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Improved in planta genetic transformation efficiency in bitter gourd (Momordica charantia L.)

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Abstract

Production of transformed bitter gourd plants through in vitro regeneration is a laborious practice, which may also result in somaclonal variations. Thereby, in the current study, we have established a less tedious in planta protocol for more efficient genetic transformation of bitter gourd var. NBGH-470 (Apurva) using the seed as a target material. Bitter gourd seeds were transformed by Agrobacterium tumefaciens strain LBA4404 bearing the pCAMBIA1301 binary vector, and the transformed plants were selected against hygromycin B. The putatively transformed bitter gourd (To) plants were examined by GUS assay. Two parameters, including vacuum infiltration and sonication improving the in planta transformation, have been investigated in the present work. Amongst several time durations examined, the highest transformation rate (37%) was achieved upon subjecting the pre-cultured bitter gourd seeds to sonication for 15 min, followed by vacuum infiltration for 6 min in LBA4404 culture suspension. The transformed bitter gourd (T1) plants were selected against hygromycin B, and the transgene integration was evaluated by polymerase chain reaction (PCR), Southern hybridization, and reverse transcriptase PCR (RT-PCR). The developed protocol in the current study is suitable to genetically engineer the bitter gourd plants with disease and pest-resistant traits.

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Acknowledgements

Dr. Sivabalan Karthik acknowledges Jawaharlal Nehru Memorial Fund, New Delhi, India, for the grant of Jawaharlal Nehru Scholarship (Ref no: SU-1/88/2016-17/79) to conduct the present work. The authors also acknowledge RUSA, 2.0-Biological Sciences, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India, for providing financial support.

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Authors and Affiliations

  1. Department of Biotechnology, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India

    Sivabalan Karthik, Gadamchetty Pavan, Arumugam Prasanth, Selvam Sathish & Markandan Manickavasagam

  2. Division of Crop Improvement, Indian Council of Agricultural Research (ICAR)-Sugarcane Breeding Institute, Coimbatore, Tamil Nadu, 641 007, India

    Chinnaswamy Appunu

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  1. Sivabalan Karthik
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  2. Gadamchetty Pavan
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  3. Arumugam Prasanth
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  5. Chinnaswamy Appunu
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  6. Markandan Manickavasagam
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Correspondence to Markandan Manickavasagam.

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Editor: Yong Eui Choi

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Karthik, S., Pavan, G., Prasanth, A. et al. Improved in planta genetic transformation efficiency in bitter gourd (Momordica charantia L.). In Vitro Cell.Dev.Biol.-Plant 57, 190–201 (2021). https://doi.org/10.1007/s11627-021-10160-w

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