Optimization of Media Formulations for Callus Induction and direct Organogenesis in Justicia Gendrussa Burm Fil

The Uses and Advantages of an Exotic Medicinal Plant in Kerala


Scientific Study, 2017
39 Pages, Grade: 1.5

Excerpt

ii
Table of contents
Table of contents ... ii
Table of figures ... iv
Table of tables ... v
List of abbreviations ... vi
Optimization of media formulations for callus induction and direct organogenesis in
Justicia gendarussa
Burm Fil., an exotic medicinal plant in Kerala ... 8
Abstract ... 8
1. Introduction ... 9
1.1 Aim ... 10
1.2 Objectives ... 10
2. Review of literature ... 10
2.1
Justicia gendarussa
Burn Fil. Common names ... 16
2.2 Taxonomical classification:
Justicia gendarussa
Burn Fil. ... 16
3. Hypothesis ... 18
4. Materials and Methods ... 18
4.1 Study area ... 18
4.2 Plant material ... 18
4.3 Equipment's used for experiment ... 18
4.4 Culture media ... 18
4.5 Compositions of MS media (Chawla H.S.) ... 18
4.6 Preparation of MS media (stock preparation) ... 19
4.7 Compositions of nutrient broth ... 23
4.8 Preparation of nutrient broth ... 23
4.9 Compositions of MHA ... 24
4.10 Preparation of MHA ... 24
4.11 Test organisms... 24
4.12 Experimental systems ... 24
4.13 Preparation of MS media ... 25

iii
4.14 Preparation of explants ... 25
4.15 Surface sterilization of explants ... 25
4.16 Inoculation procedure ... 25
4.17 Statistical analysis ... 29
5. Results and discussion ... 33
5.1 Establishment of callus cultures from nodal and leaf explants ... 33
5.2 Establishment of direct organogenesis from nodal explants ... 34
5.3 Optimization of callus initiation of
Justicia
gendarussa
... 35
5.4 Evaluation of antimicrobial activity ... 35
6. Conclusions ... 36
References ... 37

iv
Table of figures
Figure 1. Description of
Justicia gendarussa
Burm Fil. a) plant with leaves, b) plant
with flowers, c) and d) plant with leaves and flowers, e) with flowers. ... 17
Figure 2. Map of Kerala showing the sample collection point. Authors own work. ... 29
Figure 3. Details of
Justicia
gendarussa
a), explant showing the induction of callus b)
and c, callus induction on MS medium supplemented with 0.7 mg/L NAA + 0.2 mg/L
BA d) and e), callus induction on medium supplemented with 0.5 mg/L NAA and 0.1
mg/L BA (leaf explant). Authors own images. ... 30
Figure 4. Details of
Justicia
gendarussa
Burm
Fil
.
a) and b) different stages of callus
induction on MS medium supplemented with 0.1 mg/L 2, 4-D and 0.1 mg/L kinetin (leaf
explant) c) and d) different stages of callus on MS medium supplemented with 0.5
mg/L 2, 4-D and 0.5 mg/L kinetin (nodal explant). Authors own images. ... 31
Figure 5. Details of
Justicia gendarussa
Burm Fil
.
a), rhizogenesis from nodal explants
(0.7 mg/L NAA + 0.5 mg/L kinetin) b), MHA plate with
E. coli
(C-control; T-ethanolic
extract of callus c), MHA plate with
K. pneumoniae
(C-control; T-ethanolic extract of
callus d), MHA plate with
S. aureus
(C-control; T-ethanolic extract of callus. Authors
own images. ... 32

v
Table of tables
Table 1. Stock solution of macro-salts. ... 20
Table 2. Stock solution of micro-salts (100X). ... 21
Table 3. Stock solution of vitamins (100X). ... 21
Table 4. Stock solution of potassium iodide (100X). ... 22
Table 5. Stock solution of iron (10X). ... 22
Table 6. Stock solution of glycine (10X). ... 22
Table 7. Stock solution of mesoinositol (100X). ... 23
Table 8. Stock solution of hormones. ... 23
Table 9. Concentrations of combinations taken for present study. ... 27
Table 10. The concentration of hormones taken for present study ... 27
Table 11. Combinations of NAA and kinetin used for direct organogenesis ... 27
Table 12. Combinations of NAA and kinetin used for direct organogenesis ... 28
Table 13. Growth response of leaf and nodal explants of
Justicia gendarussa
on MS
medium with different hormone combinations (NAA+BA). ... 33
Table 14. Growth response of leaf and nodal explants of
Justicia gendarussa
on MS
medium with different hormone combinations. ... 34
Table 15. Growth response (rhizogenesis) of nodal explants
Justicia
gendarussa
on
MS medium with different hormones. ... 34
Table 16. Growth response (rhizogenesis) of nodal explants
Justicia
gendarussa
on
MS medium with different hormones. ... 35
Table 17. Evaluation of antimicrobial activity. ... 36

vi
List of abbreviations
%
: Percentage
°C
: Degree celsius
µl
: Microliter
mL
: Millilitre
mm
: Millimetre
sp
: Species
g
: Gram
UV
: Ultra Violet
L
: Litre
2, 4-D
: 2, 4-Dichlorophenoxy Acetic Acid
BAP
: Benzylaminopurine
NAA
: - Naphthalene acetic acid
IAA
: Indole 3-acetic acid
MS
: Murashige and Skoog
Fe EDTA
: Ferrous Ethylene diamine tetra acetic acid
mg
: Milligram
PTC
: Plant tissue culture
µg
: Microgram
nm
: Nanometre
cm
: Centimeter
FeSO4
: Iron sulfate
HCl
: Hydrochloric acid
KI
: Potassium iodide
Kn
: Kinetin
K
.
pneumoniae
:
Klebsiella
pneumoniae
MHA
: Muller hinton agar
mg/L
: Milligram per liter
NaCl
: Sodium chloride
NaOH
: Sodium hydroxide
P
.
aeruginosa
:
Pseudomonas
aeruginosa
S
.
aureus
:
Staphylococcus
aureus

vii
Na
2
EDTA
: Disodium Ethylene diamine tetra acetic acid
B
.
subtilis
:
Bacillus
subtilis

8
Optimization of media formulations for callus induction and direct
organogenesis in
Justicia gendarussa
Burm Fil., an exotic
medicinal plant in Kerala
Prem Jose Vazhacharickal
1
*, Noby Mathew
2
and Elizabeth Cherian
2
* premjosev@gmail.com
1
Department of Biotechnology, Mar Augusthinose College, Ramapuram, Kerala, India-686576
2
Department of Biotechnology and Applied Microbiology, St. Thomas College, Pala, Kerala, India-
686574
Abstract
In this study, the callus initiation and rhizogenesis of
Justicia gendarussa
from nodal
and leaf explants were established in Murashige and Skoog medium supplemented
with different hormonal concentrations and also optimization of callus initiation at
hormonal concentrations. As a result the callus was initiated on MS medium
supplemented with different hormones. The higher concentration of NAA and BA
induced callus from nodal and leaf explants. The higher concentrations of 2, 4 ­ D and
kinetin induced callus on nodal explants. Lower concentration of 2, 4 ­ D and kinetin
induce callus on leaf explants. The combination of NAA and kinetin induce
rhizogenesis from nodal explants. The antimicrobial activity of callus was evaluated by
well diffusion method.
Escherichia coli
,
Pseudomonas aeruginosa
,
Bacillus subtilus
,
Staphylococcus aureus
,
Klebsiella pneumonia
were used as test organisms. The
results showed that
Justicia gendarussa
have antimicrobial activity on
Escherichia coli
,
Klebsiella pneumonia
, and
Staphylococcus aureus
.
Keywords:
Justicia gendarussa
, callus initiation, direct organogenesis, antimicrobial
assay.

9
1. Introduction
Biotechnology is the advanced technology that is the use of biological processes,
organisms or systems to manufacture products intended to improve the quality of
human life. It has many applications in different fields such as medical, agriculture,
industry, plant and animal science too. Plant tissue culture is one of the main branches
of biotechnology (Alfermann and Petersen, 1995).
Plant tissue culture is the technique of maintaining and growing plant cells, tissues or
organs especially on artificial medium in suitable containers under controlled
environmental conditions. The part which is cultured is called as explants i.e., any part
of the plant taken as explants and grown in a test tube, under sterile conditions in
special nutrient media. This capacity of cell to explants to regenerate to a whole plant
is called cellular totipotency. Gottileb Haberlandt was first initiated the tissue culture
technique in 1902 (Alfermann and Petersen, 1995; Razdan, 2003; Pierik, 1997).
Plant tissue cultures are associated with a wide range of applications. The most
important being the production of pharmaceutical, medicinal and other industrially
important compounds (Razdan, 2003; Pierik, 1997).
Justicia gendarussa
belongs to the family Acanthaceae. It is important medicinal plant
used in the treatment of various health problems. It is commonly known as willow-
leaved justicia.
Justicia gendarussa
is commonly known as Vathamkolly in Kerala. It
is a small erect branched shrub. It has been described as rare and endemic to India.
The plant is widely used in various forms for many of its medicinal and insecticidal
properties and that it is a quick growing, evergreen forest shrub considered to be a
native of China and distributed in Sri Lanka, India and Malaysia (Rahmatullah et al.,
2010; Silja et al., 2008).
It is a shade loving, quick growing, erect, branched and evergreen shrub by measuring
0.6-1.2 m in height. The leaves are simple, opposite, lanceolate or linear-laneolate,
acute at base, tapering into rounded apex and glabrous and shining leaves 8-12.5 cm
long, 1.2-2 cm broad, with prominent purple veins beneath. The stem is quadrangular,
thickened at and above the nodded and internodes measure 2-7 cm long (Rahmatullah
et al., 2010; Silja et al., 2008).
The flowers are in terminal or axillary spikes and are irregular, bisexual, sessile, and
white with pink or purple spots inside and red in the throat and lip (Rahmatullah et al.,
2010; Silja et al., 2008).
Excerpt out of 39 pages

Details

Title
Optimization of Media Formulations for Callus Induction and direct Organogenesis in Justicia Gendrussa Burm Fil
Subtitle
The Uses and Advantages of an Exotic Medicinal Plant in Kerala
College
Mar Augusthinose College
Grade
1.5
Authors
Year
2017
Pages
39
Catalog Number
V378387
ISBN (eBook)
9783668616318
ISBN (Book)
9783668616325
File size
2444 KB
Language
English
Tags
Optimization, Formulation, Callus, Induction, Organogenesis, Burm
Quote paper
Dr. Prem Jose Vazhacharickal (Author)Elizabeth Cherian (Author)Noby Mathew (Author), 2017, Optimization of Media Formulations for Callus Induction and direct Organogenesis in Justicia Gendrussa Burm Fil, Munich, GRIN Verlag, https://www.grin.com/document/378387

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