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Identification of growth phenotype-related genes in Aspergillus oryzae by heterologous macroarray and suppression subtractive hybridization

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Author: Biesebeke, R. te · Levin, A. · Sagt, C. · Bartels, J. · Goosen, T. · Ram, A. · Hondel, C. van den · Punt, P.
Institution: TNO Kwaliteit van Leven
Source:Molecular Genetics and Genomics, 1, 273, 33-42
Identifier: 238360
doi: doi:10.1007/s00438-004-1082-9
Keywords: Biology · Biotechnology · Aspergillus · Heterologous hybridization · Macroarray hybridization · Morphology · Polarized growth · complementary DNA · RNA · article · Aspergillus niger · Aspergillus oryzae · biosynthesis · cell wall · controlled study · DNA isolation · DNA library · DNA probe · fungus growth · gene expression · genetic transcription · liquid culture · molecular cloning · morphogenesis · morphological trait · nonhuman · Northern blotting · nucleotide sequence · phenotype · priority journal · protein expression · seed kernel · suppression subtractive hybridization · wheat · Aspergillus oryzae · Base Sequence · Blotting, Northern · Computational Biology · DNA Primers · Gene Expression Regulation, Fungal · Gene Library · Genes, Fungal · Molecular Sequence Data · Nucleic Acid Hybridization · Phenotype · Sequence Analysis, DNA · Triticum · Aspergillus · Aspergillus niger · Aspergillus oryzae · Triticum aestivum


Aspergillus oryzae requires polarized growth for colonization of solid substrates, and this growth phenotype differs from that seen in liquid medium. Various experimental approaches were used to identify genes that are differentially expressed when A. oryzae is grown on wheat kernels and in a wheat-based liquid medium. Hybridization of A. oryzae RNAs to a macroarray bearing cDNAs isolated from a library representing at least 16% of the total number of A. niger genes identified 14 differentially expressed cDNA clones, showing that heterologous macroarray analysis with an A. niger cDNA library can be used to identify regulated gene transcripts in the related species A. oryzae. Moreover, Northern analysis with a selection of eight probes for A. niger genes encoding proteins involved in morphological development and cell wall biosynthesis identified five more differentially expressed genes. A suppression subtractive hybridization procedure revealed another 12 differentially expressed genes. The results presented show that, of the 29 identified genes which are expressed at higher levels during growth on wheat kernels, six encode proteins that are functionally related to polarized growth, four encode products known to be involved in morphogenesis, three code for proteins related to cell wall composition, and nine of the cDNA clones encode novel proteins. These findings pinpoint genes associated with the changes in cellular morphogenesis seen in A. oryzae grown on wheat kernels as opposed to wheat-based liquid medium. © Springer-Verlag 2005.