Anther Culture & Production of Haploid Plants
Introduction
Anther culture and pollen culture are in-vitro techniques for producing haploid plants.
The process involves culturing anthers under controlled conditions to generate haploid plants from pollen grains.
Structure of Anther
A cross-section of an anther reveals two pollen sacs.
These sacs contain pollen grains, which are embedded in specialised loops.
Formation of Pollen Grains
During the early development of the pollen sac, it contains primary mother cells.
These cells undergo meiosis to form tetrads (clusters of four cells).
Each cell of the tetrad develops into a pollen grain (or microspore).
Haploid Nature of Pollen Grains
Pollen grains are haploid in nature (contain only one set of chromosomes).
This makes them suitable for anther culture, where all pollen grains can potentially develop into haploid plants.
Steps Involved in Anther Culture
Collection of Immature Flower Buds
Immature flower buds are carefully collected for the process.
Sterilization
The collected buds are sterilized using:
70% alcohol,
A surface sterilizer, and
Sterile distilled water.
Isolation of Anther
The flower buds are slit open with a scalpel to release the anthers.
The filament is discarded, and only the anther is retained.
Placement in Liquid Medium
The anther is placed in a liquid medium that facilitates callus formation (a mass of undifferentiated cells).
Transfer to Nutrient Media
The callus is transferred to MS medium (Murashige and Skoog medium) to support its differentiation into a plantlet.
Subculturing and Hardening
The plantlets are subcultured into fresh media multiple times to enhance hardening.
Proper humidity is maintained using polythene coverings.
Transfer to Soil
Hardened plantlets are transplanted into small pots with sterilized soil.
After a few days, they are moved to larger pots.
Final Production of Haploid Plants
Over time, the plants adapt to the environment, completing the process of haploid plant production.