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Bowman-Birk Inhibitor Mutants of Soybean Generated by CRISPR-Cas9 Reveal Drastic Reductions in Trypsin and Chymotrypsin Inhibitor Activities.
Kim, Won-Seok; Gillman, Jason D; Kim, Sunhyung; Liu, Junqi; Janga, Madhusudhana R; Stupar, Robert M; Krishnan, Hari B.
Afiliación
  • Kim WS; Division of Plant Science and Technology, University of Missouri, Columbia, MO 65211, USA.
  • Gillman JD; Plant Genetics Research Unit, US Department of Agriculture-Agricultural Research Service, Columbia, MO 65211, USA.
  • Kim S; Division of Plant Science and Technology, University of Missouri, Columbia, MO 65211, USA.
  • Liu J; Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA.
  • Janga MR; Institute of Genomics for Crop Abiotic Stress Tolerance, Texas Tech University, Lubbock, TX 79409, USA.
  • Stupar RM; Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN 55108, USA.
  • Krishnan HB; Plant Genetics Research Unit, US Department of Agriculture-Agricultural Research Service, Columbia, MO 65211, USA.
Int J Mol Sci ; 25(11)2024 May 21.
Article en En | MEDLINE | ID: mdl-38891766
ABSTRACT
Despite the high quality of soybean protein, raw soybeans and soybean meal cannot be directly included in animal feed mixtures due to the presence of Kunitz (KTi) and Bowman-Birk protease inhibitors (BBis), which reduces animal productivity. Heat treatment can substantially inactivate trypsin and chymotrypsin inhibitors (BBis), but such treatment is energy-intensive, adds expense, and negatively impacts the quality of seed proteins. As an alternative approach, we have employed CRISPR/Cas9 gene editing to create mutations in BBi genes to drastically lower the protease inhibitor content in soybean seed. Agrobacterium-mediated transformation was used to generate several stable transgenic soybean events. These independent CRISPR/Cas9 events were examined in comparison to wild-type plants using Sanger sequencing, proteomic analysis, trypsin/chymotrypsin inhibitor activity assays, and qRT-PCR. Collectively, our results demonstrate the creation of an allelic series of loss-of-function mutations affecting the major BBi gene in soybean. Mutations in two of the highly expressed seed-specific BBi genes lead to substantial reductions in both trypsin and chymotrypsin inhibitor activities.
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Glycine max / Inhibidor de la Tripsina de Soja de Bowman-Birk / Edición Génica Idioma: En Revista: Int J Mol Sci Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Glycine max / Inhibidor de la Tripsina de Soja de Bowman-Birk / Edición Génica Idioma: En Revista: Int J Mol Sci Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos