Morphological and Histological Observation of Embryogenic Calli Derived from Immature Embryo of BRRI Dhan28 (Oryza sativa L.) Variety

Somatic embryogenesis is the most common method for regeneration in rice. In vitrostudies of indica rice ( Oryza sativa L.) variety BRRI dhan28 was used for obtainingembryogenic calli from immature embryo culture on Murashige & Skoog mediumsupplemented with 2.5 mg/l dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/lnaphthaleneacetic acid (NAA) showed the highest percentage (91%) of callus induction. Inthis combination 80% embryogenic calli were formed uneven with a crisp texture, loosestructure and salient multicellular structures on the surface while non embryogenic calliwere compact with a smooth surface. Under microscopic observation, embryogenic cells weresmaller, globular and abundant in cytoplasm with one or two big nuclei. Non embryogeniccells were little cytoplasm and few large vacuoles with no or only a small nucleus and wideintercellular spaces. Non embryogenic cells had a very low cell division capability whileembryogenic cells had a high capability for cell division and continued to divide andproduced somatic pro-embryos with a well–defined protodermis which could developfurther through the typical globular, heart, torpedo and cotyledonary stages. Only 80% ofembryogenic cells were induced high differentiation rate and developed 65 globular, 52heart-shape, 43 torpedo and 37 cotyledonary embryos of embryogenic cells after 30 to 45 daysof induction.

[1]  A. M. Espinoza,et al.  Effect of 2,4-d, hydric stress and light on indica rice (Oryza sativa) somatic embryogenesis. , 2014, Revista de biologia tropical.

[2]  N. Haq,et al.  Regeneration efficiency and genotypic effect of some indica type Bangaldeshi rice (Oryza sativa L.) landraces , 2005 .

[3]  G. Hahne,et al.  Histological analysis of somatic embryogenesis induced in leaf explants ofHelianthus smithii Heiser , 1997, Protoplasma.

[4]  M. Rajam,et al.  Efficient plant regeneration from long-term callus cultures of rice by spermidine , 1995, Plant Cell Reports.

[5]  C. Fauquet,et al.  An improved rice transformation system using the biolistic method , 1993, Plant Cell Reports.

[6]  T. Davis,et al.  Zeatin-induced shoot regeneration from immature chickpea (Cicer arietinum L.) cotyledons , 2004, Plant Cell, Tissue and Organ Culture.

[7]  R. Xu,et al.  Agrobacterium-mediated transformation of élite indica and japonica rice cultivars , 1997, Molecular biotechnology.

[8]  J. Schel,et al.  Cytodifferentiation during the development of friable embryogenic callus of maize (Zea mays) , 1991 .

[9]  N. Michaux-ferrière,et al.  Histology of Somatic Embryogenesis from Leaf Explants of the Oil Palm Elaeis guineensis , 1988 .

[10]  I. Vasil,et al.  Somatic Embryogenesis in Sugarcane (Saccharum officinarum L.): Growth and Plant Regeneration from Embryogenic Cell Suspension Cultures , 1983 .

[11]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[12]  F. Skoog,et al.  A revised medium for the growth and bioassay with tobacco tissue culture , 1962 .