Tissue Culture Technique for Callus Induction

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Sarhad J. Agric. Vol. 23, No. 1, 2007
* Department of Plant Breeding and Genetics, NWFP Agricultural University, Peshawar – Pakistan. ** Agricultural Biotechnology Institute, National Agricultural Research Center, Islamabad – Pakistan. TISSUE CULTURE TECHNIQUES FOR CALLUS INDUCTION IN RICE

Hidayat Ullah*, Iltaf Ullah*, Sultan Akbar Jadoon* and Hamid Rashid** ABSTRACT
Seeds of two rice varieties Basmati-370 and Basmati-385 were evaluated for invitro callus induction at Agricultural Biotechnology Institute in National Agricultural Research Center (NARC), Islamabad during 2001-02, using M.S and N6 media supplemented with 2, 4-D (2,4-Dichlorophenoxi acetic acid) and BAP (6-benzylaminopurine) at @ 2.0, 2.5 and 0.1, 0.5 mgl-1 respectively. Objective of our study is to know the actual procedure for seed sterilization and to provide a well-adopted technique for callus induction in these varieties by using M.S and N6 media along with different concentrations of growth hormones. Seeds were used as explant source and to avoid chance of contamination sodium hypochlorite and ethanol at the rate of 50% and 70% were used respectively. Best response toward callus induction were observed for Bas-385 on both MS and N6 media however N6 media was proved to be best. Calli obtained from Bas-385 were friable and vigorous as compare to Bas-370. Bas-370 respond moderately on MS and N6 media at various combinations of 2,4-D and BAP. Bas-370 on MS media supplemented with 2,4-D and BAP @ 2.0 and 0.1 mgl-1 gave good performance towards callus induction. Over all results indicated that best callus were induced on MS and N6 when supplemented with 2,4-D and BAP @2.0 and 0.0 mgl-1 for variety Bas-385.

INTRODUCTION
In Pakistan rice is the second leading crop after
wheat. In 2000-2001, the world annual production
of rice was 598 million tones (FAO 2002). Asia is
the largest producer of rice, with Bangladesh,
China, Philippines, Thailand and Vietnams as the
leading rice producing nations. In Pakistan rice
covered an area of 2.52 million ha, with the
production of 5.16 million tones in 1999-2000. It
also play important role in our national economy.
During 1999-2000 about 1.92 million tones of rice
was exported and earn about 465.8 million US$.
Rice is susceptible to a range of diseases and pests,
which annually destroy about 55 percent of rice
crops. The most common diseases are caused by
the fungi sheath blight and rice blast, and the stalk
borer is a common insect pest. Rice is composed
of essential food components, therefore more than
two billions people in the globe depend on rice for
more than half of the proteins and calories they
consume (Khan et al. 2000). Due to its increasing
importance in nutrition and economy, it is now felt
that new varieties of rice, having good agronomic
characters, should be evolved.
Crop improvement through tissue culture
techniques is easier and more often in use as
compared to conventional plant breeding
(Yamada, 1986). Somaclonal variations commonly
appear after tissue culture, which involve a callus
stage (Larkin and Scow Croft 1981). Callus is
undifferentiated mass of rapidly proliferating cells,
can be obtained by culturing explants source (seed,
node, bud, leaves, meristem and root tips etc) on
nutrient medium containing specific growth
regulators along with a standard recipe of
chemicals. Rashid et al. (2000) studied that rice
seeds have more potential for callogenesis as
compared to node or tip. Successful callus
induction from rice seed has been reported by
several researchers (Gonalz 2000; Navraj et al.
1999; Marrassi 1996; Valdez et al. 1997; Xie et al.
1995). But an improved method for callogenesis
was reported by Rashid et al. 2000).
The present study is based on tissue culture
techniques carried out in ABI laboratory for callus
induction in rice varieties viz. Basmati-370 and
Basmati-385. Two types of basal media M.S
(Murashige and Skoog 1962) and N6 (Nitsch and...
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