Hi, I want to do my research work on doubled haploid production in Tagetus (Marigold). I couldn't find any successful protocol for the same. Hence, I am looking for stage of pollen collection, media and other procedures for developing haploid lines.
Ravindra Kumar K
For developing doubled haploid plants, haploid plants are to be developed first by culturing anther or unfertilized female flower/ inflorescence under in vitro conditions through andro or gynogenesis. To induce andro or gynogensis, various hormone PGR combinations are to be supplemented in culture media and the exact requirement has to be standardized, which will vary plant to plant. Then, the regenerated haploid shoots has to be sub-cultured with medium supplemented with colchicine (concentration has to be standardized) for doubling (diploidization) of chromosomes in haploid shoots to get doubled haploid shoots. Then the doubled haploid shoots can be cultured for rooting and plantlets established after proper hardening/ acclimation. Haploidy of the gynogenic or androgenic shoots must be confirmed through chromosome study (cytology) and the same can be repeated for doubled haploid plants also.
Thank you Dr.Subramaniyam and Goumi for your reply. I am starting with anther culture for producing DH.
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