Plant growth-promoting fungi play an important role in development of sustainable agriculture. In the current study, 13 fungal strains were isolated from the rhizosphere of healthy (wheat) plant and screened for their indolic auxin production potential. strain PGFW, strain BFW and strain DGFW were amongst the most efficient indolic auxin-producing strains. Indolic auxins such as indole 3 acetate (IAA), indole 3 butyrate (IBA) and indole 3 propionate (IPA) are produced by fungi. The conventional method to assess the IAA production is through a spectrophotometric assay using Salkowski's reagent, which quantifies all indolic auxins and not individual auxins. Moreover, it was also observed that the absorption maxima () of the samples, when compared to that of standard indole-3-acetic acid, showed deviation from the latter, indicative of production of a mixture of indolic derivatives by the fungi. Hence, for further profiling of these indolic compounds, high-performance thin layer chromatography (HPTLC) based protocol was standardized to precisely detect and quantify individual indolic auxins like IAA, IBA and IPA in the range of 100-1000 ng per spot. HPTLC analysis also showed that the fungal strains produce different indolic auxins in media with and without fortification of tryptophan, with the production of indolic auxins being enhanced in presence of tryptophan. Thus, this standardized HPTLC protocol is an efficient and sensitive methodology to separate and quantify the indolic derivatives.
Since December 2019, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been spreading worldwide, triggering one of the most challenging pandemics in human population. In the light of reporting of this virus in domestic and wild animals from several parts of world, a systematic surveillance study was conceptualized to detect the SARS-CoV-2 among species of Veterinary importance. Nasal and/or rectal samples of 413 animals (Dog=195, cattle=64, horse=42, goat=41, buffalo=39, sheep=19, cat=6, camel=6 and monkey=1) were collected from different places of Gujarat state of India. RNA was extracted from samples and subjected to RT-qPCR based amplification of target sequences in viral nucleoprotein (N), spike (S) and ORF1ab genes. A total of 95 (23.79 %) animals were found positive, comprised of 67 (34.35 %) dogs, 15(23.43 %) cattle and 13(33.33 %) buffaloes. Overall, nasal samples (N=80/412, 19.41 %) gave more positive results than rectal samples (N=70/407, 17.19 %) in RT-qPCR. The whole SRAS-CoV-2 genome sequencing was done on one sample (ID-A4N; from dog) where 32 mutations, including 29 single nucleotide variation (SNV) and two deletions, were detected. Among them, 9 mutations were located in the receptor binding domain of the spike (S) protein. The consequent changes in amino acid sequence revealed that T19R, G142D, E156-, F157-, A222V, L452R, T478K, D614G, P681R mutation in S protein and D63G, R203M and D377Y in N protein. The lineage assigned to this SARS-CoV-1 sequence is B.1.617.2. Thus, the present study highlights the importance of SARS-CoV-2 surveillance in non-human host.
Hot springs are of great importance due to their unique physicochemical properties. Due to unique selection pressure in this habitat, a diverse microbial community is prevailing and can be analyzed by high throughput sequencing technology. Present study focuses on metagenomic sequencing of two hot springs from Gujarat, India namely Tuwa and Unnai through both, culturable and culture independent approach. Sequence analysis from both the water reservoirs depicted higher species richness and diversity based on various diversity indices. The microbial community structure at both the hot springs was distinct and dependent on physicochemical factors like temperature, pH, mineral content etc. Enrichment by cultivation before metagenome sequencing revealed the abundance of Firmicutes (up to 96%) representing cultivable organisms in hotsprings. The bacterial phyla Firmicutes, Proteobacteria, Bacteroidetes, Thermotogae, Deinococcus-Thermus, and Chloroflexi dominate the thermoalkaline spring at Unnai and Tuwa in different proportion. Economically important microorganisms belonging to genera Thermus, Brevibacillus, Anoxybacillus, Bacillus, Pseudomonas, and Geobacillus were prevalent in hot springs. The analysis of functional potential by KEGG revealed pathways for metabolism of carbohydrates, amino acids, vitamins, cofactors and xenobiotics. Annotation with Carbohydrate Active EnZymes (CAZy) revealed the presence of four major classes of enzymes: glycosyl transferase, glycoside hydrolase, polysaccharide lyase and carbohydrate-binding modules. The study provides insight into the microbial community structure and their untapped functional potential for various biotechnological and environmental applications.
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