Anther or Pollen culture
What is Anther or Pollen culture? – Definition
Pollen culture (microspore culture) is a technique in which haploid plants are obtained from isolated pollen grains while in anther culture those are obtained from pollens, by placing anthers on a suitable, synthetic culture medium.
The technique was discovered by Guha and Maheshwari (1964). It is one of the various tissue culture techniques or methods used. Progeny developed by this technique contains a single set of chromosomes.
Procedure for In-vitro Anther culture
Collection of anthers – Preferably should be taken from plants grown under controlled conditions or may be taken from plants grown in the field.
Transfer of anthers to the culture medium – Flower buds of the appropriate developmental stage are collected and surface sterilized. Anthers from the buds are excised and placed horizontally on surface (or partially embed) on culture medium.
Important to note is, injury to anthers may induce callus formation from anther walls. To avoid this, small anthers cultured as such and sometimes entire inflorescence is cultured.
Procedure for In-vitro Pollen culture
Pollens are isolated either by squeezing or float culturing of anthers.
- Squeezing method – Anthers (about 50 to 60) are placed in 20 ml of medium. Squeezed gently with a glass rod to separate pollens from anther walls. The prepared solution is filtered through a nylon mesh of suitable pore size. Filtrate is centrifuged for about 5 min at 500-800 rpm. The resulting pollen dense pellet is collected, washed twice. Final concentration of pollens is adjusted to 103-104 pollens/ml.
- Float culture – Anthers are excised. Those are placed in petri dishes containing liquid medium. Anthers float in liquid medium. The anthers release their pollen grains into the medium in a few days.
Culture Medium for anther or pollen culture
Medium used depends on various factors like
- Genotype of donor plants and anthers
- Age of donor plants and anthers
- Growing conditions of donor plants
Isolated pollen grains are cultured on a synthetic medium, the important ingredients of which are glutamine, L-serine and inositol. For anther cultures sucrose is essential ingredient.
Culture conditions are different according to certain conditions as we have listed under culture medium. Alternate light (12¬ to 18 hr; 5,000-10,000 lux) and dark periods (12-6 hr) at 22°C.
On the page Pathways of development in pollens, four pathways of pollen development and factors affecting to it are provided. Applications – Advantages of pollen and anther culture enlist applications, merits and demerits of the concept.