Establishment of uracil auxotrophic dikaryotic strains of Lentinula edodes by crossbreeding

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  • Zhou Chenli
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Xi Liping
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Mao Wenjun
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Wan Jianing
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Li Yan
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Wang Ying
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi
  • Bao Dapeng
    Laboratory of Edible Fungi Genetics, National Engineering Research Center of Edible Fungi; Key Laboratory of Applied Mycological Resources and Utilization, Ministry of Agriculture; Shanghai Key Laboratory of Agricultural Genetics and Breeding; Institute of Edible Fungi

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  • Establishment of uracil auxotrophic dikaryotic strains of <i>Lentinula edodes</i> by crossbreeding

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<p>The uracil auxotrophic monokaryotic strain 423-9 of Lentinula edodes was crossed with nine monokaryons (cro2-2-9, W66-1, xd2-3-2, QingKe 20A, 241-1-1, 9015-1, L66-2, 241-1-2, and Qing 23A) derived from wild type strains of L. edodes. Nine dikaryotic hybrids were established from these crosses. These hybrids were fruited and 496 single spore isolates were obtained. Among these single spore isolates, 166 were identified as monokaryons under a microscope. We screened these monokaryons on selective medium and obtained 19 uracil auxotrophic monokaryons. By using the Monkaryon-monkaryon crossing method among the uracil auxotrophic monokaryons, 56 uracil auxotrophic dikaryotic strains were established on selective medium. These dikaryotic strains were unable to grow on minimal medium without uracil and exhibited slow growth rates on PDA plates compared to the wild type strain. The uracil auxotrophic dikaryotic strains also showed more vigorous growth on sawdust cultivation medium containing uracil than that without uracil. The fruiting tests showed that they formed normal fruiting bodies on the sawdust medium containing uracil. The results show that the uracil auxotrophic dikaryotic strain of L. edodes could be produced by mating, and will provide a valuable resource for future genetic studies and for spawn protection and identification.</p>

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