Biotech approach to neutralize the invasiveness of burning bush (Euonymus alatus), a progress report on development of its genetic transformation system and functional analysis of sterile genes

摘要

We have recently developed an in vitro plant regeneration method and an Agrobacterium tumefaciens-mediated genetic transformation method for Euonymus alatus. More than 60% cotyledon explants from 10-day old E. alatus seedlings produced 3 shoots on average with woody plant medium (WPM) in the presence of 5.0 mg/L 6-benzylaminopurine (BA) and 0.2 mg/L alpha -naphthalene acetic acid (NAA). Approximately 80% of shoots produced roots with 0.3 mg/L NAA and 0.5 mg/L IBA. We have constructed a "super sterile" gene cassette that should lead to a male and female sterility and production of normal sized fruits when expressed in plants. We have demonstrated that in tobacco and tomato plants the sterile gene cassette is effective to abolish pollen or seed formation but capable of producing normal sized fruit. The "super sterile" gene cassette was also used to generate transgenic E. alatus plants. Because an intron containing gusplus gene was incorporated into the "super sterile" gene cassette, the E. alatus plants that were transformed with the sterile gene cassette have been confirmed based on the expression of the gusplus gene and an additional molecular analysis.