Unraveling the mystery of commercial cultivation of Agaricus bisporus: plant biomass utilization and its effect on mushroom production

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1 Unraveling the mystery of commercial cultivation of Agaricus bisporus: plant biomass utilization and its effect on mushroom production Supplementary data Chapter 5 Uncovering the abilities of Agaricus bisporus to degrade plant biomass throughout its life cycle Table S2 Figures S and S2

2 Table S5. Primers used for qrt-pcr. Table S2 Primers used for qrt-pcr. Gene ID Putative function Primers (5' - 3') 9269 endo-β-,4-mannanase 942 endo-β-,4-glucanase 939 cellobiohydrolase / endoglucanase 9452 cellobiohydrolase / chitosanase 3354 endo-β-,4-xylanase 968 endo-β-,4-xylanase 2649 α-glucuronidase xyloglucan endo- β-,4-glucanase / endoβ-,4-glucanase α-galactosidase / α-nacetylgalactosaminidase β-,4-xylosidase / α-arabinofuranosidase / endoarabinanase β-,4-xylosidase / α-arabinofuranosidase / endoarabinanase α-arabinofuranosidase lytic polysaccharide monooxygenase 2467 xyloglucan endo-β-,4-glucanase laccase manganese peroxidase 9928 Glucose-Methanol-Choline oxidoreductase 939 glyoxal oxidase 3677 acetylxylan esterase β-xylosidase 3863 glyceraldehyde-3-phosphate dehydrogenase F: TTTCTACACAAACACCGCAATCA R: ACTCACGAAGACCCGGACAT F: TCTGACGGTTCCGGCACTA R: GGAGTCGTTCGCCACCAAT F: GCTCGTCCCAGGCTTTGTC R: GCCATTACCACCGCATTGA F: GGCGTAACTTGTGCCGAGAA R: ACCGTATGTGCCTGCGTAGTC F: TCAAGCGCCAGAATGCAA R: AAGATGCATTTGTGTACCGATACC F: TCAACGTCGGCAACCACTT R: TGTGCGAACCCAACCTAAGAC F: CGGTTATGGTTGGAATGCTACTC R: CCAAGCAGAGACGAAATCGTT F: TCCTCTTACTCGCTCCATTTGTT R: CCCTGGCGCTGTTCGA F: GCCGCCTGGGCTTTCT R: TTGCGAAAGATCGGTTCCA F: GTCACCGCTCTTTCTCAACGT R: AAAATGACGCTGGCTTCGA F: TGGACTCCGTTCCAGACGAT R: CAAAACCGTCCCATGCCTTA F: CGGGTCCCCTGCCAAT R: GCCTCCCCAGCAGCACTT F: TCGCCGCTCTTGCATTTC R: GATGAGGTTGTTCCAGATGTAGTGA F: CGACGAGTTCGGGTAATGGA R: ACCTTGACCACCGCATTGAC F: AGACAGGAACCCCAATCTAAACAT R: CACCATGGTAGCGCAGGAT F: GCTGGTTTTACCCCCACTGAA R: CCGAAGCTGCGACAGTATGA F: TTCTTGCCAGCCGTCTTTCT R: GGGCCTGCTTCAATAAGTAAGACT F: TGGTGCGCAGTCAGGTTATG R: TTGCCGACCGGATCTTTC F: TCGGTGCCCGTGAAACTAC R: TTGACGACGGGTCCTGATGT F: TGCGTCCGACTTTAGCCATT R: TGGATGGAATAGGCAGTGTTGT F: TCGATCTTGTTGTTCGTCTTGAG R: GCGCATGACCTCCTTGATTT Optimized primer concentration (nm)

3 Figure S Expression patterns and validation of RNA seq analysis by of twenty selected genes involved in plant biomass degradation by A.bisporus. Columns and bars represent the means and standard errors of. Lines represent value (GH5_5 β-,4-endoglucanase-cbm) st 2nd (AA9 lytic polysaccharide monooxygenase) st 2nd (GH6 cellobiohydrolase-cbm) 9452 (GH7 cellobiohydrolase-cbm) st 2nd st 2nd (GH2 β-,4-endoglucanase / xyloglucan β-,4-endoglucanase) 9269 (GH5_7 β-,4-endomannanase-cbm) st 2nd st 2nd (GH β-,4-endoxylanase-cbm) 968 (GH β-,4-endoxylanase) st 2nd st 2nd 3

4 28425 (GH43 α-arabinofuranosidase) (GH43 α-arabinofuranosidase) st 2nd st 2nd (GH5 α-glucuronidase) (GH5 α-arabinofuranosidase) st 2nd st 2nd (GH27 α-galactosidase) 2467 (GH74 xyloglucan β-,4-endoglucanase -CBM) st 2nd st 2nd (AA_ laccase) (AA2 manganese peroxidase) st 2nd st 2nd

5 9928 (AA3_2 Glucose-Methanol-Choline oxidoreductase) 939 (AA5_ glyoxal oxidase) st 2nd st 2nd (CE5 acetylxylan esterase -CBM) (GH5_22 β-xylosidase) st 2nd st 2nd 5

6 Figure S2 Persentage distribution of KOG functional classification related genes. Distribution of the genes which are highly expressed ( > 3) during the growth of A. bisporus in compost (A); are diffrentially expressed (all the growth s compared to spawning day 6): upregulated (B) and downregulated (C). A After 2 nd 2nd 2 nd After st st % % 2% 3% 4% 5% 6% 7% 8% 9% % B % 2% 4% 6% 8% % CELLULAR PROCESSES AND SIGNALING INFORMATION STORAGE AND PROCESSING METABOLISM POORLY CHARACTERIZED No KOG After 2 nd / 2 nd / After st / st / / After 2 nd / C % 2% 4% 6% 8% % 2 nd / After st / st / / % 2% 4% 6% 8% % 6

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