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Effect of Media Components on Nature of Callus and Subsequent Regeneration of Spring Wheat (triticum Aestivum L.)

M. Z. I. Talukder
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

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

H. Rahman
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

L. Hassan
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

Abstract/ Executive Summary:

The experiment was conducted to investigate the nature of callus and regeneration potential of immature embryos of wheat (Triticum aestivum L.) of five varieties: Protiva, Pavon, Kheri, Sonalika and Kanchan on MS supplemented with different concentrations and combinations of auxins and cytokinins. Calli developed from immature embryos by culturing them in MS with different concentrations of 2,4-D. There was a clear variation among the varieties and also among the different concentrations and combinations of auxin and cytokinin. The varieties Pavan and Protiva were found to be the best for producing good, compact and organized calli. MS containing 1.5 mg/l Kn gave good, compact and organized calli which turned yellow to brown over the period of incubation. Plant regeneration was best from compact, off-white, smaller calli although shoot regeneration was higher from calli of larger size.

Keywords: Callus nature, Media effect, Regeneration, Spring wheat

Location: Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Wheat

Objectives:

To evaluating the effects of different combinations of growth regulators in culture medium and also the culture period on regeneration potentiality of callus from five varieties of Spring wheat.

Methodology:

Seeds of five varieties, namely Kanchan, Protiva, Kheri, Pavon and Sonalika of Spring wheat (Triticum aestivum L.) were sown in earthen pots containing well tilled soil collected from the field laboratory of the Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh in November 2001. When the plants attained booting stage, the pots were transferred to a greenhouse where 25/16⁰C day/night temperature with 16 h photoperiod was maintained. Immature 15-day-old caryopses were harvested from the main spike. They were surface sterilized with 70% (v/v) ethanol for 3 min, rinsed with water and then with 6% sodium hypochlorite for 15 minutes. They were washed again several times with sterile distilled water. Immature embryos were aseptically excised from the caryopsis and those measuring 0.5 - 2.0 mm in dia were placed with the scutellum upwards on a solid agar MS supplemented with three concentrations of 2,4-D (2, 4 and 6 mg/l) in Petri dishes. In the control embryos were placed on basal MS. They were incubated for ten days at 25 ± 1_C in continuous darkness. Compact and nodular calli were transferred to the regeneration media. The calli were kept for six weeks at 25 ± 1⁰C in a 16 h/8 h light/dark cycle provided by white fluorescent tubes (3000 lux). Plantlets regenerated from the calli. When roots and shoots were established and the plantlets were 3 - 4 cm tall, they were transferred to culture tubes containing hormone free MS to enable further shoot and root elongation. After 20 - 25 days regenerated plants developed roots. About 7 - 10 cm tall plantlets were transplanted to soil in pots. Initially the pots were kept in a growth chamber under a controlled environment (25 ± 1⁰C and 4500 lux light intensity). Thereafter the plants were transferred to soil in earthen pots containing cowdung and fertilizers and kept in the greenhouse where temperature regime was 25/16⁰C (day/night), a 16 h photoperiod and 45 to 60% relative humidity. Finally, after 10 - 15 days the plants were transferred to the field and grown to maturity. In the present study data were recorded at 15, 30 and 45 days after inoculation of immature embryos on the regeneration medium. The calli were graded according to their colour in a scale of 3 to 1 (scale : 3 = off-white /greenish, 2 = yellow, 1 = brown). The nature of callus was measured by the callus compactness and graded into two categories: compact (c) and friable (f). Abundance of callus was measured by a transparent measuring ruler and graded according to their length (scale : 3 = 5.1 mm and above, 2 = 3.1 to 5.0 mm and 1 = 3.0 mm and below). Ten immature embryos were cultured per Petri dish and they were sealed with parafilm. The effects of these qualitative characters with duration of time for regeneration were estimated in percentage. The per cent of shoots and plantlets were estimated on the basis of the number of calli.

Source of Information: Plant Tissue Cult. 14 (2) : 113-121, 2004 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, after the inoculation of callus in the regeneration media, the callusing behavior changed progressively. After 45 days of inoculation, the varieties Pavon and Protiva gave the best results on MS + Kn and produced maximum off-white, compact and organized calli. The highest plant regeneration was found from immature embryos derived from off-white, compact and organized calli. Plant regeneration was best from compact, off-white, smaller calli although shoot regeneration was higher from calli of larger size.

Knowledge Management: Journal