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.     

Antimicrobial Action of Compounds from Marine Seaweed

Seaweed produces metabolites aiding in the protection against different environmental stresses. These compounds show antiviral, antiprotozoal, antifungal, and antibacterial properties. Macroalgae can be cultured in high volumes and would represent an attractive source of potential compounds useful for unconventional drugs able to control new diseases or multiresistant strains of pathogenic microorganisms. The substances isolated from green, brown and red algae showing potent antimicrobial activity belong to polysaccharides, fatty acids, phlorotannins, pigments, lectins, alkaloids, terpenoids and halogenated compounds. This review presents the major compounds found in macroalga showing antimicrobial activities and their most promising applications.     

Induction of Defense Gene Expression and the Resistance of Date Palm to Fusarium oxysporum f. sp. Albedinis in Response to Alginate Extracted from Bifurcaria bifurcata

In many African countries, the Bayoud is a common disease spread involving the fungus Fusarium oxusporum f. sp. albedinis (Foa). The induction of plant natural defenses through the use of seaweed polysaccharides to help plants against pathogens is currently a biological and ecological approach that is gaining more and more importance. In the present study, we used alginate, a natural polysaccharide extracted from a brown algae Bifurcaria bifurcata, to activate date palm defenses, which involve phenylalanine ammonia-lyase (PAL), a key enzyme of phenylpropanoid metabolism. The results obtained showed that at low concentration (1 g·L⁻¹), alginate stimulated PAL activity in date palm roots 5 times more compared to the negative control (water-treated) after 24 h following treatment and 2.5 times more compared to the laminarin used as a positive stimulator of plant natural defenses (positive control of induction). Using qRT-PCR, the expression of a selection of genes involved in three different levels of defense mechanisms known to be involved in response to biotic stresses were investigated. The results showed that, generally, the PAL gene tested and the genes encoding enzymes involved in early oxidative events (SOD and LOX) were overexpressed in the alginate-treated plants compared to their levels in the positive and negative controls. POD and PR protein genes selected encoding β-(1,3)-glucanases and chitinases in this study did not show any significant difference between treatments; suggesting that other genes encoding POD and PR proteins that were not selected may be involved. After 17 weeks following the inoculation of the plants with the pathogen Foa, treatment with alginate reduced the mortality rate by up to 80% compared to the rate in control plants (non-elicited) and plants pretreated with laminarin, which agrees with the induction of defense gene expression and the stimulation of natural defenses in date palm with alginate after 24 h. These results open promising prospects for the use of alginate in agriculture as an inducer that triggers immunity of plants against telluric pathogens in general and of date palm against Fusarium oxysporum f. sp. albedinis in particular.     

Seaweed Protein Hydrolysates and Bioactive Peptides: Extraction,Purification, and Applications

Seaweeds are industrially exploited for obtaining pigments, polysaccharides, or phenolic compounds with application in diverse fields. Nevertheless, their rich composition in fiber, minerals, and proteins, has pointed them as a useful source of these components. Seaweed proteins are nutritionally valuable and include several specific enzymes, glycoproteins, cell wall-attached proteins, phycobiliproteins, lectins, or peptides. Extraction of seaweed proteins requires the application of disruptive methods due to the heterogeneous cell wall composition of each macroalgae group. Hence, non-protein molecules like phenolics or polysaccharides may also be co-extracted, affecting the extraction yield. Therefore, depending on the macroalgae and target protein characteristics, the sample pretreatment, extraction and purification techniques must be carefully chosen. Traditional methods like solid–liquid or enzyme-assisted extraction (SLE or EAE) have proven successful. However, alternative techniques as ultrasound- or microwave-assisted extraction (UAE or MAE) can be more efficient. To obtain protein hydrolysates, these proteins are subjected to hydrolyzation reactions, whether with proteases or physical or chemical treatments that disrupt the proteins native folding. These hydrolysates and derived peptides are accounted for bioactive properties, like antioxidant, anti-inflammatory, antimicrobial, or antihypertensive activities, which can be applied to different sectors. In this work, current methods and challenges for protein extraction and purification from seaweeds are addressed, focusing on their potential industrial applications in the food, cosmetic, and pharmaceutical industries.     

茶尺蠖为害提高邻近茶苗对茶尺蠖幼虫的防御能力

本文研究了茶树虫害诱导挥发物(Herbivore-induced plant volatiles,HIPVs)对邻近茶苗防御能力的影响。将健康茶苗放在茶尺蠖取食的茶苗附近,作为HIPVs的处理,测定了茶树防御相关基因在不同处理下的表达水平、抗性防御物质的水平,并检测了茶尺蠖危害茶苗邻近植株对茶尺蠖幼虫生长的影响。研究结果表明,与对照相比,茶尺蠖幼虫为害茶苗释放的HIPVs在处理后24 h和12 h内分别显著诱导邻近茶苗Csi LOX1和Csi ACS1基因的表达水平,这些挥发物还能增强茶尺蠖取食诱导的防御基因的表达。HIPVs处理3 d后,茶树的重要防御物质多酚氧化酶(Polyphenol oxidases,PPOs)活性显著高于对照,是对照的1.36倍,同时,取食这些茶苗的茶尺蠖幼虫体重也显著低于对照。上述结论表明茶尺蠖取食诱导的挥发物能够作为信号物质在茶树中传递,并能够通过增强茉莉酸和乙烯抗虫途径提高对茶尺蠖的抗性。     

Antiviral Activity and Mechanisms of Seaweeds Bioactive Compounds on Enveloped Viruses—A Review

In the last decades, the interest in seaweed has significantly increased. Bioactive compounds from seaweed’s currently receive major attention from pharmaceutical companies as they express several interesting biological activities which are beneficial for humans. The structural diversity of seaweed metabolites provides diverse biological activities which are expressed through diverse mechanisms of actions. This review mainly focuses on the antiviral activity of seaweed’s extracts, highlighting the mechanisms of actions of some seaweed molecules against infection caused by different types of enveloped viruses: influenza, Lentivirus (HIV-1), Herpes viruses, and coronaviruses. Seaweed metabolites with antiviral properties can act trough different pathways by increasing the host’s defense system or through targeting and blocking virus replication before it enters host cells. Several studies have already established the large antiviral spectrum of seaweed’s bioactive compounds. Throughout this review, antiviral mechanisms and medical applications of seaweed’s bioactive compounds are analyzed, suggesting seaweed’s potential source of antiviral compounds for the formulation of novel and natural antiviral drugs.     

Foliar Application of Benzothiadiazole and Salicylic Acid to Combat Sheath Blight Disease of Rice

A field study was undertaken to ascertain the effects of elicitors viz. benzothiadiazole (BTH) and salicylic acid (SA) on defense related enzymes viz. peroxidase, phenylalanine ammonia lyase, superoxide dismutase, chitinase and β-1,3-glucanase, and phenols in rice (Pusa Basmati I) plants. First foliar spray of BTH (50 mg/kg) and SA (50 mg/kg) was done at the maximum tillering stage and inoculation with Rhizoctonia solani was carried 24 h after elicitor treatment. Elicitors were further sprayed at every growth stage. Time course analysis showed peak accumulation of defense related enzymes and phenols in the rice leaves treated with BTH and SA, and accumulation was the highest at the flowering stage. Higher enzymatic activity was observed in elicitor treated plants inoculated with R. solani. Compared to the untreated control plants, application of elicitors before R. solani inoculation significantly elicited the defense related enzymes and phenols. Moreover, application of elicitors had a positive effect on yield and disease reduction. It is suggested that pretreatment of rice leaves with BTH and SA could be used to enhance the level of protection against sheath blight and to improve rice yield in the fields.     

Depolymerized Fractions of Sulfated Galactans Extracted from Gracilaria fisheri and Their Antibacterial Activity against Vibrio parahaemolyticus and Vibrio harveyi

Various seaweed sulfated polysaccharides have been explored for antimicrobial application. This study aimed to evaluate the antibacterial activity of the native Gracilaria fisheri sulfated galactans (NSG) and depolymerized fractions against the marine pathogenic bacteria Vibrio parahaemolyticus and Vibrio harveyi. NSG was hydrolyzed in different concentrations of H₂O₂ to generate sulfated galactans degraded fractions (SGF). The molecular weight, structural characteristics, and physicochemical parameters of both NSG and SGF were determined. The results revealed that the high molecular weight NSG (228.33 kDa) was significantly degraded to SGFs of 115.76, 3.79, and 3.19 kDa by hydrolysis with 0.4, 2, and 10% H₂O₂, respectively. The Fourier transformed spectroscopy (FTIR) and ¹H− and ¹³C−Nuclear magnetic resonance (NMR) analyses demonstrated that the polysaccharide chain structure of SGFs was not affected by H₂O₂ degradation, but alterations were detected at the peak positions of some functional groups. In vitro study showed that SGFs significantly exerted a stronger antibacterial activity against V. parahaemolyticus and V. harveyi than NSG, which might be due to the low molecular weight and higher sulfation properties of SGF. SGF disrupted the bacterial cell membrane, resulting in leakage of intracellular biological components, and subsequently, cell death. Taken together, this study provides a basis for the exploitation and utilization of low-molecular-weight sulfated galactans from G. fisheri to prevent and control the shrimp pathogens.     

不同玉米品种对大斑病的抗性与相关防御酶活性的关系研究

以吉林省玉米早熟、中早熟区种植的4个玉米品种及其亲本为材料,对不同玉米品种对大斑病的抗性与苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）和多酚氧化酶（PPO）等3种防御酶活性的关系进行研究。结果表明,3种防御酶PAL、POD和PPO与玉米对大斑病的抗性关系非常密切,玉米植株受大斑病菌侵染后,防御酶活性比对照明显增高。防御酶的作用受品种遗传背景的影响,不同品种中不同防御酶对其大斑病抗性的贡献有差异。受大斑病菌侵染后,抗病性强的品种,PAL和PPO或PAL和POD酶活性发生协同增强,且从抽丝期到灌浆期酶活性协同增强的防御酶不发生酶活性衰减或衰减微弱;感病的品种,从抽丝期到灌浆期存在2种或3种防御酶活性同时衰减现象。     

The Structural Characteristics of Seaweed Polysaccharides and Their Application in Gel Drug Delivery Systems

In recent years, researchers across various fields have shown a keen interest in the exploitation of biocompatible natural polymer materials, especially the development and application of seaweed polysaccharides. Seaweed polysaccharides are a multi-component mixture composed of one or more monosaccharides, which have the functions of being anti-virus, anti-tumor, anti-mutation, anti-radiation and enhancing immunity. These biological activities allow them to be applied in various controllable and sustained anti-inflammatory and anticancer drug delivery systems, such as seaweed polysaccharide-based nanoparticles, microspheres and gels, etc. This review summarizes the advantages of alginic acid, carrageenan and other seaweed polysaccharides, and focuses on their application in gel drug delivery systems (such as nanogels, microgels and hydrogels). In addition, recent literature reports and applications of seaweed polysaccharides are also discussed.     

牛角瓜根的抑菌活性研究

以牛角瓜根为实验材料,采用索氏提取法,以4种不同溶剂(石油醚、氯仿、乙酸乙酯、甲醇)对牛角瓜的根进行提取,并采用纸片琼脂扩散法对粗提物的抑菌活性进行了评价,供试菌包括白色念珠菌等6种真菌和金黄色葡萄球菌等7种细菌。结果表明:石油醚粗提物对枯草芽孢杆菌、白色念珠菌、黑曲霉菌、大肠埃希氏菌都表现出良好的抑菌活性,且抑菌效果接近或优于阳性对照氟康唑,对尖孢镰刀菌和鲍曼不动杆菌也表现出良好的抑菌活性;氯仿粗提物对白色念珠菌及粪链球菌表现出良好的抑菌活性;乙酸乙酯粗提物对粪链球菌表现出最好的抑菌活性;甲醇粗提物的抑菌活性较弱。     

Biodegradation and Prospect of Polysaccharide from Crustaceans

Marine crustacean waste has not been fully utilized and is a rich source of chitin. Enzymatic degradation has attracted the wide attention of researchers due to its unique biocatalytic ability to protect the environment. Chitosan (CTS) and its derivative chitosan oligosaccharides (COSs) with various biological activities can be obtained by the enzymatic degradation of chitin. Many studies have shown that chitosan and its derivatives, chitosan oligosaccharides (COSs), have beneficial properties, including lipid-lowering, anti-inflammatory and antitumor activities, and have important application value in the medical treatment field, the food industry and agriculture. In this review, we describe the classification, biochemical characteristics and catalytic mechanisms of the major degrading enzymes: chitinases, chitin deacetylases (CDAs) and chitosanases. We also introduced the technology for enzymatic design and modification and proposed the current problems and development trends of enzymatic degradation of chitin polysaccharides. The discussion on the characteristics and catalytic mechanism of chitosan-degrading enzymes will help to develop new types of hydrolases by various biotechnology methods and promote their application in chitosan.     

水稻对穗枯病的抗病机理初步研究

【目的】水稻在孕穗期比苗期更容易感染穗枯病(Bacterial panicle blight of rice)并出现病症。本研究旨在探究水稻不同时期对穗枯病的抗性机理,为培育抗病水稻品种奠定基础。【方法】采用喷雾法和注射法分别对苗期和孕穗期的抗、感病水稻品种接种颖壳伯克氏菌(Burkholderia glumae),测定处理组与对照组的3种抗氧化酶(POD、CAT、SOD)活性的差异,并利用实时荧光定量PCR测定5种防卫反应基因(PR1a、PR10b、Rcht、LOX、PAL)的表达量。【结果】B.glumae的侵染能引起水稻活性氧的积累,提高抗氧化酶活性,使部分防卫反应基因大量表达,但是苗期和孕穗期的应答有较大区别。水稻孕穗期的抗氧化酶(SOD、CAT和POD)活性和防卫反应基因(PR10b、Rcht和PAL)的表达量比苗期高,但PR1a和LOX的表达量却低于苗期。【结论】B.glumae能诱导孕穗期的水稻产生更多抗病反应,参与抗病反应的主要是水杨酸信号传导途径。     

The Mucus of Actinia equina (Anthozoa,Cnidaria): An Unexplored Resource for Potential Applicative Purposes

The mucus produced by many marine organisms is a complex mixture of proteins and polysaccharides forming a weak watery gel. It is essential for vital processes including locomotion, navigation, structural support, heterotrophic feeding and defence against a multitude of environmental stresses, predators, parasites, and pathogens. In the present study we focused on mucus produced by a benthic cnidarian, the sea anemone Actinia equina (Linnaeus, 1758) for preventing burial by excess sedimentation and for protection. We investigated some of the physico-chemical properties of this matrix such as viscosity, osmolarity, electrical conductivity, protein, carbohydrate, and total lipid contents. Some biological activities such as hemolytic, cytotoxic, and antibacterial lysozyme-like activities were also studied. The A. equina mucus is mainly composed by water (96.2% ± 0.3%), whereas its dry weight is made of 24.2% ± 1.3% proteins and 7.8% ± 0.2% carbohydrates, with the smallest and largest components referable to lipids (0.9%) and inorganic matter (67.1%). The A. equina mucus matrix exhibited hemolytic activity on rabbit erythrocytes, cytotoxic activity against the tumor cell line K562 (human erythromyeloblastoid leukemia) and antibacterial lysozyme-like activity. The findings from this study improve the available information on the mucus composition in invertebrates and have implications for future investigations related to exploitation of A. equina and other sea anemones’ mucus as a source of bioactive compounds of high pharmaceutical and biotechnological interest.     

A Lymphoid Organ Specific Anti-Lipopolysaccharide Factor from Litopenaeus vannamei Exhibits Strong Antimicrobial Activities

Different shrimp species are known to possess apparent distinct resistance to different pathogens in aquaculture. However, the molecular mechanism underlying this finding still remains unknown. One kind of important antimicrobial peptides, anti-lipopolysaccharide factors (ALF), exhibit broad-spectrum antimicrobial activities. Here, we reported a newly identified ALF from the shrimp Litopenaeus vannamei and compared the immune function with its counterpart in the shrimp Fenneropenaeus chinensis. The ALF, designated as LvALF8, was specifically expressed in the lymphoid organ of L. vannamei. The expression level of LvALF8 was apparently changed after white spot syndrome virus (WSSV) or Vibrio parahaemolyticus challenges. The synthetic LBD peptide of LvALF8 (LvALF8-LBD) showed strong antibacterial activities against most tested Gram-negative and Gram-positive bacteria. LvALF8-LBD could also inhibit the in vivo propagation of WSSV similar as FcALF8-LBD, the LBD of LvALF8 counterpart in F. chinensis. However, LvALF8-LBD and FcALF8-LBD exhibited apparently different antibacterial activity against V. parahaemolyticus, the main pathogen causing acute hepatopancreatic necrosis disease (AHPND) of affected shrimp. A structural analysis showed that the positive net charge and amphipathicity characteristics of LvALF8-LBD peptide were speculated as two important components for its enhanced antimicrobial activity compared to those of FcALF8-LBD. These new findings may not only provide some evidence to explain the distinct disease resistance among different shrimp species, but also lay out new research ground for the testing and development of LBD-originated antimicrobial peptides to control of shrimp diseases.     

海藻精对玉米种子萌发及苗期水分胁迫的影响

盆栽条件下,以珠玉甜1号、珠玉糯1号为材料,研究重度干旱、中度干旱、轻度干旱、正常供水、轻度淹水和重度淹水等6个不同水分条件下,4个不同浓度的海藻精对两个鲜食玉米品种种子萌发和幼苗生长的影响。结果表明,在水分胁迫下,海藻精对两个鲜食玉米种子萌发无显著效果;在玉米苗期施用海藻精0.50 g/L可促进根长增长、根条数增加,提高保护酶活性。在重度干旱和重度淹水下,珠玉甜1号幼苗叶片丙二醛(MDA)含量较未施用分别减少8.6%、22.3%,可溶性糖分别增加33.2%、42.1%。表明施用海藻精可减轻水分胁迫对细胞膜的损害,并积累可溶性糖进而增强鲜食玉米苗期的抗逆性。在水分胁迫频繁发生的背景下,华南地区种植鲜食玉米时施用海藻精有利于其稳产和高产。     

Seaweed-Derived Polysaccharides Attenuate Heat Stress-Induced Splenic Oxidative Stress and Inflammatory Response via Regulating Nrf2 and NF-κB Signaling Pathways

With global warming, heat stress (HS) has become a worldwide concern in both humans and animals. The ameliorative effect of seaweed (Enteromorpha prolifera) derived polysaccharides (SDP) on HS-induced oxidative stress and the inflammatory response of an immune organ (spleen) was evaluated using an animal model (Gallus gallus domesticus). In total, 144 animals were used in this 4-week trial and randomly assigned to the following three groups: thermoneutral zone, HS, and HS group supplemented with 1000 mg/kg SDP. Dietary SDP improved the antioxidant capacity and reduced the malondialdehyde (MDA) of the spleen when exposed to HS, regulated via enhancing nuclear factor erythroid 2-related factor-2 (Nrf2) signaling. Furthermore, the inclusion of SDP reduced the levels of pro-inflammatory cytokines and alleviated HS-induced splenic inflammatory response by suppressing the nuclear factor-kappa B (NF-κB) p65 signaling. These findings suggest that the SDP from E. prolifera can be used as a functional food and/or feed supplement to attenuate HS-induced oxidative stress and inflammatory responses of the immune organs. Moreover, the results could contribute to the development of high-value marine products from seaweed for potential use in humans and animals, owing to their antioxidant and anti-inflammatory effects.     

Chemical Synthesis and Structure-Activity Relationship Study Yield Desotamide a Analogues with Improved Antibacterial Activity

Desotamides A, a cyclohexapeptide produced by the deep-sea-derived Streptomyces scopuliridis SCSIO ZJ46, displays notable antibacterial activities against strains of Streptococcus pnuemoniae, Staphylococcus aureus, and methicillin-resistant Staphylococcus epidermidis (MRSE). In this study, to further explore its antibacterial potential and reveal the antibacterial structure-activity relationship of desotamides, 13 cyclopeptides including 10 new synthetic desotamide A analogues and wollamides B/B1/B2 were synthesized and evaluated for their antibacterial activities against a panel of Gram-positive and -negative pathogens. The bioactivity data reveal that residues at position II and VI greatly impact antibacterial activity. The most potent antibacterial analogues are desotamide A4 (13) and A6 (15) where l-allo-Ile at position II was substituted with l-Ile and Gly at position VI was simultaneously replaced by d-Lys or d-Arg; desotamides A4 (13) and A6 (15) showed a 2–4-fold increase of antibacterial activities against a series of Gram-positive pathogens including the prevalent clinical drug-resistant pathogen methicillin-resistant Staphylococcus aureus (MRSA) with MIC values of 8–32 μg/mL compared to the original desotamide A. The enhanced antibacterial activity, broad antibacterial spectrum of desotamides A4 and A6 highlighted their potential as new antibiotic leads for further development.     

Effects of initial air removal methods on microorganisms and characteristics of fermented plant beverages

The effects of 3 different methods for removing the initial air on the properties of fermented plant beverages produced from phom-nang seaweed (Gracilaria fisheri) and wild forest noni (Morinda coreia Ham.) were investigated. Only method M which covered the space above the fermentation liquid with a water filled plastic bag produced no surface film of yeast, had the highest acidity and also antibacterial activity from both plants after 90 days of fermentation. However, the yeast count still exceeded the standard guidelines for plant beverages. The fermented beverage from wild forest noni showed more antibacterial activity against 3 of 4 pathogenic bacteria tested than that from the phomnang seaweed, probably for its higher levels of acidity and ethanol content. Lactic Acid Bacteria (LAB) isolated from the fermentation samples from days 1-5 using the method M from both fermented plant beverages were Leuconostoc mesenteroides supsp. mesenteroides and Leu. mesenteroides subsp. dextranicum while presence of Lactobacilus plantarum was only recorded at days 4-5 in the wild forest noni beverage. From days 6-14 the isolates were Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus brevis from wild forest noni beverage, whereas only L. brevis was not detected in the seaweed beverage. During days 21-45 both beverages had a similar LAB population of L. plantarum and L. brevis while L. coryniformis was only found in the wild forest noni beverage. Between days 60-90 in both plant beverages only L. plantarum and Lactobacillius sp. were detected.