Seaweed-Based Compounds and Products for Sustainable Protection against Plant Pathogens.

Sustainable agricultural practices increasingly demand novel, environmentally friendly compounds which induce plant immunity against pathogens. Stimulating plant immunity using seaweed extracts is a highly viable strategy, as these formulations contain many bio-elicitors (phyco-elicitors) which can significantly boost natural plant immunity. Certain bioactive elicitors present in a multitude of extracts of seaweeds (both commercially available and bench-scale laboratory formulations) activate pathogen-associated molecular patterns (PAMPs) due to their structural similarity (i.e., analogous structure) with pathogen-derived molecules. This is achieved via the priming and/or elicitation of the defense responses of the induced systemic resistance (ISR) and systemic acquired resistance (SAR) pathways. Knowledge accumulated over the past few decades is reviewed here, aiming to explain why certain seaweed-derived bioactives have such tremendous potential to elicit plant defense responses with considerable economic significance, particularly with increasing biotic stress impacts due to climate change and the concomitant move to sustainable agriculture and away from synthetic chemistry and environmental damage. Various extracts of seaweeds display remarkably different modes of action(s) which can manipulate the plant defense responses when applied. This review focuses on both the similarities and differences amongst the modes of actions of several different seaweed extracts, as well as their individual components. Novel biotechnological approaches for the development of new commercial products for crop protection, in a sustainable manner, are also suggested.

Overexpression of a novel SbMYB15 from Salicornia brachiata confers salinity and dehydration tolerance by reduced oxidative damage and improved photosynthesis in transgenic tobacco.

MAIN CONCLUSION: SbMYB15, R2R3-type MYB was induced by the different stresses, and conferred stress tolerance in transgenic tobacco by regulating the expression of stress-responsive genes. MYBs are the master regulators of various metabolic processes and stress responses in plants. In this study, we functionally characterised a R2R3-type SbMYB15 transcription factor (TF) from the extreme halophyte Salicornia brachiata. The SbMYB15 acts as a transcriptional activator. Transcriptional analysis showed that SbMYB15 transcript was strongly upregulated in red shoots and was also induced by different stresses; however, its expression remained unchanged with ABA. Overexpression of SbMYB15 in tobacco significantly improved salinity and dehydration tolerance. The enhanced tolerance of the transgenic plants was defined by the changes in chlorophyll, malondialdehyde (MDA), proline, total soluble sugar and total amino acid contents. The transgenic plants exhibited a higher membrane stability and reduced electrolyte leakage, H2O2 and O 2 (-) content compared to the wild type (WT). With ionic stress, transgenics showed a low Na(+) and a high K(+) content. In the transgenic plants, the expression of stress-responsive genes such as LEA5, ERD10D, PLC3, LTP1, HSF2, ADC, P5CS, SOD and CAT was enhanced in the presence of salinity, dehydration and heat. Exposure to gradual salinity and dehydration resulted in an increased stomatal conductance, water use efficiency, photosynthesis rate, photochemical quenching and reduced transpiration rate. Thus, SbMYB15 served as an important mediator of stress responses regulating different stress signalling pathways, leading to enhanced stress tolerance.

A red seaweed Kappaphycus alvarezii-based biostimulant (AgroGain®) improves the growth of Zea mays and impacts agricultural sustainability by beneficially priming rhizosphere soil microbial community.

The overuse of chemical-based agricultural inputs has led to the degradation of soil with associated adverse effects on soil attributes and microbial population. This scenario leads to poor soil health and is reportedly on the rise globally. Additionally, chemical fertilizers pose serious risks to the ecosystem and human health. In this study, foliar sprays of biostimulant (AgroGain/LBS6) prepared from the cultivated, tropical red seaweed Kappaphycus alvarezii increased the phenotypic growth of Zea mays in terms of greater leaf area, total plant height, and shoot fresh and dry weights. In addition, LBS6 improved the accumulation of chlorophyll a and b, total carotenoids, total soluble sugars, amino acids, flavonoids, and phenolics in the treated plants. LBS6 applications also improved the total bacterial and fungal count in rhizospheric soil. The V3-V4 region of 16S rRNA gene from the soil metagenome was analyzed to study the abundance of bacterial communities which were increased in the rhizosphere of LBS6-treated plants. Treatments were found to enrich beneficial soil bacteria, i.e., Proteobacteria, especially the classes Alphaproteobacteria, Cyanobacteria, Firmicutes, Actinobacteriota, Verrucomicrobiota, Chloroflexi, and Acidobacteriota and several other phyla related to plant growth promotion. A metagenomic study of those soil samples from LBS6-sprayed plants was correlated with functional potential of soil microbiota. Enrichment of metabolisms such as nitrogen, sulfur, phosphorous, plant defense, amino acid, co-factors, and vitamins was observed in soils grown with LBS6-sprayed plants. These results were further confirmed by a significant increase in the activity of soil enzymes such as urease, acid phosphatase, FDAse, dehydrogenase, catalase, and biological index of fertility in the rhizosphere of LBS6-treated corn plant. These findings conclude that the foliar application of LBS6 on Z. mays improves and recruits beneficial microbes and alters soil ecology in a sustainable manner.

A biostimulant prepared from red seaweed Kappaphycus alvarezii induces flowering and improves the growth of Pisum sativum grown under optimum and nitrogen-limited conditions.

Nitrogen (N) is one of the critical elements required by plants and is therefore one of the important limiting factors for growth and yield. To increase agricultural productivity, farmers are using excessive N fertilizers to the soil, which poses a threat to the ecosystem, as most of the applied nitrogen fertilizer is not taken up by crops, and runoff to aquatic bodies and the environment causes eutrophication, pollution, and greenhouse gas emissions. In this study, we used LBS6, a Kappaphycus alvarezii-based biostimulant as a sustainable alternative to improve the growth of plants under different NO3 - fertigation. A root drench treatment of 1 ml/L LBS6 significantly improved the growth of Pisum sativum plants grown under optimum and deficient N conditions. No significant difference was observed in the growth of LBS6-treated plants grown with excessive N. The application of LBS6 induced flowering under optimum and deficient N conditions. The total nitrogen, nitrate and ammonia contents of tissues were found to be higher in treated plants grown under N deficient conditions. The LBS6 treatments had significantly higher chlorophyll content in those plants grown under N-deficient conditions. The root drench application of LBS6 also regulated photosynthetic efficiency by modulating electron and proton transport-related processes of leaves in the light-adapted state. The rate of linear electron flux, proton conductivity and steady-state proton flux across the thylakoid membrane were found to be higher in LBS6-treated plants. Additionally, LBS6 also reduced nitrogen starvation-induced, reactive oxygen species accumulation by reduction in lipid peroxidation in treated plants. Gene expression analysis showed differential regulation of expression of those genes involved in N uptake, transport, assimilation, and remobilization in LBS6-treated plants. Taken together, LBS6 improved growth of those treated plants under optimum and nitrogen-limited condition by positively modulating their biochemical, molecular, and physiological processes.

Understanding the mode of action of AgroGain®, a biostimulant derived from the red seaweed Kappaphycus alvarezii in the stimulation of cotyledon expansion and growth of Cucumis sativa (cucumber).

Seaweed-based biostimulants are sustainable agriculture inputs that are known to have a multitude of beneficial effects on plant growth and productivity. This study demonstrates that Agrogain® (Product code: LBS6), a Kappaphycus alvarezii-derived biostimulant induced the expansion of cucumber cotyledons. Seven days treatment of LBS6-supplementation showed a 29.2% increase in area of expanded cotyledons, as compared to the control. LBS6-treated cotyledons also showed higher amylase activity, suggesting starch to sucrose conversion was used efficiently as an energy source during expansion. To understand the mechanisms of LBS6-induced expansion, real time gene expression analysis was carried out. This revealed that LBS6-treated cotyledons differentially modulated the expression of genes involved in cell division, cell number, cell expansion and cell size. LBS6 treatment also differentially regulated the expression of those genes involved in auxin and cytokinin metabolism. Further, foliar application of LBS6 on cucumber plants being grown under hydroponic conditions showed improved plant growth as compared to the control. The total leaf area of LBS6-sprayed plants increased by 19.1%, as compared to control. LBS6-sprayed plants efficiently regulated photosynthetic quenching by reducing loss via non-photochemical and non-regulatory quenching. LBS6 applications also modulated changes in the steady-state photosynthetic parameters of the cucumber leaves. It was demonstrated that LBS6 treatment modulated the electron and proton transport related pathways which help plants to efficiently utilize the photosynthetic radiation for optimal growth. These results provide clear evidence that bioactive compounds present in LBS6 improved the growth of cucumber plants by regulating the physiological as well as developmental pathways.

The SbSOS1 gene from the extreme halophyte Salicornia brachiata enhances Na(+) loading in xylem and confers salt tolerance in transgenic tobacco.

BACKGROUND: Soil salinity adversely affects plant growth and development and disturbs intracellular ion homeostasis resulting cellular toxicity. The Salt Overly Sensitive 1 (SOS1) gene encodes a plasma membrane Na(+)/H(+) antiporter that plays an important role in imparting salt stress tolerance to plants. Here, we report the cloning and characterisation of the SbSOS1 gene from Salicornia brachiata, an extreme halophyte. RESULTS: The SbSOS1 gene is 3774 bp long and encodes a protein of 1159 amino acids. SbSOS1 exhibited a greater level of constitutive expression in roots than in shoots and was further increased by salt stress. Overexpressing the S. brachiata SbSOS1 gene in tobacco conferred high salt tolerance, promoted seed germination and increased root length, shoot length, leaf area, fresh weight, dry weight, relative water content (RWC), chlorophyll, K(+)/Na(+) ratio, membrane stability index, soluble sugar, proline and amino acid content relative to wild type (WT) plants. Transgenic plants exhibited reductions in electrolyte leakage, reactive oxygen species (ROS) and MDA content in response to salt stress, which probably occurred because of reduced cytosolic Na(+) content and oxidative damage. At higher salt stress, transgenic tobacco plants exhibited reduced Na(+) content in root and leaf and higher concentrations in stem and xylem sap relative to WT, which suggests a role of SbSOS1 in Na(+) loading to xylem from root and leaf tissues. Transgenic lines also showed increased K(+) and Ca(2+) content in root tissue compared to WT, which reflect that SbSOS1 indirectly affects the other transporters activity. CONCLUSIONS: Overexpression of SbSOS1 in tobacco conferred a high degree of salt tolerance, enhanced plant growth and altered physiological and biochemical parameters in response to salt stress. In addition to Na(+) efflux outside the plasma membrane, SbSOS1 also helps to maintain variable Na(+) content in different organs and also affect the other transporters activity indirectly. These results broaden the role of SbSOS1 in planta and suggest that this gene could be used to develop salt-tolerant transgenic crops.

Ascophyllum nodosum Biostimulant Improves the Growth of Zea mays Grown Under Phosphorus Impoverished Conditions.

Phosphorous is one of the major limiting factors determining plant growth. Current agricultural practices mainly rely on the use of chemical fertilizers posing threat to the ecosystem. In this study, the application of an Ascophyllum nodosum extract (ANE) in phosphorous (P)-limited conditions improved the fresh and dry weight of shoots and roots of Zea mays. ANE-treated Z. mays grown under P-limited conditions showed a higher P content than the control. ANE activated simultaneous responses, at multiple levels, in Z. mays grown under P-limited conditions as seen from the regulation of gene expression at the whole-plant level to specific biochemical responses on a subcellular level. ANE-supplemented Z. mays grown under P-limited conditions also showed reduced electrolyte leakage and lipid peroxidation by an improved membrane stability. ANE treatment reduced P-limitation-induced oxidative damage in Z. mays by reducing H2O2 and O 2 - accumulation. Furthermore, ANE also induced the accumulation of the total contents of soluble sugars, amino acids, phenolics, and flavonoids. Gene expression analysis suggested that ANE differentially modulated the expression of P-starvation responsive genes involved in metabolic, signal transduction, and developmental pathways in Z. mays. ANE also modulated the expression of genes involved in sugar, lipid, and secondary metabolism. Thus, this study illustrated the role of ANE in improving the productivity of Z. mays, an important crop, in P-limited conditions. Furthermore, it sets the framework to increase agricultural productivity in nutrient deficient soils using a sustainable, eco-friendly strategy.

A Biostimulant Preparation of Brown Seaweed Ascophyllum nodosum Suppresses Powdery Mildew of Strawberry.

Strawberry, an important fruit crop, is susceptible to a large number of pathogens that reduce fruit quality and productivity. In this study, the effect of a biostimulant prepared from Ascophyllum nodosum extract (ANE) (0.1%, 0.2%, and 0.3%) was evaluated on powdery mildew progression under greenhouse and field conditions. In the greenhouse, application of 0.2% ANE showed maximum reduction in powdery mildew progression as compared to the control. Forty-eight hour post-inoculation, foliar spray of 0.2% ANE reduced spore germination by 75%. Strawberry leaves sprayed with ANE showed higher total phenolic and flavonoid content in response to powdery mildew infection. Furthermore, application of ANE elicited defense response in strawberry plants by induction of defense-related enzymes, such as phenylalanine ammonia lyase, polyphenol oxidase, and peroxidase activity. In field conditions, foliar spray of 0.2% ANE showed a reduction of 37.2% of natural incidence of powdery mildew infection as compared to the control. ANE sprayed plant also reduces the severity of powdery mildew infection under natural conditions. These results indicate that application of ANE induces the strawberry plant's active defense against powdery mildew infection by induction of secondary metabolism and regulating the activities of defense-related enzymes.

Ascophyllum nodosum-Based Biostimulants: Sustainable Applications in Agriculture for the Stimulation of Plant Growth, Stress Tolerance, and Disease Management.

Abiotic and biotic stresses limit the growth and productivity of plants. In the current global scenario, in order to meet the requirements of the ever-increasing world population, chemical pesticides and synthetic fertilizers are used to boost agricultural production. These harmful chemicals pose a serious threat to the health of humans, animals, plants, and the entire biosphere. To minimize the agricultural chemical footprint, extracts of Ascophyllum nodosum (ANE) have been explored for their ability to improve plant growth and agricultural productivity. The scientific literature reviewed in this article attempts to explain how certain bioactive compounds present in extracts aid to improve plant tolerances to abiotic and/or biotic stresses, plant growth promotion, and their effects on root/microbe interactions. These reports have highlighted the use of various seaweed extracts in improving nutrient use efficiency in treated plants. These studies include investigations of physiological, biochemical, and molecular mechanisms as evidenced using model plants. However, the various modes of action of A. nodosum extracts have not been previously reviewed. The information presented in this review depicts the multiple, beneficial effects of A. nodosum-based biostimulant extracts on plant growth and their defense responses and suggests new opportunities for further applications for marked benefits in production and quality in the agriculture and horticultural sectors.

Ascophyllum nodosum extract mitigates salinity stress in Arabidopsis thaliana by modulating the expression of miRNA involved in stress tolerance and nutrient acquisition.

Ascophyllum nodosum extract (ANE) contains bioactive compounds that improve the growth of Arabidopsis in experimentally-induced saline conditions; however, the molecular mechanisms through which ANE elicits tolerance to salinity remain largely unexplored. Micro RNAs (miRNAs) are key regulators of gene expression, playing crucial roles in plant growth, development, and stress tolerance. Next generation sequencing of miRNAs from leaves of control Arabidopsis and from plants subjected to three treatments (ANE, NaCl and ANE+NaCl) was used to identify ANE-responsive miRNA in the absence and presence of saline conditions. Differential gene expression analysis revealed that ANE had a strong effect on miRNAs expression in both conditions. In the presence of salinity, ANE tended to reduce the up-regulation or the down-regulation trend induced caused by NaCl in miRNAs such as ath-miR396a-5p, ath-miR399, ath-miR2111b and ath-miR827. To further uncover the effects of ANE, the expression of several target genes of a number of ANE-responsive miRNAs was analyzed by qPCR. NaCl, but not ANE, down-regulated miR396a-5p, which negatively regulated the expression of AtGRF7 leading to a higher expression of AtDREB2a and AtRD29 in the presence of ANE+NaCl, as compared to ANE alone. ANE+NaCl initially reduced and then enhanced the expression of ath-miR169g-5p, while the expression of the target genes AtNFYA1 and ATNFYA2, known to be involved in the salinity tolerance mechanism, was increased as compared to ANE or to NaCl treatments. ANE and ANE+NaCl modified the expression of ath-miR399, ath-miR827, ath-miR2111b, and their target genes AtUBC24, AtWAK2, AtSYG1 and At3g27150, suggesting a role of ANE in phosphate homeostasis. In vivo and in vitro experiments confirmed the improved growth of Arabidopsis in presence of ANE, in saline conditions and in phosphate-deprived medium, further substantiating the influence of ANE on a variety of essential physiological processes in Arabidopsis including salinity tolerance and phosphate uptake.

Molecular characterization of an MYB transcription factor from a succulent halophyte involved in stress tolerance.

Abiotic stresses like drought, salinity and extreme temperature significantly affect crop productivity. Plants respond at molecular, cellular and physiological levels for management of stress tolerance. Functional and regulatory genes play a major role in controlling these abiotic stresses through an intricate network of transcriptional machinery. Transcription factors are potential tools for manipulating stress tolerance since they control a large number of downstream genes. In the present study, we have isolated SbMYB44 from a succulent halophyte, Salicornia brachiata Roxb. SbMYB44 with an open-reading frame of 810 bp encodes a protein of 269 amino acids, with an estimated molecular mass of 30.31 kDa and an isoelectric point of 6.29. The in silico analysis revealed that the SbMYB44 protein contains the conserved R2R3 imperfect repeats, two SANT domains and post-translational modification sites. The SbMYB44 transcript showed up-regulation in response to salinity, desiccation, high temperature, and abscisic acid and salicylic acid treatments. The SbMYB44 recombinant protein showed binding to dehydration-responsive cis-elements (RD22 and MBS-1), suggesting its possible role in stress signalling. Overexpression of SbMYB44 enhanced the growth of yeast cells under both ionic and osmotic stresses.

Bioengineering for salinity tolerance in plants: state of the art.

Genetic engineering of plants for abiotic stress tolerance is a challenging task because of its multifarious nature. Comprehensive studies for developing abiotic stress tolerance are in progress, involving genes from different pathways including osmolyte synthesis, ion homeostasis, antioxidative pathways, and regulatory genes. In the last decade, several attempts have been made to substantiate the role of "single-function" gene(s) as well as transcription factor(s) for abiotic stress tolerance. Since, the abiotic stress tolerance is multigenic in nature, therefore, the recent trend is shifting towards genetic transformation of multiple genes or transcription factors. A large number of crop plants are being engineered by abiotic stress tolerant genes and have shown the stress tolerance mostly at laboratory level. This review presents a mechanistic view of different pathways and emphasizes the function of different genes in conferring salt tolerance by genetic engineering approach. It also highlights the details of successes achieved in developing salt tolerance in plants thus far.