A novel strain exhibiting entomopathogenic and chitinolytic activity was isolated from mangrove marsh soil in India. The isolate was identified as Brevibacillus laterosporus by phenotypic characterization and 16S rRNA sequencing and designated Lak1210. When grown in the presence of colloidal chitin as the sole carbon source, the isolate produced extracellular chitinases. Chitinase activity was inhibited by allosamidin indicating that the enzymes belong to the family 18 chitinases. The chitinases were purified by ammonium sulfate precipitation followed by chitin affinity chromatography yielding chitinases and chitinase fragments with 90, 75, 70, 55, 45, and 25 kDa masses. Mass spectrometric analyses of tryptic fragments showed that these fragments belong to two distinct chitinases that are almost identical to two putative chitinases, a 89.6-kDa four-domain chitodextrinase and a 69.4-kDa two-domain enzyme called ChiA1, that are encoded on the recently sequenced genome of B. laterosporus LMG15441. The chitinase mixture showed two pH optima, at 6.0 and 8.0, and an optimum temperature of 70 °C. The enzymes exhibited antifungal activity against the phytopathogenic fungus Fusarium equiseti. Insect toxicity bioassays with larvae of diamondback moths (Plutella xylostella), showed that addition of chitinases reduced the time to reach 50 % mortality upon infection with non-induced B. laterosporus from 3.3 to 2.1 days. This study provides evidence for the presence of inducible, extracellular chitinolytic enzymes in B. laterosporus that contribute to the strain's antifungal activity and insecticidal activity.
Cysteine-rich peptides such as metallothioneins (MTs) are involved in metal homeostasis and detoxification in many eukaryotes. We report the characterization and expression of two MT genes, LbMT1 and LbMT2 from the ectomycorrhizal fungus Laccaria bicolor under metal stress conditions. LbMT1 and LbMT2 differ with respect to the length of the encoded peptides (58 versus 37 aa, respectively) and also by their expression patterns in response to metals. The expression levels of both LbMT1 and LbMT2 increased as a function of increased external Cu concentration, the expression levels for LbMT2 were always significantly higher compared with those of LbMT1. Only LbMT1, but not LbMT2, responded to Cd supply in the range of 25-100 mM while Zn did not affect the transcription of either LbMT1 or LbMT2. Both genes also responded to oxidative stress, but to a lesser extent compared to their responses to either Cu or Cd stress. Heterologous complementation assays in metal-sensitive yeast mutants indicated that both LbMT1 and LbMT2 encode peptides capable of conferring higher tolerance to both Cu and Cd. The present study identified LbMTs as potential determinants of the response of this mycorrhizal fungus to Cu and Cd stress.
The concentration of heavy metals such as Ba, Co, Cr, Cu, Ni, Pb, Rb, Sr, V, Y, Zn, Zr were studied in soils of Balanagar industrial area, Hyderabad to understand heavy metal contamination due to industrialization and urbanization. This area is affected by the industrial activities like steel, petrochemicals, automobiles, refineries, and battery manufacturing generating hazardous wastes. The assessment of the contamination of the soils was based on the geoaccumulation index, enrichment factor (EF), contamination factor, and degree of contamination. Soil samples were collected from Balanagar industrial area from top 10-50 cm layer of soil. The samples were analyzed using X-ray fluorescence spectrometer for heavy metals. The data revealed that the soils in the study area are significantly contaminated, showing high level of toxic elements than normal distribution. The ranges of concentration of Cr (82.2-2,264 mg/kg), Cu (31.3-1,040 mg/kg), Ni (34.3-289.4 mg/kg), Pb (57.5-1,274 mg/kg), Zn (67.5-5819.5 mg/kg), Co (8.6-54.8 mg/kg), and V (66.6-297 mg/kg). The concentration of abovementioned other elements was similar to the levels in the earth's crust pointed to metal depletion in the soil as the EF was \1. Some heavy metals showed high EF in the soil samples indicating that there is a considerable heavy metal pollution, which could be correlated with the industries in the area. A contamination site poses significant environmental hazards for terrestrial and aquatic ecosystems. They are important sources of pollution and may results in ecotoxicological effects on terrestrial, groundwater and aquatic ecosystems.
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