Evaluation of Factors Influencing In Vitro Regeneration and Transformation Protocols to Produce Salinity Tolerant Tomato (Solanum lycopersicum L.)
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Evaluation of Factors Influencing In Vitro Regeneration and Transformation Protocols to Produce Salinity Tolerant Tomato (Solanum lycopersicum L.). / Ferdous, Manzur-E-Mohsina; Datta, Anamika; Islam, Aparna.
In: Journal of Applied Biotechnology Reports, Vol. 9, No. 3, 2022, p. 726-739.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Evaluation of Factors Influencing In Vitro Regeneration and Transformation Protocols to Produce Salinity Tolerant Tomato (Solanum lycopersicum L.)
AU - Ferdous, Manzur-E-Mohsina
AU - Datta, Anamika
AU - Islam, Aparna
N1 - Publisher Copyright: © 2022 The Author(s).
PY - 2022
Y1 - 2022
N2 - Introduction: Tomato (Solanum lycopersicum L.) is one of the essential vegetables worldwide, for consumption and mitigating malnutrition. Genetic transformation conceivably overcomes its yield challenges due to salinity, a crucial constraint for the economical use of 30% of cultivable lands in the coastal region of Bangladesh. Therefore, a robust and reproducible protocol has been established for in vitro regeneration and transformation to develop transgenic salt-tolerant tomato plants. Materials and Methods: During micropropagation, cotyledonary leaf explants were excised and cultured on MS media containing different combinations and concentrations of plant growth regulators. In transformation, the pre-cultured explants were inoculated and co-cultivated with Agrobacterium. Then they were transferred to the antibiotics-supplemented media to achieve salt-tolerant putative transformed plants. The transformation was confirmed by P-glucuronidase (GUS) assay and PCR for the antiporter gene. Results: Maximum regeneration response was achieved from the explants abaxially positioned at a 1.5 cm distance apart. BARI Tomato 14 and BINA Tomato 3 showed the highest shoot regeneration response (%) on MS media supplemented with 2 mg/L BAP and with 0.1 mg/L IAA for BARI Tomato 2 and BARI Tomato 15. Bacterial culture of OD600 0.68 for 30 min and a Co-cultivation period of 48 h resulted in the highest transformation frequency (47%) in Agrobacterium-mediated transformation with pBI 121 in BARI Tomato 3. The highest regeneration frequency (20.5%) was obtained in transformation with pH7WG2_OsNHX1_1.6. Conclusions: The optimized procedure is simple, efficient, and can be used for micro-propagation and the production of tolerant varieties to increase yield in saline areas.
AB - Introduction: Tomato (Solanum lycopersicum L.) is one of the essential vegetables worldwide, for consumption and mitigating malnutrition. Genetic transformation conceivably overcomes its yield challenges due to salinity, a crucial constraint for the economical use of 30% of cultivable lands in the coastal region of Bangladesh. Therefore, a robust and reproducible protocol has been established for in vitro regeneration and transformation to develop transgenic salt-tolerant tomato plants. Materials and Methods: During micropropagation, cotyledonary leaf explants were excised and cultured on MS media containing different combinations and concentrations of plant growth regulators. In transformation, the pre-cultured explants were inoculated and co-cultivated with Agrobacterium. Then they were transferred to the antibiotics-supplemented media to achieve salt-tolerant putative transformed plants. The transformation was confirmed by P-glucuronidase (GUS) assay and PCR for the antiporter gene. Results: Maximum regeneration response was achieved from the explants abaxially positioned at a 1.5 cm distance apart. BARI Tomato 14 and BINA Tomato 3 showed the highest shoot regeneration response (%) on MS media supplemented with 2 mg/L BAP and with 0.1 mg/L IAA for BARI Tomato 2 and BARI Tomato 15. Bacterial culture of OD600 0.68 for 30 min and a Co-cultivation period of 48 h resulted in the highest transformation frequency (47%) in Agrobacterium-mediated transformation with pBI 121 in BARI Tomato 3. The highest regeneration frequency (20.5%) was obtained in transformation with pH7WG2_OsNHX1_1.6. Conclusions: The optimized procedure is simple, efficient, and can be used for micro-propagation and the production of tolerant varieties to increase yield in saline areas.
KW - Agrobacterium-mediated Transformation
KW - Explants Spacing
KW - In Vitro Regeneration
KW - Orientation
KW - Phytohormones
KW - Salt Tolerance
KW - Tomato
U2 - 10.30491/JABR.2022.336979.1519
DO - 10.30491/JABR.2022.336979.1519
M3 - Journal article
AN - SCOPUS:85139455647
VL - 9
SP - 726
EP - 739
JO - Journal of Applied Biotechnology Reports
JF - Journal of Applied Biotechnology Reports
SN - 2322-1186
IS - 3
ER -
ID: 344732545