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Chapter 6 Impact of salinity stress on medicinal and aromatic plant biotechnology 233
Table 6.1: Impact of salinity on the growth and development of medicinal plants.
Parameter Observation Example species
Seed germination Reduced germination rate under high salinity levels Catharanthus roseus
Chlorophyll Content and biomass
Allocation Shift toward root biomass; reduction in shoot biomass Moringa oleifera
Flowering Delayed or inhibited flowering due to ionic and osmotic stress Trachyspermum
Decreased chlorophyll concentration and photosynthetic efficiency
St. John’s wort
ammi
of reproductive success and medicinal compound production, reduced flowering under salinity stress can severely affect yield, particularly in species that derive me­dicinal properties from their flowers [41].
6.3.2 Impact on photosynthesis and water relations
Salinity stress disrupts the photosynthetic machinery of MAPs, reducing both the chlo­rophyll content and photosynthetic efficiency. For example, salinity-stressed St. John’s wort plants displayed an 18.9% reduction in chlorophyll levels compared to un­stressed controls [42]. Furthermore, salinity affects critical parameters related to pho­tosynthetic performance. Reductions in F
(maximum quantum yield of PSII), Fv/F
v/Fm
(maximum primary yield of PSII photochemistry), and PI (performance index) under salinity stress highlight significant losses in photosynthetic capacity, leading to re­duced biomass and growth [43].
Salt stress disrupts plant water uptake by altering soil solution dynamics and cre­ating artificial drought conditions, despite water availability [44, 45]. To counteract this osmotic challenge, plants synthesize osmoprotectants like amino acids and carbo­hydrates, which help maintain cellular equilibrium in saline environments [46]. This adaptive response enables plants to mitigate some of the harmful effects of salt expo­sure, showcasing their resilience to environmental stressors.
6.3.3 Ionic stress and nutrient imbalance
Salt stress in plants leads to an overaccumulation of sodium and chloride ions, disrupt­ing key cellular processes in Figure 6.1. Studies on moringa have shown increased Na and Cl- levels under saline conditions, accompanied by reduced potassium uptake [47, 48]. This ionic imbalance interferes with the absorption of essential nutrients like potas-
o
+
Figure 6.1: Effect of soil salinity on photosynthesis and water retention.
234 Fatemeh Ahmadi
sium, calcium, and magnesium [49]. Consequently, plants experience impaired growth, metabolic disturbances, and decreased productivity due to the combined effects of ion toxicity and nutrient deficiencies [50].
6.3.4 Oxidative stress and antioxidant response
Salt stress triggers excessive production of reactive oxygen species (ROS) in plants, threatening cellular components. Plants respond by activating antioxidant defenses, in­cluding enzymes like superoxide dismutase and catalase, as well as nonenzymatic com­pounds such as ascorbic acid and flavonoids [51, 52]. These mechanisms help neutralize ROS, protecting cellular structures and enhancing plant resilience under saline condi­tions. Studies on medicinal plants have shown increased antioxidant activity correlating with salt exposure levels, demonstrating plants’ adaptive strategies against oxidative stress [53, 54].
6.3.5 Impact on secondary metabolite production
Salt stress paradoxically affects medicinal plants, often stunting growth, while boost­ing the production of valuable secondary metabolites [55]. Under saline conditions,
Chapter 6 Impact of salinity stress on medicinal and aromatic plant biotechnology 235
these plants frequently increase their synthesis of compounds like tannins, saponins, and phenols [56]. This response involves changes in key biosynthetic pathways that are sensitive to environmental stress [57]. Research has shown that salt exposure can enhance essential oil production in various aromatic plants, potentially serving as a survival mechanism [58]. This phenomenon has significant implications for the medic­inal plant industry, suggesting that controlled salinity could be used to stimulate the production of high-value bioactive compounds [59]. By integrating stress management into cultivation practices, growers may enhance both the therapeutic efficacy and eco­nomic value of medicinal plants. This approach represents a promising strategy for optimizing medicinal plant production in challenging environments [60].

6.4 Molecular responses to salinity stress

Medicinal plants adapt to salt stress through complex molecular mechanisms, involv­ing changes in gene expression, protein activity, and metabolite production. Proteo­mic studies have revealed stress-induced alterations in proteins crucial for tolerance, photosynthesis, and secondary metabolite synthesis [61]. Salt exposure triggers the up­regulation of genes linked to osmolyte production, ion transport, and antioxidant de­fenses. This molecular plasticity not only enhances salt tolerance but also promotes the accumulation of valuable secondary metabolites. Understanding these intricate molecular responses provides insights into how medicinal plants adapt to saline con­ditions, while potentially increasing their therapeutic and economic value [62].
6.5 Salt stress and primary metabolites
in medicinal plants
6.5.1 Amino acids
Salt stress triggers significant shifts in plant amino acid metabolism. Many amino acids, including alanine, arginine, and glycine, increase under saline conditions, with proline showing the most dramatic rise. Nonprotein amino acids and amides also ac­cumulate. Proline’s buildup, observed in various medicinal plants, results from re­duced proline oxidase activity and serves as a key osmoprotectant [60]. This helps maintain cellular balance and structure under salt stress. The overall increase in free amino acids, seen in plants like Catharanthus roseus, aids in osmotic adjustment and provides resources for energy and biosynthesis during stress. These metabolic changes highlight plants’ adaptive strategies for surviving in saline environments, demonstrating the crucial role of amino acids in stress resilience [57].
236 Fatemeh Ahmadi
6.5.2 Proteins
The elevation of free amino acids in plants experiencing salt stress is partially ex­plained by the breakdown of proteins. For example, protein degradation has been ob­served in Catharanthus roseus exposed to salinity [63]. In chamomile and sweet mar­joram, salt stress reduced the levels of soluble proteins, likely due to protein aggregation within the cells [64]. In Achillea fragratissima, a decline in crude protein synthesis was noted at 4,000 ppm salinity [65]. However, some studies indicate an in­crease in protein synthesis under higher salinity levels, suggesting that plants may store nitrogen in proteins for use during recovery from stress [66].
6.5.3 Carbohydrates
Salinity disrupts carbohydrate metabolism in plants, typically leading to imbalances due to reduced photosynthesis and nutrient availability. For instance, fennel plants exhibited a decrease in carbohydrate content under saline conditions [67]. In contrast, plants such as Salvia officinalis and Satureja hortensis showed an increase in carbohy­drates as salinity levels rose, highlighting species-specific responses to salt stress [68, 69].
6.5.4 Lipids
Salt stress significantly impacts the lipid profile of plants, influencing fatty acid and oil synthesis. For example, in Ricinus communis, salinity reduced oil yield in roots but increased oil content in shoots [70]. In Coriandrum sativum, salinity stress led to a no­table decrease in total fatty acid content, with reductions in key fatty acids such as α­linolenic and linoleic acids as NaCl concentrations increased [71–75].

6.6 Study of alkaloids through proteomic and other approaches

Proteomic studies have enhanced our knowledge of secondary metabolite production in medicinal plants, revealing complex mechanisms behind the synthesis of therapeu­tically valuable compounds [76, 77]. Researchers are using advanced techniques like cell cultures and metabolic engineering to increase the yield of these naturally limited substances. Studies on plants such as Catharanthus roseus have shown how various factors, including phytohormones and salt stress, affect protein expression and alka­loid production [78, 79]. This research has identified key enzymes and proteins in-
Chapter 6 Impact of salinity stress on medicinal and aromatic plant biotechnology 237
volved in metabolite synthesis, providing insights into how plants respond to stress by altering their biochemical pathways. These findings open new possibilities for en­hancing the production of medicinal compounds for pharmaceutical and commercial use [80]. In Chelidonium majus, two-dimensional gel electrophoresis revealed 21 pro­teins associated with stress signaling, nucleic acid binding, and defense responses, shedding light on the molecular mechanisms underpinning salinity tolerance [81]. Studies on Papaver somniferum identified codeinone reductase as a central enzyme in morphine biosynthesis, emphasizing its critical role in stress-induced secondary me­tabolite production [82].
These advancements in proteomics provide invaluable insights into the complex molecular pathways that regulate secondary metabolite production, paving the way for optimized cultivation practices and biotechnological innovations to maximize the medicinal and economic potential of these compounds [83, 84].

6.7 Phenolic compounds during stress

Phenolic compounds have emerged as important indicators of salt stress in plants. These diverse molecules, numbering around 9,000, play crucial roles in plant defense, particularly in neutralizing ROS generated during stress conditions [85]. Salt stress dis­rupts photosynthetic processes, leading to increased ROS production and oxidative damage. In response, plants synthesize various phenolic compounds, including phenolic acids, flavonoids, and proanthocyanidins, which act as antioxidants [85]. Research on crop plants consistently shows elevated phenolic content under saline conditions. For example, spearmint and Achillea fragratissima exhibit higher levels of phenolic acids and tannins when exposed to salt stress. Matricaria chamomilla demonstrates increased production of specific phenolic acids like protocatechuic, chlorogenic, and caffeic acids. Similarly, Nigella sativa and Mentha pulegium show a positive correlation between phe­nolic accumulation and salinity levels [86]. In Nigella, cultivated in saline soils, com­pounds such as quercetin, apigenin, and trans-cinnamic acid are found in higher con­centrations, illustrating the plant’s adaptive response to salt stress through enhanced phenolic synthesis.

6.8 Strategies for improving salt tolerance in MAPs

The growing issue of soil salinity poses a major threat to the cultivation of MAPs, which are valued for their therapeutic and economic significance. To address this, various strat­egies have been developed, ranging from traditional agronomic methods to advanced bio­technological approaches, aimed at mitigating the impact of salinity on MAPs [86]
238 Fatemeh Ahmadi
Table 6.2: Strategies to improve salt tolerance in medicinal plants.
Strategy Mechanism Example
plants
Use of plant growth regulators
Application of Mycorrhizae
Genetic engineering Overexpression of salt-tolerance genes like SOS and PCS Artemisia annua
Exogenous osmoprotectants
Enhances stress tolerance through hormonal regulation and antioxidant activity
Improves nutrient uptake and water relations under saline conditions
Protects cellular structures and maintains osmotic balance Chamomile and
Basil and chamomile
Marjoram
sage
(Table 6.2). This section explores these strategies, focusing on their mechanisms and effec­tiveness in enhancing salt tolerance [87].
6.8.1 Exogenous application of plant growth regulators
Plant growth regulators (PGRs) have shown promise in enhancing the salt tolerance of MAPs by influencing key physiological processes [88]. Among these, salicylic acid (SA) has demonstrated particular efficacy in alleviating salt stress effects. Research on St. John’s wort, exposed to saline conditions, revealed that SA treatment significantly boosted growth parameters and photosynthetic performance [89]. SA application led to marked improvements in chlorophyll levels, photosystem efficiency, and electron transport. Addi­tionally, SA treatment reduced stress hormone levels, while increasing antioxidant en­zyme activity, thereby improving the plant’s ability to manage oxidative stress under sa­line conditions. These findings underscore the potential of PGRs, especially SA, as a practical approach to enhancing salt tolerance in medicinal plants, offering a promising strategy for maintaining crop productivity in salt-affected areas [90].
Gibberellic acid (GA3): Gibberellic acid has shown considerable potential in allevi­ating the detrimental effects of salinity on MAPs. For instance, in basil (Ocimum basili- cum), GA3 application under saline conditions significantly improved growth metrics such as plant height, leaf area, and biomass (both fresh and dry weight). Moreover, GA3 enhanced the production of essential oils and their primary constituents, includ­ing linalool and methyl chavicol. These benefits are attributed to GA3’s role in pre­serving membrane stability, enhancing antioxidant enzyme activity, and regulating osmolyte accumulation, which collectively contribute to improved stress resilience [91, 92].
Brassinosteroids (BRs): Brassinosteroids have also demonstrated promising ef­fects on salt tolerance in MAPs. For example, foliar application of 24-epibrassinolide in chamomile (Matricaria chamomilla) under saline conditions enhanced growth,
Chapter 6 Impact of salinity stress on medicinal and aromatic plant biotechnology 239
photosynthetic efficiency, and essential oil content. BRs improve salt tolerance by acti­vating the plant’s antioxidant defense mechanisms, enhancing water relations, and promoting the accumulation of compatible solutes. These combined effects enable plants to maintain physiological stability and adapt to saline environments more ef­fectively [93].
The application of PGRs, including SA, GA3, and BRs, underscores their potential as valuable tools for mitigating salt stress in MAPs, ultimately supporting improved growth, productivity, and secondary metabolite synthesis in challenging environ­ments.
6.8.2 Use of beneficial microorganisms
Utilizing beneficial microorganisms, such as arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR), offers an eco-friendly approach to en­hance salt tolerance in MAPs [94]. These microorganisms improve plant resilience to salinity stress by enhancing nutrient uptake, increasing water absorption, balancing hormones, and activating stress-related biochemical pathways. AMF enhances root–soil interactions, facilitating phosphorus uptake, while PGPR promotes root growth, produ­ces beneficial phytohormones, and bolsters antioxidant defenses 95]. For instance, inoc­ulating Origanum majorana with Glomus mosseae helps mitigate salinity effects by im­proving growth and essential oil production. Similarly, in basil (Ocimum basilicum), PGPR strains like Pseudomonas putida and Bacillus lentus enhance growth and essential oil yield under salt stress by improving antioxidant enzyme activity and nutrient ab­sorption [96]. Overall, these beneficial microbes play a crucial role in supporting plant health and productivity in saline environments, aligning with sustainable agricultural practices.
6.8.3 Genetic approaches
Genetic strategies hold great potential for improving salt tolerance in MAPs by com­bining traditional breeding with advanced biotechnological methods. Traditional breeding allows for the selection and crossing of salt-tolerant varieties, while modern techniques such as genetic engineering and marker-assisted selection enable precise targeting of genes linked to salinity resistance [97]. These approaches can lead to the development of MAPs that are more resilient to saline conditions, enhancing their me­dicinal and economic value. Research has identified specific genes that contribute to salt tolerance, such as the Na introduced into Artemisia annua, resulting in improved growth and artemisinin pro­duction under salt stress [98]. Overall, these genetic advancements provide promising
+/H+
antiporter gene (NHX1), which has been successfully
240 Fatemeh Ahmadi
pathways for enhancing the resilience of MAPs against salinity, supporting sustain­able agricultural practices.
RNA Interference (RNAi) technology: RNAi has been used to suppress genes that hinder salt tolerance [99]. For example, silencing the SmMYB39 gene in Salvia miltior- rhiza enhanced the plant’s tolerance to salt stress. This was achieved by increasing the production of phenolic acids and tanshinones, which are important medicinal compounds [100].
6.8.4 CRISPR/Cas9 gene editing
The CRISPR/Cas9 genome editing system offers precise and efficient modifications, making it a promising approach for enhancing salinity tolerance in MAPs. While its application in MAPs is still developing, this tool shows great promise for creating salt­tolerant plants with improved medicinal properties [101].
6.8.5 Agronomic practices
Agronomic interventions are essential for managing the effects of salinity on MAPs by improving soil conditions and ensuring better plant growth in saline environ­ments [102].
Irrigation management: Efficient irrigation techniques can control soil salinity levels. Drip irrigation, for example, has been shown to maintain lower salinity in the root zone compared to traditional methods like furrow irrigation. Studies on Rosmar- inus officinalis (rosemary) revealed that drip irrigation not only improved plant growth but also increased the yield and quality of essential oils under saline condi­tions [103].
6.8.6 Use of mulches
Mulching is an effective agronomic strategy to mitigate salinity stress. Organic mulches help reduce water loss through evaporation and prevent salt buildup in the root zone. For example, straw mulch applied to Salvia officinalis (sage) significantly enhanced growth and essential oil yield under saline conditions. This practice also improved soil moisture retention, minimized temperature fluctuations, and boosted soil biological ac­tivity [104, 105].
Chapter 6 Impact of salinity stress on medicinal and aromatic plant biotechnology 241
6.8.7 Application of organic amendments
Organic amendments, such as compost and biochar, have proven effective in improv­ing soil health and mitigating salinity-induced stress in MAPs. These amendments en­hance soil structure, promote water retention, and alleviate the negative effects of salt stress [106]. For instance, in Foeniculum vulgare (fennel), vermicompost applica­tion significantly enhanced plant-growth-boosted essential oil yield, and improved an­tioxidant activity under saline conditions. These improvements were attributed to en­hanced soil properties, increased nutrient availability, and stimulation of beneficial microbial activity [107, 108].
6.8.8 Silicon supplementation
Silicon (Si) supplementation has emerged as an effective method for enhancing plant tolerance to salinity [109]. While not essential for growth, Si plays a vital role in miti­gating abiotic stresses, including salt stress. In basil, Si treatment has been shown to alleviate the negative impacts of salinity, resulting in improved growth, enhanced photosynthesis, and increased essential oil production [110]. These benefits are linked to heightened antioxidant enzyme activity, better water absorption, reduced sodium accumulation, and improved potassium balance. Similarly, in peppermint (Mentha pi- perita), Si application leads to better growth and essential oil yield, while reinforcing antioxidant defenses and maintaining membrane integrity under saline conditions. Overall, Si supplementation proves beneficial for promoting resilience in plants facing salinity stress [111].
6.8.9 Application of polyamines
Polyamines, including putrescine, spermidine, and spermine, are crucial for regulat­ing plant growth and stress responses, especially under saline conditions [112]. Their application has been recognized for enhancing salt tolerance in MAPs by stabilizing cell membranes, reducing oxidative damage from ROS, and balancing ion levels. Ad­ditionally, polyamines promote the expression of stress-related genes, improve nu­trient uptake, and assist in osmotic adjustment, thereby strengthening plants’ ability to cope with salinity [113]. For example, in chamomile, putrescine application im­proved growth, flower yield, and essential oil content under salt stress by increasing antioxidant enzyme activity and enhancing photosynthetic efficiency [114]. In sage, spermidine application helped mitigate salinity effects by improving water reten­tion and promoting compatible solute accumulation, which supported growth and essential oil production. Overall, polyamines present a promising strategy for sus-
242 Fatemeh Ahmadi
tainable MAP cultivation in saline environments due to their multifunctional roles in enhancing plant resilience [115].
6.8.10 Nanofertilizers and nanoparticles
Nanofertilizers and nanoparticles are innovative tools for enhancing nutrient uptake efficiency and improving plant resilience to salinity. In Mentha piperita (peppermint), zinc oxide nanoparticles (ZnO NPs) significantly enhanced growth, essential oil yield, and antioxidant activity under saline conditions. These improvements were linked to increased accumulation of compatible solutes, better photosynthesis, and more stable nutrient levels [116]. Similarly, in basil, iron oxide nanoparticles (Fe salt stress by improving growth, increasing essential oil production, and boosting anti­oxidant enzyme activity. These nanoparticles also supported secondary metabolite synthesis and helped maintain membrane stability in plants subjected to salin­ity [117].
NPs) alleviated
₃O₄
6.8.11 Application of melatonin
Melatonin is recognized as an effective enhancer of salt tolerance in various plants, in­cluding medicinal and aromatic species. Acting both as a growth regulator and antioxi­dant, melatonin helps mitigate salinity stress, which has garnered significant research interest. In lemon balm (Melissa officinalis), applying melatonin has been shown to im­prove growth, essential oil yield, and antioxidant activity under saline conditions [118]. This enhancement is linked to melatonin’s ability to promote the accumulation of com­patible solutes like proline, which aids in osmotic regulation, stabilizes chloroplast structures, and maintains ionic balance by reducing sodium uptake and increasing po­tassium retention. These mechanisms collectively improve the plant’s resilience to sa­linity [84].
Melatonin’s potential to enhance salt tolerance, combined with its ability to boost secondary metabolite production, makes it a promising solution for the sustainable cultivation of MAPs in saline environments, offering both therapeutic and economic benefits [119]. Similarly, in Lavandula angustifolia (lavender), melatonin treatment al­leviated the adverse effects of salt stress on growth and essential oil production. The protective mechanism involved heightened antioxidant enzyme activity, improved water relations, and stabilization of membrane integrity [120, 121].
The strategies for improving salt tolerance in MAPs are diverse, ranging from basic agronomic practices to advanced biotechnological techniques. Exogenous appli­cations of PGRs, beneficial microorganisms, silicon, polyamines, and melatonin have shown great potential for enhancing salt tolerance in MAPs by modulating physiologi-