Biotechnological
Communication
Biosci. Biotech. Res. Comm. 11(2): 208-215 (2018)
Comparative analysis of plant growth regulators on
bud break in
Prosopsis
and
Tecomella
for sustainable
agriculture in arid and semi-arid regions of India
Sunita Khatak*, Archit Sharma and Rashi Saini
Department of Biotechnology, University Institute of Engineering and Technology, Kurukshetra University,
Kurukshetra, Haryana, India
ABSTRACT
Prosopsis cineraria commonly known as Khejri and Tecomella undulata known as Rohida are Golden Trees of Thar
Desert in India belonging to the family Fabaceae and Begnoniaceae respectively. Both the plants are multipurpose tree
whose almost all parts are used in pharmaceutical industry for preparing medicines. The medicinal uses of this plant
has necessitated large scale production and as raw material to medicinal industry, leading to its over-exploitation.
Both the trees are important component of desert Ecosystem of India as biomass producer and enrich desert soil, x
atmospheric nitrogen and provide a green coverage. Both contribute to ecological stability of the region and provid-
ing extensive support to human beings, livestock and the nutrient de cient soils. The plant tissue culture techniques
can play an important role in propagation and qualitative improvement of plants of medicinal aspects. Axilary nodes,
and shoot tips were aseptically cultured on MS basal medium forti ed with different concentrations of cytokinins
(BAP, Kinetin) and auxins (NAA, IAA,IBA) along with sucrose as energy source. The plant growth regulators act in
synergistic way to proliferate shoots of almost 1-5cm lengths which were rooted on rooting media to regenerate the
whole plant. In vitro developed complete seedlings were acclimatized and propagated in vitro for mass cultivation.
KEY WORDS: BUD BREAK; BAP; MICRO PROPAGATION;
TECOMELLA, PROSOPSIS
208
ARTICLE INFORMATION:
*Corresponding Author: sunkhatak@rediffmail.com
Received 10
th
March, 2018
Accepted after revision 11
th
June, 2018
BBRC Print ISSN: 0974-6455
Online ISSN: 2321-4007 CODEN: USA BBRCBA
Thomson Reuters ISI ESC / Clarivate Analytics USA and
Crossref Indexed Journal
NAAS Journal Score 2018: 4.31 SJIF 2017: 4.196
© A Society of Science and Nature Publication, Bhopal India
2018. All rights reserved.
Online Contents Available at: http//www.bbrc.in/
DOI: 10.21786/bbrc/11.1/3
Sunita Khatak, Archit Sharma and Rashi Saini
INTRODUCTION
Woody trees are vital to arid environments because of
their ecofriendly and multipurpose nature, and the fact
that they are well able to tolerate drought situations.
Prosopis and Tecomella are the principal genera in these
regions, and both have great biological diversity and
ecological plasticity. These are used worldwide in arid
regions to improve the local economy. These tree species
are biologically diverse and are well adapted to stress
as a result of multiple interbreeding species. Legumi-
nous tree products are economically important sources
of food, fodder, rewood, and timber. Improvement in
quality attributes through selection, modi cation and
mass production of germplasm is desirable. Propagation
through seeds is the most common practice for raising
quality trait seedlings for new plantations in arid areas.
such as cuttings, suckers, air layering and tissue cultures
are available but more efforts towards their re nement
are still required, particularly with regards nursery and
laboratory techniques, before commercial cultivation
(Baksha et al., 2007; Biswas et al., 2009; Roy, 2008).
Plant cell tissue culture has offered a very novel tech-
nique to mass multiply, true to type and providing dis-
ease resistant plants in controlled conditions.
Prosopis cineraria commonly known as khejri belongs
to pea family, Fabaceae and is a multipurpose tree of
desert in Western Rajasthan. It is also called kalptaru,
‘wonder tree’ and the ‘king of desert. (Singh et al. 2013,
Tarachand et. al. 2012). It is native to arid portions of
Western and the Indian subcontinent, including Afghan-
istan, Iran, India, Oman, Pakistan, Saudi Arabia, the
United Arab Emirates, and Yemen. In India it is found
in the various parts of Rajasthan, Gujarat, Haryana,
Uttar Pradesh and Tamil Nadu (Rathore et al., 1991).
It is regarded as a backbone of rural economy being a
good biomass producer and xes atmospheric nitrogen
and provides a green coverage and in turn helps in the
enrichment of desert soil. It contributes to ecological
stability of the region and providing extensive support
to human beings, livestock and the nutrient de cient
soils (Chaudhry 2011, Panwar et al., 2014 and Hua et al.
(2015).
The tree is well adapted to arid and semi-arid condi-
tions of the Indian desert, due to their well-developed
and expansive tap root system which reach up to a
length of 20 m, often reaching out the ground water
resources (Gehlot et al., 2008). Pods of this plant locally
called “Sangri” are considered as dry fruits of desert.
Pods contain various phytoconstituents like tannins
(gallic acid), steroids (stigma sterol, campestral, sitos-
terol, etc.), Flavones derivatives (prosogerin A, B, C, D,
and E), alkaloids (spicigerine, prosophylline), etc. have
been isolated from the sangri pods (Gehlot et al., 2008).
The ashes of bark are rubbed over the skin to remove
hair. Fresh Leaves juice mixed with lemon juice is used
for dyspepsia; extract of crushed pods is used for ear-
ache, toothache, pain relief from fractured bones (Garg
and Mittal 2013).
The whole plant is used in the Indigenous System of
Medicine as a folk remedy for various ailments like lep-
rosy, dysentery, bronchitis, asthma, leucoderma, piles,
muscular tremor and wandering of the mind. It is also
known to possess anthelmintic, antibacterial, antifungal,
antiviral and anticancer activities. Tecomella undulata
belongs to family Begnoniaceae is an important medici-
nal plant. Wide range of therapeutic activities has been
attributed to this plant. The plant is excellent blood puri-
er hence rewarding in hepatitis. Bark forms a major
constituents of various herbal formulations like Livo-
plus, Liv-52, Livosan,Herboliv, Amylcure for curing
in ammatory hepatic disease. The leaves have oleanolic
acid, ursolic acid and betulinic acid which are strong
prohibitors of HIV (Nandwani et al., 1995; 1996). It
has been reported to be used in phyto remediation of
soil contaminated with crude petroleum oil its common
agro forestry tree in arid and semi arid regions used for
research, including bio fertilizer aspect as well as affor-
estation programmes (Rathore et al., 1991 and Shekha-
wat et al., 1993).
The plant is propagated mainly using seeds and the
tree is slow growing species. Till date the woody plants
lack suitable methods for vegetative propagation on
mass scale. Seeds are on prime importance for extensive
plantation. The seeds are available from month of April
to June. Freshly harvested seeds have more potential for
germination as compared to unripe fruits or ripe fruits
collected in June. So present investigation was under-
taken with an objective to devise one suitable protocol
for bud initiation and plant propagation using nodal tis-
sues in both Tecomella and Prosopsis (Bhansali, 1993).
MATERIAL AND METHODS
The seedlings of Prosopsis and Tecomella were used in
the present study for bud break and shoot prolifera-
tion, which were procured from Tau Devi Lal Herbal
Park, Near Khizrabaad Highway, Yamunanagar District,
Haryana, India. Axillary nodes were selected as explants
to avoid genetic alterations and somaclonal variations
observed using indirect regeneration. The disease free
axillary buds were collected from 4 weeks old healthy
plant. The explants were excised and the contaminants
were washed under running tap water for 4-5 minutes,
followed by washing in the liquid detergent Tween -20
(few drops / 100 ml solution) and then rinsed with run-
ning tap water. The cleaned explants were surface steri-
BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS 209
Sunita Khatak, Archit Sharma and Rashi Saini
lized with variable concentration of mercuric chloride
(0.1%-0.5%) under laminar air ow. The explants were
subjected to different time intervals to optimize the
sterilization procedure and then rinsed 4-5 times with
sterilized distilled water. After trimming the cut ends,
equal sized, surface sterilized explants were cultured on
the culture medium de ned by Murashige and Skoog,
(1962). Among all experiments, a treatment of (0.1%
and 0.3%) HgCl
2
for 3 minutes proved to be the best for
sterilization in Prososopsis and Tecomella, a large per-
centage (85%) of explants. The surface sterilization was
optimized and this helped in preventing blackening of
tissue on exposure to mercuric chloride and establish-
ment of clean cultures.
Axillary nodes were excised from the young seed-
lings grown in vivo conditions and were cultured on
different shoot proliferation media (Table 1,2) consist-
ing of different concentration and combination of cyto-
kinin and auxin as plant growth regulator. Axillary node
explants were implanted vertically. Test tubes and ask
containing the explants were sealed with sterilized cot-
ton plugs and incubated for 4 weeks at 25±2C under
a 16 hour photoperiod. Radiation source was supplied
by soft white uorescence tubes. For each treatment, a
minimum of 3 replicates were carried out. Percent shoot
proliferation and multiple shoot formation was recorded
after four weeks of culture.
RESULTS AND DISCUSSION
The bud break, shoot proliferation and exploring the
potential for multiple shoots regeneration in Prosopsis
and Tecomella is an utmost requirement for propagation
of this plant.Sterilization of explants is a crucial step in
plant tissue culture and to achieve 100 percent steriliza-
tion explants were subjected to various concentration
of mercuric chloride (0.1%-0.5%) along with antifungal
supplement and a prior ethanol treatment. Maximum
sterilization (80%) was observed using a concentration
of 0.3% of mercuric chloride for 3 min. A higher con-
centration of 0.5% of mercuric chloride burn the tis-
sue and cause blackening at the edges resulting in 23
% sterilization while a lower concentration of 0.1% of
mercuric chloride resulted in only 40% sterilization. It
was found that mercuric chloride alone at a concentra-
tion of 0.3% for 3 minutes is suf cient to sterilize the
explants without a prior treatment of ethanol and anti-
fungal supplements.
The axillary nodes were cut in equal size and surface
sterilized using standard procedure. A concentration of
0.1% HgCl
2
for 3 minutes resulted in 85% of sterilization
of explants while as compared a lower concentration
of HgCl
2
was less effective in sterilization process. Even
a concentration of 0.05 for 5 minutes resulted in 51%
decontamination of explant. However a pretreatment of
ethanol (rinse) and Bavistin (45min) resulted in higher
percentage of healthy sterilized tissue.
The axillary nodes excised from in vivo grown young
seedlings were sterilized using standard mercuric chlo-
ride solution at 0.1% concentration for 3 min which
was suf cient to obtain 80% sterilization. The prolif-
eration of shoots from cultured explant was remarkably
in uenced by the type of concentration of the growth
regulator used. Axillary node cultured on MS medium
supplemented with different concentration of BAP alone
showed the best results.
TECOMELLA UNDULATA
Complete plant development through tissue culture
strongly relies on synergistic effects of growth hormone
or say auxin and cytokinin which play a signi cant role
in cell differentiation and whole plant growth in Teco-
mella. Different concentrations of cytokinin and auxins
were used alone or in combination for initiation of shoot
proliferation. BAP was taken alone without any auxin
in combination at different concentrations (0.1,0.5,1.0
mg/L) resulted in bud break and shoot proliferation
of up to 2cm from sterilized axillary nodes cultured .
Further increase in BAP concentration from 1.5 to 2.0
mg/L resulted in shoot proliferation 1-2 cm in length
with a regeneration frequency of 50% in approximately
10 days. Similar results were obtained using axillary
nodes in Prosopsis by Pareek et al., 2012 and Kumar and
Singh (2009) who supported the effectiveness of BAP
and KIN in regeneration of multiple shoots in Prosopsis.
Similar concentration of BAP alone at 0.2 mg/l found to
be effective in axillary bud break in Baccopa monnieri,
(Sharma et al., 2010).
While on further increase in BAP concentration from
2 to 3 and 3.5mg/l found to be inhibitory. No bud break
or initiation in shoot proliferation was noticed on higher
concentration which clearly demonstrates that there is
decrease in plant cell differentiation with much higher
concentration of hormones. With an increase in con-
centration of growth regulators the days required to bud
break also increased which is further supported by our
present studies. The results totally contradict the nding
of Kumar and Singh (2009) who observed one to multi-
ple shoots on medium containing higher concentration
of BAP (5.0mg/l) along with IAA at (1.0mg/l). Indole ace-
tic acid was used as auxin in combination with BAP as
cytokinin. The concentration of BAP was kept constant
(1.0mg/l) and IAA as auxin was used in combination
at four different concentrations (0.01,0.02,0.05,0.1mg/l)
resulted in bud break and shoot proliferation of 1-2cm
in length with an increase in regeneration frequency
210 COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
Sunita Khatak, Archit Sharma and Rashi Saini
from 50%to 65% and bud break initiated in lesser time
period of 8 days. This combination reveals the synergistic
action of cytokinin to act and promote cell differentiation
along with auxin. After getting positive response of BAP
along with IAA, the concentration of BAP was raised to
1.5mg/l and was kept constant while the concentration of
IAA was altered (0.01,0.02,0.05,0.1mg/l).The shoot length
increased in size to 3 cm as compared to earlier with bud
break and shoot proliferation in 7 days.
The regeneration frequency also get increased to
95%.Our results well corroborate with the ndings of
Lal and Singh(2010) in Celastrus paniculatus using dif-
ferent cytokinin BAP and Kinetin(0.5,1.0,2.0mg/L) along
with auxins (IAA, NAA, and 2,4-D) using nodal explants
from mature tree of this species and observed less num-
ber of shoots per explants and 100% bud break on MS
medium supplemented with BAP.(1.0mg/L). Yadav et al
in 2011 reported that season of collection of explants
showed direct in uence on bud break in Celastrus pan-
iculatus using shoot tip explants obtaining highest
percentage (90%) bud break and multiple shoot forma-
tion(4/explant)on MS medium containing 1.0 mg/l BAP.
On further increase in BAP concentration from 1.5
to 2.0mg/l with IAA in combination at four different
concentrations (0.01, 0.02, 0.05, 0.1mg/l) no signi cant
increase was observed. Although the bud break and
shoot proliferation was observed which was almost same
resulting in 1-2 shoots / explants of 1-2 cm in length
with a regeneration frequency of 60% only .The results
clearly depicts the synergy between auxin and cytokinin
to act in combination for bud break and shoot prolifera-
tion but a higher concentration of BAP was found to be
inhibitory whether used alone or in combination with
IAA. Present results strongly contradict to that observed
by Warrier et al in 2010 in Aegle marmelos where higher
concentration of BAP (2.5mg/l) was bene cial for induc-
ing multiple shoots. (Table-1 and gure 1)
PROSOPSIS CINERARIA
Different cytokinins and auxins were employed alone
or in combination for bud break and shoot proliferation
in Prosopsis. Eighteen different media combinations
were used for Prosopsis micropropagation which was
recalcitrant to grow taking different concentrations of
growth hormones. Six different medium were used hav-
ing BAP (0.5.1.0,1.5, 2.0mg/l) along with NAA as auxin
at two different concentrations of (0.5 and 1.0 mg/l).
Four medium consisted of BAP along with IBA where
IBA was kept constant at a concentration of (1.0mg/l)
while the BAP concentration was raised from (1.0 mg
/l to 4.0 mg/l). Further six medium consisted of kinetin
in combination with two different auxins ie. NAA and
IBA. NAA was kept constant while kinetin was used at
two different concentrations of (2.0 and 3.0 mg/l),while
with IBA the concentration was kept constant at 1.0
mg/l and kinetin was used at very high concentration
of(5.0,7.0,9.0,10.0mg/l)to search out the possibility of
multiple shoots at much higher concentration.
Six more combinations of auxin and cytokinin
includes Zeatin in combination with IBA,where IBA con-
centration was kept constant at 1.0 mg/l but Zeatin con-
centration was altered (1.0,1.5,2.0,2.5,3.0,4.0). Although
a higher concentration of Zeatin was also used similar to
kinetin (5.0,7.0,9.0,10.0mg/l) but the higher concentra-
tion as observed earlier in case of Tecomella were found
to inhibitory and no bud break or shoot proliferation
was observed in Prosopsis.
On increasing the concentration of BAP step by step
from 0.5to2.0mg/l and keeping the NAA concentration
constant at 0.5mg/l. At lower concentrations bud break
was observed in 15 to 20 days at 0.5 and 1.0mg/l of
BAP while on further increase in concentration (1.5and
2.0mg/L)1-2 shoots per explants were observed of
1-1.5cm in length with higher regeneration frequency
as compared to lower concentrations used within same
time frame (Figure 2). When NAA concentration was
raised from 0.5 to 1.0mg/l in combination with BAP at
two different concentrations of 0.5and 1.0mg/l no sig-
ni cant change was observed, moreover the medium
resulted in bud break only.
Similar ndings were reported by Singh et al. (2014)
in Shorea robusta a woody valuable tree species using
nodal explants. They reported that BAP at 1.0mg/l along
with NAA at a concentratons of 0.5mg/l found to be the
best medium for shoot initiation and proliferation. Simi-
lar ndings observed by Tyagi et al (2010) in Capparis
deciduas using axillary shoots observed shoots of 2cm in
length when cultured on MS medium supplemented with
2mg/l BAP along with 0.5mg/l NAA.in 2010.
Girijashanker (2011) in Acacia auriculiformis a mul-
tipurpose tree of medicinal forestry observed higest
percentage of shoots induction on BAP (2mg/l )along
with NAA at (0.1mg/L). In combination of Zeatin with
IBA, IBA was kept constant at 1.0mg/l while the zeatin
concentration at different concentration resulted in 1-2
shoots per explants of 1-5cm in length in 45 days with
a regeneration frequency of 55%. Similar combinations
have been reported by Hua et al. (2015) to be effective
in Pitaya for shoot proliferation, they used 3.0uM Zea-
tin in combination with IBA at 0.5mg/l resulting more
vigorous multiplication of shoots. With an increase in
Zeatin concentration has positive effects on bud break
and shoot length elongation as a maximum of 5cm
length shoots were observed on 4.0mg/l concentration,
while an further increase in concentration of Zeatin
has inhibitory effects on cell differentiation same as
kinetin.
BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS 211
Sunita Khatak, Archit Sharma and Rashi Saini
Table 1. Composition of MS medium supplemented with different growth regulators and result
obtained after incubation of Tecomella undulata nodal culture.
S.No.
Media
Code
Growth Hormones (mg/L) No. of
shoots
Shoot
length
% of
regeneration
Days of
initiation
BAP IAA ABA NAA
1 MS - 1 --- --- --- --- Control
2 MS - 2 0.1 --- --- --- Bud break --- 16 9
3 MS - 3 0.5 --- --- --- Bud break --- 30 10
4 MS - 4 1.0 --- --- --- Bud break --- 25 9
5 MS - 5 1.5 --- --- --- 1 1.5 50 13
6 MS - 6 2.0 --- --- --- 1 1.0 45 12
7 MS - 7 2.5 --- --- --- Bud break --- 35 11
8 MS - 8 3.5 --- --- --- --- --- --- 11
9 MS - 9 1.0 0.01 --- --- --- --- --- 8
10 MS - 10 1.0 0.02 --- --- 1 1.5 60 7
11 MS - 11 1.0 0.05 --- --- 1 1.0 55 8
12 MS - 12 1.0 0.1 --- --- 2 1.0 65 7
13 MS - 13 1.5 0.01 --- --- 2 1.5 80 7
14 MS - 14 1.5 0.02 --- --- 3 2.5 95 7
15 MS - 15 1.5 0.05 --- --- 1 1.5 80 9
16 MS - 16 1.5 0.1 --- --- Bud break --- 50 11
17 MS - 17 2.0 0.01 --- --- Bud break --- 54 7
18 MS - 18 2.0 0.02 --- --- Bud break --- 70 8
19 MS - 19 2.0 0.05 --- --- Bud break --- 65 7
20 MS - 20 2.0 0.1 --- --- 1 1.0 60 9
21 MS - 21 2.5 0.01 --- --- 1 1.5 65 10
22 MS -22 2.5 0.02 --- --- 2 1.0 55 9
23 MS - 23 2.5 0.05 --- --- 2 1.0 60 11
24 MS -24 2.5 0.1 --- --- 2 1.0 55 10
25 MS -27 1.5 --- --- 0.01 Bud break --- 70 8
26 MS -28 1.5 --- --- 0.1 Bud break --- 65 9
27 MS -29 2.0 --- --- 0.01 1 1.5 55 7
28 MS - 30 2.0 --- --- 0.1 --- --- --- 8
The results depicts that higher concentrations of hor-
mone should not be taken as alternative to mass propa-
gate plants in shorter period of time. BAP was used at
different concentrations (1.0,2. 0,3.0,4.0), while IBA con-
centration was kept constant at 1.0mg/l resulted in 1-2
shoots per explants of 2-3cm in length in 35-40 days
approximately. NAA was kept constant at a concentra-
tion of (1.5mg/l) and using kinetin at two different con-
centration (2.0 and 3.0mg/l) the shoot length increased
in size from 2 to 4cm almost double while the initiation
process of shoot proliferation also shorten by 10 days
from 40 to 30 days. The results well corroborate with
that of Dhabhai et al. (2010) in Acacia nilotica a nitrogen
xing tree through direct regeneration using nodal seg-
ments cultured on MS medium supplemented with Kine-
tin (1.0mg/l along with NAA at 0.6mg/l they observed
highest no of shoots of 2-3 cm in 15-20 days of culture.
Multiple shoots were not observed on any of the media
used only one or two shoots were observed otherwise
the shoots just proliferated to 2-4 cm in 15 days. Simi-
lar ndings were observed in Vitex negundo, where MS
medium supplemented with BAP individually enhanced
the induction of multiple shoots within an average time
of 8 to 12 days, while BAP (2.0mg/L) in combination
212 COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
Sunita Khatak, Archit Sharma and Rashi Saini
FIGURE 1. Bud break on MS medium supplemented with different growth regulators in Tecomella undulate. A)
BAP (0.1mg/L),B) (0.5mg/L)andC) (1.0mg/l).
Table 2. Composition of MS medium supplemented with different growth regulators and result obtained after incubation
of Prosopis cineraria nodal culture.
S. No Media
code
Growth hormones (mg/L) No. of shoots Shoot
length (cm)
% of Regeneration Day of
initiation
Zeatin IBA BAP NAA
1 MS - 1 1.0 1.0 --- --- --- --- 30 30
2 MS - 2 1.5 1.0 --- --- 2 1.5 30 30
3 MS - 3 2.0 1.0 --- --- 1 1.5 50 30
4 MS - 4 2.5 1.0 --- --- 1 5 50 30
5 MS - 5 3.0 1.0 --- --- 1 5 60 40
6 MS - 6 4.0 1.0 --- --- 1 4.5 60 40
7 MS - 7 --- 1.0 1.0 --- 2 2.5 30 40
8 MS - 8 --- 1.0 2.0 --- 1 2.5 40 40
9 MS - 9 --- 1.0 3.0 --- 1 3.0 25 30
35 30
10 MS - 10 --- 1.0 4.0 --- 1 2.5
11 MS - 11 --- --- 0.5 0.5 Bud break --- 15 15
12 MS - 12 --- --- 1.0 0.5 Bud break --- 20 20
13 MS - 13 --- --- 1.5 0.5 2 1.5 20 20
14 MS - 14 --- --- 2.0 0.5 1 1.0 20 20
15 MS - 15 --- --- 0.5 1.0 Bud break --- 20 10
16 MS - 16 --- --- 1.0 1.0 Bud break --- 10 15
17 MS - 17 --- --- --- 1.5 1 2 10 30
18 MS - 18 --- --- --- 1.5 1 4 15 40
with NAA (0.5 mg/L) resulted in a higher percentage
(93%) of multiple shoot formation. The results were fur-
ther supported in Gloriosa superba, Rauvol a serpen-
tine
and Boerrhavia diffusa (Baksha et al., 2007; Hassan
and Roy, 2005 Biswas et al., 2009). MS media without
growth regulators failed to induce bud break from nodal
segments of Tecomella which is probably due to insuf -
cient level of endogenous growth regulators in explants
to induce bud break and it required an exogenous
supply.
Our present investigation focused on initial bud
break. Supplementation of cytokinins like BAP at dif-
BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS 213
Sunita Khatak, Archit Sharma and Rashi Saini
FIGURE 2. Shoot propagation on MS medium supplemented with growth regulators (BAP 1.5mg/l + NAA 0.5mg/l)
ferent concentration induces bud break as well as initia-
tion of shoot. Cytokinin supplemented media nulli es
the effect of apical dominance and promote axillary bud
proli ration into shoots. This is due to exogenous appli-
cations of cytokinins disturbs the internal polarity and
change the genetically physiology of explants, result-
ing in organogenesis. Similar effectiveness of cytokinins
in promoting in vitro auxillary bud break have been
reported in many forest plants. Auxins and cytokinins
have synergistic action in plant cells proliferation and
differentiation. The ratio of auxin to cytokinin has major
impact on growth and bud break. The release or absorp-
tion of one growth regulator is regulated by the presence
of other.
CONCLUSION
Multiplication of germplasm through plant tissue cul-
ture methods is one of the applications of biotechnol-
ogy via which elite trees can be mass produced rapidly.
MS medium without growth regulators failed to induce
bud break from nodal segments of Tecomella and Pros-
opsis probably due to insuf cient level of endogenous
growth regulators in explants to induce bud break and it
requires an exogenous supply of growth hormones. Our
present investigation was merely focused on initial bud
break and shoot proliferation with axillary buds in Teco-
mella and Prosopsis. Both the plants have importance as
medicinal value as reported in ancient literature. Both
can withstand the adverse conditions prevalent in semi-
arid and arid regions. With temperature 500ºF in winters
and 110-114ºF in summers. The tree provides grazing
material to livestock and plays an important role in
livelihood and ecosystem preservance. Both are excel-
lent soil binders and important constituents of arid veg-
etation system. The National Research Centre Institute
for Arid Horticulture, Bikaner (CIAH), Central Arid Zone
Research Institute (CAZRI), Jodhpur and Arid Zone For-
estry Research Institute (AFRI), Jodhpur are working to
collect germplasm, and to improve and propagate fruit
and forest trees found in hot arid zones. Still there is
a need to cope up with overgrazing and exploitation
of both trees as important structure for livelihood of
local people residing in deserts areas. New protocols are
indeed a basic requirement to propagate such trees on
214 COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS
Sunita Khatak, Archit Sharma and Rashi Saini
large scale multiplication through tissue culture tech-
nology .Conclusively this study has established the bud
break and shoot initiation from axillary node of both
woody taxa. Further research in order to develop plant-
lets and their acclimatization to eld conditions will be
next stepping stone .Meanwhile this protocol offers an
ef cient method to initiate shoot proliferation and also
paved the path to improve vegetation coverage in arid
and semi-arid regions of India.
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BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS COMPARATIVE ANALYSIS OF PLANT GROWTH REGULATORS 215