Hits:

Indexed by:期刊论文

Date of Publication:2009-02-01

Journal:BIOTECHNOLOGY LETTERS

Included Journals:SCIE、EI、PubMed

Volume:31

Issue:2

Page Number:295-303

ISSN No.:0141-5492

Key Words:Linear smGFP cassette; Marker-free cassette; Ovary-drip transformation; Soybean; Soybean transformation; Vector-free cassette

Abstract:A tissue culture-independent plant transformation method, called ovary-drip transformation, was established in which a minimal linear gene cassette [35S CaMV promoter, open reading frame of soluble modified green fluorescent protein (smGFP), and NOS terminator] was transformed into soybean. The method is characterized by directly dripping a DNA solution, which is supplemented with a surfactant, onto the ovary wound 6-8 h after self-pollination. The growth of the pollen tube was measured after self-pollination. The movement of smGFP across the passageway toward the embryo sac was monitored using fluorescein isothiocyanate-labeled DNA. The transformation frequency reached 3.2% by PCR analysis. Southern analysis of the primary transformants denoted the integration of a single site smGFP. The transgenic plants exhibited a high level of smGFP expression which was visible in the immature embryos of the transgenic soybean.