Title: Establishment of in planta transformation protocol of tomato (Solanum lycopersicum L.) through antiporter gene for improved salinity tolerance

Biography:

Anamika Datta is currently working as a Doctoral Student in Kyushu University, Department of Stem Cell Biology under Faculty of Arts and Science, Graduate School of System Life Sciences, Kyushu University, Japan. She has been receiving scholarship from Japan Science and Technology Agency. She completed her MS in Biotechnology from BRAC University, Dhaka, Bangladesh and worked as a MS fellow under a project titled “Production of tomato tolerant to environmental stresses like salinity”; by BAS-USDA PLAS Endowment fund (Li-03). She developed her skill on plant tissue culture and Agrobacterium-mediated transformation during her working period. She has been worked as faculty member in BRAC University, Dhaka Bangladesh for 3 years. During her research career she has published 2 original research paper, 2 review papers, with 6 conference papers and 2 poster publications in international conferences. Currently she is pursuing her doctoral studies in Neurobiology and Stem Cell related research.

Abstract

Salinity has been accounted for global production loss of the tasteful and nutritious vegetable “Tomato” lately. In this backdrop, a simple and efficient Agrobacterium mediated in planta transformation protocol has been established, to transform and overexpress Na+/H+ antiporter gene into five Bangladeshi tomato varieties named as BARI tomato 2, BARI tomato 3, BINA tomato 2, BINA tomato 3 and Bahar, to improve their salt tolerance, through optimization of crucial transformation factors like optical density, infection time, co-cultivation period etc. Two vectors were constructed by cloning Na+/H+ antiporter gene from Arabidopsis (pK7WG2_AtNHX1_1.6) and Rice (pK7WG2_OsNHX1_1.6) individually to gateway vector pENTR/D-TOPO and electroporated to Agrobacterium while another vector pBI121 was used as control. During transformation using the three vectors, non- pricked seeds infected under conditions of OD600 1.1-1.4 with 30 min of infection time followed by 24h co-cultivation period were found optimum for achieving more than 90% efficiency for GUS expression and germination percentages. Screening of putatively transformed plantlets were carried out through resistance to Kanamycin 50mg/l in germination medium while Cefotaxime 100mg/l was applied to prevent Agrobacterium overgrowth during co-cultivation. Next, salt stress tolerance has been observed in putatively transformed plants through Leaf Disc Bioassay. Here, tolerance of 100mM salt for 14 days was noted. Meanwhile, no significant morphological changes were observed during acclimatization of putatively transformed plantlets. These findings can be considered as the foundation for further improvement of saline-tolerant tomato development and an essential step toward solving the problem of ensuring global nutritional demands.