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Plant Regeneration from Mature Embryo Derived Callus of Basmati Rice

M. S. Ferdous
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. A. Samad
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2200, Bangladesh

M. S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

S. A. Mony
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

The experiment was conducted with mature embryos of an aromatic rice variety (Basmati 370) to investigate the appropriate growth condition for callus induction as well as plant regeneration. Different concentrations of 2,4-D (0.0, 1.5, 2.5, 3.5, 4.5 mgL^-1) in MS medium were used for callus induction. Among the concentrations of 2,4-D, 2.5 mgL^-1 was the best concentration for callus induction (95.83%). Different concentrations of Kinetin (0.0, 8.0, 10.0, 12.0, 14.0 mgL^-1) with a constant concentration of NAA (0.5 mgL^-1) were used for plant regeneration from callus. The combination of 10.0 mgL-1 Kinetin and 0.5 mgL^-1 NAA was found to be the best for shoot regeneration (77.50%). Different concentrations of MET (1.5, 2.5, 3.5 mgL^-1) in MS medium with a constant concentration of NAA (0.5 mgL^-1) were used for root induction. A combination of 2.5 mgL^-1 MET and 0.5 mgL^-1 NAA was found most efficient for root induction (85.00%) with higher number of roots per plant (19.50) and better root elongation (1.88 cm). The plantlet was survived well in the plastic pot as well in the earthen pot or in the field.

Keywords: Rice, Plant regeneration, Mature embryo, Callus induction

Location: Tissue Culture Laboratory of Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh, Bangladesh

Start Date: 00-01-2007

End Date: 00-11-2007

Program Area: Variety and Species

Commodity: Rice

Objectives:

a. To develop to observe the effect of different concentrations of hormone on callus induction and plant regeneration and

b. To develop a suitable and reproducible protocol for in vitro plant regeneration using embryogenic calli from mature embryos of an aromatic rice.

Methodology:

The experiment was conducted during the period from January to November 2007 at the Tissue Culture Laboratory of Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh. Mature embryos of aromatic rice (Oryza sativa L.) Basmati 370 variety was used as experimental materials in the present investigations. For surface sterilization, the dehusked seeds were taken into a sterilized tube and immersed into 0.1% HgCl₂ solution for 20 minutes followed by 4-5 rinses with sterile distilled water to remove traces of HgCl₂. Mature embryos attached to endosperm were the explants for embryo culture. The explants were cultured directly in MS (Murashige and Skoog, 1962) medium supplemented with different combinations of hormones. MS medium supplemented with 0.0, 1.5, 2.5, 3.5 and 4.5 mgL^-1 2, 4-D were used for callus induction, while for shoot differentiation MS medium was supplemented with 8.0, 10.0, 12.0 and 14.0 mgL^-1 Kn with a constant addition of 0.5 mgL^-1 NAA and for root induction MS medium supplemented with 1.5, 2.5 and 3.5 mgL^-1 MET with a constant concentration of 0.5 mgL^-1 NAA were used. Sterilized potting mixture containing garden soil, sand and cowdung in the ratio of 1:2:1 was taken into 10 cm plastic pots for growing the plantlets at in vivo condition. The regenerated plantlets with sufficient roots were then taken out from the culture vessels and thoroughly washed in running tap water to remove all adherent culture medium. Data recorded for different parameters under study were compiled and tabulated for statistical analysis.

Source of Information: Progress. Agric. 19(2) : 45-50, 2008 ISSN 1017-8139

Article Link (Online Journal): http://www.banglajol.info/index.php/PA/article/download/16921/11880

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation, a technique for plantlet regeneration in vitro through callus induction, shoot regeneration and rooting of the shoots has been established by using mature embryos of aromatic rice genotype namely Basmati 370 as explants. The explants were inoculated on MS medium supplement with different concentrations of 2,4-D for the callus formation. In medium without growth regulator, no callus was induced. The highest percentage (95.83%) of explants showed callus induction in 2.5 mgL^-1 2,4-D followed by 1.5 mgL-1 2,4-D (79.16%) and the lowest percentage (50.00%) of callus induction was found in 4.5 mgL^-1 2,4-D.The present results were supported by the findings of Islam (2004) who observed the effect of MET (2.5 mgL^-1) and NAA (0.5 mgL^-1) on root formation in indica rice (Oryza sativa L. cv. Kataribhogs) and reported that maximum root induction was occurred from this hormonal combination.

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