Pollen Culture
Definition: Pollen culture is the process of culturing pollen grains under in-vitro conditions to produce haploid plants.
Similarity to Anther Culture:
The steps and process of pollen culture are the same as those of anther culture.
Steps Involved in Pollen Culture
Collection of Immature Flower Buds
Immature flower buds are collected at an early stage to ensure viable pollen grains (microspores).
Sterilization
The flower buds are sterilized using:
70% alcohol,
A surface sterilizer, and
Sterile distilled water.
Isolation of Pollen Grains
The buds are slit open, and pollen grains (microspores) are carefully isolated from the anthers.
Placement in Liquid Medium
The isolated pollen grains are cultured in a liquid medium under controlled conditions.
The medium promotes either direct androgenesis (embryo formation) or indirect androgenesis (callus formation).
Transfer to Nutrient Medium
If a callus is formed, it is transferred to MS medium (Murashige and Skoog medium) to support differentiation into plantlets.
For embryos, the medium encourages further growth into plantlets.
Subculturing and Hardening
The plantlets are subcultured into fresh media for further growth and hardening.
Polythene coverings are used to maintain proper humidity during this stage.
Transfer to Soil
Hardened plantlets are transferred to small pots containing sterilized soil.
After further growth, the plants are moved to larger pots for full development.
Androgenesis
Definition: Androgenesis is the process by which a haploid plant is produced from a pollen grain (microspore) under in-vitro conditions.
It is classified into two types:
Direct Androgenesis
The microspore acts directly as a zygote.
It develops into an embryo, which eventually forms the plantlet.
Indirect Androgenesis
The microspore undergoes rapid cell division to form a callus (mass of undifferentiated cells).
The callus then differentiates into plantlets.