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论文类型：期刊论文

发表时间：2009-02-01

发表刊物：BIOTECHNOLOGY LETTERS

收录刊物：SCIE、EI、PubMed

卷号：31

期号：2

页面范围：295-303

ISSN号：0141-5492

关键字：Linear smGFP cassette; Marker-free cassette; Ovary-drip transformation; Soybean; Soybean transformation; Vector-free cassette

摘要：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.