Effect of storage on plant biostimulant and bioactive properties of freeze-dried Chlorella vulgaris biomass.

Microalgae are potential plant biostimulants and biocontrol agents. A major hurdle towards their commercialization is the production of large volumes of biomass at the correct time of year. Secondary metabolites are unstable and the "shelf-life" of bioactive microalgal biomass needs to be investigated. The aim of the study was to investigate the effects of storage conditions on freeze-dried microalgae to determine how long the biomass retained its growth promoting and bioactive properties under various temperature and light conditions. Chlorella vulgaris biomass was stored in the dark at - 70 °C, 10 °C, and 25 °C and in the light at 25 °C. Samples were tested every 3-4 months for 15 months. Storage time significantly influenced the rate of change in the bioactivity in the C. vulgaris biomass with storage temperature also having some effect. Rooting activity decreased in the mungbean rooting assay over time up to 12 months and then increased slightly. Antimicrobial activity increased against Staphylococcus aureus and Escherichia coli for up to 12 months and then declined. Antioxidant activity measured in the DPPH assay remained relatively stable for up to 12 months and then significantly decreased with longer storage. The change in bioactivity over time was attributed to the gradual breakdown of the rigid cell wall of C. vulgaris, thereby improving extraction efficiency but exposing the secondary metabolites to oxygen, thus quickening their degradation. Biomass produced for commercial purposes requires preliminary validation as the results of the present study showed that bioactive compounds are susceptible to degradation over time. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10811-021-02596-9.

Interactive effects of plant growth-promoting rhizobacteria and a seaweed extract on the growth and physiology of Allium cepa L. (onion).

Detrimental effects caused by the overuse of synthetic agrochemicals have led to the development of natural biostimulants such as seaweed extracts and plant growth-promoting rhizobacteria (PGPR) being used as an alternative, environmentally-friendly technology to improve crop growth and increase agricultural yields. The present study aimed to investigate the interactions between PGPR and a commercial seaweed extract on the growth and biochemical composition of onion (Allium cepa). A pot trial was conducted under greenhouse conditions where onion plants were treated individually with the two PGPR, namely Bacillus licheniformis (BL) and Pseudomonas fluorescens (PF) and a seaweed extract Kelpak® (KEL) and combinations of KEL + BL and KEL + PF. Growth and yield parameters were measured after 12 weeks. KEL-treated plants showed the best growth response and overcame the inhibitory effects of BL treatment. KEL-treated plants also had the highest chlorophyll content. PGPR application improved the mineral nutrition of onion with these plants having the highest mineral content in the leaves and bulb. All biostimulant treatments increased the endogenous cytokinin and auxin content with the highest concentrations generally detected in the PF-treated plants. These results suggest that co-application of different biostimulant classes with different modes of action could further increase crop productivity with an improvement in both growth and nutrition content being achieved in onion with the co-application of a seaweed extract and PGPR.

Synthetic strigolactone (rac-GR24) alleviates the adverse effects of heat stress on seed germination and photosystem II function in lupine seedlings.

There is increasing experimental evidence that strigolactones, a class of carotenoid-derived sesquiterpenoid hormones, and their downstream signal components play a role in plant resilience to abiotic stress. Strigolactones positively influence plant coping mechanisms in response to abiotic stressors like drought and high salinity. In this study, we examined the effects of rac-GR24 (a synthetic strigolactone analog) and strigolactone inhibitors on the physiological and molecular responses associated with thermotolerance during seed germination and seedling development in Lupinus angustifolius under heat stress. Photosystem I & II functions were also evaluated via Chl a fluorescence transient analysis in heat stressed lupine seedlings. Our results suggest a putative role for GR24 in mediating tolerance to heat stress during seed germination and seedling development albeit these responses appeared independent of D14-mediated signalling. Seeds primed with GR24 had the highest of all germination indices, enhanced proline content and reduced peroxidation of lipids. GR24 also enhanced the activities of enzymes of the antioxidant and glyoxalase systems in lupine seedlings. The JIP-test indicated that GR24 conferred resistance to heat stress-induced damage to the oxygen evolution complex while also preventing the inactivation of PSII reaction centres thus ensuring PSII thermotolerance.

A critical review on exploiting the pharmaceutical potential of plant endophytic fungi.

The escalating demand for secondary metabolites in international markets poses a severe threat to many plant species. An unscrupulous collection is also the immediate challenge to the survival of many unthreatened as well as vulnerable plants. Fungal endophytes have emerged in recent years as a promising substitute for sources of plant secondary metabolites. Many appealing secondary metabolites with potent antibacterial, antifungal, insecticidal, antioxidant, cytotoxic and anticancer properties have been discovered from endophytic fungi. Concerning their distinctive genetic and metabolic diversity and promising activities, they hold a plausible application in medicine and industry. However, there is little success in utilizing the pharmaceutical potential of fungal endophytes. Cutting-edge research is desirable to establish and bolster in vitro biosynthetic proficiency of fungal endophytes. Modern biotechnological techniques [such as multilocus sequence typing (MLST), metabolomics, metagenomics and next-generation sequencing (NGS) technologies] and bioinformatics approaches can fill a gap in fungal endophyte research. The present review focuses on how advanced chemical, biotechnological and computational molecular biology methods can be used for robust exploitation of bioactive compounds from these microorganisms.

Role of Smoke Stimulatory and Inhibitory Biomolecules in Phytochrome-Regulated Seed Germination of Lactuca sativa.

The biologically active molecules karrikinolide (KAR1) and trimethylbutenolide (TMB) present in wildfire smoke play a key role in regulating seed germination of many plant species. To elucidate the physiological mechanism by which smoke-water (SW), KAR1, and TMB regulate seed germination in photosensitive 'Grand Rapids' lettuce (Lactuca sativa), we investigated levels of the dormancy-inducing hormone abscisic acid (ABA), three auxin catabolites, and cytokinins (26 isoprenoid and four aromatic) in response to these compounds. Activity of the hydrolytic enzymes α-amylase and lipase along with stored food reserves (lipids, carbohydrate, starch, and protein) were also assessed. The smoke compounds precisely regulated ABA and hydrolytic enzymes under all light conditions. ABA levels under red (R) light were not significantly different in seeds treated with TMB or water. However, TMB-treated seeds showed significantly inhibited germination (33%) compared with water controls (100%). KAR1 significantly enhanced total isoprenoid cytokinins under dark conditions in comparison with other treatments; however, there was no significant effect under R light. Enhanced levels of indole-3-aspartic acid (an indicator of high indole-3-acetic acid accumulation, which inhibits lettuce seed germination) and absence of trans-zeatin and trans-zeatin riboside (the most active cytokinins) in TMB-treated seeds might be responsible for reduced germination under R light. Our results demonstrate that SW and KAR1 significantly promote lettuce seed germination by reducing levels of ABA and enhancing the activity of hydrolytic enzymes, which aids in mobilizing stored reserves. However, TMB inhibits germination by enhancing ABA levels and reducing the activity of hydrolytic enzymes.

Bioactive molecules derived from smoke and seaweed Ecklonia maxima showing phytohormone-like activity in Spinacia oleracea L.

Bioactive compounds such as karrikinolide (KAR1 from smoke) and eckol (from the seaweed Ecklonia maxima) show promising effects on several important crop plants. These plant growth-stimulating organic biomolecules, along with crude extracts (smoke-water and Kelpak® product prepared from Ecklonia maxima), were tested on spinach plants. Eckol sprayed at 10-6 M significantly increased all the growth and biochemical parameters examined compared to control spinach plants. All tested plant growth biostimulants significantly increased total chlorophyll, carotenoids and protein content of spinach leaves. The cytokinin profile of spinach plants was also determined. Cis-zeatin, dihydrozeatin and isopentenyladenine types of cytokinins were promoted by both smoke- and seaweed-based biostimulants. In comparison to the control plants, the level of free sinapic acid was greater in all spinach plants treated with these biostimulants. The application of these biostimulants can help spinach crop by improving growth, yield and nutritional quality; moreover, they are organic and cost-effective.

Influence of biostimulants-seed-priming on Ceratotheca triloba germination and seedling growth under low temperatures, low osmotic potential and salinity stress.

Extreme temperatures, drought and salinity stress adversely affect seed germination and seedling growth in crop species. Seed priming has been recognized as an indispensable technique in the production of stress-tolerant plants. Seed priming increases seed water content, improves protein synthesis using mRNA and DNA and repair mitochondria in seeds prior to germination. The current study aimed to determine the role of biostimulants-seed-priming during germination and seedling growth of Ceratotheca triloba (Bernh.) Hook.f. (an indigenous African leafy vegetable) under low temperature, low osmotic potential and salinity stress conditions. Ceratotheca triloba seeds were primed with biostimulants [smoke-water (SW), synthesized smoke-compound karrikinolide (KAR1), Kelpak® (commercial seaweed extract), phloroglucinol (PG) and distilled water (control)] for 48h at 25°C. Thereafter, primed seeds were germinated at low temperatures, low osmotic potential and high NaCl concentrations. Low temperature (10°C) completely inhibited seed germination. However, temperature shift to 15°C improved germination. Smoke-water and KAR1 enhanced seed germination with SW improving seedling growth under different stress conditions. Furthermore, priming seeds with Kelpak® stimulated percentage germination, while PG and the control treatment improved seedling growth at different PEG and NaCl concentrations. Generally, high concentrations of PEG and NaCl brought about detrimental effects on seed germination and seedling growth. Findings from this study show the potential role of seed priming with biostimulants in the alleviation of abiotic stress conditions during seed germination and seedling growth in C. triloba plants.

Antifungal and antioxidant activities of Coleonema album and C. pulchellum against skin diseases.

CONTEXT: Coleonema album (Thunb) Bart. & H. L. Wendl (Rutaceae) has been used in the formulation of skincare products, and the Khoisan people rub it on their skin to add luster. Coleonema pulchellum I. Williams has received less attention in the South African traditional medicine. OBJECTIVE: This study investigates the antifungal and antioxidant activities of C. album and C. pulchellum essential oil (EO) and leaf extracts; and analyzes the chemical components of their EOs. MATERIALS AND METHODS: Antifungal activity of leaf extracts was determined using the microdilution method with griseofulvin and ketoconazole as controls. Antifungal capacity of EO was investigated using the 'Volatile release plate method'. Trichophyton rubrum (ATCC 28188) and T. mentagrophytes (ATCC 9533) mycelia (0.3 cm diameter) were placed on fresh yeast malt agar in Petri dishes with filter paper (impregnated with 20 µL of EO) on the lid for direct exposure to EO volatiles while plates without EO were used as controls. The incubation time was seven days. Antioxidant activities of the leaf extracts were determined. RESULTS: Methanol leaf extract of C. pulchellum inhibited the growth of three fungi tested with MIC values of 195, 391 and 49 µg/mL for Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum gypseum, respectively. Terpenes formed the major components of the EO. The EO from both plants inhibited the growth of T. rubrum in vitro. DISCUSSION AND CONCLUSION: This study revealed the therapeutic value of C. pulchellum. Coleonema album and C. pulchellum should be considered as potential plants for skin ointment from natural origin.

Can the use of natural biostimulants be a potential means of phytoremediating contaminated soils from goldmines in South Africa?

Biostimulants offer great potential in improving phytoremediation of contaminated soils. In the current greenhouse-based study, Brassica juncea seedlings grown on soils collected from Krugersdorp Goldmine and the adjourning areas (a Game Reserve and private farmland) were supplemented with different biostimulants (Kelpak® = KEL, vermicompost leachate = VCL, smoke-water = SW). Indole-3-butyric acid (IBA) was included in the study for comparative purposes because these biostimulants are known to enhance rooting. Prior to the pot trial, concentrations of elements in the three soil types were determined using Inductively Coupled Plasma-Optical Emission Spectroscopy. Plants were harvested after 105 days and the growth and concentrations of elements in the various plant organs were determined. TheB. juncea seedlings with and without biostimulants did not survive when growing in soil from the Krugersdorp Goldmine. The Game Reserve and private farmland soils supplemented with KEL produced the highest plant biomass and the lowest accumulation of metals in the organs of B. juncea. High concentrations (>13 000 mg kg(-1)) of zinc and aluminium were quantified in the roots of IBA-supplemented soils from the Game Reserve. Generally, IBA and SW enhanced the phytoremediation of B. juncea due to elevated levels of elements that accumulated in their different organs.

Enhancing growth, phytochemical constituents and aphid resistance capacity in cabbage with foliar application of eckol--a biologically active phenolic molecule from brown seaweed.

Although foliar application of seaweed extracts on plant growth and development has and is extensively studied, reliable knowledge and understanding of the mode of action of particular compound(s) responsible for enhancing plant growth is lacking. A brown seaweed Ecklonia maxima is widely used commercially as a biostimulant to improve plant growth and crop protection. Eckol, a phenolic compound isolated from E. maxima has recently shown stimulatory effects in maize, indicating its potential use as a plant biostimulant. Cabbage is a widely cultivated vegetable crop throughout the world, which requires high input of fertilizers and is susceptible to several aphid borne diseases. This study was conducted to evaluate the effect of foliar application of eckol on the growth, phytochemical constituents and myrosinase activity (aphid resistance capacity) of commercially cultivated cabbage. Foliar application of eckol (10(-6) M) significantly enhanced shoot and root length, shoot and root fresh and dry weight, leaf area and leaf number. This treatment also showed a significant increase in photosynthetic pigments (chlorophyll 'a', chlorophyll 'b', total chlorophyll and carotenoid) compared to the untreated plants. The levels of protein, proline and iridoid glycosides were significantly higher in cabbage leaves with eckol treatment. All the control plants were severely infested with cabbage aphid (Brevicoryne brassicae) but no infestation was observed on the eckol-sprayed plants, which can be attributed to an increase in myrosinase activity. This study reveals dual effects (plant growth promoting and insect repelling) of eckol on cabbage plants that need further investigations both under field conditions and in other brassicaceous species.

Enhancing Phytoremediation Potential of Pennisetum clandestinum Hochst in Cadmium-Contaminated Soil Using Smoke-Water and Smoke-Isolated Karrikinolide.

The use of plant growth regulators (PGRs) and biostimulants to enhance phytoextraction is gaining popularity in phytoremediation technology. This study investigated the stimulatory effects of smoke-water (SW), a smoke-derived compound karrikinolide (KAR1) and other known plant growth regulators (PGRs) [gibberellic acid (GA3), kinetin (Kin) and indole-3-butyric acid (IBA)] to enhance the phytoextraction potential of Pennisetum clandestinum. Pennisetum clandestinum seedlings were grown for 10 weeks in vermiculite using Hoagland's nutrient solution and were treated with cadmium (Cd) (2, 5, and 10 mg L(-1)) and SW, KAR1 and PGRs. KAR1 exhibited positive effects on shoot and root dry weight (140 and 137 mg respectively) at the highest concentration of Cd (10 mg L(-1)) compared to all the other treatments. KAR1 and SW treatments used in the present study significantly improved the phytoextraction potential of P. clandestinum (602 and 575 mg kg(-1) respectively) compared to the other tested PGRs. This is the first report on the use of SW and KAR1 to enhance phytoremediation potential in P. clandestinum. Further studies are needed to elucidate the exact mechanisms of smoke constituents involved in phytoextraction potential of plant species.

Advances in algal drug research with emphasis on enzyme inhibitors.

Enzyme inhibitors are now included in all kinds of drugs essential to treat most of the human diseases including communicable, metabolic, cardiovascular, neurological diseases and cancer. Numerous marine algae have been reported to be a potential source of novel enzyme inhibitors with various pharmaceutical values. Thus, the purpose of this review is to brief the enzyme inhibitors from marine algae of therapeutic potential to treat common diseases. As per our knowledge this is the first review for the potential enzyme inhibitors from marine origin. This review contains 86 algal enzyme inhibitors reported during 1989-2013 and commercial enzyme inhibitors available in the market. Compounds in the review are grouped according to the disease conditions in which they are involved; diabetes, obesity, dementia, inflammation, melanogenesis, AIDS, hypertension and other viral diseases. The structure-activity relationship of most of the compounds are also discussed. In addition, the drug likeness properties of algal inhibitors were evaluated using Lipinski's 'Rule of Five'.

A novel inhibitor of cytokinin degradation (INCYDE) influences the biochemical parameters and photosynthetic apparatus in NaCl-stressed tomato plants.

The effect of 2-chloro-6-(3-methoxyphenyl)aminopurine [inhibitor of cytokinin degradation (INCYDE)] at 10 nM on growth, biochemical and photosynthetic efficiency in sodium chloride (NaCl)-stressed (75, 100 and 150 mM) tomato plants was investigated. NaCl-induced decline in plant vigor index was slightly reversed by both drenching and foliar application of INCYDE. Foliar application of INCYDE significantly increased the flower number in the control and 75 mM NaCl-supplemented plants, while drenching was more effective in 150 mM NaCl-stressed plants. Antioxidant enzymes (peroxidase, catalase and superoxide dismutase) were enhanced in the presence of INCYDE in the control and NaCl-stressed plants. Higher concentration of malondialdehyde (MDA) associated with oxidative (lipid peroxidation) damage in leaf tissue which was evident in the presence of NaCl stress was significantly attenuated with the drenching and foliar application of INCYDE. Regardless of NaCl concentration, application of INCYDE had no significant influence on maximum quantum efficiency of photosystem II. However, the reduced quantum yield of photosystem II and coefficient of photochemical quenching under continuous illumination with actinic light at four intensities (264, 488, 800 and 1,200 µmol m(-2) s(-1)) in NaCl-stressed (100 and 150 mM) tomato plants were significantly alleviated by drenching application with INCYDE. Non-photochemical quenching of the singlet excited state of chlorophyll a and relative electron transfer rate were generally higher in INCYDE-treated plants than in the controls. From an agricultural perspective, these findings indicate the potential of INCYDE in protecting plants against NaCl stress and the possibility of enhanced productivity.

Plant growth regulators enhance gold uptake in Brassica juncea.

The use of plant growth regulators is well established and they are used in many fields of plant science for enhancing growth. Brassica juncea plants were treated with 2.5, 5.0 and 7.5 microM auxin indole-3-butyric acid (IBA), which promotes rooting. The IBA-treated plants were also sprayed with 100 microM gibberellic acid (GA3) and kinetin (Kin) to increase leaf-foliage. Gold (I) chloride (AuCl) was added to the growth medium of plants to achieve required gold concentration. The solubilizing agent ammonium thiocyanate (1 g kg(-1)) (commonly used in mining industries to solubilize gold) was added to the nutrient solution after six weeks of growth and, two weeks later, plants were harvested. Plant growth regulators improved shoot and root dry biomass of B. juncea plants. Inductively Coupled Plasma Optical Emission Spectrometry analysis showed the highest Au uptake for plants treated with 5.0 microM IBA. The average recovery of Au with this treatment was significantly greater than the control treatment by 45.8 mg kg(-1) (155.7%). The other IBA concentrations (2.5 and 7.5 microM) also showed a significant increase in Au uptake compared to the control plants by 14.7 mg kg(-1) (50%) and 42.5 mg kg(-1) (144.5%) respectively. A similar trend of Au accumulation was recorded in the roots of B. juncea plants. This study conducted in solution culture suggests that plant growth regulators can play a significant role in improving phytoextraction of Au.

Cadmium induces hypodermal periderm formation in the roots of the monocotyledonous medicinal plant Merwilla plumbea.

BACKGROUND AND AIMS: Merwilla plumbea is an important African medicinal plant. As the plants grow in soils contaminated with metals from mining activities, the danger of human intoxication exists. An experiment with plants exposed to cadmium (Cd) was performed to investigate the response of M. plumbea to this heavy metal, its uptake and translocation to plant organs and reaction of root tissues. METHODS: Plants grown from seeds were cultivated in controlled conditions. Hydroponic cultivation is not suitable for this species as roots do not tolerate aquatic conditions, and additional stress by Cd treatment results in total root growth inhibition and death. After cultivation in perlite the plants exposed to 1 and 5 mg Cd L(-1) in half-strength Hoagland's solution were compared with control plants. Growth parameters were evaluated, Cd content was determined by inductively coupled plasma mass spectroscopy (ICP-MS) and root structure was investigated using various staining procedures, including the fluorescent stain Fluorol yellow 088 to detect suberin deposition in cell walls. KEY RESULTS: The plants exposed to Cd were significantly reduced in growth. Most of the Cd taken up by plants after 4 weeks cultivation was retained in roots, and only a small amount was translocated to bulbs and leaves. In reaction to higher Cd concentrations, roots developed a hypodermal periderm close to the root tip. Cells produced by cork cambium impregnate their cell walls by suberin. CONCLUSIONS: It is suggested that the hypodermal periderm is developed in young root parts in reaction to Cd toxicity to protect the root from radial uptake of Cd ions. Secondary meristems are usually not present in monocotyledonous species. Another interpretation explaining formation of protective suberized layers as a result of periclinal divisions of the hypodermis is discussed. This process may represent an as yet unknown defence reaction of roots when exposed to elemental stress.

Effect of cadmium accumulation on anti-inflammatory activity in two Eucomis species.

Eucomis species (Hyacinthaceae) are widely used in South Africa as traditional medicine. The bulbs are used to alleviate a variety of symptoms including pain and inflammation. High levels of cyclooxygenase-1 and -2 (COX-1 and COX-2) inhibitory activity have been associated with certain Eucomis species. The aim of this study was to quantify cadmium(Cd) accumulation and examine its effect on COX-1 and COX-2 anti-inflammatory activity in Eucomis autumnalis and Eucomis humilis. Cadmium application at 2 mg L(-1) over a 6 week period revealed a substantial difference in total Cd accumulation in E. autumnalis and E. humilis (40.2 and 15.3 mg Cd kg-1, respectively). When supplied with Cd at 2 mg L(-1), E. humilis bulbous extracts showed lower inhibitory activity than the control for both COX-1 and COX-2. E. autumnalis bulbous extracts had greater COX-1 activity compared to the control. While COX-2 activity was suppressed. Researchers should be aware of the effect of environmental contaminants when reporting on biological activity of crude plant extracts.