GROWTH REGULATION ASPECTS

OF
PROPAGATION,
EMBRYOGENSIS,
SEED AND BUD DORMANCY

SUBMITTED BY SUBMITTED TO
VIKAS RAMTEKE DR. S.J. PATIL
PH.D. 1ST SEMESTER I/C PROFESSOR AND HEAD
DEPT. OF FRUIT SCIENCE DEPT. OF FRUIT SCIENCE
ACHF, NAU, NAVSARI ACHF, NAU, NAVSARI
Propagation
AUXINS
Root initiation
 In contrast to stem, the higher concentration of auxin inhibits the

elongation of roots.

 But the number of lateral roots is considerably increased i.e., higher

concentration of auxin induces more lateral branch roots.

 Application of IAA in lanolin paste (lanolin is a soft fat prepared from

wool and is good solvent for auxin) to the cut end of a young stem

results in an early and extensive rooting.

 This fact is of great practical importance and has been widely utilized to

promote root formation in economically useful plants which are

propagated by cuttings.
Callus formation
 Besides cell elongation, auxin may also be active in cell
division.
 In many tissue cultures, where the callus growth is quite
normal, the continued growth of such callus takes place
only after the addition of auxin.
 Commonly used synthetic auxins in tissue culture are 2,4-
dichlorophenoxyacetic acid (2,4-D; often used for callus
induction and suspension cultures), and 1-
naphthaleneacetic acid (NAA; when organogenesis is
required).
 Plant tissue culture
 Within plant tissues, IAA and other naturally occurring auxins
combine with small molecules (alcohols, amino acids, sugars)
to produce ester, amide, or glycoside conjugates (Bandurski
et al., 1995).
 This appears to be a mechanism for storing auxins in cells
and stabilizing the level of free auxin by metabolizing the
excess.
 Auxin in conjugated molecules is protected from oxidative
breakdown and may be later enzymatically released when
required.
Gibberellins
Seed germination

 Certain light sensitive seeds eg. Lettuce and tobacco show

poor germination in dark.
 Germination starts vigorously if these seeds are exposed
to light or red light.
 This requirement of light is overcome if the seeds are
treated with gibberellic acid in dark.
 It has been suggested that there are related or common
receptors for dormancy breaking agents within the plasma
membrane of the responsive embryonic cells.
 When triggered, these receptors then initiate a signal
transduction cascade, perhaps involving synthesis of or
sensitization to germinationpromoting gibberellins that lead
to the completion of germination.
 Changes in the phosphorylating activity of membrane-
associated, Ca+2-dependent protein kinases that lead to
dormancy or germination have been proposed as well.
 Micropropagation

 When GAs are added to plant tissue culture media, they

often diminish or prevent the formation of roots, shoots, or
somatic embryos, although the opposite has also been
seen.
 Some endogenous GAs may be necessary for normal
callus growth (Lance et al., 1976), and inhibition of GA
biosynthesis can influence development of cells in liquid
cultures (Ziv and Ariel, 1991).
 Some differences in the inhibition or promotion of
adventitious root and shoot formation by GAs might be
due to the fact that GAs inhibit meristemoid initiation
(Thorpe and Murashige, 1970) but are required for
assisting the further growth and development of preformed
organs.
 GAs may also alter the availability of endogenous auxin.
Growth of shoots in meristem and shoot cultures may be
enhanced by addition of GA (Fry and Street, 1980).
CYTOKININS (Kinetin)
In tissue culture, cytokinins play a crucial role as
promoters of cell division and act in the induction and
development of meristematic centers leading to the
formation of organs, mainly shoots (Peeters et al.,
1991).
 Kinetin (KIN) has been applied for micropropagation of many

ornamental plants (Jain and Ochatt, 2010). In vitro shoot

proliferation and multiplication are largely based on media

formulations containing cytokinins as a major plant growth

regulators (Mamidala and Nanna, 2009; Hoque, 2010).

 Some reports indicated that the presence of cytokinin in the

culture medium helped in the multiplication of shoots (Kumar

et al., 2001).
 Rooting can be initiated by transferring the regenerated

shoots to a medium containing only auxins.

 The best explants for micropropagation of most plants,

without any intervening callus phase, are shoot tips and

axillary buds (Pati et al., 2005; Hashemabadi and

Kaviani, 2010).
Seed and bud dormancy
 During the developmental cycle of the plant, at some
phase or the other certain structures like buds, tubers,
seeds, etc., go through a period of temporary suspension
of growth activity.  Such a state is called dormancy.
 Generally plants or plant structures, in order to overcome
or survive against hostile environmental conditions
undergo a period of dormancy with suitable
modifications. 
Mechanism of Induction of Dormancy in
Buds:
 The onset of short day or long dark photoperiods in
winter stimulates the synthesis of various growth
inhibiting compounds of which Abscissin dominates.
 Abscissin is a well known growth inhibiting hormone. 
By inhibiting the synthesis of proteins, RNA and other
metabolic processes, ABA imposes dormancy on
meristematic tissues of the plant body.
 Breaking Bud Dormancy:
 Cytokinins are known to be synthesized in root tips but
under cold conditions because of the snow fall, the root
meristems are very inactive and they don’t synthesize
sufficient quantities of cytokinin required for the buds to
be active. 
 That is probably one of the reasons why buds remain
dormant. 
 As soon as cytokinins are provided to dormant buds,
mitotic activity is initiated and buds start sprouting. 
Gibberellins
 Another class of phytohormones, which overcomes the bud

dormancy, is Gibberellins. 

 Now it is certain that Gibberellin synthesis takes place in

plastids.  Moreover, the synthesis of GA and ABA starts from

the same precursor called mevolonate. 

 Under short day conditions, the pathway from mevolonate is

directed towards ABA synthesis and GA synthesis is inhibited,

but during long day photoperiods it is directed towards GA

synthesis and ABA synthesis is blocked. 

 That is the reason why gibberellins under long day
conditions or not light treatment, break bud dormancy and
nullify the effect of ABA present in such dormant buds.
 Model for the regulation of dormancy and germination by
ABA and GA in response to the environment;
 According to this model ambient environmental factors
(e.g. temperature) affect the ABA/GA balance and the
sensitivity to these hormones.
 ABA synthesis and signalling (GAcatabolism) dominates
the dormant state, whereas, GA synthesis and signalling
(ABA catabolism) dominates the transition to germination.   
Somatic embryogensis
The effect of PGR is an important
factor impacting SE and plant
regeneration. In most cases,
successful plant SE needs a mixture of
the different concentration ratios of
auxin and cytokinin (CTK), both of
which are necessary for plant culture
in vitro.
Auxins
 In Angiosperm monocots, primary embryogenesis was
exclusively induced by auxin-supplemented media,
there is a large variation of growth regulators used to
induce somatic embryogenesis in dicot species.
 From a list of 65 dicot species reviewed by
Raemakers et al. (1995), somatic embryogenesis was
induced in 17 species on hormone-free media, in 29
species on auxin-containing media and in 25 species on
cytokinin-supplemented media.
 Among auxins, the most frequently used was 2,4-D (49%)
followed by naphthalene acetic acid (27%), indole-3-acetic
acid (IAA) (6%), indole-3-butyric acid (6%), Picloram (5%) and
Dicamba (5%).
 However, not all plants requires 2,4-D for SE.
 IAA is necessary to induce SE of Begonia cathayana Hemsl.
 Picloram would induce the SE of banana rather than 2,4-D.
 The somatic embryo of Gladiolus could be induced in media
containing NAA but not in media containing other PGR.
Gibberellins
 Despite the wide range of physiological effects of gibberellins,
their effect when added to culture media, primarily as
gibberellic acid (GA3), has only been minimal (Krikorian, 1995).

 Exogenous application of GA3 has been reported to inhibit

somatic embryogenesis and somatic embryo development in
several species; but it was also reported that this substance is
required for germination of the mature somatic embryos if
chilling is not applied provided (Takeno et al., 1983 and
references therein).
Gibberellins

 Gibberellic acid (GA) and Abscisic acid (ABA) are not

necessary for the induction of SE.
 GA has been shown to promote SE for Apple, Pear,
Garnetberry, and Cherry and also plays an inhibition role
in Citrus SE.
Cytokinins
 The reasonable ratio of CTK and auxin is one of the
major factors to induce horticultural plant SE.
 The high efficiency of SE in poinsettia depends on the
ratio of CTK to auxin.
 The high ratio of auxin to CTK would promote the SE of
American chestnut.
 Whereas a high ratio of CTK to auxin would promote
apple SE.
 It is a requirement of two types of auxin and CTK to
induce direct SE for Chrysanthemum morifolium ,
 But walnut and peach require two types of CTK and IAA
to induce SE.
Absassic acid
 ABA has promoting or inhibitory effects depending on the
specific plant. Wei et al. had shown that ABA inhibited the SE
of banana embryogenic suspension callus (ESC), and the
extent of callus increased with increasing ABA concentration
in the media.
 Different plants have specific requirements for ABA
concentration. For example, increased concentrations of ABA
(100 mmol·L–1) was required for high quality somatic
embryos of cherry to induce formation.However, the SE of
Papaya would form in medium containing only ABA.
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