Taiz,L. & E. Zeiger. 2010.

Plant
Physiology
Chapter 24

Brassinosteroids:
Regulators of Cell Expansion
and Development

IR. KOESRIHARTI, MS.

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PLANT HORMONES
 BRASSINOSTEROID
Hormon steroid telah dikenal pada hewan, dan
beberapa tahun terakhir ini telah diketemukan
ada dalam tanaman.
Hormon steroid hewan termasuk sex
hormones ( estrogen, androgen dan progestin)
dan adrenal cortex hormones (glucocorticoid
dan mineralocorticoid)
Brassinosteroids (BRs) merupakan kelompok
hormon steroid yang berperan vital pada
sejumlah fenomena perkembangan tanaman:
Pembelahan sel & pemanjangan sel pada batang dan
akar
Photomorphogenesis
Perkembangan reproduktif
Senescence daun
Stress responses

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 BRASSINOSTEROID
Brassinosteroids (BRs) pertama kali diisolasi dari
pollen Brassica pada tahun 1979 merupakan steroid
yang secara kimiawi mirip cholesterol dan sex hormones
pada hewan
BR-deficient mutant mengalami gangguan
pertumbuhan dwarfism, berkurangnya apical
dominance dan male fertility, serta penundaan
pembungaan dan senescence
Brassinosteroid menghidupkan gen spesifik melalui
inaktivasi suatu protein yang sebaliknya secara tidak
langsung memblokir transkripsi dari gen tersebut.

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 Brassinosteroids (BRs)

60 types, brassinolide most common

Stimulates cell elongation, leaf expansion
BR mutants extreme dwarfs, small crinkled
leaves
Dark grown BR mutants de-etiolated

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Plant Genes on Steroids
Science, Vol 307, Issue 5715, 1569-1570 , 11 March 2005

BIN2 catalyzes breakdown of BES1 & BZR2 proteins (phosphorylation)

BR regulates activity of key growth transcription factors
-BES1(activator)
-BZR1(repressor)
6
Figure 13.12 (p.290)

7
 Fig. 17.20: Signal transduction chain for the response to
brassinosteroids

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 Flow Chart

Signal
The steroid binds to the
intracellular BR1 Kinase
and starts the
phosphorylation cascade.
Transduction
Series of events occur that
lead to transcription factors
that turn on specific genes.

Response
This gene that produces a protein that
promotes cell growth in the roots,
leaves, and stems of the plant. It will
also increase plant production.

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 Struktur Brassinosteroid
Ada dua bioassay utama yang telah
digunakan untuk memurnikan
Brassinosteroid:
Bean second-internode bioassay (Gambar
24.1)
Rice lamina (leaf) inclination bioassay
(Gambar 24.2)
Bioassay ini membedakan antara BRs yang
secara biologis aktif dengan intermediate atau
metabolit inaktif dan menggambarkan jumlah
senyawa yang aktif ada (Gambar 24.3)
Struktur dasar kimiawi Brassinosteroid berupa
steroidal lactone yang dimurnikan melalui X-
ray crystallographic analysis (Gambar 24.4)
disebut: Brassinolide (BL)
Prekursor Castasterone (CS)

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 Brassinosteroid Metabolic Pathway in Arabidopsis
sterol biosynthesis

DET2

HO O O HO HO
H H HO
(24R)-ergost-4-en-3-one (24R)-5-ergostan-3-one
campesterol (CR) (4-en-3-one) (3-one) campestanol(CN) 6-oxoCN

OH OH
DWF4 OH DWF4 OH DWF4 OH DWF4 DWF4

DET2

O HO HO cathasterone(CT)
HO O H H 6-deoxoCT H
O
22-hydroxyCR 22-hydroxy-4-en-3-one 22-hydroxy-3-one CYP90C1 CYP90C1
OH OH
CYP90D1 CYP90D1
OH OH OH

CYP90C1 CYP90C1
BR6ox1, 2 teasterone (TE)
CYP90D1 CYP90D1 CYP90C1 HO HO
H 6-deoxoTE H
O
CYP90D1
HO
H
3-epi-6-deoxoCT CYP90C1
CYP90D1 OH OH
OH OH

OH OH
OH OH DET2
BR6ox1, 2 3-dehydroteasterone
O O
HO O H HO (3DT)
22,23-dihydroxyCR 22,23-dihydroxy-4-en-3-one 6-deoxo3DT
OH OH OH OH OH
OH OH
OH OH OH OH OH
HO BR6ox1, 2 HO HO

HO HO HO typhasterol(TY) HO HO O
H H H
O H
O
26-hydroxyCS HO 26-hydroxyBL
6-deoxoTY BAS1
6-deoxoCS
OH BR6ox1, 2 OH OH OH
BAS1
O UGT73C5 OH BR6ox2 OH UGT73C5 O
HO Glc HO HO HO Glc
edited by
HO HO HO O HO O
H H HO HO BL-23-O-glucoside
S. Fujioka & O O
Y. Shimada CS-23-O-glucoside castasterone(CS) brassinolide (BL)
 Figure 24.1 Bean second-internode bioassay for
brassinosteroids

Potongan dari second-internode

tanaman kacang diapungkan
dalam larutan yang mengandung
BRs pada berbagai konsentrasi
selama beberapa hari.
Kontrol yang tidak direndam (kiri)
Pada konsentrasi rendah, BRs
menginduksi pertumbuhan
memanjang
Konsentrasi yang lebih tinggi
mengakibatkan penebalan,
pelengkungan, dan pecah

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 Figure 24.2 Dwarf rice lamina inclination bioassay for
brassinosteroids

Setetes kecil sample yang

dilarutkan dalam ethanol
diberikan pada pertemuan
antara lamina dan leaf
sheath
Setelah inkubasi selama
dua hari pada kelembapan
tinggi, sudut lengkung theta
() antara lamina dan leaf
sheath diukur
Sudut proporsional dengan
jumlah brassinosteroid dalam
sample
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 Figure 24.3 Doseresponse curves for three active BRs in the
rice lamina inclination bioassay Sudut lengkung meningkat
fungsi dari konsentrasi BR, sampai 10 ng
Aktivitas relatif tinggi dari BL; 24-epiBL, 24-epibrassinolide; 28-
homoBL, 28-homobrassinolide; BL, brassinolide

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 Figure 24.4 Structures of brassinosteroids

1. BRs are polyhydroxylated steroid hormones.

2. Brassinolide (BL) is the most active BR.
3. Hydroxyl pada rantai samping pada carbon 22 dan 23 essensial untuk
aktivitas
4. Senyawa C27 , C28 , atau C29 dapat dibedakan dari struktur rantai samping
5. 24-epiBL disintesis lebih murah daripada brassinolide sering digunakan
dalam eksperimen

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 Brassinosteroid

Bioassays distinguish active BRs from

inactive intermediates and permit
quantitation
BRs are a group of polyhydroxylated
steriod hormones, with brassinolide
(BL) being the most widespread and
active BR in plants
BRs have been detected in all tissues
examined with greatest activity in the
apical shoot
BRs are ubiquitous plant hormones
that predate the evolution of land
plants

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 Figure 24.5 Phenotypes of Arabidopsis BR mutants
(A) Homozygous bri1
mutant(kiri) umur 3 minggu
light-grown sangat kerdil
dibandingkan dengan
heterozygous bri1 mutant
(kanan), yang menunjukkan
morfologi wild-type
(B) Homozygous cpd
mutant(kiri) umur 3 minggu
light-grown juga
menunjukkan phenotype
kerdil; heterozygous mutant
dengan phenotype wild-
type(kanan)
(C) det2 dewasa light-grown
(kiri) lebih kerdil
dibandingkan dengan
tanaman wild-type (kanan)
(D) det2 dark-grown (kiri)
pendek, hypocotyl tebal
dan cotyledon membesar;
dark grown wild-type
(kanan)
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 Figure 24.6 BL and intermediates of BL biosynthetic
pathway restore normal growth to cpd mutant
Bibit Wild-type dan cpd mutant ditanam
selama 14 hari dengan tanpa steroid (-)
atau dengan 0,2 M senyawa BR
intermediate.
Catatan: apakah campesterol (CL) atau
cathasterone (CT) mempunyai efek
pada phenotype cpd mutant, karena
intermediate ini terjadi sebelum reaksi
yang dikatalisis oleh CPD
Sebaliknya, teasterone (TE), 3-
dehydroteasterone (DT), typhasterol
(TY), catasterone (CS), dan
brassinolide (BL) semua phenotype
selamat karena terjadi sesudah reaksi
CPD-catalyzed
Wild-type telah mengandung taraf
optimal dari intermediate ini dan oleh
karena itu sedikit terhambat oleh BL
dan immediate precursor
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 Photomorphogenesis BR deficient mutant
terganggu

Morfologi det2 and cpd

diidentifikasi pada screening untuk
bibit Arabidopsis yang ditanam
pada light-grown sesdudah
ditanam selama beberapa hari
dalam gelap total
BR-deficient mutant menunjukkan
photomorphogenesis abnormal,
yang dapat dicegah dengan
aplikasi BL eksogen (atau
intermediates downstream of the
steps catalyzed by the mutated
enzymes).
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 Figure 24.7 Domain structure of the BR receptor BRI1

Receptor BRI1
bertempat pada plasma
membran
Daerah extracellular
terdiri dari bidang
leucine-rich repeat
sequences (LRRs) yang
mengandung island
domain yang berfungsi
sebagai bagian dari
brassinolide (BL)
binding site
Bagian intracellular
mengandung
juxtamembrane domain,
kinase domain, dan C-
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 Figure 24.8 A model for BR signaling
BAK1: BRI1-associated receptor kinase 1
BKI1: BRI1-kinase inhibitor 1
BSK: BR-signaling kinase

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 Figure 24.8 A model for BR signaling

BL terikat pada BRI1 receptor yang

ditemukan dalam plasma membrane
dan endosomal membranes
Pengikatan BL mengaktivasi BRI1,
yang menjadi terfosforilasi pada
multiple sites
Aktivasi dari BRI1/BAK1
menginisiasi pemancaran signal
yang menuntun pada transkripsi gen
yang diatur-BR
Bentuk de-phosphorylated dari
BES1 and BZR1 mengaktivasi atau
menekan BR target genes

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Figure 24.9 BL inhibits the phosphorylation of BES1

Highly
phosphorylated form

De-phosphorylated
form

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Figure 24.10 Simplified pathways for brassinolide (BL)
biosynthesis and catabolism

All the enzymes

converting campesterol
to BL are cytochrome
P450 monoxygenases
located on the ER.

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 Figure 24.12 Structure of brassinazole, a triazole compound
that inhibits brassinosteroid biosynthesis

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Figure 24.11 Brassinosteroid levels are controlled by both
negative and positive feedback

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 Figure 24.13 Effect of brassinazole on light-grown 14-day-old
Arabidopsis seedlings

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 Figure 24.14 Overexpression of the BR biosynthetic gene
DWF4 in Arabidopsis

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 Figure 24.15 Effects of reciprocal grafting between wild type and
BR-deficient mutant of pea

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 BRs

Taraf BR diatur melalui multiple control

mechanisms, yaitu: catabolism,
conjugation, dan negative feedback dari
signaling pathway

Tanaman yang ditanam pada Brz

menunjukkan BR-deficient phenotype, yang
dapat dibalikkan melalui penambahan BL
pada medium pertumbuhannya

BRs berperan dekat synthesis-sites dan

tidak mengalami long-distance transport;
setiap organ mensintesis dan merespon
BRs aktif yang dimiliknya

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 Figure 24.16 The kinetics of BR stimulation of soybean
epicotyl elongation

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 Figure 24.17 BRs increase the plastic wall extensibility of
soybean epicotyls

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Figure 24.18 Effect of BR on microtubule organization in
Arabidopsis seedlings

Wild-type parenchyma cell

Green: microtubules
(b) BR deficient mutant parenchyma cell
Yellow: chloroplasts
(c) BR-deficient mutant treated with BR
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Figure 24.19 BL and IAA act synergistically to promote lateral root
development

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 Figure 24.20 BR is required for a normal vascular
development

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Figure 24.21 Zinnia leaf mesophyll cell before and after
differentiation into a tracheary element

BRs are essential for this differentiation process

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 Figure 24.22 BR stimulates germination of Arabidopsis
seeds

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 BRs: Effects on Growth and
Development
Involved in development of fiber, lateral
roots, and vasculature, as well as
maintenance of apical dominance,
pollen tube growth, seed germination,
leaf senescence, and plant defenses
Promote both cell proliferation and cell
elongation
Maintain normal microtubule abundance
and organization needed for cell wall
growth
Promote root growth at low
concentrations and inhibit root growth at
high concentrations
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 BRs: Effects on Growth and Development

Promote lateral root development by

altering polar auxin transport
Promote differentiation of the xylem
and suppress that of the phloem
Promote seed germination by
interacting with other hormones, such
as GA and ABA
BR application to crop plants is most
effective under stress conditions
BRs are useful in plant propagation

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 BRs: effects on growth and development
fiber development in cotton
development of lateral roots
maintenance of apical dominance
vascular differentiation
male sterility (pollen tube growth)
plant defense (resistance to infection)
seed germination
leaf senescence

Small- scale studies and large-scale trials using BRs

are bound to emerge,
is most beneficial to growth under stress conditions

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 Summary:

1. Brassinosteroids (BRs) are polyhydroxylated steroid

hormones. Brassinolide (BL) is the most active BR.

2. Bioassays can be used to distinguish BRs from other

plant hormones.

3. Deficiency in BR biosynthesis results in dwarfism.

4. BRs promote both cell elongation and cell division.

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Brassinosteroids such as brassinolide

Brassinosteroids such as
brassinolide
are found in seeds, fruits, shoots,
leaves, and flower buds.
Functions include:
a. inhibiting root growth at high
concentrations (promote it at low
concentrations) and inhibiting early
germination
b. promote cell expansion and cell
division in shoots
c. promoting xylem differentiation
d. promote pollen tube elongation
Types of Hormones
Auxin to grow or increase; apical dominance
Cytokinin cytokinesis (cell division)
Abscisic acid abscission/leaf senescence; dormansi;
stomata regulation (closes)
Jasmonic acid found in jasmine oil
Gibberellic acid pathogen Gibberella
Ethylene pematangan buah
Brassinosteroids derived from Brassica spp.
Each are present in cells in varying concentrations
Jasmonic Acid:
First identified in jasmine oil
Response to biotic stress:
Wounding induces JA biosynthesis
Microbial and fungal invasion
 Jasmonic Acid

http://park.itc.u-tokyo.ac.jp/biotec-res-
ctr/kampo/eng/research_plant.html
44
Herbivores are organisms that feed on plants.

Responses to these
predators include:
1. Physical defenses such as
thorns
2. Chemical defenses such as
noxious chemicals and toxic
chemicals
3. Production of chemoattractant
for a parasitoid wasp that lays
eggs in a herbivorous caterpillar.
The physical damage to the leaf
facilitates the release of this
chemoattractant.
Plant Defences

Alarm pheromones that are produced by a

damaged plant that pass this information
onto other individuals (disclaimer- I am not
sure that alarm pheromone is appropriate
term but it is a term that is used to describe
chemical cues that are produced by injured
animals that are detected and responded to
by conspecifics)
Pathogens are also predators in a sense and they also cause damage
or death to plants

1. Like in animals, there is a first line defense that is composed of dermal

tissue that prevents pathogenic organisms from getting into the plant
body
2. Like in animals, there is also a second line of defense that involves:
a. a recognition of pathogens
b. the production of products that destroy or attack these pathogens
3. Damage to cells by pathogens facilitates the production of
antimicrobial compounds called phytoalexins.
4. Infection also activates genes that produce PR proteins.
5. Plants also produce alarm hormones that are transported from
the sight of damage throughout the plant stimulating the production
of phytoalexins and PR proteins. This response is referred to as
systemic acquired resistance (SAR) and it is nonspecific and
provides protection against various pathogens for days
EGG database:
tp://www.genome.jp/kegg/
49