Transcriptome of different fruiting stages in the cultivated mushroom Cyclocybe aegerita suggests a complex regulation of fruiting and reveals enzymes putatively involved in fungal oxylipin biosynthesis

- Transcriptome of different fruiting stages in the cultivated mushroom Cyclocybe.
- To elucidate the so far barely understood biosynthesis of fungal volatiles, alterations in the transcriptome during different developmental stages of C.
- aegerita were analyzed and combined with changes in the volatile profile during its different fruiting stages..
- Combining transcriptome and volatilome data, enzymes putatively involved in the biosynthesis of C8 oxylipins in C.
- Furthermore, we were able to localize the mycelium as the main source for sesquiterpenes predominant during sporulation in the headspace of C.
- In contrast, changes in the C8 profile detected in late stages of development are probably due to the activity of enzymes located in the fruiting bodies..
- The formation of fruiting bodies (FBs, basidiomes, basi- diocarps) that are in particular formed by species from the Basidiomycota class Agaricomycetes [1] is one of the most complex developmental processes in the fungal life cycle.
- The branches in the hyphal knots intertwine successively to initials, being for e.g.
- Usually, as in the model agaric Cyclocybe aegerita (V.
- with G-protein coupled receptors being involved in the signal transduction [18].
- Recently, the changes of the volatilomes in the head- space (HS) of Cyclocybe aegerita (syn.
- This study revealed drastic changes in the volatile profile across developmental stages.
- In early stages, alcohols and ketones, including oct-1-en-3-ol and cyclopenta- none, were the main substances in the HS of the dikar- yon.
- cubebene and δ-cadinene, were the dominant substances detected in the HS at this stage.
- aegerita AAE-3, chosen to be similar with stages sam- pled in the volatilome study of C.
- Combining the volatilome and trancriptome data sets and comparing volatile profiles in the HS of C..
- aegerita with transcription patterns of selected genes, we were able to identify enzymes putatively involved in the formation of VOCs in fungi.
- To identify changes in the transcriptome during mush- room tissue formation and FB maturation in C.
- Thus, we focus on the FRGs as well as on genes that are involved in the biosynthesis of volatile compounds mainly produced during fructification and sporulation..
- In the genome sequence of C.
- In total, four larger cohorts can be distinguished from another: two in the mycelium and two in the mushroom tissue.
- aegerita AAE-3, despite its sub-threshold expression values in the RNA-seq ana- lysis above.
- Genes coding for enzymes of the mevalonate path- way were identified in the C.
- Generally, the maximum transcription levels of the STSs were noticeably lower in the examined FB stages than in the mycelial samples, never exceeding 50 NRC in the FB samples.
- When comparing the occurrence of Δ 6 -protoil- ludene, the most dominant VOC in the HS of C.
- 4), with the gene expression values of the two known Δ 6 - protoilludene synthases Agr6 and Agr7 [59], the tran- scription pattern of AGR6 (AAE3_04120) in the myce- lium perfectly reflects the occurrence of Δ 6 - protoilludene.
- In contrast, AGR7 (AAE3_10454) showed the highest transcription levels in the mycelium after sporulation at day 28 p.i.
- aegerita under the applied ex- perimental conditions, neither in the mycelium nor in the FB samples (Additional file 4: Figure S6)..
- Interestingly, the composition of the three volatile oxylipins detected in the HS of C..
- aeger- ita were noticeably higher in FBs than in the mycelium (Fig.
- In contrast, in the mycelium the tran- scription of LOX4 (Fig.
- aegerita as well as the relative concentrations of the corresponding sesquiterpenes (red) in the HS of C.
- in the mycelium revealing a 10-fold higher maximum expression at day 10 p.i.
- Besides the activity of LOXs on linoleic acid, DOXs might also play a crucial role in the formation of fungal volatile oxylipins [49] (Additional file 6: Figure S8).
- aegerita as well as the relative concentrations of volatile oxylipins (red) in the HS of C.
- putative DOXs genes were found in the genome of C..
- Interestingly, the transcription of the putative DOX gene AAE3_00407 was upregulated in the mycelium of early fruiting stages (2-fold change), whereas the transcription in the FBs remained comparably low (Fig.
- Interestingly, in FB stages as well as in the mycelium, the course of expression of AAE3_09203 was comparable to the transcription of the putative DOX AAE3_13098.
- In contrast, AAE3_06380 revealed quite constant transcription levels (about 1000 NRC) in the mycelium as well as in FB stages..
- In this context, ADHs and ene-reductases might play an im- portant role, explaining the observed decrease of oct-1- en-3-ol during sporulation and, thereafter, the increase of octan-3-one in the HS of C.
- Putative ADHs and ene-reductases were identified in the genome of C.
- The transcription levels of putative ADHs and ene-reductases were analyzed in the mycelium and in FB stages (Additional file 6: Figures S10 and S11).
- In general, most genes coding for putative ADHs showed transcription levels under 500 NRC in the mycelium and in FB tissue samples.
- aegerita as well as the relative concentrations of volatile oxylipins (red) in the HS of C..
- Of the differentially expressed genes rele- vant to fruiting initiation in the related agaric S.
- com- mune [10, 12], the transcription factor-encoding genes BRI1 and FST4 [56] showed a clear transcriptional in- duction in primordia and, in the case of the latter gene, also in immature FBs of C.
- Our observation that BRI1 and FST4 also get strongly induced during the sporulation/post-sporulation stage in the mycelium might relate to the phenomenon that C.
- aegerita AAE-3 in the fruiting regime of Herzog et al.
- Grayed-out genes did not show differential expression in the present study in the respective developmental stage(s) of C.
- aegerita AAE-3 but have previously been shown to trigger relevant fruiting body developmental processes in the model mushroom S.
- cinerea implying a role also for later stages of FB development in the plectenchyma of C.
- The para- logized C2H2, which is relevant to the transition from FB initials to primordia in the related agaric S.
- Being involved in the FB maturation-associated process of stipe elongation in C..
- might also contribute to the high diversity of sesquiter- penes observed in the HS of C.
- coli clones were also present during sporulation in the HS of C..
- Interestingly, genes of the mevalonate pathway as well as of the STSs showed generally higher expression levels in the mycelium than in FB samples during the late phase of the fruiting process.
- This indi- cates that the mycelium rather than the FB tissue is the origin of the high amounts of sesquiterpenes observed in the HS of C.
- Generally, we ob- served a higher expression of genes involved in the fatty acid synthesis and further processing to linoleic acid in FB stages than in mycelial stages.
- The role of 13-HPOD in the formation processes of fun- gal VOCs is still largely unknown.
- However, no LOX activity was detected in the mycelium, excluding LOXs as a source for oct-1-en-3-ol present in the mycelium [49].
- Our transcriptomic data indicates similar with only little pronounced expression of LOX genes in the mycelium despite the presence of oct-1-en-3-ol during all mycelial stages [54]..
- This is due to the presence of a NXXQ motif in the I-helix of the CYP (cytochrome P450) domains, proven to be in- volved in the hydroperoxide isomerase activities of 7,8- and 5,8-LDS (linoleate diol synthase) [91], and the oc- currence of the YRWH sequence.
- solani regarding leu- cine and valine residues in the DOX domain (AAE3_.
- In the C-terminal CYP domains of AAE3_13098 and AAE3_00407, the NXXQ motif is absent.
- nidulans resulted in the produc- tion of oct-1-en-3-ol, oct-2-en-1-ol, oct-2-enal and.
- In this context, the putative DOX AAE3_13098, which also shows comparably high transcription in the mycelium and similarities of its expression course to the oct-1-en- 3-ol pattern in the HS of C.
- In FB samples as well as in the mycelial stages, the course of expression of the putative HPL gene AAE3_09203 was comparable to the transcription of the putative DOX gene AAE3_13098.
- Both genes showed similarities to the oct-1-en-3-ol pattern in the HS of C.
- thaliana, shared similarities in the expression pattern with the gene coding for the characterized Lox4.
- In this context, ADHs and ene-reductases might play an important role, explaining the observed decrease of oct- 1-en-3-ol during sporulation and, thereafter, the increase of octan-3-one in the HS of C.
- Using the amino acid sequence of this ADH (Q9P6C8), we identi- fied putative ADHs in the genome of C.
- This high expres- sion came along with a decreasing amount of oct-1-en- 3-ol and an increasing amount of octan-3-one in the HS of C.
- Overall, it seems that the first occurrence of octan-3- one in the HS of C.
- aegerita in late FB developmental stages is probably due to enzymatic activities in the FB tissue and not in the mycelium.
- This would also ex- plain why octan-3-one was not observed in the HS of C..
- origin of the sesquiterpenes appearing in the HS of C..
- In this work, we investigated the changes in the tran- scriptome of C.
- The here-observed differential expression patterns of partially highly paralogized FRGs during fruiting in contrast to the situation in the other model agarics C.
- Generally, proteins were identified in the genome of C..
- For identification of enzymes involved in the meva- lonate pathway analogues in S.
- Putative DOXs were identified in the genome of C..
- Putative ene-reductases were identified in the genome of C.
- aegerita AAE-3 chiefly congruent to the ones of the RNA-seq analysis in the present study, deviating only by these: fruiting body initials (FBi) at day 15 to 16 post inoculation (p.i.
- 5 fold decrease between day 22 and day 22 in the mycelium.
- 5 fold increase between day 22 and day 22 in the mycelium..
- Expression of genes involved in the mevalonate pathway and of the sesquiterpene synthases Agr1 to Agr9 in C.
- Expression of genes putatively involved in the fatty acid biosynthesis of C.
- Transcription levels of putative HPLs as well as the relative concentrations of volatile oxylipins in the headspace of C.
- Transcription levels of putative ADHs as well as the relative concentrations of volatile oxylipins in the headspace of C.
- Transcription levels of putative ene-reductases as well as the relative concentrations of volatile oxylipins in the headspace of C.
- Life history and developmental processes in the basidiomycete Coprinus cinereus.
- Fruiting in the higher fungi.
- Cloning and sequencing of the Aa-Pri1 gene specifically expressed during fruiting initiation in the edible mushroom Agrocybe aegerita, and analysis of the predicted amino-acid sequence.
- Nucleus-specific expression in the multinuclear mushroom-forming fungus Agaricus bisporus reveals different nuclear regulatory programs.
- Comparative genomics reveals unique wood-decay strategies and fruiting body development in the Schizophyllaceae.
- Transcriptome data reveal conserved patterns of fruiting body development and response to heat stress in the mushroom-forming fungus Flammulina filiformis.
- Difference in the volatile composition of pine- mushrooms (Tricholoma matsutake sing.) according to their grades.
- Genome expansion and lineage-specific genetic innovations in the forest pathogenic fungi Armillaria.
- Comparative transcriptome analysis of dikaryotic mycelia and mature fruiting bodies in the edible mushroom Lentinula edodes.
- Diversity of sesquiterpene synthases in the basidiomycete Coprinus cinereus

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