Isolation, Regeneration and PEG-Induced Fusion of Protoplasts of Pleurotus pulmonarius and Pleurotus florida

Author: mycolabadmin
2006-06-30
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Summary

This research focused on developing improved strains of oyster mushrooms through a cellular fusion technique. Scientists successfully combined genetic material from two different mushroom species to create a hybrid strain with beneficial characteristics from both parents. This work has important implications for agriculture and food production. Impacts on everyday life: • Could lead to more productive mushroom crops for farmers • May result in more resilient mushroom varieties that are easier to grow • Could help develop mushroom strains with enhanced nutritional properties • Demonstrates sustainable ways to improve food production • Could lead to more affordable mushroom products for consumers

Background

Protoplast fusion technology enables the development of inter-specific, intra-specific, and inter/intra generic hybrids with enhanced capabilities compared to parent strains. This technique has potential applications in improving strains for antibiotic production, enzyme production, and mushroom yields. While oyster mushrooms are commonly cultivated in Tamil Nadu, India due to their simple cultivation requirements, strain improvement through protoplast fusion remains underdeveloped as a technique.

Objective

To standardize optimal conditions for protoplast release and regeneration in Pleurotus florida and Pleurotus pulmonarius, and to isolate a fusion product through PEG-induced protoplast fusion that exhibits desirable characteristics from both parent strains.

Results

The optimal conditions for maximum protoplast yield included using a mixture of cellulase, chitinase and pectinase enzymes, 0.6M KCl as osmotic stabilizer, pH 6.0, and 3-hour incubation of 3-day-old mycelia. This yielded 5.3-5.75 × 10⁷ protoplasts/g for P. florida and 5.6-6 × 10⁷ protoplasts/g for P. pulmonarius. Regeneration efficiency was 3.3% for P. florida and 4.1% for P. pulmonarius. PEG-induced fusion resulted in 0.28% fusion frequency.

Conclusion

The study successfully developed a protocol for protoplast isolation, regeneration, and fusion between P. pulmonarius and P. florida. The stable fusant strain exhibited characteristics of both parents, including carbendazim tolerance and vanillin utilization ability, though it required lower temperatures for fruiting than its parents.

Published in:Mycobiology,
Study Type:Laboratory Research,
Source: 10.4489/MYCO.2006.34.2.073