Abstract
Understanding the molecular processes of seed development is important especially in agronomic crops that produce large amounts of nutrient reserves. Because soybean is a vital source of vegetable protein worldwide, producers are concerned about increasing the total amount of protein in the seed without substantially lowering the amount of oil, another economically important product. Here we describe a transgenic soybean line with increased protein and protein/oil ratio, containing an average of 42.2% protein vs. 38.5% in controls and with a protein/oil ratio of 2.02 vs. 1.76 in controls over several generations of greenhouse growth. Other phenotypic data show that the seeds are heavier, although there are overall lower yields per plant. We postulate these effects result from insertion site mutagenesis by the transgenic construct. As this line never achieves homozygosity and appears to be embryo lethal when homozygous, one functional copy of the gene is most likely essential for normal seed development. Global transcript analyses using RNA-Seq for 88,000 gene models over two stages of cotyledon development revealed that more genes are over-expressed in the transgenic line including ribosomal protein related genes and those in the membrane protein and transporters families. Localization of the insertion site should reveal the genes and developmental program that has been perturbed by the transgenic construct, resulting in this economically interesting increase in protein and the protein/oil ratio.
Highlights
Soybean is one of the most important sources of vegetable protein and oil in the world
A seed-targeted transgenic construct based on RNA interference (RNAi) was produced (S1 Fig) and introduced into soybean tissue in an attempt to down-regulate the pyruvate dehydrogenase kinase (PDHK) gene
Soybean embryogenic culture pieces were transformed with this construct and some regenerated plants that were positive for the hygromycin gene of the construct via PCR and Southern blots were selected [15]
Summary
Soybean is one of the most important sources of vegetable protein and oil in the world. A total of 236 Glyma models were significantly over-expressed in the non-transgenic segregant (Neg) plants compared to the transgenic (Pos) plants (Fig 6D–6F and S9 Table). When the transgenic plants were compared to the non-transformed Jack control plants, 463 Glyma models were significant and over-expressed in the transgenics (Pos) (S7 Fig and S10 Table).
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