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AGROBACTERIUM-MEDIATED TRANSFORMATION AND DIRECT 

SHOOT REGENERATION IN IRANIAN TOMATO 

(SOLANUM LYCOPERSICUM L.) CULTIVAR FALAT- CH.

NAHEED KAUSER1,4, SAIFULLAH KHAN1*, SEYED ABOLGHASEM MOHAMMADI2, BEHZAD GHAREYAZIE3*, EBRAHIM DORANI ULIAIE2 AND BANAFSHEH DARVISHROHANI3

1Biotechnology Wing, H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi- 75270, Pakistan,

2Genomics & Molecular Breeding Lab, Department of Plant Breeding & Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, 51666 Iran,

3Agricultural Biotechnology Research Institute of Iran (ABRII), Karaj, Iran.

4Jinnah University for Women, Nazimabad No. 5, Karachi, Pakistan.

*Corresponding authors: Email: drsaif65@gmail.com Phone # 92-21-99261693

ghareyazie@yahoo.com Phone#98-2133272234

Abstract

Falat CH is an important commercial tomato cultivar being used in Iran. In this article an optimized protocol with increased transformation and regeneration rate for this tomato variety is reported. Several explants including cotyledon, leaf and hypocotyl were evaluated for direct shoot formation and the effect of various combinations of BAP, Zeatin, IAA and IBA were studied. It is the first report on two cytokinins BAP and Zeatin in various combinations to evaluate the synergetic effect of cytokinins on direct shoot regeneration. The synergetic combination of 1.5mg/l BAP, 0.5 mg/l Zeatin and 0.2 mg/l IAA was considered as the best treatment which resulted in higher plant regeneration rates from all of the explants over previous reported methods. Using the best regeneration treatment obtained, the HBsAg gene was transferred into the tomato explants using Agrobacterium mediated transformation technique Percent of the putative transgenic plants regenerated was 68%. PCR of putative transformed plants showed that 87.1% of regenerated plants amplified nptII and HBsAg gene when specifically designed primers were used; giving a final transformation rate of 34.85%.

Key words: Agrobacterium tumefaciens, GV3101, BAP, IAA, IBA, Transformation escapes, Synergetic effect of cytokinins, Zeatin.