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Effects of Medium Composition and Genetic Background on Agrobacterium-mediated Transformation Efficiency of Lentinula edodes

Summary

This research developed an improved method for genetically modifying shiitake mushrooms, making it easier to study and potentially enhance these commercially important fungi. The study found that both the growing medium and genetic makeup of different mushroom strains significantly affect how well they can be modified. Impacts on everyday life: • Could lead to improved shiitake mushroom varieties with better growth or nutritional properties • Advances our ability to study and understand how mushrooms grow and develop • May help develop more efficient production methods for commercial mushroom farming • Could contribute to developing mushrooms with enhanced medicinal properties • Provides tools for creating more disease-resistant mushroom strains

Background

Filamentous fungi, particularly basidiomycetes like Lentinula edodes (shiitake), play important roles in agriculture, industry, and medicine. While genome sequences of many filamentous fungi are now available, functional genomics research requires efficient genetic transformation methods. Existing transformation methods for L. edodes have shown insufficient efficiency to support large-scale functional genomics research.

Objective

To establish and optimize an efficient Agrobacterium-mediated transformation method for both monokaryon and dikaryon strains of L. edodes by evaluating the effects of medium composition and genetic background on transformation efficiency.

Results

The transformation efficiency varied significantly with medium composition and strain genotype. For dikaryon strain W1, 1/4 sawdust medium produced the most hygromycin-resistant colonies. For monokaryon strain W1-26, only millet medium yielded transformants with 75.48% positive frequency. Among different strains, YS3334 showed the highest transformation efficiency with 85.12% PCR positive frequency. Southern blot analysis confirmed successful T-DNA integration, mostly as single copies.

Conclusion

The study successfully established an efficient Agrobacterium-mediated transformation method for both monokaryon and dikaryon strains of L. edodes. Medium composition and genetic background significantly influenced transformation efficiency. The optimized protocol provides a practical approach for functional genomics research in L. edodes and potentially other filamentous fungi.
  • Published in:Genes (Basel),
  • Study Type:Laboratory Research,
  • Source: 10.3390/genes10060467
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