Evidence of the change in ploidy levels in the plantlets established from endosperm culture of Persian shallot ( Allium hirtifolium )

Allium hirtifolium (Amaryllidaceae), called “Mooseer” or Persian shallot in Iran, is a bulbous plant with high medicinal values. It is native and endemic to the North West to central and South West of Iran. In the present study, regeneration plants with chromosomal changes are produced through in vitro mature and immature endosperm culture. In our study, both cultured endosperms formed yellow- green calli induced in the MS medium supplemented with (1.5 or 2 mg·L − 1 ) 2,4-dichlorophenoxyacetic acid (2,4- D), (0.5 or 1 mg·L − 1 ) α-naphthalene acetic acid (NAA) plus (0.5 or 1 mg·L − 1 ) 6-furfurylamino purine (Kin) and 6-benzylaminopurine (BAP). The highest callus induction in mature (87.62%) and immature (62.49%) endosperms was achieved on a medium with 1 mg·L − 1 NAA and 1 mg·L − 1 BAP. Regenerated shoots were transferred for bulblet formation to investigate their potential for bulblet production. Regeneration of plantlets from endosperm tissue shows high chromosome variations and polyploidy. Flow cytometry analysis and chromosome number confirmed that diploid (2n = 2x = 16), triploid (2n = 3x = 24), tetraploid (2n = 4x = 32) and aneuploid plantlets were obtained. The comparison of stomatal density and size in complete plantlets has revealed that with the increase ploidy level, stomatal density decreased because of enlargement in stomatal size.