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All Journal Biotika: Jurnal Ilmiah Biologi Indonesian Journal of Biotechnology
RIZKITA RACHMI ESYANTI
Institut Teknologi Bandung
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

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PENGARUH KERAPATAN AGROBACTERIUM TERHADAP EFISIENSI TRANSFORMASI PADA PISANG CAVENDISH (Musa acuminata colla) Apriyani, Ria Khoirunnisa; Esyanti, Rizkita Rachmi; Dwivany, Fenny Martha
BIOTIKA Jurnal Ilmiah Biologi Vol 17, No 1 (2019): BIOTIKA JUNI 2019
Publisher : Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24198/bjib.v17i1.21898

Abstract

Transformasi genetik melalui perantara Agrobacterium tumefaciens merupakan salah satu carauntuk meningkatkan kualitas buah Pisang Cavendish (Musa acuminata colla (AAA Group))yang memiliki susunan gen triploid. Efisiensi transformasi melalui A. tumefaciens terhadapembrio Pisang Cavendish dilakukan dengan mengoptimasi berbagai kerapatan optik kulturbakteri. A. tumefaciens, yaitu strain AGL1 dan GV3101 (yang masing-masing membawaplasmid ganda pBI121), mengandung gen gusA (pengode enzim β-Glucuronidase) digunakandalam proses transformasi ini. Perbedaan kerapatan optik (0,6; 0,8; 1,0) suspensi bakteri digunakan untuk proses transformasi. Persentase efisiensi transformasi dihitung berdasarkanjumlah embrio pisang yang positif pada uji GUS dibagi dengan jumlah total embrio pisang yangdiuji, dikali 100%. Efisiensi transformasi tertinggi diperoleh dari embrio pisang yangdiinokulasi oleh A.tumefaciens strain AGL1/pBI121 pada OD600nm = 1 (96%) dan efisiensitransformasi terendah diperoleh dari embrio pisang yang diinokulasi oleh A.tumefaciens strainGV3101/pBI121 pada OD600nm = 0.6 (40%).
A robust in planta Agrobacterium‐mediated transformation in red chili (Capsicum annuum L.) Hamdani, Anti Damayanti; Nugroho, Syarul; Esyanti, Rizkita Rachmi; Faizal, Ahmad; Suhandono, Sony
Indonesian Journal of Biotechnology Vol 29, No 4 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.94653

Abstract

Plant improvement through in vitro culture and genetic engineering is a significant aspect of breeding programs aimed at producing disease‐resistant cultivars of disease‐prone red chili (Capsicum annuum L.). However, the Capsicum genus is recalcitrant to genetic transformation and in vitro regeneration. Moreover, developing a universal transformation protocol is difficult due to its highly genotype‐dependent nature. Therefore, this study aimed to develop an Agrobacterium‐mediated in planta transformation method applicable to various red chili cultivars. Two open‐pollinated varieties, Tanjung 2 and Ciko, were subjected to transformation. The young seedlings were immersed in transformation medium containing Agrobacterium tumefaciens strain GV3101 harboring the binary vector pCAMBIA1301, which carries the β‐glucuronidase (GUS) gene. GUS histochemical analysis revealed that all the primary transformants of Tanjung 2 and Ciko were identified as chimeric. The average staining in the body of the seedlings was 88.63 + 26.33% in Tanjung 2, and 90.65 + 16.77% in the Ciko variety. More than 50% of the seedlings continued to express GUS in their shoot areas 10 days after Agrobacterium infection, indicating the possibility of transgene inheritance in the following generation. The in planta transformation approach is notably genotype independent, making it a promising standard transformation protocol for different red chili varieties.
Co-Authors Ahmad Faizal Apriyani, Ria Khoirunnisa FENNY MARTHA DWIVANY Hamdani, Anti Damayanti Nugroho, Syarul SONY SUHANDONO