Agricultural uses of plant biostimulants

Background

Plant biostimulants are diverse substances and microorganisms used to enhance plant growth. The global market for biostimulants is projected to increase 12 % per year and reach over $2,200 million by 2018. Despite the growing use of biostimulants in agriculture, many in the scientific community consider biostimulants to be lacking peer-reviewed scientific evaluation.

Scope

This article describes the emerging definitions of biostimulants and reviews the literature on five categories of biostimulants: i. microbial inoculants, ii. humic acids, iii. fulvic acids, iv. protein hydrolysates and amino acids, and v. seaweed extracts.

Conclusions

The large number of publications cited for each category of biostimulants demonstrates that there is growing scientific evidence supporting the use of biostimulants as agricultural inputs on diverse plant species. The cited literature also reveals some commonalities in plant responses to different biostimulants, such as increased root growth, enhanced nutrient uptake, and stress tolerance.     

Plant biostimulants: a review on the processing of macroalgae and use of extracts for crop management to reduce abiotic and biotic stresses

A biostimulant is an organic material that, when applied in small quantities, enhances plant growth and development such that the response cannot be attributed to the application of traditional plant nutrients. This review is aimed at highlighting developments in the processing of macroalgae for agricultural biostimulants (AB), summarising the biologically active components of brown macroalgae and examining the factors supporting the use of macroalgal AB for managing abiotic and biotic stresses in crop plants. The policy drivers supporting the use of macroalgal-derived ABs in agriculture are also emphasised. We examine the use of macroalgal ABs in crop production and evaluated the benefits of seed priming, foliar application, soil drenches and hydroponic treatments. The use of macroalgal ABs on crop plants can generate multiple benefits with reported effects including enhanced rooting, higher crop and fruit yields, freezing, drought and salt tolerance, enhanced photosynthetic activity and resistance to fungi, bacteria and virus. ABs can be applied as an alternative, or used in conjunction with synthetic crop protection products and plant growth regulators, and may have a role in maintaining crop production levels, health and quality in the future when many active ingredients will be lost to the industry due to changes in European Union regulations. Worldwide, macroalgae remain largely unexploited, we highlight some of the future research and development priorities.     

Supercritical fluid extraction of algae enhances levels of biologically active compounds promoting plant growth

The aim of this research was to screen plant growth biostimulant properties of supercritical CO₂ macroalgal extracts. To this end secondary metabolites were isolated from the biomass of marine macroalgae from the Baltic Sea (species of Polysiphonia, Ulva and Cladophora). Chemical characteristics of the algal extracts were determined by inductively coupled plasma atomic emission spectroscopy for inorganic constituents and high-performance liquid chromatography and spectrophotometry for organic constituents. Inorganic (macro- and microelements) and organic (plant hormones: auxins and cytokinins; polyphenols) compounds were detected in the extract. Algal extracts were tested primarily on garden cress (Lepidium sativum L.; Brassicaceae) and wheat (Triticum aestivum L.; Poaceae). The extracts enhanced chlorophyll and carotenoid content in plant shoots, as well as root thickness and above-ground biomass. The most effective method of application of the extract was by foliar feed on cress and seed maceration for wheat. Algal extracts obtained by supercritical fluid extraction (SFE) were found to be a novel natural source of compounds, stimulating growth of cultivated plants. However, field trials will be required to show that the extracts can act as plant biopesticides and growth biostimulants.     

Biostimulant prevents yield loss and reduces oxidative damage in tomato plants grown on reduced NPK nutrition*

Plant biostimulants are substances which have the capacity to modify physiological processes in plants in a way that provides potential benefits to growth, development or stress response. Effects of biostimulant application on two tomato hybrids (Ombeline F1 and Bostina F1) submitted to reduced nitrogen, phosphorus and potassium (NPK) nutrition aiming at prevention of oxidative stress generation as well as yield and fruit quality loss were investigated in this study. According to obtained results, foliar applied Viva® biostimulant decreased superoxide dismutase (SOD, EC 1.15.1.1) and peroxidase (POD, EC 1.11.1.7) activity in tomato leaves even when recommended NPK nutrition was reduced at 40%. Fruit quality parameters (total soluble solids, total acidity, ascorbic acid and lycopene content) and yield were also maintained in reduced macronutrient fertilization when biostimulant was added. Combination of biostimulant with reduced NPK fertilizer enabled stability of cell homeostasis in tomato plants and their better adaptation to stress conditions. The possibility of biostimulant being used as environmental friendly tool in the reduction of mineral fertilizers without negative consequences regarding yield and fruit quality was discussed.     

Plant Biostimulant Regulatory Framework: Prospects in Europe and Current Situation at International Level

This review analyses the current regulatory procedures for plant biostimulants, both at an EU level and beyond the EU, and explores the future regulation of these substances in the EU. Plant biostimulants are defined as products that stimulate plant nutritional processes regardless of the product’s nutrient content, with the sole aim of improving one or more of the following characteristics of the plant and the plant rhizosphere or phyllosphere: the efficiency of nutrient use, tolerance to abiotic stresses, crop quality traits, availability of confined nutrients in the soil and rhizosphere, humification and degradation of organic compounds in the soil. This definition is reported in the proposals for new rules to regulate making CE-marked fertiliser products available on the market. This regulation, which includes a plant biostimulants category, will repeal the existing Fertilisers Regulation (EC) No. 2003/2003. This category of compounds is also used in non-European countries. Currently, as there are different market conditions and different national regulation requirements for plant biostimulants in different countries, the non-harmonised regulatory processes can lead to unfair competition between operators. The assessment of plant biostimulants should be harmonised as far as possible, to avoid fragmentation and ensure a level, reliable playing field. It is essential that a common market is created for these substances.

     

Polar Constituents,Essential Oil and Antioxidant Activity of Marsh Woundwort (Stachys palustris L.)

Stachys palustris, well‐known as marsh woundwort, is a perennial herb growing in wet environments of Europe. Its tubers, leaves and seeds are eaten raw or cooked. Alike other Stachys species, the plant is also used as a traditional remedy. Despite S. palustris has been consumed for centuries, little is known about its chemical constituents. In this work the main secondary metabolites of S. palustris from Hungary and France have been analysed. From the plant ethanolic extracts, ethanoid glucosides, isoscutellarein derivatives, caffeoyl‐quinic acids and iridoids have been isolated and structurally characterized by NMR. The essential oils were analysed by GC/MS and showed (E)‐phytol, fatty acids and carbonylic compounds as the most abundant compounds. The radical scavenging capacity of plant ethanolic extracts, as evaluated by the DPPH assay, was noteworthy, with IC₅₀ values of 92.08 – 105.42 μg/ml.     

Deciphering the mechanisms involved in Portulaca oleracea (C4) response to drought: metabolic changes including crassulacean acid‐like metabolism induction and reversal upon re‐watering

Portulaca oleracea is a C₄ plant; however, under drought it can change its carbon fixation metabolism into a crassulacean acid metabolism (CAM)‐like one. While the C₃‐CAM shift is well known, the C₄‐CAM transition has only been described in Portulaca. Here, a CAM‐like metabolism was induced in P. oleracea by drought and then reversed by re‐watering. Physiological and biochemical approaches were undertaken to evaluate the drought and recovery responses. In CAM‐like plants, chlorophyll fluorescence parameters were transitory affected and non‐radiative energy dissipation mechanisms were induced. Induction of flavonoids, betalains and antioxidant machinery may be involved in photosynthetic machinery protection. Metabolic analysis highlights a clear metabolic shift, when a CAM‐like metabolism is induced and then reversed. Increases in nitrogenous compounds like free amino acids and urea, and of pinitol could contribute to withstand drought. Reciprocal variations in arginase and urease in drought‐stressed and in re‐watered plants suggest urea synthesis is strictly regulated. Recovery of C₄ metabolism was accounted by CO₂ assimilation pattern and malate levels. Increases in glycerol and in polyamines would be of importance of re‐watered plants. Collectively, in P. oleracea multiple strategies, from induction of several metabolites to the transitory development of a CAM‐like metabolism, participate to enhance its adaptation to drought.     

Warming and drought differentially influence the production and resorption of elemental and metabolic nitrogen pools in Quercus rubra

The process of nutrient retranslocation from plant leaves during senescence subsequently affects both plant growth and soil nutrient cycling; changes in either of these could potentially feed back to climate change. Although elemental nutrient resorption has been shown to respond modestly to temperature and precipitation, we know remarkably little about the influence of increasing intensities of drought and warming on the resorption of different classes of plant metabolites. We studied the effect of warming and altered precipitation on the production and resorption of metabolites in Quercus rubra. The combination of warming and drought produced a higher abundance of compounds that can help to mitigate climatic stress by functioning as osmoregulators and antioxidants, including important intermediaries of the tricarboxylic acid (TCA) cycle, amino acids including proline and citrulline, and polyamines such as putrescine. Resorption efficiencies (REs) of extractable metabolites surprisingly had opposite responses to drought and warming; drought treatments generally increased RE of metabolites compared to ambient and wet treatments, while warming decreased RE. However, RE of total N differed markedly from that of extractable metabolites such as amino acids; for instance, droughted plants resorbed a smaller fraction of elemental N from their leaves than plants exposed to the ambient control. In contrast, plants in drought treatment resorbed amino acids more efficiently (>90%) than those in ambient (65–77%) or wet (42–58%) treatments. Across the climate treatments, the RE of elemental N correlated negatively with tissue tannin concentration, indicating that polyphenols produced in leaves under climatic stress could interfere with N resorption. Thus, senesced leaves from drier conditions might have a lower nutritive value to soil heterotrophs during the initial stages of litter decomposition despite a higher elemental N content of these tissues. Our results suggest that N resorption may be controlled not only by plant demand, but also by climatic influences on the production and resorption of plant metabolites. As climate–carbon models incorporate increasingly sophisticated nutrient cycles, these results highlight the need to adequately understand plant physiological responses to climatic variables.     

Chemical composition ofSpirulina and eukaryotic algae food products marketed in Spain

ThreeSpirulina and five eukaryotic algal food products available in the Spanish market have been extensively studied. Results are given for their gross chemical composition (water content, crude protein, total carbohydrates, lipids, nucleic acids etc.) and contents of macrominerals, trace elements, fatty acids, amino acids and neutral sugars. The results are compared to those from other studies on natural or laboratory-produced populations. An overall nutritional and toxicological evaluation of these products is included.     

A review of <Emphasis Type="Italic">Piper</Emphasis> spp. (Piperaceae) phytochemistry,insecticidal activity and mode of action

The tropical plant family Piperaceae has provided many past and present civilizations with a source of diverse medicines and food grade spice. The secondary plant compounds that produce these desired qualities function also as chemical defenses for many species in the genus Piper. The compounds with the greatest insecticidal activity are the piperamides. Many studies have shown the effectiveness of Piper spp. extracts for the control of stored products pests and recently studies from our laboratory group have tested the extracts of Piper. nigrum, P. guineense and P. tuberculatum against insect pests of the home and garden. These results and those from investigations that examined the biochemical and molecular modes of action of the piperamides singly or in combination will be the focus of this review. The conclusions of our current work with Piperaceae are that Piper extracts offer a unique and useful source of biopesticide material for controlling small-scale insect out-breaks and reducing the likelihood of resistance development when applied as a synergist with other botanical insecticides such as pyrethrum.     

Metabolic Fate of Depsides and Alkaloid Constituents in Aqueous Extracts from Mercurialis perennis L. during Fermentation

Dog's mercury (Mercurialis perennis L.) is an old medicinal plant, nowadays used in complementary medicine. Aqueous fermented extracts of the plant are being mainly applied in remedies to treat external inflammations, but a thorough phytochemical characterization is still lacking. Therefore, the conversion of characteristic compound classes from M. perennis extracts during fermentation and storage was investigated. The microbial transformation of the two main depsides phaselic acid (=(2R)‐O‐[(E)‐caffeoyl]malic acid; 1 ) and mercurialis acid (=(2R)‐[(E)‐caffeoyloxy]glutaric acid; 2 ) was monitored by HPLC‐DAD. The degradation followed a second‐order kinetic, and the calculated half‐life periods of both constituents were 67 and 30 months, respectively. Several depside metabolites were detected by GC/MS in AcOEt extracts as ^tBuMe₂Si (TBDMS) derivatives after derivatization, mainly dihydrocinnamic acids. Moreover, numerous α‐hydroxy acids were found, allegedly as degradation products from amino acids or peptides. The microbial alteration of the main alkaloid hermidin was also examined. After three days of fermentation, three novel N‐metabolites were formed and thoroughly assigned in CH₂Cl₂ extracts as a mixture of 3‐ethylhermidin, 3‐ethylhermidin quinone, and (E/Z)‐3‐ethylidenehermidin by GC/MS and NMR methods, as well as by means of total synthesis. A mechanism for the formation of these N‐metabolites starting from dimeric hermidin oxidation products is proposed. The obtained results reveal the complex pathways plant constituents may undergo during the fermentation of the extracts.     

Biostimulant activity of humic-like substances from agro-industrial waste on Chlorella vulgaris and Scenedesmus quadricauda

The use of microalgae in a number of sectors, including biodiesel, feed and food production, is proving to be of great interest. An evaluation was made of the possible biostimulant effects on Chlorella vulgaris and Scenedesmus quadricauda of humic-like substances (HLs) extracted from agro-industrial wastes. These included digestate from the waste of an agro-livestock farm (D-HL), oil extraction residues from rape (B-HL, Brassica napus L.) and tomato residues (T-HL). The microalgal growth medium (BG11) was supplemented with HLs to evaluate their effect on biomass yield as well as carbohydrate, chlorophylls a and b, lipid and fatty acid contents. Our results showed that the HLs used in the test are effective biostimulants of C. vulgaris and S. quadricauda. The biostimulant effect seems to depend on the type of extract used for cultivating the microalgae, the concentration and the species treated. Among the extracts applied to the growth medium, D-HL and T-HL seem to have a significant effect on microalgal biomass and lipid production. Although B-HL showed no significant effect on the biomass and lipid content of C. vulgaris and S. quadricauda, its presence in the growth medium increased the saturated:unsaturated fatty acid ratio (SFA/UFA) and stimulated the sugar metabolism of the microalgae by increasing their carbohydrate and chlorophyll content.     

Algae as production systems of bioactive compounds

Algal extracts are gaining increasing interest due to their unique composition and possibilities of wide industrial applications. Various extraction techniques are used for conversion of algal biomass into extracts. Recently, attention of scientists has been paid to novel methods, such as enzyme‐assisted extraction, microwave‐assisted extraction, pressurized liquid extraction, supercritical fluid extraction, and ultrasound‐assisted extraction, which enable the extraction of biologically active compounds without their degradation. In this review, the properties of biologically active compounds extracted from the biomass of algae reported in the literature are presented in a structured way. Algal extracts contain compounds such as carbohydrates, proteins, minerals, oil, fats, polyunsaturated fatty acids as well as bioactive compounds such as antioxidants (polyphenols, tocopherols [vitamin E], vitamin C, mycosporine‐like amino acids), and pigments, such as carotenoids (carotene xanthophyll), chlorophylls, and phycobilins (phycocyanin, phycoerythrin), which possess antibacterial, antiviral, antifungal, antioxidative, anti‐inflammatory, and antitumor properties. Finally, we assemble a list of applications of algal extracts in different developing branches of agriculture (biostimulants, bioregulators, feed additives) and in pharmaceutical industry.     

UPLC‐HRMS‐based untargeted metabolic profiling reveals changes in chickpea (Cicer arietinum) metabolome following long‐term drought stress

Genetic improvement for drought tolerance in chickpea requires a solid understanding of biochemical processes involved with different physiological mechanisms. The objective of this study is to demonstrate genetic variations in altered metabolic levels in chickpea varieties (tolerant and sensitive) grown under contrasting water regimes through ultrahigh‐performance liquid chromatography/high‐resolution mass spectrometry‐based untargeted metabolomic profiling. Chickpea plants were exposed to drought stress at the 3‐leaf stage for 25 days, and the leaves were harvested at 14 and 25 days after the imposition of drought stress. Stress produced significant reduction in chlorophyll content, F_v/F_m, relative water content, and shoot and root dry weight. Twenty known metabolites were identified as most important by 2 different methods including significant analysis of metabolites and partial least squares discriminant analysis. The most pronounced increase in accumulation due to drought stress was demonstrated for allantoin, l ‐proline, l ‐arginine, l ‐histidine, l ‐isoleucine, and tryptophan. Metabolites that showed a decreased level of accumulation under drought conditions were choline, phenylalanine, gamma‐aminobutyric acid, alanine, phenylalanine, tyrosine, glucosamine, guanine, and aspartic acid. Aminoacyl‐tRNA and plant secondary metabolite biosynthesis and amino acid metabolism or synthesis pathways were involved in producing genetic variation under drought conditions. Metabolic changes in light of drought conditions highlighted pools of metabolites that affect the metabolic and physiological adjustment in chickpea that reduced drought impacts.     

Accumulation,selection and covariation of amino acids in sieve tube sap of tansy (Tanacetum vulgare) and castor bean (Ricinus communis): evidence for the function of a basic amino acid transporter and the absence of a γ‐amino butyric acid transporter

Sieve tube sap was obtained from Tanacetum by aphid stylectomy and from Ricinus after apical bud decapitation. The amino acids in sieve tube sap were analyzed and compared with those from leaves. Arginine and lysine accumulated in the sieve tube sap of Tanacetum more than 10‐fold compared to the leaf extracts and they were, together with asparagine and serine, preferably selected into the sieve tube sap, whereas glycine, methionine/tryptophan and γ‐amino butyric acid were partially or completely excluded. The two basic amino acids also showed a close covariation in sieve tube sap. The acidic amino acids also grouped together, but antagonistic to the other amino acids. The accumulation ratios between sieve tube sap and leaf extracts were smaller in Ricinus than in Tanacetum. Arginine, histidine, lysine and glutamine were enriched and preferentially loaded into the phloem, together with isoleucine and valine. In contrast, glycine and methionine/tryptophan were partially and γ‐amino butyric acid almost completely excluded from sieve tube sap. The covariation analysis grouped arginine together with several neutral amino acids. The acidic amino acids were loaded under competition with neutral amino acids. It is concluded from comparison with the substrate specificities of already characterized plant amino acid transporters, that an AtCAT1‐like transporter functions in phloem loading of basic amino acids, whereas a transporter like AtGAT1 is absent in phloem. Although Tanacetum and Ricinus have different minor vein architecture, their phloem loading specificities for amino acids are relatively similar.     

Production of a carob enzymatic extract: potential use as a biofertilizer

In this paper, we describe a biological process that converts carob germ (CG), a proteinic vegetable by-product, into a water-soluble enzymatic hydrolyzate extract (CGHE). The chemical and physical properties are also described. The conversion is done using a proteolytic enzyme mixture. The main component of CGHE extracted by the enzymatic process is protein (68%), in the form of peptides and free amino acids, having a high content of glutamine and arginine, and a minor component of phytohormones, which are also extracted and solubilized from the CG. We have also compared its potential fertilizer/biostimulant capacity on growth, flowering, and fruiting of tomato plants (Licopericon pimpinellifolium cv. Momotaro) with that of an animal enzymatic protein hydrolyzate. CGHE had a significantly beneficial impact, most notably regarding the greater plant height, number of flowers per plant, and number of fruits per plant. This could be due primarily to its phytohormonal action.     

Phenotypic selection on nectar amino acid composition in the Lepidoptera pollinated orchid species Gymnadenia conopsea

Plant–pollinator interactions are potential drivers of evolution in floral traits. Because nectar chemical composition is known to mediate both plant–pollinator interactions and plant reproductive success, it can be expected that chemical composition of nectar is subjected to strong pollinator‐mediated selective forces. However, the extent of natural selection on different nectar components has not been studied so far. Using the Lepidoptera pollinated fragrant orchid Gymnadenia conopsea as a model species, we used high‐performance anion‐exchange chromatography (HPAEC) to characterize the sugar and amino acid composition of floral nectar in three calcareous grassland populations of G. conopsea. We then measured phenotypic selection on nectar composition and on other plant and floral traits through applying both linear regression and structural equation modelling. We demonstrate phenotypic selection on plant height, inflorescence height and on specific nectar amino acids, whereas spur length, total sugar and amino acid concentration were not direct targets of selection. Chemical nectar composition is thus indeed under selective pressure but nectar amino acids are much more important to fitness of G. conopsea, as compared to nectar sugars. Furthermore, as we found no evidence of selection on the total amino acid concentration, it is unlikely that amino acids increase pollinator attraction because they are a pollinator nitrogen source. To further unravel the evolutionary ecology of floral nectar, behavioural experiments with pollinators exposed to different nectar components and studies experimentally identifying the selective agents are recommended.