Agrobacterium tumefaciens mediated production of transgenic plants expressing carnation P4H1 RNAi construct
2008
Ahou, A.
The prolyl 4-hydroxylase is a large family of genes encoding proteins that catalyze the conversion of proline to 4-hydroxyproline using 2 oxoglutarate as substrate and Fe, oxygen and ascorbate as cofactors. The 4-hydroxyproline residues are widespread in plant proteins and particularly in cell wall glycoproteins while their functional significance is directly related to arabinogalactans and extensins functions in plant growth and development. Previous work in our lab resulted in the identification of 2 carnation P4Hs, DcP4H1 and DcP4H1 which are expressed throughout the progression of petal senescence. The aim of this study was the construction of the right P4H1 RNAi cassette to transform three different varieties of carnation plant, White Sim (2002), Emotion (2005) and White 116R (2016) using Agrobacterium- mediated transformation protocols. The next step was the confirmation for the insertion of the construct using PCR analysis. For this reason, the binary recombinant transformation vector pROK2 was used to produce hpRNA which has P4H1 specific sequence in the sense and antisense orientation. This dsRNA expressing construct was able to induce post-transcriptional gene silencing (PTGS). The results obtained in this work indicate the possible transformation of carnation plants using RNAi method and further research is required to thoroughly investigate the carnation p4h1 function in petal senescence.
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