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1/25/2019

Summary of the talk California State University East Bay Department of Biological Sciences Almeida Lab - Introduction: - The - morphology in the Zingiberales - Hypothesis Comparative analysis of whole flower transcriptomes in the Zingiberales - Methods: data collection and analysis Ana Almeida; Alma Piñeyro-Nelson; Roxana Yockteng; Chelsea Specht - Results and discussion - OrthoFinder Ana Almeida - Blastn to oil palm - Transcription factors

- Concluding remarks

January 2019

Diversification of flower morphology The Zingiberales order

Marantaceae g inger clade inger Cannaceae

Zingiberaceae

Costaceae b

Lowiaceae groupanana

Strelitziaceae

Heliconiaceae

(Saas et al. 2016, PeerJ)

Flower morphology in the Musaceae The Zingiberales order

Musa basjoo

Marantaceae g inger clade inger Cannaceae

Zingiberaceae

Costaceae b

Lowiaceae groupanana

Strelitziaceae Bird-of-paradise

Heliconiaceae Musaceae

1 cm (Saas et al. 2016, PeerJ)

1 1/25/2019

The Zingiberales order Flower morphology in the Zingiberales

Canna Reduction to 1 fertile

Marantaceae g Petaloid inger clade inger Cannaceae Fusion of Reduction to ½ staminodes fertile stamen

Costus Tumeric Zingiberaceae

Costaceae b

Lowiaceae groupanana Strelitziaceae spicatus sp. Heliconiaceae Musaceae

(Saas et al. 2016, PeerJ) (modif from Specht et al. 2012, Bot Rev)

Flower morphology in the Zingiberales Working hypothesis

Mechanisms of floral

Marantaceae g diversification inger clade inger Cannaceae Depp conservation of ? developmental mechanisms (organ identity) Zingiberaceae Distinct lineage-specific

Costaceae developmental programs b

Lowiaceae groupanana Diversification of floral shape Strelitziaceae (Chanderbali et al. 2010, PNAS)

Heliconiaceae Musaceae

(Saas et al. 2016, PeerJ)

Flower morphology in the Zingiberales Working hypothesis

Mechanisms of floral ginger clade diversification

Depp conservation of Marantaceae developmental mechanisms (organ identity) Cannaceae Distinct lineage-specific developmental programs

staminodes stamen gyn Diversification of floral shape (Chanderbali et al. 2010, PNAS) Zingiberaceae

Costaceae Tinkering of conserved Evolution of lineage- Co-option of non-flower mechanisms specific mechanisms mechanisms

2 1/25/2019

Working hypothesis Results and discussion

Mechanisms of floral - Assembly metrics diversification

Depp conservation of developmental mechanisms (organ identity)

Distinct lineage-specific developmental programs

Diversification of floral shape (Chanderbali et al. 2010, PNAS)

Tinkering of conserved Evolution of lineage- Co-option of non-flower mechanisms specific mechanisms mechanisms AG-AP1 ? Ab-ad netowork (Almeida et al. 2015, EvoDevo) (Almeida et al. 2014, SciRepots) (Almeida et al 2018, PeerJ)

Methods: data collection and analysis Results and discussion

- used in this study: - Assembly metrics

Marantaceae

Cannaceae Canna sp.

Zingiberaceae officinale

Costaceae

Lowiaceae Orchidantha fimbriata

Strelitziaceae

Heliconiaceae Musaceae Musa basjoo

Outgroup Dichorisandra thyrsiflora (Almeida et al 2018, PeerJ)

Methods: data collection and analysis Results and discussion

- Protocol Developing *** - OrthoFinder results Total flower RNA extraction  41,557 orthogroups between transcriptome and CDS from sequenced genomes; cDNA libraries for Illumina (HS2000) Adapters Low quality  17,418 orthogroups had counterparts in at least on of the sequenced genomes; Sequence clean-up and trimming Low complexity Duplicates  6,916 orthogroups with representatives of all species studied; De novo assembly with Contaminants Trinity v2.1.0  Of those, only 28 were single copy. Prediction of long ORFs with  11,511 orthogroups had counterparts in Arabidopsis thaliana; TransDecoder v4.1.0 Blastp against Uniprot  142 orthogroups were Arabidopsis specific. Contig filtering (>300bp) HMM search against Pfam Arabidopsis thaliana Orthology between AGAMOUS-like (AGL) genes: ALG6, ALG12. AGL20, AGL26, AGL29, AGL44, AGL58, genomes and filtered CDS with AGL61, AGL65, AGL104; Phoenix dactylifera Orthofinder v2.2.3 Elaeis guineensis MADS-box genes: APETALA3 (AP3), PISTALLATA (PI), SEPALLATA3 (SEP3), CAULIFLOWER Blastn to Elaeis guineensis (CAL), SHATTERPROOF2 (SHP2), CRABS CLAW (CRC), various TCP. Transcription factors were identified based on PlantTFDB v4.0

3 1/25/2019

Results and discussion Results and discussion

- Blastn against Elaeis guineensis - Blastn against Elaeis guineensis  Contigs blasted to 22,959 E. guineensis CDS  Contigs blasted to 22,959 E. guineensis CDS

- Developmental genes: AP2-like homologs AGL6-like homologs HUA2-like homologs LEUNIG-like homologs KNOTTED1-like homologs GATA-like homologs MYB-like homologs TCP-like homologs CONSTANS-like homologs WUSHEL-like homologs

- Stress and pathogen response: (WRKY, NAC)

Results and discussion Results and discussion

- Blastn against Elaeis guineensis - Blastn against Elaeis guineensis  Contigs blasted to 22,959 E. guineensis CDS  Paralogues of the same gene/gene in different floral transcriptomes:

Marantaceae LATERAL ORGAN BOUNDARIES LOB18-like (LOB)-domain homologs Cannaceae found in all lineages

47% - enzymes Zingiberaceae LOB4-like 26% - structural proteins Costaceae LOB40-like, 15% - uncharacterized LOB41-like ~12% - regulatory Lowiaceae

Strelitziaceae LOB6-like, LOB36-like 508 proteins could be assigned Heliconiaceae to known transcription factor families Musaceae Outgroup LOB15-like

Results and discussion Results and discussion

- In terms of Transcription Factors - Blastn against Elaeis guineensis  Paralogues of the same gene/gene family in different floral transcriptomes:

Marantaceae LATERAL ORGAN BOUNDARIES LOB18-like (LOB)-domain homologs Cannaceae found in all lineages

Zingiberaceae LOB4-like LOB40-like, LIM domain Bromodomain Bromodomain Costaceae LOB41-like family family (GTE4-like, family (GTE9- (SEUSS-like) GTE6-like), like), YL1 Lowiaceae PUR-A domain domain family NCBI Conserved Domain family (PURA1-like) (SWR1) Strelitziaceae LOB6-like, Dataset Other lineage-specific LOB36-like paralogues: Bromodomain-family (GTE7- Heliconiaceae like), TFIIS-domain family DIVERICATA-like (IWS1-like), SAND-domain Musaceae TGA-like family (ULTRAPETALA1-like) ASIL-like Outgroup LOB15-like

4 1/25/2019

Results and discussion Concluding remarks

- Blastn against Elaeis guineensis Mechanisms of floral diversification  Lineage-specific regulatory genes:

Marantaceae Tinkering of conserved Evolution of lineage- Co-option of non-flower mechanisms mechanisms Cannaceae specific mechanisms

 A clear, well-established phylogenetic framework is essential - “model clades” Zingiberaceae (Chanderbali et al. 2016);  Duplications are a widespread phenomenon; Costaceae

Lowiaceae FLORICAULA/LEAFY-like Strelitziaceae ODORANT1-like JUNGBRUNNEN-like Heliconiaceae RAX-like DPB-like Musaceae SOMBRERO-like among others. Outgroup

Results and discussion Concluding remarks

- Blastn against Elaeis guineensis Mechanisms of floral diversification  Lineage-specific regulatory genes:

Marantaceae Tinkering of conserved Evolution of lineage- Co-option of non-flower mechanisms Cannaceae specific mechanisms mechanisms

 A clear, well-established phylogenetic framework is essential - “model clades” Zingiberaceae (Chanderbali et al. 2016);  Duplications are a widespread phenomenon; Costaceae  The set of Commelinid shared genes (~4,000) - conserved regulators of floral Lowiaceae CCA1-like development in monocots. It is yet unclear what their roles might be in flower FLX2-like development and diversification in the monocots; Strelitziaceae KT112-like  Lineage-specific gene sets provide interesting candidate gene lists for further studies; YABBY4-like  Heliconiaceae CPC-like Addition of other outgroup species might influence the Zingiberales specific SPATULA-like datasets; Musaceae  Expression levels an analysis of the non-coding sequences might provide Outgroup additional layers of developmental regulation.

Results and discussion Acknowledgements

- Blastn against Elaeis guineensis  Lineage-specific regulatory genes:

Marantaceae Holly Forbes (UCBG) AS1-like Cannaceae DROOPING -like CUC2-like EXORDIUM-like Zingiberaceae FAF-like Chelsea Specht Roxana Yockteng Alma Piñeyro SPX-like Cornell U CORPIOCA UAM Costaceae

Lowiaceae Strelitziaceae Thank you! Heliconiaceae Musaceae

Outgroup

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