Agrobacterium tumefaciens Mediated Transformation of the Proteolysis 6 (PRT6) Gene into Cotyledons of Tomato cv. Micro Tom

作者：Suka Intan Elya; Roslan Nur Farhana; Chew Bee Lynn; Goh Hoe Han^*; Zainal Zamri; Isa Nurulhikma Md^*

来源：Sains Malaysiana, 2018, 47(7): 1465-1471.

DOI：10.17576/jsm-2018-4707-14

• Antibiotic
• A. tumefaciens
• callus
• co-cultivation
• Proteolysis 6

Proteolysis 6 (PRT6) gene plays an important role in the N-end rule pathway which functions as an E3 ligasc enzyme. PRT6 functions to recognise target proteins for degradation. The main objective of this study is to transform the PRT6 RNAi construct through Agrobacterium tumefaciens into tomato. The purpose of this study was to understand the role of the N-end rule pathway during fruit ripening. Several factors affecting transformation efficiency such as co-cultivation time and concentration of antibiotics were optimised. The results from this study showed that pre-cultured cotyledons incubated for 48 h in co-cultivation medium increased the callus formation to 61% while using 500 mg/L carbenicillin antibiotic in the shoot regeneration medium reduced the contamination of A. tumefaciens to 52%. Besides, A. tumefaciens strain C58 was shown to be the most suitable A. tumefaciens strain to be used in this study. Polymerase Chain Reaction (PCR) was performed on the regenerated shoots to confirm integration of the PRT6 fragment into the tomato genome . Based on the PCR analysis, all putative transgenic shoots were positive transformants.

• 出版日期2018-7