This presentation gives details about the organ culture in plant tissue culture and its basic applications, also this provide an detailed information about the technique of root culture and gives small view about its appilications.

1. Introduction to organ culture in Plant tissue
culture and root culture.
By, Mithil Shetty
M.Sc biotechnology ,
D.Y. Patil biotechnology & bioinformatics
Insistute, Pune

2. Organ culture:
 It is in vitro culture and maintenance of an excised
organ primordia or whole or part of a organ in a way
that may allow differentiation and preservation of the
architecture and function.

3. HISTORY:
 W. Kotte and W. J. Robbins (1922)- reported the 1st culture of excised
root tips from germinated wheat seedlings.
 P. R. White (1930)- reported the successful culture of root segments of
ascetically germinated tomato seedlings.
 C. D. LaRue (1942)- reported first the in
vitro culture of excised flower buds of
Kalanchoe globulifera and
Nemesia strumosa.

4.  S. W. Loo (1945)- reported the culture of shoot tips of
Asparagus seedlings on a medium.
 L. L. Jansen and J. Bonper (1949)- grew the ovaries
of Lycopersicon pimpinellifolium on medium.
 J. P. Nitsch (1949-51)- successfully cultured the ovaries of Lycopersicon
esculentum, cucunis anguria, Phaseolusvulgaris, Fragaris sp. and
Nicotinia tabacum.

5.  N. Maheshwari (1958)- isolated ripe pollen and ovules of
Papaver somniferum and cultured them together and
thus observed all stages from pollen germination,
through fertilization to developments of mature seeds.
 E. Galun, Y. Yung and A. Lang(1962-63)- tested the effect of IAA and
gibberellin acid 3 upon the expression of culture of isolated floral buds of
Cucumis sativus.
 G. Morel(1952-53)- showed that through aseptic culture of stem tips the
certain virus infections can be eliminated from potato and dahlia, which
allowed recovery of healthy plants and also discovered the rapid
multiplication of tropical orchid Cymbidium using apical meristem culture.

6.  T. A. Steeves and I. M. Sussex (1966)- first said that culture of excerised
leaf primordia would provide an experimental system to study the complete
development of leaves under controlled conditions.

7. Types of organ culture:
 Vegetative organs.
1. Root culture.
2. Leaf culture.
3. Shoot-tip culture.
 Reproductive organs
1. Complete flower culture.
2. Isolated ovary culture.
3. Isolated ovule and embryo
culture.

8. Importance of organ culture:
 Provide an excellent experimental system to define the
nutrients and growth factors.
 Study of morphogenesis.
 Study of secondary metabolites and its biosynthesis.
 Lead to new developments in agriculture and horticulture.

9. ROOT CULTURE:
Root culture is culturing of excised radical tip of
aseptically germinated seed in the liquid medium to
grow independently under controlled conditions.

10. “
”
Figure: Root and its important parts

11. Important parts and function of plant root system:
 Root hair - extension of specialized root epidermal cells increasing
the surface area for adsorption of water and minerals.
 Epidermis - outer layer of root.
 Cortex - region between epidermis and vascular cylinder.
 Pericycle - origin for secondary roots.
 Xylem - carry water and minerals throughout the plants.
 Phloem – carry dissolved sugar and organic components through the
plants

12. Principle:
 The root culture is initiated from aseptically germinated seeds, which is
maintained in an moving liquid medium, which induce the growth of roots
system as in the natural condition.
 Also the clone of the excised roots can be established from single root
culture by repeatedly cutting and transferring the main root tips or of lateral
tips into fresh medium in very subculture at the interval of definite period .
 The growth of exercised roots can be expressed in terms of fresh and dry
weight, increase in length of main axis, number of emergent laterals and
total length of laterals per culture.

13. Protocol:
 Initiation of isolated root cultures:
Seeds are first surface sterilized by proper method
Seeds are germinated on whites basal medium at 25⁰C in dark
As the seedlings roots are 20 to 40 mm in length, 10 mm apical tip are exercised and
transferred into the liquid medium.

14.  Initiation of clones:
The root culture is established and maintained for 10 days.
Then the main axis of the root is cut into number of pieces, each bearing four
or five young laterals.
Then each individual sector inoculum is transferred into medium and
incubated.
Again after some maintaince, the growing sector is cut into 4 to 5 sectors to
initiate the sector culture again.

15.  Figure: Procedure for root culture

16. Importance of root culture:
1. Give the knowledge of carbohydrate metabolism and role of minerals,
ions and vitamins etc in root growth.
2. Provided the basic information regarding the dependence of roots on
shoots for growth hormones.
3. Root clones are used in study of effect of various compounds on root
growth.
4. Study of nodulation of leguminous roots in culture.
5. For regeneration of shoots on roots.
6. Study of synthesis of secondary metabolites from root culture.

17.  Ref: Plant tissue culture by Kalyan Kumar De.

18. Thank you.