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Plant regeneration through Somatic Embryogenesis from leaf Sheath derived callus of Sugarcane (saccharum officinarum l.) Var. isd -16

Mohashweta Roy*
Institute of Biological Sciences, Rajshahi University, Rajshahi-6205, Bangladesh.

M. Hossain
Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh.

M. K. Biswas
Institute of Biological Sciences, Rajshahi University, Rajshahi-6205, Bangladesh

A. Biswas
Institute of Biological Sciences, Rajshahi University, Rajshahi-6205, Bangladesh.

R. Islam
Institute of Biological Sciences, Rajshahi University, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

Leaf sheath explants of an indigenous variety Isd ?16 of sugarcane ( Saccharum officinarum L.) produced light yellow friable callus after culturing on to MS with 2,4? D (2 ? 4 mg/l) and NAA (3 ? 5 mg/l) singly. Callus formation was the maximum on MS + 3 mg/l 2,4? D. Callus underwent embryogenesis producing huge number of somatic embryos when subcultured on MS with 15 ? 30 mg/l L? proline, 3 mg/l 2,4? D + 5 ? 10% coconut water (v/v) and 3 mg/l 2,4? D + 10% CW (v/v) + 300 ? 500 mg/l CH. L? proline significantly enhanced somatic embryo? genesis and 25 mg/l L? proline in MS was the best culture medium formulation. Most of the somatic embryos germinated and developed plantlets after 1 ? 2 weeks of incubation in proline? supplimented medium. On the other hand, maturation and germination of embryos were achieved on half ? strength MS with or without 0.25 ? 1.0 mg/l L? proline, and 5% coconut water (v/v). Somatic embryos derived plantlets were then successfully transferred to natural condition through successive phages of acclimation.

Keywords: Plant regeneration, Leaf sheath, Somatic embryogenesis, Sugarcane

Location: Institute of Biological Sciences, Rajshahi University, Rajshahi-6205, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Sugarcane

Objectives:

To develop efficient and stable plant regeneration through somatic embryo genesis for a popular sugarcane var. Isd 16 of Bangladesh.

Methodology:

Immature leaf sheath from 6 months old field grown plants were used as explant. Apical portion of healthy shoots were stripped to the terminal bud and attached immature leaf rolls were surface sterilized with 0.1% HgCl₂ for 9 min. Approximately 3 slices (5 × 3 mm) were taken from each cylindrical leaf roll and cultured on MS supplemented with 2 ? 4 mg/l 2,4?D and 3 ? 5 mg/l NAA alone and incubated in dark for callus induction. Sub ? cultures were done at every 3 weeks after incubation. In order to induce somatic embryogenesis, the proliferated friable callus was transferred to MS containing 15 ? 30 mg/l L? proline, 15?30 mg/l L? glutamine, 3 mg/l 2,4? D + 5 ? 10% (v/v) coconut water (CW) and 3 mg/l 2,4? D + 10% (v/v) CW + 300 ? 500 mg/l casein hydrolysate (CH) with 3% (w/v) sucrose (BDH, England). The pH of the medium was adjusted to 5.6 and gelled with 0.7% (w/v) agar. The cultures were also incubated in a growth chamber at 25 ± 1ºC in dark. Number of somatic embryos was counted by the help of Stereo Microscope from about 150 mg embryogenic callus. For germination, the embryogenic calli with developing somatic embryos were transferred to MS with or without L? proline (0.25 ? 1.00 mg/l) and 5% CW (v/v) and incubated in 16/8h light/dark condition. Regenerated plantlets with well ? developed root systems were transferred to pots containing soil : manure : sand (1 : 2 : 1) mixture through successive phases of acclimation. Data collected on different quantitative characters were analyzed for mean and standard error. Each experiment was performed thrice using 15 replicates. Analysis of variance was carried out and the test of significance was done by DMRT. The data in percentage were transformed using arc? sine/square root transformation before analysis and were transformed back to percentage for presentation. All statistical analyses were conducted using MSTAT? C software.

Source of Information: Plant Tissue Cult. & Biotech. 21(2): 143-149, 2011 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The present study elucidated an efficient protocol for plant regeneration via embryogenesis of sugarcane ( Saccharum officinarum L. ) var. Isd ?16 and this will be helpful in conducting genetic transformation.

Knowledge Management: Journal