A yellow Gram-stain-positive, non-motile, non-endospore -forming, spherical endophytic actinobacterium, designated strain AE-6T, was isolated from the inner fleshy leaf tissues of Aloe barbadensis (Aloe vera) collected from Pune, Maharashtra, India. Strain AE-6T grew at high salt concentrations [10 % (w/v) NaCl], temperatures of 15–41 °C and a pH range of 5–12. It showed highest (99.7 %) 16S rRNA gene sequence similarity with Micrococcus yunnanensis YIM 65004T followed by Micrococcus luteus NCTC 2665T (99.6 %) and Micrococcus endophyticus YIM 56238T (99.0 %). Ribosomal protein profiling by MALDI-TOF/MS also showed it was most closely related to M. yunnanensis YIM 65004T and M. luteus NCTC 2665T. Like other members of the genus Micrococcus , strain AE-6T had a high content of branched chain fatty acids (iso-C15 : 0 and anteiso-C15 : 0). MK-8(H2) and MK-8 were the predominant isoprenoid quinones. Cell wall analysis showed an ‘A2 l-Lys-peptide subunit’ type of peptidoglycan and ribose to be the major cell wall sugar. The DNA G+C content was 70 mol%. Results of DNA–DNA hybridization of AE-6T with its closest relatives from the genus Micrococcus produced a value of less than 70%. Based on the results of this study, strain AE-6T could be clearly differentiated from other members of the genus Micrococcus . We propose that it represents a novel species of the genus Micrococcus and suggest the name Micrococcus aloeverae sp. nov., with strain AE-6T ( = MCC 2184T = DSM 27472T) as the type strain of the species.
An endophytic species of was isolated from leaf (syn. ) and screened for protease production with five other species of. Data indicated that endophytic AE-6 MCC 2184 and DSM 21948 showed efficient protease production potential and secreted active protease at high salt (10%), temperature (40 °C) and in wide range of pH 8-10. Unlike . DSM 21948, protease production by . AE-6 MCC 2184 was stringently controlled by pH. Protease induction study using different group of peptides, peptide carbohydrates and peptide macronutrient combinations showed variable response with both the organisms. Result indicated that the amount of protease was not directly related to cell biomass but it depends on nature of inducible peptides. In this study we also developed a modified agar-well assay for semi-quantitative data from large number of replicates.
A freshwater dwelling cyanobacterium (strain MKW3) was isolated from a sample collected from a water logged sugarcane field located in Malkapur, Karad, Maharashtra, India, and was characterized using a polyphasic approach. In the 16S rRNA gene phylogenetic analysis, strain MKW3 clustered with two misidentified strains—Nostoc sp. CENA239 and Calothrix sp. NIES2100. The phylogenetically related members included strains identified as Nostoc, Aulosira, Calothrix, Tolypothrix, Camptylonemopsis and Microchaete. The phylogenetic and the morphological analysis of the strain MKW3 indicated that it does not belong to any of the above mentioned genera. Furthermore, the 16S-23S ITS secondary structure analysis provided clear evidence indicating that strain MKW3 is different from Nostoc sp. CENA239 and Calothrix sp. NIES2100. Based on the morphological, phylogenetic and 16S-23S ITS secondary structure analysis we describe our strain as Constrictifilum karadense gen. et sp. nov. in accordance with the International Code of Nomenclature for algae, fungi and plants.
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