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ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Research Project #443591

Research Project: Development of Soybean Lines Without Both Trypsin and Chymotrypsin Inhibitors

Location: Plant Genetics Research

Project Number: 5070-21000-044-016-A
Project Type: Cooperative Agreement

Start Date: Oct 1, 2022
End Date: Mar 31, 2024

Soybean meal cannot be directly used for animal feed mixtures due to the presence of various anti-nutritional compounds which reduce animal weight gain. There are two major proteinaceous anti-nutritional factors (ANF) in soybean seeds: 1) Kunitz-trypsin inhibitors (KTi) which inactivate the animal digestive enzyme trypsin; and 2) Bowman-Birk inhibitors (BBi) which inactivates both trypsin and chymotrypsin animal digestive enzymes. Here, we are proposing a project which will culminate in development of adapted soybean germplasm which contains drastically reduced (or eliminated) anti-nutritionals.

We will employ gene editing to delete the abundant KTi gene expressed in seeds. The transgenic events generated by the gene editing system will be planted in a greenhouse and seeds harvested from these plants will be utilized for measuring the KTi and BBi activities following well-established protocols. We have raised antibodies against soybean KTi and BBi which will be used to quantify the amount of these proteins and activities in transgenic events by western blot analysis and/or ELISA. Non-transformed soybean plants will also be used for measuring progress. Later, KTi gene edited transgenic soybeans will be crossed with BBI gene edited soybean lines to create a soybean line that lacks both KTi and BBI.