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Sandaruvini PLPS and Ratnasekera D*
1Department of Agricultural Biology, Faculty of Agriculture, University of Ruhuna, Kamburupitiya, Sri Lanka.
Abstract
Doubled-haploid plant production through anther culture provides an efficient and appropriate system for
rapid production of homozygous lines. Various culture conditions influence callus induction and plant regeneration efficiency.
In this study, the optimization of media requirements and culture conditions for high-frequency callus induction and plant
regeneration for selected indica rice varieties were evaluated using improved anther culture media. Panicles of ten selected
rice cultivars (Bg 358, Bg 379-2, Bw 361, At 362, Bw, 364, Bw267-3, Bw 272-6b, At 306, Bg 357 and Kahata wee) were cold
pre-treated at 10ºC for 7-10 days. After that anthers were cultured in agar solidified N6 and L8 media where hormonal
combinations were modified. Calli were transferred to three different Murasinge and Skoog media for plant regeneration.
Green and Albino shoots were observed. Best callus induction frequencies (6.9-11.1%) were acquired in the L8 medium containing
6% (w/v) maltose, over the N6 (3.6—5.7%) medium. Highly responsive genotypes for callus initiation were AT362, BG 379-2, BW 267-3
and AT 306. BG357 and BW 364 were the least responsive genotypes. The best callus induction frequency of 11.1% was observed in the
L8 medium supplemented with 2.0mgl-1 2, 4-D and 0.5mgl-1Kinetin (KN) for AT 362. Maximum green plant regeneration could be observed
in Kahata wee (29.2%) and AT362 (18.27%) varieties in MS media supplemented with 2mg/l BAP, 1mg/l NAA and 0.5mg/l KN. After 7-10days
of callus initiation, they were transferred to MS medium to regenerate plants. Plant regeneration was observed after two weeks after
the transfer. This information could be used in the development of homozygous plants with high yields and good grain quality from the
best hybrid varieties.
Keywords: Anther culture, Indica, N6&L8 media, Plant regeneration
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* - Corresponding Author
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