Development of an Efficient in Vitro Regeneration System for Endangered Wild Orange Citrus Chrysocarpa L.

Shamsul H. Prodhan, Md. Nazmul Hasan, Hammadul Hoque, S. S. Alam, Mohammed Raqibul Hasan, Anup Gupta, Most. Ummay Salma Khatun, Afroza Parvin, Ziaul Faruque Joy


A method for in-vitro propagation of wild type Indian orange (Citrus chrysocarpa L.) was developed by shoot organogenesis from seed. Mature seed embryos were used as explants and treated with different hormones and plant growth regulators on MS medium for callus, shoots and roots induction. For callus induction seed embryos were treated with different concentration of 2, 4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and a-naphthalene acetic acid (NAA) and maximum 90.90% callus was observed on MS medium supplemented with 2, 4 -D at 16.0 M concentration.

Approximately 85% of the callus was granular, while 15% was smooth and compact. Maximum 83.33% shoot regeneration were observed on MS medium supplemented with BA at 13M concentration. Regenerated shoots were rooted on MS medium supplemented with different hormones and maximum 80% roots were observed in MS medium supplemented with 10 M Indole-3-butyric acid (IBA). The regenerated plantlets were successfully acclimatized in ex-situ condition for further study.


Citrus chrysocarpa L.; In-vitro; Callus induction; Shoots formation; Explants, Rooting; acclimatization.

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