Abstract
Background: Production and cultivation of edible mushrooms are one of the commercial applications of microbial technologies in order to biodegrad agricultural waste into valuable food products. One of the edible mushrooms that uses these agricultural wastes for its growth and development is Pleurotus florida. In P. florida, various enzymes are responsible for the decomposition of lignin compounds in the compost environment from the beginning of mycelial growth to the end of the fruiting period. One of the most important of these enzymes is manganese peroxidase (MnP), which plays a major role in the breakdown of lignin compounds. In addition, the most important compost-degrading mushroom that produces the enzyme MnP is Agaricus bisporus. Objectives: The aim of this study was to produce hybrids of P. florida by increasing the production efficiency through hyphae fusion of P. florida and the A. bisporus. Methods: The hyphae fusion and hybridization were performed for two strains P. florida and A. bisporus, in PDA culture media. Samples were isolated from the fusion areas and were transferred to the culture media, and then spawn was produced. Performance testing was carried out with hybrids and P. florida. We designed a specific primer pair of P. florida MnP gene by Primer Premier software (V.7.0). Polymerase chain reaction (PCR) was used to confirm the MnP gene. For this purpose, RNAs were extracted, and their cDNA was synthesized by reverse transcriptase enzyme. PCR with specific primers was performed, and electrophoresis was loaded on 1% agarose gel. Results: This study performed hybridization with formation clamp connections at the junction line. Hybrid N09 strain with biological efficiency (82.92) produced the highest performance in all evaluations compared to other hybrids and the parent strains. Based on the results of PCR product electrophoresis, banding pattern N09 (hybrid N09 strain) indicated that this hybrid, in addition to the P. florida MnP gene of P. florida, also received the amino acid sequences of this gene in A. bisporus. Conclusions: According to the obtained results, the use of hyphae fusion increased the traits as a result of mixing the genetic contributions of parent strains, followed by the diversity of hybrid strains and increased production yield.