2021 Vol.27 No.1 PP 42-48
https://doi.org/10.33451/florafauna.v27i1pp42-48
Induction of nucellar embryogenesis in nucellar tissue of Citrus aurantifolia for their clonal multiplication
*Manjula Singh, S.P.Paliwal and Shailendra Singh
Department of Botany,
Narain College,
SHIKOHABAD- 283 135 Dist. FIROZABAD (UP) INDIA.
*Corresponding Author :
Email : manjulapundhir@yahoo.co.in
ABSTRACT
Citrus aurantifolia (lime) has been selected as explant for nucellar embryogenesis. Nucellus is a non-vascularized tissue
being true-to-type same as mother plant, meristematic cells have no plasmodesmata connection, no virus can pass through
nucellus, thus it seems to be a good material for production of virus freeplantlet.Putrescine at 0.25 or 0.5 mg1-1 and
anapthaleneacetic acid at 0.10 mg1-1 supplemented to nutrient formulation were most effective in alleviating cotyledonary
proliferation and fasciation while promoting embryo-to-embryo proliferation producing numerous whitish globular embryos
were formed. For further development of globular embryos to well-differentiated cotyledonary embryos, additional presence
of 2-isopentenyladenine at concentrations of 0.10 or 0.25 mg 1-1 was essential, contrary to incorporation of 0.10 or 0.25 mg
1-1 6benzylaminopurine, which promoted excessive proliferation of cotyledonary structures and their fasciation while zeatin
at the same concentrations produced intermediate response. In the optimum treatment containing 0.25 mg l-1 putrescine, 0.10
mg 1-1 isopentenyladenine, 0.10 mg 1-1 indole-3-acetic acid and 100 mg l-1 malt extract, an average 10 well-developed embryos
per culture were formed, besides some abnormal cotyledonary structures. Well-developed embryos measuring ca. 2 cm. in length
(leaving the root) germinated 100% into plantlets, during 60 days, in the additional presence of amino acid supplement
comprising, 5 mg 1-1 each of L-arginine, L-asparagine, L-histidine, L-cysteine, L-lysine and 10 mg l-1 L-glutamine. Such
plantlets nurtured in a different medium attained a height of ca. 4 cm in 45 days before they were taken out for ex vitro
growth. There was 100% transplant success and the plants grew normally.
Key words : C.aurantifolia, Cloning, Embryo-to-embryo proliferation, Nucellarembryogenesis.