PJB-2018-1654
Estabglishment of an efficient protocol for plantlets regeneration via direct and indirect organogenesis Citrus reticulata (Kinnow Mandarin
Mubashir Hussain, Naveed Iqbal Raja, Hamid Rashid, Zia-ur-Rehman Mashwani and Muhammad Iqbal
Abstract
Studies were initiated to explore the role of plant growth regulators and explant types on efficient plantlets regeneration via direct and indirect organogenesis in Kinnow mandarin [Citrus reticulata L. (Blanco)]. Explants were cultured on MS medium containing varying concentrations of phytohormones. The best callus induction response was obtained in MS medium containing 5 mg/l 2,4-D and 1 mg/l BAP where 90% from nucellus tissue, 58% from shoot apical meristem and 56% from nodal segments explants showed callogenic response after 2 weeks of inoculation.Best shoot induction medium via indirect organogenesis was found for nucellus tissue when MS medium was supplemented with 1.5 mg/l Kin and 500 mg/l malt extract which was 46% whereas for SAM and nodal segments, best response was obtained when MS medium was supplemented with BAP and NAA (3.0 + 0.5) mg/l which was 40% and 48% from SAM and nodal segments respectively after 11 weeks of inoculation.The shoot induction as well as multiplication were also studied by using different combinations and different concentrations of phytohormones in growth medium. The best medium for shoot induction percentage via direct organogenesis was [M3= MS + BAP (1.0 mg/l)] whereas 92%, 90% and 82% from nucellus tissue, SAM and nodal segments respectively showed organogenic response after 4 weeks of inoculation, whereas maximum number of shoots per explants was obtained in medium (M9)containing MS + Kin where 4, 7 and 6 average number of shoots were obtained from nucellus tissue,SAM and nodal segments respectively after 11 weeks of inoculation.Best rooting medium was found when MS medium was supplemented with 2 mg/l NAA where 88% from nucellus tissue and 80% from the nodal segments showed rooting response whereas for SAM best rooting response was obtained when supplemented with 1.5 mg/l NAA i.e 78%. The plantlets were successfully acclimatized in different potting mixtures and highest survival rate (100%) was achieved in potting mixture containing sand and peat moss (2:1).
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