This document discusses plant tissue culture techniques, including the types of explants used and sterilization protocols. It describes that meristem culture uses meristematic tissues like shoot apical meristems. Seed and ovule culture uses fertilized ovules from plants with small or minute seeds. Callus culture produces an unorganized cell mass. Haploid culture generates haploid cells from tissues like pollen and anthers. Different sterilization chemicals like sodium hypochlorite and calcium hypochlorite are used depending on the explant type. Protocols vary in chemical concentration and duration to sterilize tissues without loss of viability. Cells in culture go through growth phases including exponential growth, before requiring sub-cult

1. PREPARATION AND CHOICE OF EXPLANT FOR PLANT
TISSUE CULTURE AND TECHNIQUES
S. No. CONTENT
1 Types of Culture
2 Types of Explant
3 Sterilization
4 Sterilization Protocols
5 Cell Phases
6 Sub- culturing

2. TYPES OF CULTURE
Meristem Culture
Seed Culture
Somatic Embryo Culture
Callus Culture
Haploid Culture
Organ Culture
Protoplast Culture

3. PLANT PARTS USED IN TISSUE CULTURE

4. TYPES OF EXPLANT- Meristem Culture
Meristem Culture comprised of meristematic regions like SAM (Shoot
Apical Meristem), RAM (Root Apical Meristem) and auxiliary shoots.

5. SEED/ OVULE CULTURE
Here the explant is fertilized ovule of those plants that produce low level
of viable seeds or whose seeds are minutes as well as non albuminous
type.

6. EMBRYO CULTURE

7. CALLUS CULTURE
Callus is an unorganized mass of cell that contain only parenchymatous
cells

8. HAPLOID CULTURE
Haploid can be generated from pollen, anther and unfertilized ovum and
majorly used for pure line production.

9. ORGAN CULTURE

10. PROTOPLAST CULTURE

11. STERILIZATION OF EXPANTS
Sodium
hypochlorite
Calcium
hypochlorite

12. STERILIZATION PROTOCOLS
• Protocols of sterilization are
specific for different explants.
• Harsh explants requires long
duration as well as high
concentration of chemicals
• While soft tissues like embryos,
anther and ovules require
particular types of chemicals.
• Sterilization process are
followed in outer area as well as
in laminar air flow hood.
• Increased concentration of
chemicals will affect the
viability of explant.

13. PHASES OF CELLS
Growth
of
cells

14. EXPONENTIAL GROWTH

15. •When the cells in culture are reaching at stationary
state it should be sub-cultured.
•Sub-culturing may be in similar or dissimilar type of
media with respect to the requirements.