Transcriptional control of floral

development

© 2017 American Society of Plant Biologists

 Why floral structure matters

Gideon Pisanty Bharathimohanr

• Plant reproductive strategy and genetic diversity

• Pollen dispersal
• Pollinator interactions
• Reproductive isolation
USDA • Benefits to people
• Food crops for human consumption
• Aesthetics

MDF See Hileman, L.C. (2014). Trends in flower symmetry evolution revealed through phylogenetic and
developmental genetic advances. Philos. Trans. R. Soc. Lond. B. Biol. Sci. 369: 20130348..

© 2017 American Society of Plant Biologists

 Most plants have whorls of floral
organs arranged in concentric
circles
sepal

From out to in petal

Whorl 1: sepals
Whorl 2: petals
Whorl 3: stamen
Whorl 4: carpel stamen carpel

Arabidopsis flower Arabidopsis flower

Side view Top view

In this model, the outer two whorls (sepal and petal) form the perianth, the non-
reproductive structures. The inner two whorls (stamen and carpel) represent the male
and female reproductive organs.
Guo, S., Sun, B., Looi, L.S., Xu, Y., Gan, E.S., Huang, J., and Ito, T. (2015). Co-ordination of flower development through epigenetic
regulation in two model species: Rice and Arabidopsis. Plant Cell Physiol. 56: 830–841 by permission from Oxford University Press.

© 2017 American Society of Plant Biologists

 Within that pattern, flowers occur in
many different forms, with varying
organ number and shape
Phalaenopsis Serracenia Epiphylum
oxypetalum

Hibiscus
schizopetalus Senecio confusus Image credits: Fleet

© 2017 American Society of Plant Biologists

 Amid that variation, all flowers share a perianth
(non-reproductive structures) and male and/ or
female reproductive structures
Campanula medium Lily Campanula medium

The perianth can include Oryza sativa The reproductive organs

structures such as sepals can include male
and petals (top left), tepals stamens, female carpels,
(top right), or lemma, palea or both
and lodicule (right),
depending on the species.

Image credits: Top: Fleet. Bottom: Lombardo, F. and Yoshida, H. (2015). Interpreting lemma and palea homologies: a point of view from rice floral mutants. Front. Plant Sci. 6: 1–6. CC-
BY.

© 2017 American Society of Plant Biologists

ABC(DE) model of flower
development

Dornelas, M. C. and Dornelas, O. (2005). From leaf to flower: revisiting Goethe's concepts on the ¨metamorphosis¨ of plants. Braz. J. Plant Physiol. 17: 335-344 CC-BY.

© 2017 American Society of Plant Biologists

Floral development models were first built from
studies in Arabidopsis thaliana and
Antirrhinum majus

Sepal
Petal
Stamen
Carpel

Arabidopsis thaliana A. majus (snapdragon)

Arabidopsis image reproduced with permission from Parcy, F., Bomblies, K., and Weigel, D. (2002). Interaction of LEAFY, AGAMOUS and
TERMINAL FLOWER1 in maintaining floral meristem identity in Arabidopsis. Development 129: 2519–2527.; Antirrhinum: image credit Fleet

© 2017 American Society of Plant Biologists

 Early studies used scanning electron
microscopy (SEM) to observe changes in the
development of floral organs

SEM of different stages of development of a wild-type Arabidopsis flower

100 μm
10 μm 10 μm

MS=medial stamen primordium, LS=lateral stamen primordium, G=gynoecial primordium

Republished with permission from Bowman, J.L., Smyth, D.R., and Meyerowitz, E.M. (1991). Genetic interactions among floral homeotic genes of Arabidopsis. Development 112: 1–20.

© 2017 American Society of Plant Biologists

 In the classic ABC model, key transcription
factors promote identity of floral organs
Within the floral meristem, the key
transcription factors for specifying
floral organs are known as the ABC
genes (and sometimes D and E – more
on those later). Different
combinations of these ABC genes
result in different whorls of organs.

B Whorl 1=sepal, A genes

Whorl 2=petal, A+B genes
A C Whorl 3=stamen, B+C genes
whorl 1 2 3 4 Whorl 4=carpel, C genes

Guo, S., Sun, B., Looi, L.S., Xu, Y., Gan, E.S., Huang, J., and Ito, T. (2015). Co-ordination of flower development through epigenetic
regulation in two model species: Rice and Arabidopsis. Plant Cell Physiol. 56: 830–841 by permission from Oxford University Press.

© 2017 American Society of Plant Biologists

Homeotic mutations convert identity
of one floral organ to another
Loss-of-function of one or more
of the ABC genes can result in
homeotic transformations, in
which one organ is changed into
another organ type.

WT Arabidopsis

ag mutant showing
homeotic conversion
to form extra petals

Reproduced with permission from Parcy, F., Bomblies, K., and Weigel, D. (2002). Interaction of LEAFY, AGAMOUS
and TERMINAL FLOWER1 in maintaining floral meristem identity in Arabidopsis. Development 129: 2519–2527.

© 2017 American Society of Plant Biologists

 Missing B gene function: homeotic conversion
of petals to sepals and stamen to carpels

Snapdragon
deficiens mutant
Arabidopsis lacking B gene function
pistillata mutant A C
lacking B gene function

Republished with permission from Bowman, J.L., Smyth, D.R., and Meyerowitz, E.M. (1991). Genetic interactions among floral homeotic genes of Arabidopsis. Development 112: 1–20 and Masiero, S., Li, M.-A.,
Will, I., Hartmann, U., Saedler, H., Huijser, P., Schwarz-Sommer, Z., and Sommer, H. (2004). INCOMPOSITA: a MADS-box gene controlling prophyll development and floral meristem identity in Antirrhinum.
Development 131: 5981–5990. See also Sommer, H., Beltrán, J.P., Huijser, P., Pape, H., Lönnig, W.E., Saedler, H., and Schwarz-Sommer, Z. (1990). Deficiens, a homeotic gene involved in the control of flower
morphogenesis in Antirrhinum majus: the protein shows homology to transcription factors. EMBO J. 9: 605–613.

© 2017 American Society of Plant Biologists

 Missing C gene function results in homeotic
conversion of stamen and carpel to produce
repeating whorls of sepals and petals

B Snapdragon
plena mutant
Arabidopsis A A
agamous mutant

Republished with permission from Bowman, J.L., Smyth, D.R., and Meyerowitz, E.M. (1991). Genetic interactions among floral homeotic genes of Arabidopsis. Development 112: 1–20;
Keck, E., McSteen, P., Carpenter, R., and Coen, E. (2003). Separation of genetic functions controlling organ identity in flowers. EMBO J. 22: 1058–1066. See also Bradley, D., Carpenter, R.,
Sommer, H., Hartley, N., and Coen, E. (1993). Complementary floral homeotic phenotypes result from opposite orientations of a transposon at the plena locus of Antirrhinum. Cell 72: 85–95.

© 2017 American Society of Plant Biologists

 Missing A gene function results in missing
petals, and can result in sepals converted to
carpel-like organs

• A genes seem to have a

role in specifying
flower formation as
well as identity of
specific whorls.
• Their functions are
less well conserved
across species, with
some species having no
B A gene identified to
date.
Arabidopsis C C
apetala2 mutant

Republished with permission from Krogan, N.T., Hogan, K., and Long, J.A. (2012). APETALA2 negatively regulates multiple floral organ identity genes in Arabidopsis by recruiting the co-repressor
TOPLESS and the histone deacetylase HDA19. Development 139: 4180–4190 and Bowman, J.L., Smyth, D.R., and Meyerowitz, E.M. (1991). Genetic interactions among floral homeotic genes of
Arabidopsis. Development 112: 1–20. See also Keck, E., McSteen, P., Carpenter, R., and Coen, E. (2003). Separation of genetic functions controlling organ identity in flowers. EMBO J. 22:
1058–1066

© 2017 American Society of Plant Biologists

ABC plus D and E

D genes play a role in ovule formation,

and E genes work with ABCD genes to
specify the whorls.
In the absence of E genes, only leaf-like
organs are formed.

While much of ABC function was learned

from forward genetics, E function was
largely learned through reverse genetics
(working from gene sequence to
determine function).

Image credit: Guo, S., Sun, B., Looi, L.S., Xu, Y., Gan, E.S., Huang, J., and Ito, T. (2015). Co-ordination of flower development through
epigenetic regulation in two model species: Rice and Arabidopsis. Plant Cell Physiol. 56: 830–841 by permission from Oxford University
Press

© 2017 American Society of Plant Biologists

 Summary of ABCDE genes

 Different combinations of transcription factors promote the identity of

different whorls of floral organs.
 A function alone promotes sepal formation.
 A plus B functions promote petal formation.
 B plus C functions promote stamen formation.
 C function alone promotes carpel formation.
 D function promotes ovule formation.
 E genes pair with each of the others to promote flower formation.
 When one of these key genes is missing, the wrong organ can form in the
affected whorl by homeotic transformation.

© 2017 American Society of Plant Biologists

The floral ABCDE proteins bind DNA
to regulate transcription
MADS box
• Most of the ABC genes belong to a
DNA-binding
motif transcription factor family known
as MADS box genes.
• The proteins encoded by these
genes share a common amino acid
DNA motif that allows them to bind to
DNA and to other MADS domain
proteins.
• By binding promoter regions with
common DNA sequences, they
activate transcription of a set of
downstream genes necessary to
produce each floral organ type.

Gramzow, L. and Theissen, G. (2010). A hitchhiker’s guide to the MADS world of plants. Genome Biol. 11: 214.

© 2017 American Society of Plant Biologists

MADS domain proteins share a conserved
amino acid sequence motif

Alignment of the MADS domain of floral development proteins

Gramzow, L. and Theissen, G. (2013). Phylogenomics of MADS-Box Genes in Plants - Two Opposing Life Styles in One Gene Family. Biology (Basel). 2: 1150-1164.

© 2017 American Society of Plant Biologists

 ABCDE proteins function in
combinations
This is known as the “floral quartet”
model. CArG indicates the DNA
motif bound by MADS box proteins
(shown as colored circles)

Different combinations of MADS box

proteins bind to each other and bind
to DNA in order to activate
transcription of the genes needed to
specify each floral organ type.
Type E genes (SEP in Arabidopsis)
interact with the other MADS box
proteins for each whorl.

Image credit Gramzow, L. and Theissen, G. (2010). A hitchhiker’s guide to the MADS world of plants. Genome Biol. 11: 214; See also Honma, T. and Goto, K. (2001). Complexes of MADS-box proteins are sufficient
to convert leaves into floral organs. Nature 409: 525–529 and Theissen, G. and Saedler, H. (2001). Floral Quartets. Nature 409: 469–471 used by permission.

© 2017 American Society of Plant Biologists

 MADS box genes are present in all
land plants
• MADS box genes have been
Angiosperms Gymnosperms Ferns found in all land plants, and
Eudicot Monocot Basal angiosperm Conifers Gnetales genes of ABCDE type (the
MIKC MADS) group are found
in both gymnosperms and
Photo: Roar Joha
angiosperms.
Katorisi nsen BotBln
• Some angiosperms have nearly
100 different MADS box
proteins, and this diversity is
proposed to play a role in
differences in flower form.
• Differences in MADS box genes
and where they are expressed
may be part of the answer to
how flowers have diversified.

Adapted from Theissen, G., Becker, A., Rosa, A. Di, Kanno, A., Kim, J.T., Winter, K., and Saedler, H. (2000). A short history of MADS-box genes in plants. Plant Mol. Biol. 42: 115–149.

© 2017 American Society of Plant Biologists

 Amplification of MADS box genes in flowering
plants may lead to new functions and altered
flower morphology

Species # Type II MADS box genes

Selaginella moellendorffi (moss) 6
Picea abies (spruce) 249
Musa acuminata (banana) 68
Oryza sativa (rice) 42
Malus domesticata (apple) 113
Glycine max (soybean) 96
Arabidopsis thaliana 46

Flowering plants have many more MADS box genes than non-flowering plants

Adapted from Gramzow, L. and Theissen, G. (2013). Phylogenomics of MADS-Box Genes in Plants - Two Opposing Life Styles in One Gene Family. Biology (Basel). 2: 1150-1164 and Gramzow, L., Weilandt, L., and
Theißen, G. (2014). MADS goes genomic in conifers: Towards determining the ancestral set of MADS-box genes in seed plants. Ann. Bot. 114: 1 407–1429, by permission of Oxford University Press.

© 2017 American Society of Plant Biologists

Downstream targets of floral
development genes

Image credit Fleet and Bowman, J.L., Smyth, D.R., and Meyerowitz, E.M. (1991). Genetic interactions among floral homeotic genes of Arabidopsis. Development 112: 1–20.

© 2017 American Society of Plant Biologists

 ABC proteins activate transcription of downstream
genes to promote floral organ development

A B C

Targets include:
Downstream transcription factors for specific organ development
Genes for hormone synthesis and response
Other signaling molecules
Protein degradation pathways

Organ formation

See O’Maoileidigh, D.S., Wuest, S.E., Rae, L., Raganelli, A., Ryan, P.T., Kwasniewska, K., Das, P., Lohan, A.J., Loftus, B., Graciet, E., and Wellmer, F. (2013). Control of reproductive floral organ identity specification
in Arabidopsis by the C function regulator AGAMOUS. Plant Cell 25: 2482–2503.Thomson, B., Zheng, B., and Wellmer, F. (2017). Floral organogenesis: When knowing your ABCs is not enough. Plant Physiol. 173:
56–64.

© 2017 American Society of Plant Biologists

 Plant growth hormone pathways are
targets of ABCDE genes
A B C E C

gibberellin (GA) auxin GA

auxin GA
brassinosteroids
Hormones play important roles upstream in regulation of flower meristem formation.
In addition, techniques such as ChIP-seq and gene expression analysis identify
hormones as downstream targets of the ABC genes. Genes related to cytokinin,
jasmonic acid and abscisic acid have also been implicated as downstream targets.

Chandler, J.W. (2011). The Hormonal Regulation of Flower Development. J. Plant Growth Regul. 30: 242–254; Kaufmann, K., Muiño, J.M., Jauregui, R., Airoldi, C.A., Smaczniak, C., Krajewski, P., and Angenent,
G.C. (2009). Target genes of the MADS transcription factor sepallata3: Integration of developmental and hormonal pathways in the Arabidopsis flower. PLoS Biol. 7: 0854–0875; Thomson, B., Zheng, B., and
Wellmer, F. (2017). Floral Organogenesis: When Knowing Your ABCs Is Not Enough. Plant Physiol. 173: 56–64.

© 2017 American Society of Plant Biologists

Future directions
 What additional genetic changes
account for floral diversity across
species?
 What are the roles of hormones as
downstream targets of the ABCDE
genes?
 What other components are associated
with the known gene regulatory
networks for flowering?
 How do we develop predictive models
to describe flower morphogenesis and
predict gene functions?

Thomson, B., Zheng, B., and Wellmer, F. (2017). Floral Organogenesis: When Knowing Your ABCs Is
Not Enough. Plant Physiol. 173: 56–64; Boudon, F., Chopard, J., Ali, O., Gilles, B., Hamant, O.,
Boudaoud, A., Traas, J., and Godin, C. (2015). A Computational Framework for 3D Mechanical
Modeling of Plant Morphogenesis with Cellular Resolution. PLoS Comput. Biol. 11: e1003950.

© 2017 American Society of Plant Biologists