Establishment of a simple efficient in-vitro regeneration protocol for locally grown tomato (Lycopersicon esculentum Miller) cultivers of Bangladesh

Abstract

A standardized in- vitro regeneration protocol was developed by reducing complexity and increasing in- vitro regeneration efficiency of Bangladeshi three most popular tomato cultivars namely, Bina tomato-3 (B-3), Bina tomato-5(B-5), Bahar (BR) and Indian cultivar Pusa Ruby (PR). In the present study cotyledonary leaves were used as explant and were collected from 8-10 days old in vitro germinated seedling. To increase germination, agitation of seeds following sterilization was found to be effective and addition of Tween- 20 helped to get contamination free germination. Throughout the study different concentrations of 6-benzylaminopurin (BAP) and indol-3 acetic acid (IAA) were used alone and in combination. Significant differences for regeneration percentages, time taken to regenerate and number of shoot/explant were observed for different treatments and different cultivars. Maximum regeneration % was obtained with 2mg/l BAP as well as it required less time and produced highest number of shoot/explant. Addition of IAA in the best regeneration media were found to have negative effect on regeneration %, number of shoot/explants and time to get regenerated shoots in all four cultivars. Dorsal placement of explants in the best responsive media showed best response regarding regeneration responses for all cultivars. Rooting was best in half strength MS media supplemented with 0.2 mg/l IAA. The regenerated plantlets successfully acclimatized in soil, where they flowered and formed fruits. Seeds collected from these fruits were found to be viable in germination tests.