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Effects of Growth Regulators on Induction of Callus and Regeneration of Plants in Spring Wheat (triticum Aestivum L.)

Farhana Sharmeen
Department of Genetics and Plant Breeding, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

A. K. M. Shamsuddin
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Abstract/ Executive Summary:

Effects of growth regulators on induction of callus and regeneration of plants were studied in five varieties of wheat namely: Chyria 3, Sonalika, Shatabdi, Mayoor and Kanchan. There were significant variations among the varieties for callus size, number of bud forming calli and regeneration of plants from the callus. The varieties Mayoor and Chyria 3 performed better for callusing in callus induction media. These two varieties also showed good performance for regeneration of plants in regeneration media. The lower concentration (1.0 mg L^-1) of 2, 4-D and Picloram was found suitable for induction of callus. The regeneration media containing NAA at 0.5mgL^-1 produced higher number of plants per callus. The variety Chyria 3 showed significantly better performance for regeneration of plants in the media containing either NAA at 0.5 mgL^-1 or BAP at 1.0 mgL^-1 or Casein hydrolysate at 100.0 mgL^-

Keywords: Spring wheat, Callus, Regeneration, Growth regulators

Location: Mymensingh

Start Date: 00-11-2004

End Date: 00-06-2005

Program Area: Variety and Species

Commodity: Wheat

Objectives:

The present study aims to develop of protocol as well as assessment of varietal differences of wheat for their regeneration efficiency through tissue culture.

Methodology:

The experiment was conducted in the Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, during the period from November, 2004 to June, 2005. Five varieties of spring wheat viz. Chyria 3, Sonalika, Shatabdi, Mayoor and Kanchan were grown in earthen pots. At booting stage the pots were transferred to a green house provided with 25/17⁰C day/night temperature and 16h photoperiod regime. When the wheat spikes were at the age of 16 to 18 days after anthesis, they were harvested and the caryopses were collected. The caryopses were surface sterilized by treating with 70% ethyl alcohol for 3 minutes and commercial bleach (chlorox containing 4.7% chlorine) supplemented with a few drops of Tween 20 for 15 minutes. Then caryopses were thoroughly washed in double distilled water. The embryos were excised in sterile condition under a laminar airflow cabinet and plated in the following four callus induction media. MS+ 2,4-D at 1.0 mgL^-1, MS+ 2,4-D at 2.0 mgL^-1, MS+ picloram at 1.0 mg L^-1_, MS+ picloram at 2.0 mg L-1.

The embryos were incubated at 25±1⁰C in continuous dark for 20 days. Incubation gave development of calli, and the compact and noduller calli were transferred to petridishes containing following plant regeneration media. MS (used as control), MS+ casein hydrolysate at 100 mgL^-1, MS+ casein hydrolysate at 150 mgL^-1, MS+ benzylaminopurine at 0.5 mgL^-1, MS+ benzylaminopurine at 1.0 mgL^-1, MS+ naptheleneacetic acid at 0.5 mgL^-1 MS+ naptheleneacetic acid at 1.0 mgL^-1.

The calli were kept in regeneration media at 25±1⁰C and 16h photoperiod regime in an incubation room for 6 to 7 weeks. After incubation, when the plants grew upto 3 to 4 cm in height, they were transferred to culture tubes containing MS medium free of any hormone. The regenerated plants at the age of 20 to 25 days, developed sufficient roots and were transferred to small cubicles containing soil and organic matter. The plants were hardened in a growth chamber where the temperature was 25 ±1⁰C and photoperiod was 16h with 45000-lux light intensity. The plants were kept in growth chamber for one week and then they were transferred in earthen pots containing normal soil and organic matter. The plants were grown in green house under 25±1⁰C with 16h photoperiod regime. The experiment was carried out in two phases, (i) induction of callus (ii) regeneration of plants. After induction of callus data were taken on size of callus (measured by diameter of the callus) and number of bud forming calli. In the regeneration phase number of regenerated plants from each callus were recorded. The data were analyzed for variance following the principles of randomized block design.

Source of Information: Bangladesh J. Pl. Breed. Genet., 19(1) : 01-06, 2006

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Overall results of the experiment suggest that the varieties Chyria 3 and Mayoor were the responsive varieties to tissue culture and MS medium supplemented with NAA would give high regeneration of plants in wheat.

Knowledge Management: Journal