Use of an anthocyanin production phenotype as a visible selection marker system in transgenic tobacco plant

• Published in: Plant biotechnology reports Vol. 6; no. 3; pp. 203 - 211
• Main Authors: Lim, Sun-Hyung, Sohn, Seong-Han, Kim, Dong-Hern, Kim, Jae Kwang, Lee, Jong-Yeol, Kim, Young-Mi, Ha, Sun-Hwa
• Format: Journal Article
• Language: English
• Published: Japan Springer-Verlag 01.07.2012; Springer Japan; Springer Nature B.V; 한국식물생명공학회
• Subjects: Agriculture; Agrobacterium radiobacter; Agrobacterium tumefaciens; anthocyanins; Antibiotics; Arabidopsis; Biomedical and Life Sciences; Biotechnology; Cell Biology; Cloning; color; Corn; Genes; genotype; Genotype & phenotype; herbicide resistance; Herbicides; Infiltration; Leaves; Life Sciences; Original Article; phenotype; Pigmentation; Plant Biochemistry; Plant Sciences; progeny; regulator genes; reverse transcriptase polymerase chain reaction; Tobacco; transcription factors; Transgenic plants; 농학; Anthocyanin; Co-transformation; Transcription factor; Selectable marker
• ISSN: 1863-5466; 1863-5474
• DOI: 10.1007/s11816-012-0215-6

To develop a potentially alternative method for the selection of transgenic plants instead of antibiotic and herbicide resistance, anthocyanin pigmentation phenotype was examined to provide a visible selection marker system. Two regulatory genes of the anthocyanin biosynthetic pathway, the R2R3 MYB mPAP1 gene from Arabidopsis and the basic helix loop helix B-Peru gene from maize, were amplified by RT-PCR and then individually cloned into a plant expression vector. The requirement of these two genes for anthocyanin pigmentation was pre-confirmed via an in vivo assay using tobacco agro-infiltration. The mPAP1 and B-Peru vectors were further stably co-transformed into tobacco plants using Agrobacterium tumefaciens strain LBA4404. Tobacco plants harboring both genes could be readily selected through the manifestation of a red color due to anthocyanin accumulation in the whole plant body. The T1 segregants showed red or green phenotypes depending on the genotype. The need for both the mPAP1 and B-Peru genes for a red color phenotype due to anthocyanin pigmentation was further confirmed by genotyping of the T1 generation by genomic PCR analysis and an in vivo assay using agro-infiltration. From these results, we conclude that co-transformation with two individual vectors harboring a critical anthocyanin transcriptional factor has potential utility as an alternative visible selectable marker system for transgenic progeny selection in plants.