PJB-2016-294
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 DARVISHROHANI
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%.
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