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

Antimicrobial activity of leaf extracts of Indian medicinal plants against clinical and phytopathogenic bacteria

The ethnobotanical efficacy of Indian medicinal plants; Achyranthes aspera, Artemisia parviflora, Azadirachta indica, Calotropis gigantean, Lawsonia inermis, Mimosa pudica, Ixora coccinea, Parthenium hysterophorus and Chromolaena odorata were examined using agar disc diffusion method against clinical bacteria (Escherichia coli and Staphylococcus aureus) and phytopathogenic bacteria (Xanthomonas vesicatoria and Ralstonia solanacearum). Leaves were extracted using different solvents such as methanol, ethanol, ethyl acetate and chloroform. Among treatments, maximum in vitro inhibition was scored in methanol extracts of C. odorata which offered inhibition zone of 10, 9, 12 and 12 mm against E. coli, S. aureus, X. vesicatoria and R. solanaccearum, respectively, followed by chloroform extract of the same plant leaf with inhibition zone of 8, 4, 4 and 4 mm, respectively. A significant inhibition of E. coli was found in aqueous and in all tested solvent extracts of A. indica. In case of S. aureus, maximum inhibition of 8 mm was obtained in aqueous extracts of A. indica and 6 mm from methanol extract of L. inermis. The minimum inhibitory concentration (MIC) value for the clinicalbacteria ranged between 0.35 to 4.0 mg/ml and 0.25 to 4.0 mg/ml for phytopathogenic bacteria when tested with all four solvents extracts of C. odorata. Whereas, extracts of A. aspera, A. parviflora, C. gigantean, L. inermis, M. pudica and I. coccinea were found to be ineffective or showed poor inhibition on tested human and phytopathogenic bacteria

Screening for plant growth promoting fungi and their ability for growth promotion and induction of resistance in pearl millet against downy mildew disease

A total of forty nine plant growth promoting fungi (PGPF) were successfully isolated from the rhizosphere of various grass species in Karnataka State, India. All the PGPF isolates were tested for their ability to enhance pearl millet seed quality parameters and to induce resistance against downy mildew disease in pearl millet. Susceptible pearl millet seeds 7042S were treated with PGPFs conidial suspension (1 x 108 cfu ml-1) and barley grain inocula (BGI) at 5%, 10% and 20% concentrations. Only six isolates among the forty nine tested recorded significant (P < 0.001) enhancement of seed germination and vigor when compared with the untreated control. Of the PGPF, Penicillium sp. (UOM PGPF 27) at 5% (w/w) concentration recorded highest seed germination of 92% and 1701.9 seedling vigor. The in planta colonization of the six PGPF isolates determined successfully in re-isolating the fungus from the basal root segments of 6 cm and 4 cm plated on PDA plates and also from the rhizosphere serial dilution of 10 3 to 10 5. Among the PGPFs tested in two modes, in BGI treatments, Penicillium sp. (UOM PGPF 27) at 5% (w/w) and Pythium sp. (UOM PGPF 41) at 10% (w/w) showed maximum disease protection of 67% and 61% respectively against downy mildew disease of pearl millet In case of conidial suspension treatments Penicillium sp. (UOM PGPF 27) and Trichoderma sp. (UOM PGPF 37) recorded highest disease protection of 71% and 66%, respectively under greenhouse conditions. Thus, the present study suggests that the tested PGPF, both as BGI inocula and conidial suspensions, can be used for pearl millet downy mildew disease management and also for plant growth. &nbsp

Beneficial microbes promote plant growth and induce systemic resistance in sunflower against downy mildew disease caused by Plasmopara halstedii

Seven plant growth promoting fungi (PGPF) native to the rhizosphere soil were screened for their potential to promote growth and to induce resistance in sunflower against Plasmopara halstedii, an incitant of sunflower downy mildew disease. Effect of seed priming with each of these seven PGPF isolates on seed quality parameters and resistance induction were studied under in vitro and green house conditions. Root colonization ability of these PGPF was also studied under green house conditions. The results showed that seed priming with conidial suspension of PGPF at 1 x 108 cfu ml-1 significantly increased seed germination and seedling vigor compared to non primed control. Plants raised with primed seeds demonstrated a significant reduction in downy mildew disease severity and provided a maximum of 61% protection under green house conditions when compared to the untreated control. The experimental results proved that the tested PGPF, promoted growth and induced systemic resistance (ISR) in sunflower plants against the downy mildew disease caused by P. halstedii, due to their growth promoting and biocontrol abilities. Present study has revealed the fact that there is a strong correlation between root colonization and resistance induction in PGPF treated plants

Context Matters: Intertextuality and Voice in the Early Modern English Controversy about Women

This dissertation examines three clusters of works from the early modern English controversy about women--the debate about the merits and flaws of womankind--in order to argue that authors in the controversy took advantage of the malleability of women's voices to address issues beyond the worth of women. I depart from standard treatments of the controversy by giving priority to the intertextual contexts among works that engage with one another. Attending to the intertextual elements of this genre reveals the metapoetic concerns of the authors and the way such authors fashion their feminine apologists as discursive agents in order to express those concerns. Chapter 1 examines Edward Gosynhyll's sixteenth-century works in tandem with Geoffrey Chaucer's The Legend of Good Women and "The Wife of Bath's Prologue and Tale," arguing that Gosynhyll's revisions of Chaucer--revisions embodied by the feminine apologists in the texts--are integral to his project of establishing the controversy genre as multivalent and dialectical. The resulting metacommentary examines in a new light the age-old rhetorical tradition of exemplarity, a persuasive tool used in diverse literary genres. Chapter 2 considers the way the anonymous play Swetnam the Woman-Hater uses cross-voicing and cross-dressing to establish the performative nature of controversy conventions. In doing so, the play argues for the social benefits of abandoning essentialist logic in favor of gender performance, as such performance makes the role of apologist available to men and women alike. This cluster reconsiders the very processes by which a person--male or female--can be known to others. Finally, I trace John Taylor's use of the marginal woman in his controversy works in order to demonstrate the extent to which Taylor makes these women instrumental in establishing his own poetic and social identity. This project contributes to studies on the English controversy as well as to the field of early modern women and women's writing by arguing that authors found the genre generally and the woman's voice specifically to be fit vehicles for articulating poetic agendas beyond the immediate task of debating the nature of womankind

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

Anti-oomycete compounds from ganoderma appalantum, a wood rot basidiomycete

Solvent extracts of 17 different basidiomycetes were tested for their ability to inhibit Sclerospora graminicola. Among those tested, only three basidiomycete crude extracts exhibited significant inhibition on pathogen sporulation, zoospore release and zoospore motility. In vitro, the chloroform extract of Ganoderma appalantum (3 mg mL-1) recorded a maximum sporangial inhibition of 52.5 and 82.0% zoospore release and 92.5% motility. Crude extracts of three different basidiomycete fungi were treated to pearl millet seeds and assessed for seed germination, seedling vigour and effectiveness against downy mildew disease under greenhouse conditions. None of the solvent extracts were found to be phytotoxic. Chloroform and petroleum ether extracts of G. appalantum (3 mg mL-1) proved to be the best by offering disease protection of 55.6 and 43.7%, respectively, followed by chloroform and petroleum ether extracts of Polyphorus spp., with 42.5 and 39.1% disease protection. The thin layer chromatography (TLC) spots that developed were eluted and tested for inhibition of S. graminicola zoosporangia. The partially purified compound from TLC chloroform extract of G. appalantum consistently showed good inhibitory effect against S. graminicola, which exhibited inhibition of sporangia (42.3%), zoospore release (76.7%) and zoospore motility (86.7%), compared to the chloroform control which offered only 1.9, 2.9 and 1.7% inhibition of sporangia, zoospore release and zoospore motility, respectively. The partially purified compound from petroleum ether extract of G. appalantum resulted in 38.0, 63.0 and 81.6% inhibition of sporangia, zoospore release and motility, respectively, compared to petroleum ether control. However, azoxystrobin 250 SC (2 g mL-1) and apron 35 SD (0.015 mg mL-1) treated on sporangial suspension showed the highest inhibition of S. graminicola pathogen compared to chloroform and petroleum ether TLC fractions

Anti-oomycete compounds from Ganoderma appalantum, a wood rot basidiomycete

Solvent extracts of 17 different basidiomycetes were tested for their ability to inhibit Sclerospora graminicola. Among those tested, only three basidiomycete crude extracts exhibited significant inhibition on pathogen sporulation, zoospore release and zoospore motility. In vitro, the chloroform extract of Ganoderma appalantum (3 mg mL<SUP>−1</SUP>) recorded a maximum sporangial inhibition of 52.5 and 82.0% zoospore release and 92.5% motility. Crude extracts of three different basidiomycete fungi were treated to pearl millet seeds and assessed for seed germination, seedling vigour and effectiveness against downy mildew disease under greenhouse conditions. None of the solvent extracts were found to be phytotoxic. Chloroform and petroleum ether extracts of G. appalantum (3 mg mL<SUP>−1</SUP>) proved to be the best by offering disease protection of 55.6 and 43.7%, respectively, followed by chloroform and petroleum ether extracts of Polyphorus spp., with 42.5 and 39.1% disease protection. The thin layer chromatography (TLC) spots that developed were eluted and tested for inhibition of S. graminicola zoosporangia. The partially purified compound from TLC chloroform extract of G. appalantum consistently showed good inhibitory effect against S. graminicola, which exhibited inhibition of sporangia (42.3%), zoospore release (76.7%) and zoospore motility (86.7%), compared to the chloroform control which offered only 1.9, 2.9 and 1.7% inhibition of sporangia, zoospore release and zoospore motility, respectively. The partially purified compound from petroleum ether extract of G. appalantum resulted in 38.0, 63.0 and 81.6% inhibition of sporangia, zoospore release and motility, respectively, compared to petroleum ether control. However, azoxystrobin 250 SC (2 μg mL<SUP>−1</SUP>) and apron 35 SD (0.015 mg mL<SUP>−1</SUP>) treated on sporangial suspension showed the highest inhibition of S. graminicola pathogen compared to chloroform and petroleum ether TLC fractions

Seed priming with plant gum biopolymers enhances efficacy of metalaxyl 35 SD against pearl millet downy mildew

`Priming' the plant and seed induces a physiological state in which plants are able to activate defense responses. Plant-based exudates are excellent gum biopolymers which contain plant growth-regulating hormones with priming potential without any side effects. In this study, gum exudates of Acacia arabica, Moringa oleifera, Carica papaya and Azadirachta indica were evaluated for synergistic effects of seed priming with exuded gum biopolymer combined with metalaxyl (Apron 35 SD) on pearl millet seed quality, growth parameters, and resistance to Sclerospora graminicola. Seeds of 7042S were primed with gum biopolymers and metalaxyl 35 SD and evaluated under laboratory and greenhouse conditions. Seed germination and vigor were synergistically enhanced using gum biopolymers solution (1:2 w/v) with 3 g kg(-1) metalaxyl 35 SD. A. arabica and A. indica gum biopolymers alone or with 3 g kg(-1) of metalaxyl 35 SD resulted in seed germination of > 91%. Seed priming with 6 g kg(-1) of metalaxyl 35 SD gave 89% seed germination and was not significantly different from control. A similar trend in vigor was observed among treatments. Seed priming with gum biopolymers alone provided varied disease protection levels when compared with the control. A. arabica or A. indica gum with 3 g kg(-1) of metalaxyl 35 SD was the superior treatment, offering significant 86% disease reduction while exhibiting a growth-promoting effect. Synergistic use of gum biopolymers and metalaxyl 35 SD by seed priming is highly effective in growth promotion and management of pearl millet downy mildew disease