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Wheat Information Service
Number 72: 95-97 (1991)


Effect of culture condition on plant regeneration capacity of mature embryo derived callus in wheat (Triticum aestivum L.)

K. Kato, S. H. Chowdhury and S. Harada

Faculty of Agriculture, Kochi University, Nankoku 783, Japan


Immature embryo has so far been the best explant for efficient regeneration of euploid plant in wheat. However it is required to exploit another explant, because it is usually difficult to get immature embryo throughout the year. In addition, the suitable stage of immature embryo is strictly limited (Shimada and Yamada 1979). This study was, therefore, attempted to establish an efficient culture method of callus derived from mature embryo as an alternative explant.

An Ethiopian local wheat cultivar 'IL 68' was used for embryo culture. To kow the effect of pre-treatment with 2,4-D in the course of embryo development, detached ears cultured after Kato et al (1990) were dipped in 2,4-D solution (2mg/l) for 3 minutes. This treatment was conducted at one of the seven developmental stages, that is, 8, 11, 14, 17, 20, 23 and 26 days after anthesis. Then treated car was again cultured on liquid medium until their maturation. After ripening, caryopses were harvested and used for culture as well as control caryopses harvested from non-treated detached ear and from intact ear of field grown plant. These seeds were first soaked in hormone solution at 5C for 3 hours. The solution contained MS inorganic salts, 30g/l sucrose and 2mg/l 2,4-D, combined with or without 1mg/l BAP as described by Sasakuma and Tei (1989). After seed sterilization, excised embryos were placed on agar medium with the scutellum uppermost. The medium contained MS inorganic salts, 30g/l sucrose and 8g/l agar, supplemented with 2,4-D and BAP as shown in
Table 1. Callus induction and maintenance were conducted under a 26C and contiunous light (1500-2000 lux) regime.

Callus induction rate was not different between the two cultures, being 83.3% in culture No.1 and 85.7% in culture No.3. It was also independent of the pre-treatment stage during detached ear culture. In culture No.1 and No.2, shoot bud appeared frequently at green spot region after 90 days of culture (
Fig. 1). On the other hand, in culture No.3, it appeared after 130 days of culture. This result indicated that BAP added to pre-soaking medium and to callus induction medium enhanced the speed of shoot differentiation.

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