Generation of Arabidopsis early flowering plants by destruction of CLF transcripts via RNA silencing
2014
Didar, Neda | Pazhouhandeh, Maqsoud | Mahmoudi, Fatemeh
The control of flowering time requires activation of a cascade of successive genes which are affected by several internal and external factors. Some of these genes, such as Curly Leaf (CLF), have inhibitory effect on flowering. Thus, by silencing of these genes, initiation of flowering can be accelerated. In this research, the RNA Silencing technology is used to investigate the possibility of production of early-flowering plants via CLF silencing. First, in order to produce hairpin structure, a fragment of CLF gene of Arabidopsis thaliana was amplified by PCR and then cloned in two sense and antisense directions, into pFGC5941 vector. The resulted recombinant vector was then transferred into Arabidopsis plants via Agrobacterium method. As expected, initiation of flowering was accelerated in transgenic plants in comparison with the wild-type plants. In agreement to the obtained results, RT-qPCR analysis showed that CLF expression in transgenic plants was decreased in comparison with the wild-type plants. In contrast, FT (a gene which induces flowering) expression was increased in transgenic plants. These results show that RNA silencing as a novel and time-saving biotechnological method could be applied to generate early-flowering plants.
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