Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions

Trypsin inhibitors (TIs) in soybean seeds reduce the availability of processed soybean foods and animal feed. This study aimed to evaluate the trypsin inhibitor activity (TIA) in 999 Korean soybean accessions and conduct molecular characterization of soybean accessions with low TIA. TIA was evaluate...

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Main Authors: Aron Park, Se-Hee Kang, Byeong-Hee Kang, Sreeparna Chowdhury, Seo-Young Shin, Won-Ho Lee, Jeong-Dong Lee, Sungwoo Lee, Yu-Mi Choi, Bo-Keun Ha
Format: Article
Language:English
Published: MDPI AG 2023-10-01
Series:Agriculture
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Online Access:https://www.mdpi.com/2077-0472/13/11/2070
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author Aron Park
Se-Hee Kang
Byeong-Hee Kang
Sreeparna Chowdhury
Seo-Young Shin
Won-Ho Lee
Jeong-Dong Lee
Sungwoo Lee
Yu-Mi Choi
Bo-Keun Ha
author_facet Aron Park
Se-Hee Kang
Byeong-Hee Kang
Sreeparna Chowdhury
Seo-Young Shin
Won-Ho Lee
Jeong-Dong Lee
Sungwoo Lee
Yu-Mi Choi
Bo-Keun Ha
author_sort Aron Park
collection DOAJ
description Trypsin inhibitors (TIs) in soybean seeds reduce the availability of processed soybean foods and animal feed. This study aimed to evaluate the trypsin inhibitor activity (TIA) in 999 Korean soybean accessions and conduct molecular characterization of soybean accessions with low TIA. TIA was evaluated using colorimetric analysis through a substrate–enzyme reaction. The average TIA of the 999 soybean accessions was 90.31%, ranging from 43.70% to 99.51%. Kunitz trypsin inhibitor 3 (KTi-3) gene target sequencing analysis was performed on seven soybean germplasm accessions (IT105782, IT170889, IT273590, IT274513, IT274515, IT276197, and IT022891) showing less than 60% TIA. Four soybean accessions (IT274513, IT274515, IT276197, and IT022891) exhibited the same mutations (a G-to-T transversion and an AG deletion) in the KTi-3 gene (<i>Glyma.08g341500</i>) in PI542044. In addition, whole-genome re-sequencing was performed on three soybean accessions with no mutations in the KTi-3 gene. Compared with the reference soybean genome, an in-frame insertion and five missense mutations were identified in the coding sequencing of the KTi-1 gene (<i>Glyma.01g09500</i>) in IT105782. A RT-qPCR analysis showed that the mRNA expression level of KTi-1 was reduced by approximately 17% in IT105782 during seed development. In this study, we identified a previously unreported mutation in the KTi-1 gene and developed a KASP marker using this new KTi-1 variant.
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spelling doaj-art-e7f99234d6ba41f5bf7987b3ba3ac37b2024-12-16T13:35:15ZengMDPI AGAgriculture2077-04722023-10-011311207010.3390/agriculture13112070Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean AccessionsAron Park0Se-Hee Kang1Byeong-Hee Kang2Sreeparna Chowdhury3Seo-Young Shin4Won-Ho Lee5Jeong-Dong Lee6Sungwoo Lee7Yu-Mi Choi8Bo-Keun Ha9Department of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaDepartment of Applied Biosciences, Kyungpook National University, Daegu 41566, Republic of KoreaDepartment of Crop Science, College of Agriculture and Life Sciences, Chungnam National University, Daejeon 34134, Republic of KoreaNational Agrobiodiversity Center, National Institute of Agricultural Sciences, RDA, Jeonju 54874, Republic of KoreaDepartment of Applied Plant Science, Chonnam National University, Gwangju 61186, Republic of KoreaTrypsin inhibitors (TIs) in soybean seeds reduce the availability of processed soybean foods and animal feed. This study aimed to evaluate the trypsin inhibitor activity (TIA) in 999 Korean soybean accessions and conduct molecular characterization of soybean accessions with low TIA. TIA was evaluated using colorimetric analysis through a substrate–enzyme reaction. The average TIA of the 999 soybean accessions was 90.31%, ranging from 43.70% to 99.51%. Kunitz trypsin inhibitor 3 (KTi-3) gene target sequencing analysis was performed on seven soybean germplasm accessions (IT105782, IT170889, IT273590, IT274513, IT274515, IT276197, and IT022891) showing less than 60% TIA. Four soybean accessions (IT274513, IT274515, IT276197, and IT022891) exhibited the same mutations (a G-to-T transversion and an AG deletion) in the KTi-3 gene (<i>Glyma.08g341500</i>) in PI542044. In addition, whole-genome re-sequencing was performed on three soybean accessions with no mutations in the KTi-3 gene. Compared with the reference soybean genome, an in-frame insertion and five missense mutations were identified in the coding sequencing of the KTi-1 gene (<i>Glyma.01g09500</i>) in IT105782. A RT-qPCR analysis showed that the mRNA expression level of KTi-1 was reduced by approximately 17% in IT105782 during seed development. In this study, we identified a previously unreported mutation in the KTi-1 gene and developed a KASP marker using this new KTi-1 variant.https://www.mdpi.com/2077-0472/13/11/2070soybeantrypsin inhibitor (TI)trypsin inhibitor activity (TIA)KASP marker
spellingShingle Aron Park
Se-Hee Kang
Byeong-Hee Kang
Sreeparna Chowdhury
Seo-Young Shin
Won-Ho Lee
Jeong-Dong Lee
Sungwoo Lee
Yu-Mi Choi
Bo-Keun Ha
Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
Agriculture
soybean
trypsin inhibitor (TI)
trypsin inhibitor activity (TIA)
KASP marker
title Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
title_full Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
title_fullStr Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
title_full_unstemmed Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
title_short Identification of a Novel KTi-1 Allele Associated with Reduced Trypsin Inhibitor Activity in Soybean Accessions
title_sort identification of a novel kti 1 allele associated with reduced trypsin inhibitor activity in soybean accessions
topic soybean
trypsin inhibitor (TI)
trypsin inhibitor activity (TIA)
KASP marker
url https://www.mdpi.com/2077-0472/13/11/2070
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