TITLE:
In-Vitro Regeneration of Citrus sinensis (L.) Osbeck from Mature Seed Derived Embryogenic Callus on Different Solid Basal Media
AUTHORS:
Md. Nazmul Hasan, Mohammed Raqibul Hasan, Shakhawat Hossain Foysal, Hammadul Hoque, Md. Fahim Khan, Md. Fahmid Hossain Bhuiyan, Shamsul H. Prodhan
KEYWORDS:
Citrus sinensis (L.) Osbeck, PGRs, Basal Media, Regeneration
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.10 No.2,
February
26,
2019
ABSTRACT: In-vitro callus
induction and regeneration method was developed using different plant growth
regulators (PGRs), and basal media (Murashige and Skoog (MS), CHU (N6) and
Gamborg (B5) media) of Citrus sinensis (L.)
Osbeck. Observations of the effect of PGRs were carried out using different
concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D),1-naphthalene acetic
acid (NAA) and combinations of 2,4-D and NAA using different basal media. This
study found Citrus sinensis (L.)
Osbeck exhibited a high frequency of callus induction on MS medium supplemented
with 3 mg/L 2,4-D and callus induction frequency was 86.7% ± 3.4% whereas N6
and B5 showed lower callus induction frequency of 83.3% ± 8.8% and 82.2% ± 1.9%
respectively compared to that of MS media with supplementation of the same
hormone. Among the induced calli, the morphological analysis showed only 40% - 50% was embryogenic calli.
Regeneration of plantlets from calli was done using different concentrations
and combinations of auxin and cytokinin. The study showed that 3 mg/L
6-benzylaminopurine (BAP) supplemented medium has the maximum potential to
promote regeneration of Citrus sinensis (L.)
Osbeck from embryogenic calli with the frequency of 89.3% ± 8.8% but no
regeneration occurred from the non-embryogenic calli. The regenerated plantlets
were rooted on MS medium with supplementation of 5 mg/l NAA. These observations
in Citrus sinensis (L.)
Osbeck regeneration will be helpful for genetic improvement with desired
traits.