Molecular and phenotypic characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycin phosphotransferase II gene

Abstract

Abstract. Irsyadi MB, Sawitri WD, Purwantoro A. 2022. Molecular and phenotypic characteristics of T1 transgenic yellow cosmos (Cosmos sulphureus) carrying neomycin phosphotransferase II gene. Biodiversitas 23: 6097-6105. Yellow cosmos (Cosmos sulphureus) is a tropical ornamental flower that contains secondary metabolites useful fornatural pesticide application. Plant genetic transformation is one genetic engineering method used to increase secondary metabolite accumulation. However, published reports on this issue have yet to be made available. This is the first report of genetic transformation in transgenic yellow cosmos using neomycin phosphotransferase II (nptII) encoding gene. This study aimed to determine the efficiency transformation and phenotypic character of the T1 yellow cosmos transgenic nptII. This genetic transformation was carried out by the floral dip method mediated with Agrobacterium tumefaciens and characterized based on quantitative and qualitative observations, as well as a cluster analysis of transgenic plants. The T1 transgenic yellow cosmos was confirmed using PCR with a transformation efficiency of 73.33% based on the total plants resistant to Kanamycin and 10.57% based on the total seeds transformed. The presence of the nptII encoding gene was shown in transgenic plant samples with a size of 550 bp. In general, the introduction of the nptII gene generated no novel traits except the resistance to the kanamycin antibiotic. However, there was a decrease and increase in the number of stomata and the size of stomata in transgenic plants, respectively. In addition, there was a change in the type of ray floret to a mixture of ligulate and tubular, indicating that the nptII gene affected the phenotype of yellow cosmos. The result revealed that the nptII gene was inserted fairly randomly into the plant genome.

Efficient floral dip transformation method using Agrobacterium tumefaciens on Cosmos sulphureus Cav.

“…Thus, the seedlings germinated had kanamycin resistance. The NPT II gene cannot inhibit protein synthesis and ribosome binding in transformant plants ( Irsyadi et al, 2022 ). However, the selected plants can be affected by their resistance properties.…”

“…Thus, the seedlings escaped at 50 ppm kanamycin concentration. Irsyadi et al (2022) reported that in 50 ppm kanamycin concentration, selected putative transformants of C. sulphureus that escaped kanamycin reached 3.84%. Dalton et al, (1995) reported the extremely low concentration of selective agents, which allowed the undesirably high numbers of escape and loss of resistant transformants.…”