Cultivating a greener future:Exploiting trichoderma derived secondary metabolites for fusarium wilt management in peas

This study aimed to identify efficient Trichoderma isolate(s) for the management of Fusarium wilt in peas. Four different pea germplasms (Sarsabz, Pea-09, Meteor and Supreme) were evaluated for resistance against Fusarium oxysporum in pot assay. Resistant germplasm exhibits a varying range of disease severity (23%) and percent disease index (21%), whereas susceptible and highly susceptible germplasm exhibit maximum disease severity (44–79%) and percent disease index (47–82%). The susceptible germplasm Meteor was selected for in vivo experiment. Five different Trichoderma spp. (Trichoderma koningii, T. hamatum, T. longibrachiatum, T. viride, and T. harzianum) were screened for the production of hydrolytic extracellular enzymes under in vitro. In-vitro biocontrol potential of Trichoderma spp. was assayed by percentage inhibition of dry mass of Fusarium oxysporum pisi (FOP) with Trichoderma spp. metabolite filtrate concentrations. Maximum growth inhibition was observed by T. harzianum (50–89%). T. harzianum metabolites in filtrate conc. (40%, 50%, and 60%) exhibited maximum reduction in biomass and were thus used for in vivo management of the disease. The pot experiment for in-vivo management also confirmed the maximum inhibition of FOP by T. harzianum metabolites filtrate at 60% by reducing disease parameters and enhancing growth, yield, and physiochemical and stress markers. Trichoderma strains led to an increase in chlorophyll and carotenoids (34-26%), Total phenolic 55%, Total protein content 60%, Total Flavonoid content 36%, and the increasing order of enzyme activities were as follows: CAT > POX > PPO > PAL in all treatments. These strains demonstrate excellent bio-control of Fusarium wilt in pea via induction of defense-related enzymes. The present work will help use Trichoderma species in disease management programme as an effective biocontrol agent against plant pathogens

Characterizing indigenous plant growth promoting bacteria and their synergistic effects with organic and chemical fertilizers on wheat (Triticum aestivum)

The excessive use of chemical fertilizers is deteriorating both the environment and soil, making it a big challenge faced by sustainable agriculture. To assist the efforts for the solution of this burning issue, nine different potential native strains of plant growth-promoting bacteria (PGPB) namely, SA-1(Bacillus subtilis), SA-5 (Stenotrophomonas humi),SA-7(Azospirillum brasilense), BH-1(Azospirillum oryzae), BH-7(Azotobacter armeniacus), BH-8(Rhizobium pusense), BA-3(Azospirillum zeae), BA-6(Rhizobium pusense), and BA-7(Pseudomonas fragi) were isolated that were characterized morphologically, biochemically and molecularly on the basis of 16S rRNA sequencing. Furthermore, the capability of indigenous PGPB in wheat (Triticum aestivum, Chakwal-50) under control, DAP+FYM, SA-1,5,7, BH-1,7,8, BA-3,6,7, DAP+ FYM + SA-1,5,7, DAP+FYM+ BH-1,7,8 and DAP+FYM+ BA-3,6,7 treatments was assessed in a randomized complete block design (RCBD). The results of the study showed that there was a significant increase in plant growth, nutrients, quality parameters, crop yield, and soil nutrients at three depths under SA-1,5,7, BH-1,7,8, and BA-3,6,7 in combination with DAP+FYM. Out of all these treatments, DAP+ FYM + BA-3,6,7 was found to be the most efficient for wheat growth having the highest 1000-grain weight of 55.1 g. The highest values for plant height, no. of grains/spike, spike length, shoot length, root length, shoot dry weight, root dry weight, 1000 grain weight, biological yield, and economic yield were found to be 90.7 cm, 87.7 cm, 7.20 cm, 53.5 cm, 33.5 cm, 4.87 g, 1.32 g, 55.1 g, 8209 kg/h, and 4572 kg/h, respectively, in the DAP+FYM+BA treatment. The DAP+FYM+BA treatment had the highest values of TN (1.68 µg/mL), P (0.38%), and K (1.33%). Likewise, the value of mean protein (10.5%), carbohydrate (75%), lipid (2.5%), and available P (4.68 ppm) was also highest in the DAP+FYM+BA combination. C:P was found to be significantly highest (20.7) in BA alone but was significantly lowest (11.9) in DAP+FYM+BA. Hence, the integration of strains BA-3, BA-5, and BA-7 in fertilizers can be regarded as the most suitable choice for agricultural growth in the sub-mountainous lower region of AJK. This could serve as the best choice for sustainable wheat growth and improved soil fertility with lesser impacts on the environment

Multi-locus analysis resolves the epidemic finch strain of Trichomonas gallinae and suggests introgression from divergent trichomonads

In Europe, Trichomonas gallinae recently emerged as a cause of epidemic disease in songbirds. A highly virulent and clonal strain of the parasite, first found in the UK, has become the predominant strain there and spread to continental Europe. Discriminating this epidemic strain of T. gallinae from other strains necessitated development of multi-locus sequence typing (MLST). Development of the MLST was facilitated by the assembly and annotation of a 54.7 Mb draft genome of a cloned stabilate of the A1 European finch epidemic strain (isolated from Greenfinch, Carduelis chloris, XT-1081/07 in 2007) containing 21,924 protein coding genes. This enabled construction of a robust 19 locus MLST based on existing typing loci for Trichomonas vaginalis and T. gallinae. Our MLST has the sensitivity to discriminate strains within existing genotypes confidently, and resolves the American finch A1 genotype from the epidemic European finch A1 genotype. Interestingly, one isolate we obtained from a captive black-naped fruit dove Ptilinopsus melanospilus, was not truly T. ¬¬¬gallinae but a hybrid of T. gallinae with a distant trichomonad lineage. Phylogenetic analysis of the individual loci in this fruit dove provides evidence of gene flow between distant trichomonad lineages at two of the 19 loci examined and may provide precedence for the emergence of other hybrid trichomonad genomes including T. vaginalis

Molecular detection and genetic characterization of Trichomonas gallinae in falcons in Saudi Arabia.

Avian trichomonosis is primarily caused by Trichomonas gallinae, a flagellated protozoan parasite that especially infects the upper digestive tract of columbid bird species and their avian predators. However, this parasite has recently been found to be distributed worldwide in various other avian species. This parasitic disease is common in captive falcons in Saudi Arabia and the Middle East. This study aimed to examine and identify the genetic variation of T. gallinae obtained from three species of falcons in Saudi Arabia via the sequencing analysis of the internal transcribed spacer (ITS) region. Swab samples from 97 saker falcons (Falco cherrug), 24 peregrine falcons (Falco peregrinus) and 37 gyrfalcons (Falco rusticolus) were cultured and analysed for infection between 2018 and 2019. The overall prevalence of infection by T. gallinae was 26.58% (n = 42), of which 35 (83.33%) were collected from Riyadh region and seven (16.67%) were collected from Qassim region. The results indicate the presence of four genotypes of T. gallinae in Saudi falcons: A, C, II, and KSA11. This study reports for the first time genetic diversity of T. gallinae in these falcons in Saudi Arabia

Ribosomal DNA Sequence-Based Taxonomy and Antimicrobial Activity of <i>Prorocentrum</i> spp. (Dinophyceae) from Mauritius Coastal Waters, South-West Indian Ocean

Microalgae are unicellular organisms and commonly present in the euphotic zone of marine ecosystems. From the western coast of Mauritius, three strains of Prorocentrum species were isolated from macrophytes and cultured under standard laboratory conditions. Morphologies were examined by light, fluorescence, and scanning electron microscopy, and phylogenetic analyses were based on partial large subunit LSU rDNA (D1-D2) and ITS1-5.8S-ITS2 (ITS) regions. Three Prorocentrum species, including the P. fukuyoi complex, P. rhathymum, and P. lima complex, were identified. The antimicrobial activities were assayed against potential human pathogenic bacterial strains. The highest zone of inhibition was recorded for intracellular and extracellular protein extracts of Prorocentrum rhathymum against Vibrio parahaemolyticus. The polysaccharide extracts of the Prorocentrum fukuyoi complex had a higher zone of inhibition (24 ± 0.4 mm) against MRSA at a minimum concentration of 0.625 μg/mL. The extracts from the three Prorocentrum species had different levels of activity against the pathogens used, and this can be of scientific interest in the search for antibiotics from natural marine sources

Elicitor-Driven Defense Mechanisms: Shielding Cotton Plants against the Onslaught of <i>Cotton Leaf Curl Multan Virus</i> (CLCuMuV) Disease

Salicylic acid (SA), benzothiadiazole (BTH), and methyl jasmonate (MeJA) are potential elicitors found in plants, playing a crucial role against various biotic and abiotic stresses. The systemic acquired resistance (SAR) mechanism was evaluated in cotton plants for the suppression of Cotton leaf curl Multan Virus (CLCuMuV) by the exogenous application of different elicitors. Seven different treatments of SA, MeJA, and BTH were applied exogenously at different concentrations and combinations. In response to elicitors treatment, enzymatic activities such as SOD, POD, CAT, PPO, PAL, β–1,3 glucanse, and chitinase as biochemical markers for resistance were determined from virus-inoculated and uninoculated cotton plants of susceptible and tolerant varieties, respectively. CLCuMuV was inoculated on cotton plants by whitefly (Bemesia tabaci biotype Asia II-1) and detected by PCR using specific primers for the coat protein region and the Cotton leaf curl betasatellite (CLCuMuBV)-associated component of CLCuMuV. The development of disease symptoms was observed and recorded on treated and control plants. The results revealed that BTH applied at a concentration of 1.1 mM appeared to be the most effective treatment for suppressing CLCuMuV disease in both varieties. The enzymatic activities in both varieties were not significantly different, and the disease was almost equally suppressed in BTH-treated cotton plants following virus inoculation. The beta satellite and coat protein regions of CLCuMuV were not detected by PCR in the cotton plants treated with BTH at either concentration. Among all elicitors, 1.1 mM BTH was proven to be the best option for inducing resistance after the onset of CLCuMuV infection and hence it could be part of the integrated disease management program against Cotton leaf curl virus

Role of mycorrhizas and root exudates in plant uptake of soil nutrients (calcium, iron, magnesium, and potassium): has the puzzle been completely solved?

Anthropogenic global change is driving an increase in the frequency and intensity of drought and flood events, along with associated imbalances and limitation of several soil nutrients. In the context of an increasing human population, these impacts represent a global-scale challenge for biodiversity conservation and sustainable crop production to ensure food security. Plants have evolved strategies to enhance uptake of soil nutrients under environmental stress conditions; for example, symbioses with fungi (mycorrhization) in the rhizosphere and the release of exudates from roots. Although crop cultivation is managed for the effects of limited availability of nitrogen (N) and phosphorus (P), there is increasing evidence for limitation of plant growth and fitness because of the low availability of other soil nutrients such as the metals potassium (K), calcium (Ca), magnesium (Mg), and iron (Fe), which may become increasingly limiting for plant productivity under global change. The roles of mycorrhizas and plant exudates on N and P uptake have been studied intensively; however, our understanding of the effects on metal nutrients is less clear and still inconsistent. Here, we review the literature on the role of mycorrhizas and root exudates in plant uptake of key nutrients (N, P, K, Ca, Mg, and Fe) in the context of potential nutrient deficiencies in crop and non-crop terrestrial ecosystems, and identify knowledge gaps for future research to improve nutrient-uptake capacity in food crop plants.info:eu-repo/semantics/acceptedVersio

Construction and Activity Testing of a Modular Fusion Peptide against <i>Enterococcus faecalis</i>

The emergence of antibiotic resistance in enterococci is a great concern encountered worldwide. Almost all enterococci exhibit significant levels of resistance to penicillin, ampicillin, semi-synthetic penicillin and most cephalosporins, primarily due to the expression of low-affinity penicillin-binding proteins. The development of new and novel antibacterial agents against enterococci is a significant need of the hour. In this research, we have constructed a modular peptide against Enterococcus faecalis. The enzymatic domain of the constructed peptide BP404 is from the bacteriocin BacL1 and the cell wall binding domain from endolysin PlyV12 of phage ϕ1. The protein BP404 was found to be active against two tested strains of Enterococcus faecalis, with a reduction in cell density amounting to 85% and 65%. The cell wall binding assay confirms the binding of the protein to Enterococcus faecalis, which was not seen towards the control strain Escherichia coli, invariably pointing to the specificity of BP404. To the best of our knowledge, this is one of the first instances of the development of a chimeric peptide against Enterococcus faecalis. This study points out that novel proteins can be genetically engineered against clinically relevant enterococci

An Enzybiotic Cocktail Effectively Disrupts Preformed Dual Biofilm of <i>Staphylococcus aureus</i> and <i>Enterococcus faecalis</i>

Multidrug-resistant bacterial infections are on the rise around the world. Chronic infections caused by these pathogens through biofilm mediation often complicate the situation. In natural settings, biofilms are often formed with different species of bacteria existing synergistically or antagonistically. Biofilms on diabetic foot ulcers are formed predominantly by two opportunistic pathogens, Staphylococcus aureus and Enterococcus faecalis. Bacteriophages and phage-based proteins, including endolysins, have been found to be active against biofilms. In this study, we evaluated the activity of two engineered enzybiotics either by themselves or as a combination against a dual biofilm formed by S. aureus and E. faecalis in an inert glass surface. An additive effect in rapidly disrupting the preformed dual biofilm was observed with the cocktail of proteins, in comparison with mono treatment. The cocktail-treated biofilms were dispersed by more than 90% within 3 h of treatment. Apart from biofilm disruption, bacterial cells embedded in the biofilm matrix were also effectively reduced by more than 90% within 3 h of treatment. This is the first instance where a cocktail of engineered enzybiotics has been effectively used to impede the structural integrity of a dual biofilm