Genetic transformation of common beans (Phaseolus vulgaris L.) through Agrobacterium tumefaciens carrying Cry1Ab gene

• Published in: Molecular biology reports Vol. 49; no. 7; pp. 7195 - 7203
• Main Authors: Sağlam Yılmaz, Sevil, Khawar, Khalid Mahmood, Çiftçi, Cemalettin Yaşar
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.07.2022; Springer Nature B.V
• Subjects: Agrobacterium; Agrobacterium tumefaciens; Animal Anatomy; Animal Biochemistry; Beans; Bioassays; Biomedical and Life Sciences; Cry1ab gene; Crystal proteins; Cultivars; Embryos; Explants; Genetic transformation; Genotypes; Germplasm; Histology; Kanamycin; Legumes; Life Sciences; Morphology; Original Article; Pest resistance; Pests; Phaseolus vulgaris; Plant breeding; Progress in genomics; Proteins; transcriptomics and breeding of crops; Transgenic plants; Beans; Bruchus sp; Insect resistance; Legume seeds
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-022-07637-1

Background Seed beetles are one of the most important causes of yield loss in bean production. It is essential to develop resistant varieties in the fight against these pests. Agrobacterium-based gene transformation is the most widely used breeding method worldwide to develop insect-resistant varieties. Methods and results Embryonic axes and plumule explants were obtained from Agrobacterium tumefciens treated mature zygotic embryos of low and high raw protein-based common bean cultivars Akman 98 and Karacaşehir 90. Agrobacterium tumefaciens contained a synthetic Bacillus thuringiensis insecticidal crystal protein gene (Bt Cry1Ab) controlled by the 35S promoter and NOS terminator sequences. The transformation event was genotype and explant dependent. The plumule explants could not withstand kanamycin-based selection pressure and died. It was possible to get two transgenic plants using embryonic axis explants of low protein cultivar Akman 98. These results were validated using GUS analysis, PCR, RT-PCR, bioassay analysis, and ELISA test from the samples taken from T 0 and T 1 generations. Bioassay tests showed that these plants were protected from the damage of legume seed insects ( Bruchus spp.). Conclusions The results are very encouraging and may help in producing better transgenic common bean germplasm leading to safe agriculture and reducing environmental pollutions.