Work

Agrobacterium‐mediated transformation of reed (Phragmites communisTrinius) using mature seed‐derived calli

Year: 2012

Type: article

Abstract: Reed (Phragmites communis) is a potential bioenergy plant. We report on its firstAgrobacterium‐mediated transformation using mature seed‐derived calli. TheAgrobacteriumstrainsLBA4404,EHA105, andGV3101, each harboring the binary vector pIG121Hm, were used to optimizeT‐DNAdelivery into the reed genome. Bacterial strain, cocultivation period and acetosyringone concentration significantly influenced theT‐DNAtransfer. About 48% transient expression and 3.5% stable transformation were achieved when calli were infected with strainEHA105 for 10 min under 800 mbar negative pressure and cocultivated for 3 days in 200 μmacetosyringone containing medium. Putative transformants were selected in 25 mg l−1hygromycin B.PCR, and Southern blot analysis confirmed the presence of the transgenes and their stable integration. Independent transgenic lines contained one to three copies of the transgene. Transgene expression was validated byRT‐PCRandGUSstaining of stems and leaves.

Source: GCB Bioenergy

DOI: 10.1111/j.1757-1707.2012.01200.x