I found in my species ,its very difficult to germinate the seeds .So i cultured the endosperm by removing the seed coat .Now i plan to propagate in large numbers for in vitro condition .Can any one suggest which explant will best regenerate ?
So, you want to regenerate from endosperm? But endosperm is triploid, so I would think that endosperm would not be the best starting material if you intend to regenerate normal diploid lines.
Hi Manasa,
I assume that you meant to ask: you are tissue-culturing the seed coat-less 'seed' (endosperm + embryo) to help it germinate in the tissue culture medium, and after it germinates into a seedling, which part ('explant') will be the best material to be used for regeneration? In this case, it depends on the species, as one scientist has mentioned above. I have done some species: carrot (we used hypocotyl to induce calli and regenerated into plants); tobacco (cotyledon or true leaves to regenerate 'shoots'), cotton (cotyledon to induce calli and regenerated into plants).
But be aware, people do use 'endosperm'-only tissue for tissue culture to rapidly regenerate triploid (3N) plants and bypass the tedious breeding process of obtaining triploid plants from crosses of 2N x 4N plants. Or, did you mean this?
Best,
Endosperm is not used for regeneration of diploid plants but triploid plants. for normal plant regeneration u have to germinate the seeds or u should use only embryo as an explant to induce callus. if u are getting callus from endosperm but it is not sure that it is diploid (from zygote) or triploid (from endosperm. I thine u should add gabberllic acid in the MS medium for germination of seeds or germinate the seeds in the green house in vivo and then use explants - hypocotl, stem leaf root for callus induction and further objectives.
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