Farkhondehtale Navi, Mansour, Dorani, Ebrahim, Haddad, Raheem, Ebadi, Ali Akbar (2022). Establishment of Callus Induction, Cell Suspension Culture, and Agrobacterium-mediated Transformation system for Iranian Rice Cultivars. , 24(5), 1217-1232.
Mansour Farkhondehtale Navi; Ebrahim Dorani; Raheem Haddad; Ali Akbar Ebadi. "Establishment of Callus Induction, Cell Suspension Culture, and Agrobacterium-mediated Transformation system for Iranian Rice Cultivars". , 24, 5, 2022, 1217-1232.
Farkhondehtale Navi, Mansour, Dorani, Ebrahim, Haddad, Raheem, Ebadi, Ali Akbar (2022). 'Establishment of Callus Induction, Cell Suspension Culture, and Agrobacterium-mediated Transformation system for Iranian Rice Cultivars', , 24(5), pp. 1217-1232.
Farkhondehtale Navi, Mansour, Dorani, Ebrahim, Haddad, Raheem, Ebadi, Ali Akbar Establishment of Callus Induction, Cell Suspension Culture, and Agrobacterium-mediated Transformation system for Iranian Rice Cultivars. , 2022; 24(5): 1217-1232.

Establishment of Callus Induction, Cell Suspension Culture, and Agrobacterium-mediated Transformation system for Iranian Rice Cultivars

Journal of Agricultural Science and Technology
Article 16, Volume 24, Issue 5, 2022, Pages 1217-1232    PDF (819.41 K)
Document Type: Original Research
Authors
Mansour Farkhondehtale Navi1; Ebrahim Dorani* 1; Raheem Haddad2; Ali Akbar Ebadi3
1Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Islamic Republic of Iran.
2Department of Agricultural Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Islamic Republic of Iran.
3Rice Research Institutes of Iran, Agricultural Research, Education and Extension Organization (AREEO), Rasht, Islamic Republic of Iran.
Abstract
Up to now, a large number of optimized plant systems have been established for various purposes in Japonica and Indica rice. Based on genetic diversity in rice cultivars, this study established highly efficient protocols for in vitro callus induction, cell suspension cultures, and genetic transformation for some Iranian rice cultivars using mature embryos. In this study, the effect of different concentrations of 2,4-D (0.0, 1.0, 1.5, and of 2.0 mg L-1) on callus induction were investigated in 10 cultivars. Regarding higher callus induction frequency and mean weight of fresh calli, Hashemi, Binam, and Kazemi cultivars were selected for the experiments. The effects of kinetin (0.0, 1.0, and 2.0 mg L-1) and sucrose (30.0 and 60.0 g L-1) concentrations were tested to improve the biomass yield of a cell suspension culture. The MS medium supplemented with 0.5 mg L-1 2,4-D, 2.0 mg L-1 kinetin, and 60.0 g L-1 sucrose exhibited the maximum cell growth in the selected cultivars. The transformation efficiency for different bacterial strains (EHA105, LBA4404, and AGL-1), OD600 (0.1, 0.3, 0.6), the concentration of acetosyringone (50, 100, 200 μM), and co-cultivation period (1, 2, 3 days) were evaluated. The presence and expression of gusA gene in transgenic cultivars were determined by GUS histochemical assay, PCR, and RT-PCR analysis. The result showed that Hashemi cultivar had the highest cell biomass and efficiency of genetic transformation (58%) with EHA105 at bacterial OD600= 0.3, in 100 (μM) acetosyringone and two days of co-culture time. The findings offer insights for genetic transformation studies in Iranian rice cultivars.
Keywords
Keywords: Callus induction; Genetic transformaion; GUS expression; MS media; Rice
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