Anther and Microscope Culture - One of the very popular methods for production of haploids is through culturing anthers or microspores on artificial culture medium. This leads to the growth of microspores into saprophytes.

After the initial reports of successful production of haploids from anther culture in Datura (Guha and Maheshwari, 1966, 1967), haploids have been obtained in more than 150 species belonging to 23 families of angiosperms (Maheshwari et a1., 1980). These include a wide variety of economically important species.

In either case, flower buds are brought to a laminar flow chamber and sterilized using appropriate chemical treatment before their dissection. While removing anthers from the flower buds, care is exercised to avoid injury, because injury leads to development of callus, giving a mixture of diploids, haploids and aneuploids.

The anthers are generally cultured on solid agar medium, where they may directly give rise to embryoids or may lead to callus formation, before differentiation. The embryoids develop into haploid plantlets, which are colchicine treated to get diploid homozygous plants to be field tested for selection.

Microspore culture may be preferred over anther culture, even though the degree of success is low in this case. Microspores are collected first using the following steps:

The microspores may develop directly into embryoids within 15 days or follow one of the several indirect paths to produce haploid plantlets.

In anther culture as well as in micros pore culture, spontaneous doubled haploids (SDH) are also obtained, so that no colchicine treatment will be needed in such cases. Efforts are being made to increase the frequency of SDH, so that these can be multiplied and directly field tested.