In species like maize, small pieces of cob (bearing, say, 10 ovaries) may be cultured 2-6 days after silk emergence. General1y, ovaries/ovules are cultured along with the pedicel.

In some dicot species, it may be desirable to retain the calyx, while in monocots like barley the glumes (lamma and palea) need to be retained since their removal is deleterious to embryo development. Wetting of ovules and stigma should be avoided since it may reduce pollen germination and interfere with normal pollen tube growth.

The information on GR requirements is scanty, but in some cases IAA (up to 10 μ/1) and kinetin (0.1 μ/1) significantly improve seed set. The cultures are usually kept at 25°C, but in species like Narcissus, 15°C gave the best seed set. Light is not necessary; the cultures are usually incubated either in diffuse light or, more often, in dark.

Pollen grains are collected from surface sterilized anthers that are kept in a sterile Petri dish so as to obtain contamination-free pol1en. The pollen grains are usually deposited directly onto ovules, placenta or stigma depending on the type of culture. In most cases, culture of ovules along with the placenta gives the best seed set.