Plant Physiology 

Assignment
Topic – Plant Hormone 
        (Brassinosteroids)

                         By.
                         Ishika Sharma
(7067)
                         Nandini Killa (7019)
Introduction
Historical aspects
Occurrence
Chemical Structure
Biosynthesis
Signal Transduction
Physiological Roles
Commercial Applications
Future Prospects
References
 INTRODUCTION
Brassinosteroids defined as the sixth group of  plant hormone after
the classic plant hormones auxin, gibberellins, cytokinin, abscisic acid
and ethylene.
Brassinosteroids are a class of polyhydroxylated steroidal,
endogenous phytohormones in plants with similar structures to
animals steroid hormones.
Brassinosteroids regulate a wide range of physiological processes
including plant growth, development and immunity.

Brassinosteroids have been found in varius species of plants,

including monoplast freshwater algae and brown algae, indicating
that BRs are a widespread ancient plant hormone.
These compound were first isolated in 1979 from bee collected pollen
grain of rape (Brassica napus) .

More than 60 brassinosteroids have so far been discovered from

different plant part such as pollens, seeds, leaves, stems, roots and
flower.
Source:
https://www.sciencephoto.com/media/180469/view/oilseed-rape-brassica-napus-
 HISTORICAL ASPECTS
• The first possible existence of Brassinosteroids was reported
in Distylium racemosum (Isunoki) in Japan in 1968.

• Later it was found in an oil fraction from rape pollen

Brassica napus L. where it reported to have a function in
plant growth activity. This fraction was named “brassins”
and speculations were made that it might turn out to be
another plant hormone.

• A bioassay conducted on second-internode of beans

revealed that brassins induced its elongation at an average
of 155 mm at a dose of 10 μg per plant, whereas untreated
ones grew only 12 mm.
Source: https://www.mdpi.com/1422-0067/21/5/1743/htm
HISTORICAL ASPECTS
• Initially brassins were thought to be fatty acids, it was only in 1994 after X-Ray
Crystallographic studies of bee collected rape pollen from Brassica napus L. that
isolation of a few crystals of a new compound was possible.

• X-Ray Crystallographic studies finally revealed that the isolated compound was a
steroidal lactone with the structure of brassinolide.

• The discovery of brassinolide prompted intensive studies on the isolation and

identification of new members of this group. Till date, over 60 BS have been
isolated and identified in plants.
OCCURENCE OF BRASSINOSTEROIDS
• Since the discovery of brassinolide (BL), 70 BRs (65 unconjugated and 5 conjugated) have been
isolated from 60 plant species including 51 angiosperms (12 monocotyledons and 39
dicotyledons), 6 gymnosperms, 1 pteridophyte (Equisetum arvense), 1 bryophyte (Marchantia
polymorpha) and 1 chlorophyte, the alga (Hydrodictyon reticulatum).

• BRs are widely distributed in the plant kingdom, including higher and lower plants. BRs have
been detected in all the plant organs such as pollen, anthers, seeds, leaves, stems, roots,
flowers, and grain. The galls of Castanea crenata and Distylium racemosum have higher levels
of BRs (several mg/kg) than the normal, healthy tissues.

• Another tissue having BRs is the crown gall cells of Catharanthus roseus which have higher
contents of BL and castasterone (CS) than the normal cells (30-40 mg/kg).

• Also, young growing tissues contain higher levels of BRs than mature tissues. Pollen and
immature seeds are the richest sources with a range of 1-100 ng g-1 fresh weight, while
shoots and leaves usually have lower level of 0.01-0.1 ng g-1 fresh weight.
CHEMICAL STRUCTURE OF BRASSINOSTEROIDS
• Based on the total number of carbons, BRs are divided into
C27, C28, and C29-type. The basic structure of C27-BRs is a
5α-cholestane skeleton, C28-BRs: 5α-ergostane, and C29-
BRs: 5α-stigmastane.
• Differences in the structure of these hormones are due to
the type and orientation of oxygenated functions in the A-
and B-ring, as well as the number and position of functional
groups in the side chain of the molecule. https://www.sciencedirect.com/topics/biochemistry-genetics-and-molec
ular-biology/brassinolide
• These modifications arise during oxidation and reduction
reactions. Based on the cholesterol (CR) side chain, BRs are
divided by different substituents into C-23, C-24, C-25, 23-
oxo, 24S-methyl, 24R-methyl, 24-methylene, 24S-ethyl, 24-
ethylidene, 24-methylene-25-methyl, 24-methyl-25-methyl;
without substituent at C-23, without substituent at C-24,
and without substituents at C-23, C-24. BRs can also
conjugate with glucose and fatty acids.
https://bmcbiol.biomedcentral.com/articles/10.1186/s12915-016-0340-8
BIOSYNTHESIS OF BRASSINOSTEROIDS
• Brassinosteroids are biosynthesized from a C₂8
plant sterol called campesterol by reductive
step followed by several oxidative steps.

• The biosynthesis of brassinolide from

campesterol requires approximately 12 steps.
The reductive step and one of the several
oxidative steps are catalysed by enzymes CYP
and CPD (Carboxypeptidase D) respectively.
Campesterol is in turn derived from a triterpene
called squalene.

• Experiments has shown that long distance

transport is possible and that the flow is
from the base to the tips (acropetal).
Source: https://www.scielo.br/j/bjpp/a/jt9wVShx5fgHCFbxhBhRFPn/?lang=en
 SIGNAL TRANSDUCTION
• BRs are perceived at the cell membrane by a co-
receptor complex, comprising brassinosteroid
insensitive-1 (BRI1) and BRI1 associated receptor
kinase 1 (BAK1). BRI1 acts as a kinase, but in the
absence of BR its action is inhibited. by another
protein, BRI1 kinase. inhibitor 1 (BKI1).
• When BR binds to the BRI1:BAK1 complex, BKI1 is
Negative
released, and a phosphorylation cascade is triggered Regulator
which results in the de-activation of another kinase,
brassinosteroid insensitive 2 (BIN2). BES 1

• BIN2 and its close homologues inhibit several

transcription factors. The inhibition of BIN2 by BSU1
releases these transcription factors to bind to DNA
and to enact certain developmental pathways.
Source: https://en.wikipedia.org/wiki/Brassinosteroid
 PHYSIOLOGICAL
ROLES 
BRs have been shown to be involved in numerous plant processes:

Promotion of cell expansion and cell elongation.

It has an unclear role in cell division and cell wall regeneration.

Promotion of vascular differentiation.

Is necessary for pollen elongation for pollen tube formation.

Acceleration of senescence in dying tissue cultured cells.

Can provide some protection to plants during chilling and drought
stress.
Inhibit root growth.

Enhance gravitropism.

Source: https://www.researchgate.net/figure/Effects-of-exogenous-nitrate-on-Parasponia-andersonii-plants-inoculated-with_fig1_324528068
 ROLE IN FLOWERING
https://www.nature-and-garden.com/gardening/strawberry-bush.html

• There has been very limited use of

steroids in regulating flowering. The
number of flowers in strawberry
increased by the application of BRs
at the foliage. However, in case of
grapes, the application of BRs in
autumn improved the number of
flowers but inhibited if the time of
application is delayed to late winter.

https://www.jordanwinery.com/blog/lifecycle-vineyard-grape-flowers/
 ROLE IN SENESCENCE
• Senescence is the process, which refers to, endogenously
regulated deteriorative, changes that become the natural
cause of death of cells, tissues, organs or that of the whole
organism.

• BRs also play a crucial role in regulating the processes

leading to senescence. The brassinolide promotes
senescence in Xanthium and Rumex explants. In addition to
it, BRs also accelerate senescence in leaves of mung bean
seedlings.

• However, brassinosteroid deficient Arabidopsis mutants

exhibited delayed senescence of chloroplast. Similarly, the
senescence of the leaves of mung bean and mustard was
delayed, if supplied with 28-homobrassinolide at early stage
of growth due to chlorophyll biosynthesis. Source: https://en.wikipedia.org/wiki/Plant_senescence
 ROLE IN PHOTOSYNTHESIS
• The aqueous solution of 28-homobrassinolide, applied to the foliage of wheat and mustard
or applied as seed soaking treatment to mungbean, Lycopersicon esculentum, Raphanus
sativus and dialkylaminoethylalkanoate or 24-epibrassinolide, in association with GA3 , to
spinach enhanced the photosynthetic rate.

• Application of 24-epibrassinolide and 28-homobrassinolide through root was also reported

to increase the net photosynthetic rate in Cucumis sativus and in Lycopersicon esculentum.
Foliar spray of aqueous solution of BR to wheat and mustard, 24-epibrassinolide to seedlings
of cucumber and brassinolide to rice increased the rate of CO2 assimilation.

• Likewise, the foliar application of 24-epibrassinolide enhanced the light saturated net CO 2
assimilation rate and carboxylation rate of rubisco, thereby increasing the capacity of CO 2
assimilation in the Calvin cycle.
 EFFECT ON CHLOROPHYLL CONTENT
• The total chlorophyll contents or its fractions increased in the
leaves of Vigna radiata and Brassica juncea by 28-
homobrassinolide and in Cucumis sativus by 24-
epibrassinolide, applied directly to their foliage. Similarly, the
values for the above parameters increased in the leaves of
rice, Brassica juncea.

• Vigna radiata, Raphanus sativus and Lycopersicon esculentum

raised from the seeds given presowing treatment with 28-
homobrassinolide.

• Moreover, the water stressed wheat plants treated with 28-

homobrassinolide possessed high chlorophyll level.
Source: https://in.pinterest.com/pin/745556913313259686/
 DIFFERENCIATION OF VASCULAR TISSUE
• The first report of a role for BRs in the differentiation of vascular tissues came in 1991.
Jerusalem artichoke (Helianthus tuberosus) cells transferred to the xylem differentiation
medium, in the presence of auxin and cytokinins will differentiate into xylem elements in 72 -
96 hours. Very few vascular elements develop in the first 24 hours following transfer into this
medium.

• However, nanomolar concentrations of BL included in the medium, resulted in a 10-fold

increase in xylem differentiation, this was observed in the first 24 hours. Also, significant
increases in cell numbers were observed, indicating a role for BRs in cell division and
differentiation. Homobrassinolide are also reported to increase the cell division in barley.

• BRs have been implicated in the transition between Stage II, and Stage III in the formation of
xylem/tracheary elements in Zinnia elegans, where secondary wall formation and cell death
occurs.
 COMMERCIAL
APPLICATIONS
Effects on Fruit Ripening

Effects on Biochemical Attribute

Effects on alleviation of Chilling Injury

Effects on Enzymatic Activity

Effects on Post-harvest Disease Management

Effects on Shelf-Life

Improvement in Fruit colour

Source: https://www.britannica.com/science/horticulture
 FRUIT-RIPENING
• Fruit ripening is a dramatically important physiological process which leads to the
development of physiochemical attributes for market need. Ripening process is
regulated by different hormone mediated pathways specially ethylene in climacteric
fruits.

• During ripening a series of quality changes takes place. Recently several researchers
observed that brassinosteroids also regulate the ripening process in climacteric and
non-climacteric fruits by altering ethylene related enzymes and ripening genes.

• Symons et al. (2006) conducted a study on grape berry and observed that BRs
treatment regulate the ripening process in non-climacteric fruits. In particular, they
found that BRs treated grape showed rapid veraison process.

• Exogenous application of brassinosteroids (45 and 60 ng g-1 FW) enhanced ethylene

biosynthesis thus promotes ripening in ‘Kingston Pride’ mango.
 BIOCHEMICAL ATTRIBUTES
• The quality of fruits involves TSS, acidity, pigments, sugar
contents, size and weight of fruit etc. Some previous data
showed that exogenous application of brassinosteroids
and its analogues regulate different quality attributes in
different fruits and vegetables.
• Exogenous application of BRs as aqueous solution
considerably enhances grape cluster berry weight,
length, and diameter. Even under cold storage, BRs help
in maintaining skin colour and reducing the rate of TSS
degradation.
• BRs treated litchi tree, produced fruits with higher
calcium content in pericarp and less cracking. Foliar
applications of BR also increases skin colour of fruit by
increasing anthocyanin content, organic acids, ascorbic
Source: https://www.flipkart.com/trothic-litchi-plant/p/itme3ba418c150b acid, and phenol content.
 ALLEVIATION OF CHILLING INJURY
• The potential application of BRs in agriculture to improve growth and yield under various stress
conditions is of immense significance as these stresses severely hamper the normal metabolism
of plants.

• A molecular model that shows the BRs participation in regulation of freezing tolerance was
presented by Ereminaet et al. in 2016. Cold stress is an influential environmental factor that
affects plant distribution and can strongly limit crop productivity. Plants have evolved
sophisticated signaling cascades that permit them to survive chilling or even freezing
temperatures.

• Chilling injury or cold injury is one of the major physiological disorders of several tropical and
subtropical fruits, as such fruits are more sensitive to low temperature storage conditions and
spoiled quickly affecting its quality.

• EBR (24- epibrassinolide) treatments was observed to augment osmo-regulation of material and
amount of antioxidant enzymes viz, superoxide dismutase (SOD), peroxidase (POD), catalase
(CAT) and ascorbate peroxidase in 72 juvenile grapevines, whereas decreased the damage caused
by reactive oxygen species (ROS) and lipid peroxidation.
 REGULATES ENZYMATIC ACTIVITY
• Enzymes are the key factors in quality management of fresh
produce. Each enzyme plays an important role to furnish
physiological process.
• BR treated peach fruits exhibited higher shikimate
dehydrogenase, phenylalanine ammonia lyase, cinnamate-
4-hydroxy-lase and 4-coumarate: coenzyme A ligase activity,
whereas reduced polyphenol oxidase and peroxidase
https://www.amazon.in/Amazing-Store-Grafted-Healthy-Outdoor/dp/B
activity.
084YV814M

• BRs treatment had induced the activity of pectinolytic

enzymes by rising pectin metabolism whereas it
inhibited cellulose enzyme activity in litchi fruits. EBR
treatment (10 µg) exogenously applied on eggplants
showed reduced activity of browning enzymes PPO
around 37% than the control fruits during storage.
https://www.indiamart.com/proddetail/fresh-brinjal-plant-14488046662.html
 POSTHARVEST DISEASE MANAGEMENT
• Fruits and vegetables are highly perishable commodity and highly susceptible to
fungal and bacterial diseases due to high moisture content. These pathogens
sometime cause huge losses due to severe disease development. Produce becomes
unacceptable and remains unfit for marketing. However different previous findings
elucidated the mechanism of brassinosteroids in controlling disease development
and pathogens.

• In jujube, EBR likely stimulated defense-related enzymes (PAL, PPO, Catalase, SOD
etc.) which increases resistance against fungus pathogen of blue mould rot (P.
expansum). Better postharvest quality and less incidence of Botrytis rot in grape
berries was observed on treatment with 0.8mg.L-1epibrassinolides.

• It was suggested that EBR reduced postharvest disease incidence in Satsuma

mandarin fruit that might attributed to accumulation of H2O2 , stress-related
metabolites and the induction of stress-related genes.
 INCREASE IN SHELF LIFE
• Shelf life is an important aspect which depends on
respiratory process of the fruits and vegetables. Shelf life
of the fruits and vegetables can be extended by
manipulating respiration which is affected by the
ethylene and BRs.
• It was reported that pre-harvest application of BR 0.75
mg and postharvest application of BR 0.2  mg strongly
increased the shelf  life  of  sweet cherry compared to
other treatments.
• It was observed that postharvest brassinolide treatment
helps in inducing colour and storage life of white button
mushroom. It is reported that action of BRs is strongly
dependent on concentration. Its postharvest application
delays the ethylene biosynthesis process resulting in
delayed senescence in jujube fruits.
Source:
https://tribune.com.pk/story/970615/5-fruits-and-vegetables-that-ma
y-make-you-gain-weight
 ENHANCEMENT IN FRUIT COLOUR
• Red table grapes may fail to develop adequate berry
colour. However, little is known about the possibility of
brassinosteroids (BRs) to improve berry colour. Thus, a
commercial BR formulation was tested in two commercial
vineyards of Red globe and Crimson Seedless in order to
evaluate its effectiveness for improving berry colour.
• The commercial formulation tested improved the colour
of Red globe berries and maintained the colour in the
resulting larger berries of Crimson Seedless.
• This study supports the potential of BRs as a tool to
improve berry colour in a commercial context, laying the
foundations for future work and incorporation of BRs
within table grape management programs. Source:
https://www.amazon.in/Creative-Farmer-Exotic-Garden-Healthy/dp/B07
P4X9QD2
 FUTURE PROSPECTS
• Thirty five years of research, on BRs has brought into
light several vital functions of this class of
phytohormones in the regulation of plant growth,
development and productivity. Further progress in
the investigation of mechanism of BRs action in
plants, on the one hand, and elaboration of
economically feasible schemes of synthesis of natural
BRs and their analogs, on the other hand, will surely
make a basis for the inclusion of this new class of
plant hormones in the regular package of chemicals
used for optimizing agricultural production.
Hopefully, as the research will progress, much more
knowledge will be added to the present literature.
Source: https://medium.com/@hydrogenapi/an-exciting-road-ahead-in-2018-f7d9600a57a4
 REFERENCES
 Hayat, S. H. A. M. S. U. L., Mori, M., Fariduddin, Q. A. Z. I., Bajguz, A. N. D. R. Z. E. J., & Ahmad, A.
(2010). Physiological role of brassinosteroids: an update. Indian J Plant Physiol, 15, 99-109.​
 Adam, G., & Marquardt, V. (1986). Brassinosteroids. Phytochemistry, 25(8), 1787-1799.​
 Clouse, S. D., & Sasse, J. M. (1998). Brassinosteroids: essential regulators of plant growth and
development. Annual review of plant biology, 49(1), 427-451.​
 Bajguz, A., & Tretyn, A. (2003). The chemical characteristic and distribution of brassinosteroids in
plants. Phytochemistry, 62(7), 1027-1046.​
 Meena, N. K., Asrey, R., Singh, J., Parajapati, U., Chaudhary, K., & Mani, A. (2018).
Effects of brassinosteroids application on quality and storage of fruits. In Trends & prospects in post
harvest management of horticultural crops (pp. 65-79). Today & Tomorrow’s Printers and Publishers.​
 Vergara, A., Torrealba, M., Alcalde, J. A., & Pérez‐Donoso, A. G. (2020).
Commercial brassinosteroid increases the concentration of anthocyanin in
red tablegrape cultivars (Vitis vinifera L.). Australian Journal of Grape and Wine Research, 26(4), 427-
433.
 Tang, J., Han, Z., & Chai, J. (2016). Q&A: what are brassinosteroids and how do they act in
plants?. BMC biology, 14(1), 1-5.