Silencing of seven tomato propyl 4-hydroxylases reduces AGPs and extensins content and induces alterations in growth and fruit ripening programs [Thesis (M.Sc.)]
2013
Salah Alla
Prolyl 4 hydroxylases (P4Hs) catalyze the hydroxylation of proline residues in hydroxyproline-rich glycoproteins (HRGPs). HRGPs are broadly implicated in all aspects of plant growth and development. A reverse genetics approach, Virus-Induced Gene Silencing, was used to silence several P4Hs (P4H1, P4H2, P4H3, P4H5, P4H7, P4H9) in tomato Solanum lycopersicum. Differences in the profile of AGPs and extensins between the silenced lines and the control were monitored using western blots and monoclonal antibodies recognizing AGPs and Extensins epitopes. Additionally, AGPs and extensins were quantified using the method of radial diffusion indicating lower AGPs and Extensins content in the silenced plants than in control. Moreover, preliminary phenotypic analysis of silenced P4H3 and P4H7 in micro-tom plants suggested a delay in fruit ripening and alternations in fruit growth and ethylene production. These results indicate that VIGS can be used as a tool to study the function of tomato gene families such as P4Hs in growth and development, and elucidate, in the long run, the physiological significance of substrate-proteins such as hydroxyproline-rich glycoproteins.
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