Hi, i want to correlate microspore development stage with flower bud size in Tagetes spp. (Marigold). I am currently using aceto-carmine stain for pollen staining, but i can find up to tetrad stage only. Later on released microspores are having thick exine layer with spines and i could not able to observe any nucleus inside the pollen grain. I want to standardize the pollen at late uni-nucleate stage for anther culture. Can any body suggest me how to prepare slide to observe nucleus development (late uni-nucleate, early bi-nucleate) with vacuole. Please find the enclosed pictures of tagetes pollen grain.
Thank You
Hi Younes,
I have used but i couldn't find any nucleus in the pollen grain (You can observe in the attached pictures). This may be due to thick exine wall, spiny projections and high accumulation of sporopollenin in Marigold pollen grain. I want a different technique to observe the pollen maturation stage.
Methylene blue staining is good to view uninucleate, early binucleate stages. The nucleus can be viewed in the cells.
Hello Ravinda, I think that if you would like to observe the nucleus, you may add cell lysis procedures. You may submerge them first in hypotonic solutions. If you would like to get rid of the coat totally, you may use cellulase.
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