Effect of selection agents to Chrysanthemum (Chrysanthemum morifolium) callus growth after Agrobacterium-mediated genetic transformation

1st International Conference on Food Security and Sustainable Agriculture in The Tropics

农业科学;经济学

Sjahril, R.^1 ; Jamaluddin, I.^2 ; Nadir, M.^3 ; Asman^4 ; Dungga, N.E.^5

Laboratory of Plant Biosciences and Reproduction Biotechnology, Faculty of Agriculture, Hasanuddin University, Jl. Perintis Kemerdekaan KM 10, Makassar

90245, Indonesia^1

Agro-technology, Graduate School of Agriculture, Faculty of Agriculture, Hasanuddin University, Jl. Perintis Kemerdekaan KM 10, Makassar

90245, Indonesia^2

Animal Feed Chemistry Laboratory, Faculty of Animal Husbandry, Hasanuddin University, Jl. Perintis Kemerdekaan KM 10, Makassar

90245, Indonesia^3

Plant Pest and Disease Laboratory, Faculty of Agriculture, Hasanuddin University, Indonesia^4

Department of Agronomy, Faculty of Agriculture, Hasanuddin University, Jl. Perintis Kemerdekaan KM 10, Makassar

90245, Indonesia^5

关键词: Agrobacterium mediated;    Agrobacterium tumefaciens;    Completely randomized designs;    Critical value;    Genetic transformation;    Selection methods;    Selective agents;    Standard protocols;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/157/1/012044/pdf
DOI  :  10.1088/1755-1315/157/1/012044

学科分类：农业科学（综合）

来源: IOP

【 摘 要 】

Genetic transformation mediated by Agrobacterium tumefaciens requires an efficient selection method for successful progress of transformation. This study aims to determine the concentration and kind of antibiotics and selection agents used during transformation to formulate standard protocol of chrysanthemum in the process of propagating disease resistant Chrysanthemum mediated by Agrobacterium tumefaciens EHA105 (pEKB-WD). The experiments were performed by planting chrysanthemum explants leaf cutting (5 mm diameter on NAA medium 2 mg L-1BAP 2 mg L-1) with addition of Kanamycin: 25, 50, 100, 150 and 200 (mg L-1); Hygromycin: 5, 10, 25, 50 and 75 (mg L-1); Paromomycin: 10, 25, 50, 75 and 100 (mg L-1). Experiment was arranged in a Completely Randomized Design (CRD). Each treatment was repeated five times thus 75 bottles of culture were used; each bottle consists of 5 pieces of leaf cuttings, resulted in total of 375 pieces. The results showed that selection agent had a critical value for Hygromycin 25 mg L-1and Kanamycin 100 mg L-1which can make explant experienced necrosis better than Paromomycin. Paromomycin at 100 mg L-1was only able to kill explant's periphery. Remained callus stayed fresh more than 50% so that when used as the selection agent could produce more escape cell. The optimum transformation with concentration of 10% Agrobacterium (vol/vol) with 30 minutes co-cultivation can produce more efficient transformed callus. Considering the high price of Hygromycin, it was best to use Kanamycin as selective agents.