FUNCTIONAL ANALYSIS OF MEDICINAL PLANTS USING SYSTEMS BIOLOGY APPROACHES

Plant derived medicine is an important source of life saving drugs, but the genome information of most important medicinal plants is still unavailable. The need of the hour is to identify more functional genes and enzymes that control secondary metabolite production in medical plants, develop new methods for systematics, engineer resistance to number of biotic and abiotic stresses, and develop new conservation strategies, more genomics, proteomics and metabolomics information needs to be produced. In this review, a brief overview of various omic technologies and its applications to medicinal and aromatic plants are discussed.Â

SCREENING OF ENDOPHYTIC FUNGI FOR THEIR ABILITY TO PRODUCE EXTRACELLULARCELLULASES

Objective: Screening endophytic isolates from woody perennial medicinal plants of Western Ghats for production of extracellular cellulasesMethods: Endophytes were isolated using normal microbiological methods and their colonization frequency and dominance were calculated using statistical methods. Efficiency of growth on complex cellulosic substrates was evaluated on media supplemented with specific substrates. Enzyme assays with identified endophytic fungi were carried from their secretome.Results:Forty endophytic fungal isolates were obtained using standard isolation methods from different medicinal plants from a biodiversity hotspot in the Western Ghats region of Karnataka, India. The isolated endophytic fungi were then identified based on their morphological characters. The percentage of dominant endophytes (ð·) was calculated based on the colonization frequency. Among the isolated fungi, F. solani and Talaromyces sp. was found to be highest, at5.5 and 5.6 respectively. Each of the identified fungi grown on CMC and seven among the 40 isolates were found to grow luxuriantly as measured by radial growth. The identities of these fungi were morphologically reconfirmed and were completely carbon drained by growing them on a low nutrient medium. These fungi were later evaluated for their growth on avicel and microcrystalline cellulose. Fusarium solani and Tarlaromyces sp. were significantly better in their growth when compared to other endoophytes tested. Further, the cellulosome complex of enzymes were analysed in the secretome of Fusarium solani and Talaromyces sp. Total filter paper activity of Fusarium solani was found to be FPU/ml, 76 FPU/ml and 70 FPU/ml at 24, 48 and 72 hours respectively. Similarly, Filter paper activity of Talaromyces sp. was found to be 89, 86 and 78 FPU/ml at 24h, 48h and 72h respectively. Endogluconase activity of Fusarium solani was found to be 63 CMCase, 60and 61 CMCase at 24, 48 and 72hours of incubation respectively, which was greater than Talaromyces sp. Similarly, Exogluconase and Beta-glucosidase activities were also found to be high in Talomyces sp. when compared to Fusarium solani at all the time intervals tested.Conclusion:The results from the present study reveals that Fusarium sSolani and Talaromyces sp.are extremely potent producers of cellulases and can thus be used for eco-friendly and economic hydrolysis of biomass for biofuel purposes. Â

The impact of an extreme climatic disturbance and different fertilization treatments on plant development, phenology, and yield of two cultivar groups of Solanum betaceum Cav

[EN] Changing climatic conditions impose a challenge both to biodiversity and food security. The effects of climate change affect different aspects of the plant or crop, such as morphological and phenological aspects, as well as yield. The effects of greenhouse conditions might be comparable in some cases to a permanent extreme disturbance in climate and weather, thus, contributing to our knowledge on climate change impacts on plant species. We have investigated the differences for 23 traits in two cultivar groups of an Andean traditional crop, Solanum betaceum, under two different environmental conditions that correspond to the traditional practices in the open field and three cultural managements under greenhouse conditions (no fertilization or control, organic, and mineral). We found that traditional practices in the open field are the less productive. Moreover, in warmer and drier conditions the treatment with organic fertilization was the most productive. Greenhouse conditions, however, delay production. We further identified traits that differentiate both cultivar groups and traits that are linked to either the new climate conditions or the fertilization treatments. Fruit characteristics were quite homogeneous between the two cultivar groups. Overall, our results provide insight on the consequences that climate change effects might exert on crops such as tree tomato, reveal that greenhouses can be a robust alternative for tree tomato production, and highlight the need to understand how different managements are linked to different solutions to fulfil the farmers' demands.M.X.R.-G. was funded by Secretaria Nacional de Educacion Superior, Ciencia, Tecnologia e Innovacion (SENESCYT: www.educacionsuperior.gob.ec/) with a Prometeo Fellowship. This research was co-financed by Universidad Politecnica de Madrid, http://www.upm.es/ (Ayudas para proyectos semilla de investigacion PID para Latinoamerica, proyecto AL14-PID-09: http://www.upm.es/sfs/Rectorado/Vicerrectarode%20de%20Relaciones%,20Internacionales/America%20Latina/AyudaLA_Adjud13.pdf) and Universidad Tecnica Tecnica Paticular de Loja, https://www.utpl.edu.ec/ (proyecto PROY_FIN_CCAA_ 0016). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Tandazo-Yunga, J.; Ruíz-González, MJ.; Rojas, J.; Capa-Mora, E.; Prohens Tomás, J.; Alejandro, J.; Acosta-Quezada, P. (2017). The impact of an extreme climatic disturbance and different fertilization treatments on plant development, phenology, and yield of two cultivar groups of Solanum betaceum Cav. PLoS ONE. 12(12). https://doi.org/10.1371/journal.pone.0190316Se0190316121

Activity of cyazofamid against Sclerospora graminicola, a downy mildew disease of pearl millet

The efficacy of cyazofamid was tested against pearl millet downy mildew disease caused by Sclerospora graminicola Schroet. Significant inhibition of sporangial sporulation, zoospore release and motility was observed at 0.3 mg mL−1, and this concentration also provided good fungicidal activity under in vitro conditions. Under glasshouse conditions, none of the concentrations tested, either 0.01–2 mg mL−1 as seed treatment or 1–10 mg mL−1 by foliar application, was found to be phytotoxic. The effect of cyazofamid was tested by seed treatment alone, seed treatment followed by foliar application and foliar application alone. Seed treatment with cyazofamid offered only 19.7% disease control, but seed treatment followed by a single foliar application to diseased plants provided good control over disease, seed treatment with two foliar applications was significantly superior and foliar application alone showed a high level of activity, with 10 mg mL−1 giving 97.9% disease control. Lack of systemic activity of cyazofamid was evident, root treatment giving disease levels on a par with the untreated control. The fungicide exhibited strong curative activity, but only moderate translaminar activity, with only marginal (34.8%) disease control after treatment of the adaxial leaf surface at 10 mg mL−1. Loss of cyazofamid activity over time was very low, indicating stable residual and rainfastness activity. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of downy mildew disease of pearl millet. Copyright © 2007 Society of Chemical Industr

Current trends and challenges in the synthesis and applications of chitosan-based nanocomposites for plants: A review

Chitosan, a low-cost and multipurpose polymer with numerous desired physicochemical and biological properties has been tested for various applications in agriculture, pharmacy, and biomedicine industries. The availability of functional groups along the backbone makes chitosan readily available for other polymers and metal ions to form bio-nanocomposites. Different types of chitosan-based nanocomposites have been designed and tested for the enhancement of chitosan efficiency and ultimately widening the application areas of chitosan in plants. These nanocomposites serve different purposes such as eliciting plant?s defence systems against different threats (pathogen attack), antimicrobial agent against bacteria, fungi and viruses, enhancement of nutrient uptake by plants, control release of micro/macronutrients, fungicides and herbicides. In this review, an extensive outlook has been provided (mainly in the last five years) to recent trends and advances in the fabrication and application of chitosan-based composites. Finally, current challenges and future development opportunities of chitosan-based nanocomposites for plants are discussed

Activity of cyazofamid against Sclerospora graminicola, a downy mildew disease of pearl millet

The efficacy of cyazofamid was tested against pearl millet downy mildew disease caused by Sclerospora graminicola Schroet. Significant inhibition of sporangial sporulation, zoospore release and motility was observed at 0.3 mg mL<SUP>−1</SUP>, and this concentration also provided good fungicidal activity under in vitro conditions. Under glasshouse conditions, none of the concentrations tested, either 0.01-2 mg mL<SUP>−1</SUP> as seed treatment or 1-10 mg mL<SUP>−1</SUP> by foliar application, was found to be phytotoxic. The effect of cyazofamid was tested by seed treatment alone, seed treatment followed by foliar application and foliar application alone. Seed treatment with cyazofamid offered only 19.7% disease control, but seed treatment followed by a single foliar application to diseased plants provided good control over disease, seed treatment with two foliar applications was significantly superior and foliar application alone showed a high level of activity, with 10 mg mL<SUP>−1</SUP> giving 97.9% disease control. Lack of systemic activity of cyazofamid was evident, root treatment giving disease levels on a par with the untreated control. The fungicide exhibited strong curative activity, but only moderate translaminar activity, with only marginal (34.8%) disease control after treatment of the adaxial leaf surface at 10 mg mL<SUP>−1</SUP>. Loss of cyazofamid activity over time was very low, indicating stable residual and rainfastness activity. These results indicate that cyazofamid has a high potential to be an effective fungicide for the control of downy mildew disease of pearl millet

Detection and Characterization of Antibacterial Siderophores Secreted by Endophytic Fungi from Cymbidium aloifolium

Endophytic fungi from orchid plants are reported to secrete secondary metabolites which include bioactive antimicrobial siderophores. In this study endophytic fungi capable of secreting siderophores were isolated from Cymbidium aloifolium, a medicinal orchid plant. The isolated extracellular siderophores from orchidaceous fungi act as chelating agents forming soluble complexes with Fe3+. The 60% endophytic fungi of Cymbidium aloifolium produced hydroxamate siderophore on CAS agar. The highest siderophore percentage was 57% in Penicillium chrysogenum (CAL1), 49% in Aspergillus sydowii (CAR12), 46% in Aspergillus terreus (CAR14) by CAS liquid assay. The optimum culture parameters for siderophore production were 30 degrees C, pH 6.5, maltose and ammonium nitrate and the highest resulting siderophore content was 73% in P. chrysogenum. The total protein content of solvent-purified siderophore increased four-fold compared with crude filtrate. The percent Fe3+ scavenged was detected by atomic absorption spectra analysis and the highest scavenging value was 83% by P. chrysogenum. Thin layer chromatography of purified P. chrysogenum siderophore showed a wine-colored spot with R-f value of 0.54. HPLC peaks with R(t)s of 10.5 and 12.5 min were obtained for iron-free and iron-bound P. chrysogenum siderophore, respectively. The iron-free P. chrysogenum siderophore revealed an exact mass-to-charge ratio (m/z) of 400.46 and iron-bound P. chrysogenum siderophore revealed a m/z of 453.35. The solvent-extracted siderophores inhibited the virulent plant pathogens Ralstonia solanacearum, that causes bacterial wilt in groundnut and Xanthomonas oryzae pv. oryzae which causes bacterial blight disease in rice. Thus, bioactive siderophore-producing endophytic P. chrysogenum can be exploited in the form of formulations for development of resistance against other phytopathogens in crop plants

Exogenous priming of chitosan induces upregulation of phytohormones and resistance against cucumber powdery mildew disease is correlated with localized biosynthesis of defense enzymes

In recent years, exploration of biopolymer-based materials to avoid hazardous chemicals in agriculture has gained enormous importance for sustainable crop improvement. In the present study, chitosan a biopolymer derived from crab-shell was used in different concentrations as priming agent to cucumber seeds and were evaluated for its effect to enhance plant growth parameters as well as its ability to induce resistance against powdery mildew disease. Among the treatments, seeds-primed with 2.5 mg/mL exhibited early seedling germination of 90% and vigour of 2665 and also remarkably enhanced the cucumber growth parameters which might be fairly attributed to the stimulation of phytohormones content in primed plants over the controls. More importantly, under greenhouse conditions a significant induced disease protection of 66.6% against powdery mildew disease was noticed in chitosan-pretreated plants at 2.5 mg/mL. The induced resistant plants also showed a significant deposition of lignin, callose and H2O2. Notably, polyphenol oxidase, phenylalanine ammonia-lyase, peroxidase and glucanase defense-responsive enzymes were upregulated in chitosan-primed plants. Considered together, these results determine that the susceptible cucumber cultivar elicits immunity after perception of priming with chitosan to upregulate phytohormones and synthesize defense-responsive enzymes, thereby induce resistance against powdery mildew disease and strengthen the growth-promotion of cucumber plants. (C) 2020 Elsevier B.V. All rights reserved

Exogenous trehalose treatment enhances the activities of defense-related enzymes and triggers resistance against downy mildew disease of pearl millet

In recent years, diverse physiological functions of various sugars are the subject of investigations. Their roles in signal transduction in plant responses to biotic and abiotic stresses have become apparent, and growing scientific evidence has indicated that disaccharides like sucrose and trehalose mediate plant defense responses similar to those induced by elicitors against the pathogens. Trehalose is a well-known metabolic osmoregulator, stress protectant and a non-reducing disaccharide commonly found in a wide variety of organisms, including fungi, bacteria and plants. Commercially procured trehalose was applied to seeds of susceptible pearl millet (Pennisetum glaucum) cultivar HB3, and tested for its ability to reduce downy mildew disease incidence by induction of resistance. Seed treatment with trehalose at 200 mM for 9 h recorded 70.25% downy mildew disease protection, followed by those with100 mM and 50 mM trehalose which offered 64.35% and 52.55% defense, respectively, under greenhouse conditions. Furthermore, under field conditions treatment with 200 mM trehalose for 9 h recorded 67.25% downy mildew disease protection, and reduced the disease severity to 32.75% when compared with untreated control which displayed 90% of disease severity. Trehalose did not affect either sporangial formation or zoospore release from sporangia, indicating that the reduction in disease incidence was not due to direct inhibition but rather through induction of resistance responses in the host. Additionally, trehalose was shown to enhance the levels of polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL) and peroxidase (POX), which are known as markers of both biotic and abiotic stress responses. Our study shows that osmoregulators like trehalose could be used to protect plants against pathogen attacks by seed treatment, thus offering dual benefits of biotic and abiotic stress tolerance