1: In vitro shoot multiplication in okra (Ablemonschus esculentues). 2: About Okra Plant. 3: Application of Tissue culture in Okra. 4: Objective. 5: Material and Method. 6: Methodology. 7: Treatment schedule. 8: Result of Program 9: Conclusion 10:References

1. Mr. Tajane Sachin Arunrao (KK/1658)
SHRI SHIVAJI COLLEGE OF
AGRICULTURAL BIOTECHNOLOGY,
AMRAVATI
Guided By
Dr. S.S. Chikte
(Assistant Professor)
Department
Plant Biotechnology

2. ABOUT OKRA PLANT
1) Okra is a vegetable crop plant belongs to the family Malavacaea. It is valued for its
edible green seeds pods.
2) The geographical origin is disputed, with supporters of West African, Ethiopia and South
Asian origins.
3) The plant is cultivated in tropical, subtropical and warm temperate regions around the
world.
4) It is among the most heat and drought-tolerant vegetable species in the world and will
tolerate soils with heavy clay and intermittent moisture.
5) A vegetable yield of 10 tons/ha can be considered a good harvest, but yielded are usually
low (2-4 t/ha) as result of non intestine growing method. Seed yield are in the range of 500 -
1000 kg/ha.

3. APPLICATION OF TISSUE CULTURE IN OKRA
1) The study of plant tissue culture describe a simple, rapid, efficient and optimized
system in okra (Abelmoschusns esculentus) by using nodal segment as explants.
2) Meristem culture in okra is possible for the development of diseases free plant.
3) Tissue culture in okra has been used for the development of Transgenic line or
somoclonal cultivar.

4. OBJECTIVES
1. Preparation of MS media for in vitro culture development in Okra.
2. Standardization of MS media for in vitro shoot multiplication in okra.

5. MATERIALES AND METHOD
1) Glassware's
2) Instruments
3) Sterilization of Glassware-The Glassware 's were autoclaved at 15 lbs pressure at 1210C
for 20 minutes. Followed by drying in oven 160 -180oC for 2 hrs.
4) Hormones- NAA, IBA and Kinetin
5) Chemicals- HgCl2, Tween-20 etc

6. METHODOLOGY
Media Preparation
Selection and isolation of explants
Surface sterilization of Explants
Culture of Explants
Culture condition

7. TREATMENT SCHELDULE
Treatment No.1 Treatment No. 2
Sr. No. Concentration of
growth
hormone(mg/lit)
No. Of
tubes
NAA IBA
1 0.25 1.0 5
2 0.5 1.0 5
3 1.0 1.0 5
4 1.5 1.0 5
5 2.0 1.0 5
Sr. No. Concentration of
growth
hormone(mg/lit)
No. Of
tubes
IBA NAA
1 0.5 1.0 5
2 1.0 1.0 5
3 1.5 1.0 5
4 2.0 1.0 5
5 2.5 1.0 5

8. Treatment No.3 Treatment No. 4
Sr. No.
Concentrati
on of
growth
hormone
(mg/lit)
No .Of
tubes
IBA
1 0.5 5
2 1.0 5
3 1.5 5
4 2.0 5
5 2.5 5
Sr. No.
Concentration of growth
hormone (mg/lit)
No.
Of
tubes
NAA IBA Kinetin
1 0.25 1.0 1.0 5
2 0.5 1.0 1.0 5
3 1.0 1.0 1.0 5
4 1.5 1.0 1.0 5
5 2.0 1.0 1.0 5

9. RESULT OF THE PROGRAMME
Sr. No.
Concentration of the
Growth Hormone
(mg/lit)
Shoot
Multiplication was
Observed After 15
Days
(Average±S.D)
Shoot Multiplication
was Observed After 30
Days
(Average±S.D)
Shoot
Multiplication
was Observed
After 60 Days
(Average±S.D)
NAA IBA
1 0.25 1.0 No Response No Response No Response
2 0.5 1.0 1.6±1.2 2.6±2.1 5.4±3.3
3 1.0 1.0 No Response No Response No Response
4 1.5 1.0 No Response No Response No Response
5 2.0 1.0 No Response No Response No Response
Treatment 1- Effect of NAA + IBA on Shoot Multiplication in Okra

10. Effect of MS+NAA(0.5
mg/lit)+IBA(1.0mg/lit) on shoot
multiplication After 15 days
Effect of MS+NAA(0.5
mg/lit)+IBA(1.0mg/lit) on shoot
multiplication After 30 days.
Effect of MS+NAA(0.5
mg/lit)+IBA(1.0mg/lit) on shoot
multiplication After 60 days

11. Treatment 2- Effect of MS+NAA (0.5mg/lit)+IBA (1.0 mg/lit) on Shoot
Multiplication in Okra
Sr. No.
Concentration of the
Growth Hormone
(mg/lit)
Shoot
Multiplication was
Observed After 15
Days
(Average±S.D)
Shoot Multiplication
was Observed After 30
Days
(Average±S.D)
Shoot
Multiplication
was Observed
After 60 Days
(Average±S.D)
NAA IBA
1 0.5 1.0 3.4±2.7 5.2±3.4 8.0±3.7
2 1.0 1.0 No Response No Response No Response
3 1.5 1.0 2.2±09 2.2±1.0 4.8±2.2
4 2.0 1.0 3.0±1.6 5.0±2.1 6.4±2.9
5 2.5 1.0 2.0±1.7 3.4±2.2 4.6±2.7

12. Fig. (D) Multiple shoot induction on MS+NAA
(0.5mg/lit)+ IBA(1.0 mg/lit) after 15 days
Fig. (E) Multiple shoot induction on
MS+NAA(0.5mg/lit)+IBAA(1.0mg/lit) after 30 days
Fig. (F) Multiple shoot induction on
MS+IBA(0.5mg/lit)+NAA(1.0mg/lit) after 60
days

13. Sr. No.
Concentrati
on of the
Growth
Hormone
(mg/lit)
Shoot Multiplication
was Observed After
15 Days
(Average±S.D)
Shoot Multiplication was
Observed After 30 Days
(Average±S.D)
Shoot
Multiplication was
Observed After 60
Days
(Average±S.D)
MS+IBA
1 0.5 3.2±1.5 5.2±2.6 9.8±3.6
2 1.0 2.8±1.7 3.4±2.2 6.0±3.6
3 1.5 4.2±2.0 9.2±3.6 13.2±7.8
4 2.0 2.1±1.4 4.0±3.3 6.4±4.2
5 2.5 1.6±1.2 3.4±2.7 5.2±3.9
Treatment 3- Effect of MS+IBA on Shoot Multiplication in Okra

14. Fig (G). Multiple Shoot induction on
MS+IBA(1.5mg/lit) after 15 days
Fig (H). Multiple Shoot induction on
MS+IBA(1.5mg/lit) after 30 days
Fig.(I). Multiple Shoot induction on MS+IBA
(1.5mg/lit) after 60 days

15. OUTCOMES OF THE PROGRAMME
1) In the present study “In vitro shoot multiplication in Okra (Ablemoschus esculentus) by
using nodal segment”; the explants were culture on MS media with respect to different
plant growth regulator to check the nodal segment potential to grown into number of
multiple shoots.
2) Nodal segment shows good response at IBA (2.5 mg/lit) for multiple shoot induction i.e
1.6±1.2 after 15 days, 3.4±2.7 after 30days, 5.2±3.9 after 60 days..
3) IBA (2.0 mg/lit) shows better response for inducing multiple shoots 2.0±1.7 after 15 days,
4.0±3.3 after 30 days. 6.4±4.2 after 60 days.
4) IBA (1.5 mg/lit) was significantly more effective for inducing multiple shoots 4.2±2.0
after 15 days, 9.2±3.6 after 30 days, 13.2±7.8 after 60 days.
5) IBA (1.5 mg/lit) alone shows best response for shoot multiplication 13.2±7.8 after 60 days
per culture; And it is the best combination suited for inducing multiple shoot. In
conclusion this combination describe an efficient rapid propagation system in Okra.

16. REFERENCES
1) G. A. Dhande, Vijay M. Patil, R. V. Raut, A. G. Ingle (2012) Regeneration of
Okra(Abelmoschus esculentus) via apical shoot culture system. African journal of
Biotechnology Vol.11(86),pp. 15226-15230.
2) Haider, S.A., R. Islam, A.H.M. Kamal, S.M. Rahman and O.I. Joarder, (1993). Direct and
indirect organogenesis in cultured hypocotyl explants of Abelmoschus esculentus (L.)
Moench. Plant Tissue Culture., 3: 85-89.
3) Mohammad Anisuzzaman , Ahmad H. Kabir , Kanak K. Sarker , Shamima Jarin and
Mohammad F. Alam, (2010) .Micropropagation of Abelmoschus esculentus L. (Moench.)
for disease free plantlets through meristem culture.Volume 43, Issue 5, 2010, pages 460-
466.

17. 4) Wang PJ, Hu NY (1980). Regeneration of virus-free okra(Abelmochus esculentus) plants
through in vitro culture. Advances in biochemical engineering: Plant cell culture, II.
Berlin: Springer-Verlag. pp. 61–99.