Brassinosteroids are a class of plant hormones that promote growth and development. Over 70 types of brassinosteroids have been isolated from various plant species, including angiosperms, gymnosperms, pteridophytes, bryophytes and algae. They are found in plant tissues and organs such as shoots, seeds, pollen and grains. Brassinosteroids have been shown to increase crop yields, enhance resistance to stresses, and improve plant growth. They are being commercialized for use in agriculture to boost productivity of important crops.

1. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-1

2. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-2
Advertising
 Paid, nonpersonal communication through
various media by business firms, not-for-
profit organizations, and individuals who
are identified in the advertising message
and who hope to inform or persuade
members of a particular audience
Seminar on
Brassinosteroids- New Class of Plant Hormone
Presented By
Sonali L. Bagade
Submitted To
DEPARTMENT OF AGRICULTURAL BOTANY
POST GRADUATE INSTITUTE
DR. PANJABRAO DESHMUKH KRISHI VIDYAPEETH,
AKOLA – (M.S.)

3. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-3
Introduction
Steroid-: These are the compounds which are widely
distributed in animals &
plantshave 1,2- cyclopenteno-perhrydro-
phenantherene nucleus there.
structure
1 ) It plays a very imp role as essential hormones in plants as well
as animals.
2 ) Plants produce numerous steroids& sterols, some
of which recognized as
hormones in animals (Geuns,1978; Jones and
Roddick 1988).
Brassinosteroids-: Brassinosteroids are a new group of plant growth
substance with significant
growth promoting activity (Grove et al. 1979)
. Pollen – Rape seed( Brassica nupus L ), for determine it’s
structure.
the biologically active molecule was a steroids that calls
Brassins. Since there
discovery over 70 BR have been isolated from plants
. It is biosynthesized from compesterol.
. Arabidopsis, pea & Tomato plans are used .
BRs have been shown to be involved in numerous plant processes:

4. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-4
Commercially available Brassinosteroids
Trade name country
Recommended crops
Biobras-6 Cuba
potato, sugarcane & onion
Bountee India
Tea
Brassinosteroids 1105 USA
Soybean
Combine India
Grapes
DAA-6 Cuba
Potato ,tomato & sugarcane
Double India
All major crops
Ts 303 Japan
Rice, potato, wheat, beet
Yaponiya Belarus
Barley

5. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-5
Immediately after the discovery of Brassinosteroids the
application of this substance for improving crop productivity
of diff. vegetables like radish ,lettuce, bushbean and paper was
started. Since the publication of this article “New chemicals
promise larger crops.”(Maugh1982) to “Twenty years of
Brassinosteroids.” (Kripach et al).
However ,promoted by the success stories ,several agrochemical
industries in Japan USA, Belarus, Cuba and India recently
started commercial production of BRs, at affordable level. “The
shadow of flop story- triacontanol also put a ride on the
commercial production.
Uses-
Improve resistance against envt. Stresses –
1. water stress 2. Salinity stress
3.Low temp stress 4. High temp stress (Rao et al 2002)
Ability to confer resistance against abiotic stress added advt. for
uses in agril. For improving crop
Agricultural uses of Brassinosteroids

6. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-6
Brassinosteroids-Potential to improve crop
productivity
Crops
Frit crops Grapes
Orange
Citrus
Strawberries
Watermelon
Banana
Compound
Epibrassinoloide
Brassinolide
Epibrassinoloide
Brassinolide
Brassinosteroids
Epibrassinoloide
Biobras-16
Response
Yield
Reduced abscission
of young flowers,
early maturation
Growth & yield
Retardation of
abscission
increased fruit lets
Retardation of
abscission
increased fruit lets
Yield
In vitro
multiplication
Reference
Ikekawa and zhao 1991
Xu et al. 1994
Takshashi et al. 1995
Iwahari et al.1990
Franco et al.2002
Pipattanawong et
al.1996
Ikekawa and zhao 1991
Rodriguez et al 1998

7. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-7
Brassinosteroids-Potential to improve crop
productivity
Crops
Veg.crops Lettuce
Bush bean
Paper
Soybean
Garlic
Potato
Yam
Spinach
onion
Tomato
Lettuce
Tomato
Compound
Brassinolide
Brassinolide
Epibrassinoloide
Homobrassinoloide
Brassinoloide
Epibrassinoloide
Biobras-6
Brassinosteroids
DA-6
Epibrassinoide
DA-6
Biobras-16
Biobras-6
Biobras-16
Homobrassinoide
Response
Growth & yield
Yield
Yield
Growth & yield
Yield
Tuber Yield
Tuber yield
Growth & yield
Bulb yield
Yield
Yield
Accelerating of
ripening
Reference
Meudt et al.1983
Meudt et al.1983
Meudt et al.1983
Nakaseko & Yoshida
1989
Nunez et al.1994
Nunez et al.1995
Labrada 1997
Liang et al.1998
Nunez et al.1998
Nunez et al.2000
Augustin 2001
Vardhini & Rao 2002

8. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-8
Brassinosteroids-Potential to improve
crop productivity
Crops
cereals
Wheat
Rice
Maiz
Barley
Compound
Brassinolide
Epibrassinoloide
Homobrassinoide
Brassinolide
Epibrassinoloide
Homobrassinoide
Biobras-6
Brassinolide
Epibrassinoide
Biobras-16
Epibrassinoloide
Response
Yield
Yield
Growth & yield
Grain Yield
Yield
Yield
Yield
Growth & yield
Growth & yield
Growth & yield
Reference
Braun and Wild 1984
Ramraj et al. 1997
Sairam 1994
Yokota and
Takashashi 1986
Ramraj et al. 1997
Franco et al.2002
Takshashi et al. 1995
Ikekawa and zhao
1991
Almenares et al 1999
Lapa et al 1998
Prusakova et al.1995

9. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-9
Brassinosteroids-Potential to improve
crop productivity
Crops
Oil Seed Groundnut
Mustard
Lens
Other crops
Tobacco
Sugercane
Sugerbeat
Cotton
Chickpea
Compound
Brassinolide
Epibrassinoloide
&
Homobrassinoide
Homobrassinoide
Homobrassinoide
Brassinolide
Biobras-6
Biobras-16
Brassinosteroids
Response
Growth & yield
Nodulation &
nitrogenase activity
Yield
Yield
Leaf yield
Cane propagation
Yield
Yield
Proteins and yield
Reference
Vardhini & Rao 2002
Hayath et al 2000 ,2001
Hayath and Ahmed
2003
Yokota and Takashashi
1986
De laFa-CF et al. 1998
Schilling et al 1991
Ramraj et al. 1997
Ramos 1997

10. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-10
Effect of BRs under abiotic & biotic stresses
Stress plant
Abiotic stress
Drought/water Sugar beat
stress
Wheat
Chick pea
Sorghum
Maiz
Mustard
Salinity stress Sugar cane
Rice
Wheat
Compound
Homobrassinoide
Homobrassinoide
Epibrassinoloide
Epibrassinoide
Epibrassinoloide
Brassinolide
Epibrassinoloide
Homobrassinoide
Brassinolide
Brassinosteroids
DAA-6
Brassinosteroids
Epibrassinoloide
Effect
increased growth &
Yield
Increased yield
Enhanced growth
& yield
Enhanced growth
increased growth
Alleviation of
stress impact on
seedling growth
Increased
tolerance
Increased oil yield
Enhanced growth
& yield
Improve tolerance
Growth recovery
Reference
Schilling et al.1991
Sairam 1994.1996
Prusakova et al.2000
Singh et al.1993
Xu et al 1994
Vardhini & Rao 2003
Li and standen 1998
Kumawat et al 1997
Gonzalez &
Gianza1997
Takeuchi 1992
Benrukova 1998

11. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-11
Effect of Brassinosteroids under
abiotic & biotic stresses
Stress plant
High temp. Wheat
Rape seed
Chilling stress tomato
Maize
Tomato
Rice
Wheat
Heavy metal Barley
stress
Compound
Brassinosteroids
Epibrassinoloide
Epibrassinoide
Epibrassinoloide
Brassinolide
Epibrassinoloide
Brassinosteroids
Brassinosteroids
Effect
Enhanced
photosynthetic rate
& nitrate
reeducates activity
Increased stress
tolerance
Increased stress
tolerance
Increased stress
tolerance
Alleviation of
stress impact on
seedling growth
Alleviation of
stress and
increased
membrane stability
Enhanced
Reference
Kuleva et al.1991
Dhaubhadel et al 1999
Dhaubhadel et al 1999
He et al 1991
Kamuro & Takasuto
1999
Xu et al 1994
Vardhini & Rao 2003
Wang and Zang 1993
Li and standen 1998
Kumawat et al 1997
Khripach et al 1999

12. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-12
Effect of Brassinosteroids under
abiotic & biotic stresses
Stress plant
Herbicides. Rice
Biotic stress Tobacco
Potato
Compound
Brassinosteroids
Brassinosteroids
containing leaf
extracts
Epibrassinoloide
Effect
Herbicides safener
Increased stress
tolerance
Enhanced
resistance against
fungal pathogens
Enhanced
resistance against
mixed fungal
pathogens
Reference
Kim et al.1993
Roth et al.2000
Volynets et al 1997

13. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-13
Unlimited potential of Brassinosteroids-
1. Inhibitors of nucleic acid synthesis (actinomycins) and protein
synthesis (cyclohexamide) were found to set aside the
epibrassinolide- stimulated growth in mungbean epicotyls .(Wu
and Zhou 1993)
2. Epibrassinolide stimulated growth of Chinese cabbage
mesophyll protoplast was associated with
enhanced protein levels .(Nakajima et al. 1996)
3. a) BRI 1 is a leucine rich repeat receptor kinase (LRRRK) known
to recognize protein ligands .( Li and Chory 1997).
b) The growth of brassinosteroid deficient dwarf mutant lkb of
Pisum sativum was restored with the exogenous application of
brassinolide .(Nomura et al. 1997)
c) Subsequently castasterone, another steroidal substance
was isolated from the insect galls of
chestnut (Castanea crenata) . As the 1st steroidal plant growth
regulator was isolated from
Brassica napus, a generic name ‘brassinosteroids’ has been
given to this new group of
phytohormones. Brassinosteroids are polyhydroxysteroids.
They have a common α- cholestane
skeleton and their structural varieties come from the kind and
the orientation in the A/B rings and side chain . (Yokou,1997)

14. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-14
Unlimited potential of
Brassinosteroids-
5. a) The studies conducted with brassinosteroid biosynthetic mutants
provided compelling evidence for the essentiality of this new group
of growth regulators for the normal growth and development. The
growth of BR-deficient dwarf mutant of Arabidopsis such as cpd, dwf I
rescued by brassinolide . (Bishop et al 1999)
b. Brassinosteroids are considered as plant hormones with pleiotropic
effects
as they influence wide array of developmental processes such as
growth,
seed germination, rhizogenesis, flowering, senescence, abscission,
and
maturation . ( Sasse 1999)
6. Inhibitor studies employing brassinozole, a specific inhibitor if
brassinolide in
plants, further strengthened the view that brassinosteroids are
essential for
growth and development in plants. (Assami et al. 2000)
7. The dwarf tomato mutant dumpy (dpy) , a dwarf with abnormal leaf
phenotype
(curled with dark ruggose leaves) with the suppression of axillary
shoots was
found rescued to wild type with the foliar application of
brassinosteroids.
( Bishop and Yokota 2001 )

15. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-15
Unlimited potential of
Brassinosteroids-
8. a) Brassinosteroids satisfy all the prerequisites to be considered
as plant hormones i.e.
natural in occurrence, mobility and activity in extremely
low concentrations . (Rao et al 2002 )
b) Rinrei, is a dwarf mutant of faba bean (Vicia faba) characterized by
dark green leaves
and reduced plant height, contributed by a single
recessive gene mutant and the gene
was tentatively identified as brassinosteroid defient dwarf
(bdd). (Fukuta et al. 2002)
c) Brassinolide, 24-epibrassinolide and 28-homobrassinolide
are the three active
brassinosteroids, being widely used in physiological
studies. Brassinosteroids have
been reported from 60 species which includes 51
angiosperms ,12 monocots and 39
dicots , 6 gymnosperms, 1 pteridophyte ( Equisetum
arvense strobillus ), 1 bryophyte
( Marchantia polymorpha ) and 1 alga ( Hydrodictyon
reticulatum ) . ( Rao et al. 2002;
Bajguz and Tretyn 2003 )
9. A) 65 brassinosteroids and 5 sugar fatty acid conjugates have

16. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-16
Distribution of BRs in Plant kingdom
Plant species Plant parts
Brassinosteroids
Monocot-
Oryza sativa Shoot
1, 4,,25
Triticum asestivum seed
11, 28, 29
Zea maiz grains
4, 8,9,10,11
Dicot-
Arabidopsis thaliana shoot
4,1
Bracssica napas pollen
1,4
Dolchios lablab seed
1,4,11,24-26,30,31
pisium sativum seed
1,4, 8,11,18
Gymnosperm-
pinus thunbergji pollen
4,8
picea sitchensis shoot
4,8
Algae-
hydrodictyon reticulatum collony

17. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-17
Arabidopsis phenotype
Figure 8. Phenotype of Arabidopsis
antiTRIP-1 transgenic lines. a) Two-
month-old α-TRIP-A (left) vs. wildtype
(right), showing the extreme dwarfism and
altered leaf morphology of the antisense
line, similar to BR-deficient and –
insensitive mutants. b) Six-month-old α-
TRIP-D showing delayed senescence and
bushy growth habit similar to the bri1
mutant. c) Two-month-old α-TRIP-C1
showing the abnormally high number of
rosette leaves and the initiation of multiple
floral meristems before the elongation of
the inflorescence. d) The arrow shows the
initiation of a new rosette of leaves and
multiple inflorescences at the terminal
point of a primary inflorescence in α-TRIP-
C1. e) Line α-TRIP-C1 initiated an
abnormally high number of floral buds

18. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-18
Phenotype of the bri1
mutant
Figure 6. Phenotype of the
bri1 mutant. Panel A
shows 2-month-old
mutant (left) and wild
type (right) plants grown
in 50 ml centrifuge tubes
in a 23 C growth
chamber (16 hr light / 8
hr dark). Panels B and
shows close-up view of
a 2-month-old bri1
mutant plant. Panel C
shows the same plant
after 4 months. All bars
represent 1 cm. Adapted

19. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-19
Dwarf Phenotype of bri1
Mutants in Different
Plant Species
Figure 3.
(A) Six-week-old wild-type and bri1-
1 mutant Arabidopsis plants.
(B) Twelve-day-old wild-type and lka
mutant pea plants.
(C) Seven-week-old wild-type and
cu-3 mutant tomato plants.
WT, wild type. Bars in (A), (B),
and enlarged inset photograph of
cu-3 in (C), 5 cm; bar in (C), 15
cm.
Plant Cell. 2002; 14(Supplement): s97–s110.
doi: 10.1105/tpc.001461.
Copyright © 2002, American Society of Plant Biologists

20. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-20
BRs on salinity induced growth inhibition of
groundnut seedlings
BRs, 24- epibrassinosteroide & 28-homobrassinosteroides were
produced from Beak consultants Inc:Brampton Ontarno, Canada
.Seeds of groundnut (Arachis hypogaea L.Var ICGS44) were
obtained from Internal crop research Institute Semi-Arid
Tropics,Patnacheru,Hyderabad.
seeds surface sterilization 0.01(W/V)inercuric chloride
were allowed to germinate at 20oc in dark in petriplates (15 cm
in dia)
Provided with sterile whatman No.1 filter papers & DW
After 72 hr.of incubation five uniform size seedlings were
transferred in to each petriplates Provided
with whatman No.1 filter papers
Each plate contained 10 ml soln of Dw ;500mM NaCl; 500mM NaCl
supplimented will

21. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-21
CONT………
This plates were kept in dark roomat20 ± 10c
on the 4th day (after transfer into the test solution)5 ml
more test soln. was
added to the petriplates .
Length & fresh wt of the sedlings were recorded on
the 7th day .
The seedlings were dried in oven at 1100c for 24 hr.&
dry wt also recorded.
Remark- The BRs not only removed the inhibitory
effect of salinity but also promoted the growth in terms of
fresh dry wt.

22. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-22
Effect of BRs on salinity induced growth inhibition of
groundnut seedlings
Compounds Length Fresh weight
Dry weight
(cm) (gm)
(gm)
Control (Dw) 7.26 ± 0.32 1.93 ± 0.06
0.40 ± 0.03
500mM Nacl+ 6.35±0.07 2.23 ± 0.07
0.52 ±0.03
1 µM Brassinolide
500mM Nacl+ 6.05±0.17 2.22 ± 0.07
0.48 ±0.01
3µM Brassinolide
500mM Nacl+ 6.28±0.21 2.76 ± 0.09
0.47 ±0.02
1µM 24- epibrassinoloide
500mM Nacl+ 5.94±0.15 2.25 ± 0.09
0.41±0.02
3µM 24- epibrassinoloide
500mM Nacl+ 6.37±0.16 2.42 ± 0.07

23. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-23
Effect of BRs on growth & yield of Tomato
The present work is designed to evaluate the influence of exogenously
applied brassinosteroides on the growth & economic yield of tomato
plants under field condition 24- epibrassinosteroide & 28-
homobrassinosteroides were produced from Beak technologies. Inc,
Brampton Canada seed of tomato Var. Pusa early dwarf were procured
from (NSC) Hyderabad.
Seed sown in 4thn week of july 2000 in 100 x 80 cm nursery beds.
20 days old plant were transplanted in expt. Plot 30 x 30cm spacing
1ml of BRs was dissolved in 1ml of ethanol & made up of a vol. 100ml with DW
From this stock soln.,0.5,1.0,& 3.0 µM dilution were prepared using DW
BRs. Soln. were applied to the plants from plants as a foliar sprayon 35th,
45th,& 55th day after sowing.
After 90th day 5 plants were removed for recording the growth in terms of
shoot & root length & root & shoot fresh wt.
On 120th day fruits were harvested for recording the fruit wt.
Yield parameters were considered 1) No. of fruits /plant
2) fresh wt of total fruits /plant.

24. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-24
Treatments control 28- homobrassinolide (3.0µM)
24-Epibrassinolide (3.0µM
Shoot length 95.33 ± 3.47 145.66 ± 1.65
144.33 ± 2.12
(cm)
Root length
(cm) 8.50 ± 0.47 17.60 ±0.52
16.60 ±0.43
No. of fruits /plant 7.00 ± 0.47 14.66 ±0.72
13.66 ±0.72
Total fruit wt /plant 86.66 ± 4.32 220.66 ±6.69
189.0 ±4.58
• Among the three 28- homobrassinoides was most effective in accounting
increase the yield.
• Present study revealed the ability of BRsin improving the yield of tomato under
field condition.
• Similar result have also observed in Wheat, (Sairam 1994),Grapes( Xu et
al.1994)under field condition.
Effect of BRs on growth & yield of tomato

25. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-25
Future prospectus
• Blending of two traits –Growth promotion & stress tolerance will
have great economic bearing in future agril.
Crop production.
for
• The entry of reputed agrochemical comp.in commercial prodn. of
BRs & hence it’s potentiality of improving crop yield can soon be
visualized.
• The physiological & molecular mechanism of the action of
brassinosteroids in enhancing productivity & improving
resistance to environmental stress, that’s why it needs further
elucidation.
• Further research may also bring into focus many more significant
roles to the this group of steroids.
• New discoveries in this field may allow us to employ BRs as-
Highly promising field.
Environment friendly natural substance.
Wide application in plant protection & yield enhancement.
• Availability of BRs at affordable prices will be cost effective.
• Expected that chemical promise held by BRs to boost crop
production will soon be accomplished.

26. Copyright © 2006 by South-Western, a division of Thomson Learning, Inc. All rights reserved. 16-26
nternational journal of plant physio