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A space-saving visual screening method, Glycine max FAST, for generating transgenic soybean.

ABSTRACT: Establishing homozygous transgenic lines of Glycine max is time-consuming and laborious. To overcome the difficulties, we developed a powerful method for selecting transgenic soybean plants, Fluorescence-Accumulating Seed Technology (GmFAST). GmFAST uses a marker composed of a soybean seed-specific promoter coupled to the OLE1-GFP gene, which encodes a GFP fusion of the oil-body membrane protein OLEOSIN1 of Arabidopsis thaliana. We introduced the marker gene into cotyledonary nodes of G. max Kariyutaka via Agrobacterium-mediated transformation and regenerated heterozygous transgenic plants. OLE1-GFP-expressing soybean seeds can be selected nondestructively with a fluorescence stereomicroscope. Among T₂ seeds, the most strongly fluorescent seeds were homozygous. GmFAST enables to reduce the growing space by one-tenth compared with the conventional method. With this method, we obtained the soybean line that had higher levels of seed pods and oil production. The phenotypes are presumably caused by overexpression of Glyma13g30950, suggesting that Glyma13g30950 regulates seed pod formation in soybean plants. An increase in seed pod number was confirmed in A. thaliana plants that overexpressed the Arabidopsis ortholog of Glyma13g30950, E6L1.Taken together, GmFAST provides a space-saving visual and nondestructive screening method for soybean transformation, thereby increasing the chance of developing useful soybean lines.

SUBMITTER: Iwabuchi K

PROVIDER: S-EPMC7053950 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Soybean (Glycine max) is an important leguminous crop that is grown throughout the United States and around the world. In 2016, soybean was valued at $41 billion USD in the United States alone. Increasingly, soybean farmers are adopting alternative management strategies to improve the sustainability and profitability of their crop. Various benefits have been demonstrated for alternative management systems, but their effects on soybean-associated microbial communities are not well-understood. In order to better understand the impact of crop management systems on the soybean-associated microbiome, we employed DNA amplicon sequencing of the Internal Transcribed Spacer (ITS) region and 16S rRNA genes to analyze fungal and prokaryotic communities associated with soil, roots, stems, and leaves. Soybean plants were sampled from replicated fields under long-term conventional, no-till, and organic management systems at three time points throughout the growing season. Results indicated that sample origin was the main driver of beta diversity in soybean-associated microbial communities, but management regime and plant growth stage were also significant factors. Similarly, differences in alpha diversity are driven by compartment and sample origin. Overall, the organic management system had lower fungal and bacterial Shannon diversity. In prokaryotic communities, aboveground tissues were dominated by Sphingomonas and Methylobacterium while belowground samples were dominated by Bradyrhizobium and Sphingomonas. Aboveground fungal communities were dominated by Davidiella across all management systems, while belowground samples were dominated by Fusarium and Mortierella. Specific taxa including potential plant beneficials such as Mortierella were indicator species of the conventional and organic management systems. No-till management increased the abundance of groups known to contain plant beneficial organisms such as Bradyrhizobium and Glomeromycotina. Network analyses show different highly connected hub taxa were present in each management system. Overall, this research demonstrates how specific long-term cropping management systems alter microbial communities and how those communities change throughout the growth of soybean.

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A space-saving visual screening method, Glycine max FAST, for generating transgenic soybean.

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