A Paris Polyphylla var. Yunnanensis Gene Encoding Cycloartenol Synthase Isolated and Introduced into Tomato by Agrobacterium-mediated Transformation

摘要

A CAS gene encoding cycloartenol synthase was cloned from Paris polyphylla var. yunnanensis by homology based PCR method. The comparative analysis of fulllength cDNA, using the BLASTN algorithm, showed that the cloned CAS gene shared more than 80% nucleotide sequence identity with the fragments from other plants encoding cycloartenol synthase. The CAS gene from Paris polyphylla var. yunnanensis was cloned into the plant binary vector pBl121 in the antisense orientation and transformed into tomato (Solan urn lycopersicum) by Agrobacterium tumefaciens mediated method. 8 transgenic plants were obtained. PCR (Polymerase Chain Reaction) and Southern blot analysis showed that the antiCAS gene had integrated into the genome of tomato. RT-PCR analysis of CAS gene inhibition in transgenic plants revealed that all the transgenic plants accumulated no detectable or extremely low levels of the CAS transcripts, whereas the CAS expression was obviously detectable in the non-transgenic plant These results showed that the cloned CAS gene from Paris polyphylla var. yunnanensis played an important role in plant cycloartenol synthase.