by the ascomycetous fungus Venturia inaequalis (VI). Methyl jasmonate (MeJA) is a stress induced plant hormone, shown to induce resistance against wide range of pathogens. The current study investigated the role of MeJA in promoting scab tolerance in susceptible apple varieties through exogenous application of optimized (100 μM) MeJA concentration, followed by VI infection. According to our analysis, applying MeJA exogenously onto leaf surfaces resulted in increased membrane stability and decreased malondialdehyde levels in Red Delicious, suggesting that MeJA is capable of protecting tissues against oxidative damage through its role in restoring membrane stability. In addition, the changes in the levels of key antioxidative enzymes and reactive oxygen species (ROS) showed that exogenous MeJA maintains ROS homeostasis as well. Higher phenylalanine ammonia-lyase activity and increased accumulation of phenylpropanoids in MeJA-treated VI-infected plants indicated the MeJA reprogrammed phenylpropanoid biosynthesis pathway for scab tolerance. Our study of scab tolerance in apples induced by MeJA provides new insights into its physiological and biochemical mechanisms.

Investigation of the antibacterial activity of benziothiazolinone against Xanthomonas oryzae pv. oryzae

Plant pathogenic bacteria can cause numerous diseases for higher plants and result in severe reduction of crop yield. Introduction of new bactericides can always effectively control these plant diseases. Benziothiazolinone (BIT) is a novel fungicide registered in China for the control of plant fungal diseases, however, its anti-bacterial activity is not well studied. The results of activity tests showed that BIT exhibited stronger inhibitory activity against bacteria, particularly for Xanthomonas oryzae pv. oryzae (Xoo) (EC50 = 0.17 μg/mL), which was superior than that of the tested fungi in vitro. BIT also exhibited excellent protective and curative activity against rice bacterial leaf blight (BLB) caused by Xoo with the control efficacies of 71.37% and 91.64% at 600 μg/mL, respectively. After treatment with BIT, Xoo cell surface became wrinkled and the cell shape was distorted with extruding cellular content. It was also found that BIT decreased DNA synthesis and affected the biofilm formation and motility of Xoo cells. However, no significant change in the protein content was observed. Moreover, the results of quantitative real-time PCR also showed that expressions of several genes related to DNA synthesis, biofilm formation and motility of Xoo cells were down- or up-regulated, which further proved the anti-bacterial activity of BIT in influencing the biological properties of Xoo. Additionally, BIT also enhanced the activity of phenylalanine ammonia lyase (PAL), a plant defense enzyme. Taken together, benziothiazolinone might be served as an alternative candidate for the control of BLB.

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.