AUTHOR=Liu Huan , Zhao Haixia , Wu Longhua , Xu Wenzhong 

TITLE=A Genetic Transformation Method for Cadmium Hyperaccumulator Sedum plumbizincicola and Non-hyperaccumulating Ecotype of Sedum alfredii

JOURNAL=Frontiers in Plant Science

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2017.01047

DOI=10.3389/fpls.2017.01047

ISSN=1664-462X

ABSTRACT=<p>The present study demonstrates the development of an <italic>Agrobacterium</italic>-mediated genetic transformation method for species of the <italic>Sedum</italic> genus, which includes the Cd/Zn hyperaccumulator <italic>Sedum plumbizincicola</italic> and the non-hyperaccumulating ecotype of <italic>S. alfredii</italic>. Multiple shoots were induced from stem nodes of two <italic>Sedum</italic> plants using Murashige and Skoog (MS) medium containing 0.1 mg/L cytokinin 6-benzyladenine (6-BA) and 1.0 mg/L auxin 1-naphthaleneacetic acid (NAA). The shoot primordia were used as direct targets for <italic>Agrobacterium</italic> infection. Selection on hygromycin was highly effective in generating <italic>Agrobacterium</italic>-transformed explants. This callus-free procedure allowed us to obtain transgenic plantlets after rooting hygromycin-resistant shoots on phytohormone-free MS medium containing the antibiotic. The presence and expression of the reporter genes <italic>gusA</italic> and <italic>GFP</italic> in transgenic plants were confirmed by a real-time polymerase chain reaction, histochemical GUS assays, and confocal microscopy. This reliable method for genetic transformation of <italic>Sedum</italic> plants will help us to understand gene functions and the molecular mechanisms underlying Cd hypertolerance and hyperaccumulation in these species.</p>