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Different temporal and spatial expression of two hydrophobin-encoding genes of the edible mushroom Agaricus bisporus

Piet W. J. De Groot^1,

¹ Section Molecular Genetics of Industrial Micro-organisms, Wageningen Agricultural University, Dreijenlaan 2, NL-6703 HA, Wageningen, The Netherlands
² Mushroom Experimental Station, PO Box 6042, NL-5960 AA, Horst, The Netherlands

ABSTRACT

In a search for genes that are only expressed in fruit bodies of the basidiomycete Agaricus bisporus, two cDNAs, hypA and hypB that encode hydrophobins have been isolated previously. In this study, the structure of hypB is resolved and it is shown that the two genes are differentially expressed, indicating that the encoded hydrophobins serve different functions in A. bisporus mushrooms. hypB encodes a polypeptide (HYPB) of 119 aa that shows little sequence identity with HYPA apart from the characteristic arrangement of eight cysteines found exclusively in hydrophobins. The temporal and spatial expression of the two hydrophobin-encoding genes during fruit body development was compared using Northern analysis and in situ hybridization. Accumulation of hypA mRNA was found in tissue fractions consisting of undifferentiated white hyphae. In situ hybridization showed that the highest hypA mRNA levels are not found in the outermost cell layers of the pileipellis but in the cell layers adjacent to that. The highest level of expression of hypB occurs early in development when the primordium differentiates into densely packed, randomly oriented cap hyphae and loosely packed, vertically oriented stipe hyphae. In mature mushrooms, a strong accumulation of hypB transcripts was found only in the transitional zone between cap and stipe tissue, demonstrating that transcription regulation of hypB is clearly distinct from hypA.

Author for correspondence: Peter Schaap. Tel: +31 317 485142. Fax: +31 317 484011. e-mail: Peter.Schaap@algemeen.mgim.wau.nl

Present address: Institute for Molecular Cell Biology, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands.

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