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Arunmani, M.
- Rock Phosphate Solubilization using Oxalic Acid - secreting Laccaria fraterna
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Authors
Affiliations
1 Methodist Hospital Research Institute, Houston, TX 77030, US
2 Baylor College of Medicine, Houston, TX 77030,, US
3 Morgan State University, Baltimore, MD 21251,, US
1 Methodist Hospital Research Institute, Houston, TX 77030, US
2 Baylor College of Medicine, Houston, TX 77030,, US
3 Morgan State University, Baltimore, MD 21251,, US
Source
Indian Journal of Bioinformatics and Biotechnology, Vol 3, No 1 (2014), Pagination: 16–21Abstract
Laccaria fraterna is an ectomycorrhizal fungus commonly associated with conifers and angiosperms. To check whether L. fraterna possess mechanism to solubilize rock phosphate by secreting organic acids, it was grown in Pikovskaya’s medium with rock phosphate as the main ‘P’ source in the presence of ammonium sulphate. The fungus grew profusely in the medium by secreting oxalic acid which caused a reduction in the pH of the medium. To understand the mechanism behind the rock phosphate solubilization, an attempt was made to clone the fungal genomic DNA. In the Pikovskaya’s medium amended with rock phosphate, the clone MAP 22 released 80 μg/mL of P at the end of 24 h while, L. fraterna released 65 μg/mL of P at the end of 30 days incubation. We used HPTLC to confirm that oxalic acid is the main mineral-transforming agent that was secreted. The wild and the sub-clone MAP 22 released 1,490 and 2,950 μg/mL of oxalic acid, respectively. Therefore, it is possible to use the clone MAP 22 for more efficient inoculum production in vivo.Keywords
Ectomycorrhizal Fungus, Laccaria fraterna, Phosphate Solubilization, Rock PhosphateFull Text
References
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- Isolation and Regeneration of Protoplasts from Laccaria fraterna - an Ectomycorrhizal Fungus
Abstract Views :559 |
PDF Views:374
Authors
Affiliations
1 Methodist Hospital Research Institute, Houston, TX 77030, US
2 Baylor College of Medicine, Houston, TX 77030, US
3 Morgan State University, Baltimore, MD 21251, US
1 Methodist Hospital Research Institute, Houston, TX 77030, US
2 Baylor College of Medicine, Houston, TX 77030, US
3 Morgan State University, Baltimore, MD 21251, US
Source
Indian Journal of Bioinformatics and Biotechnology, Vol 3, No 1 (2014), Pagination: 22–30Abstract
An efficient method for high yields of protoplast isolation and regeneration and viability was achieved in Laccaria fraterna, an ectomycorrhizal fungus. In this study, we standardized the optimal conditions such as mycelial age, temperature, pH and osmotic stabilizers for the release and regeneration of protoplasts in L. fraterna. Maximum number of protoplasts (5.1 × 108) was isolated from 4 day old mycelia suspended in an osmotically stabilized MMC buffer (pH 5.0) with 0.5 M man- nitol and 10 mg/mL Novozyme 234. Maximum yields of protoplasts were released from the mycelium using Novozyme 234 after 3 h. Protoplasts exhibited two kinds of regeneration patterns in liquid media and one in solid medium. Almost all the protoplasts were nucleated and viable as observed by acridine orange and fluorescein diacetate staining. The regeneration frequency was as high as 36% under optimal conditions. When colonies from regenerated protoplasts were inoculated with Eucalyptus globulus, few plants showed ectomycorrhizal association. Results of this study indicate that this fungus could be potentially used in transformation, protoplast fusion and other genetic studies.Keywords
Ectomycorrhizal Fungus, Laccaria fraterna, Novozyme 234, Protoplasts, Regeneration PatternsFull Text
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