Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean

WRINKLED1 (WRI1), an APETALA2/ethylene-responsive-element-binding protein (AP2/EREBP) subfamily transcription factor, plays a crucial role in the transcriptional regulation of plant fatty acid biosynthesis. In this study, <i>GmWRI1a</i> was overexpressed in the soybean cultivar ‘Dongnong...

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Main Authors: Zhikun Wang, Yuanzhuo Wang, Ping Shang, Chao Yang, Mingming Yang, Jinxiu Huang, Baizheng Ren, Zhaohui Zuo, Qingyan Zhang, Wenbin Li, Bo Song
Format: Article
Language:English
Published: MDPI AG 2022-05-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/23/9/5084
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author Zhikun Wang
Yuanzhuo Wang
Ping Shang
Chao Yang
Mingming Yang
Jinxiu Huang
Baizheng Ren
Zhaohui Zuo
Qingyan Zhang
Wenbin Li
Bo Song
author_facet Zhikun Wang
Yuanzhuo Wang
Ping Shang
Chao Yang
Mingming Yang
Jinxiu Huang
Baizheng Ren
Zhaohui Zuo
Qingyan Zhang
Wenbin Li
Bo Song
author_sort Zhikun Wang
collection DOAJ
description WRINKLED1 (WRI1), an APETALA2/ethylene-responsive-element-binding protein (AP2/EREBP) subfamily transcription factor, plays a crucial role in the transcriptional regulation of plant fatty acid biosynthesis. In this study, <i>GmWRI1a</i> was overexpressed in the soybean cultivar ‘Dongnong 50’ using <i>Agrobacterium</i>-mediated transformation to generate three transgenic lines with high seed oil contents. PCR and Southern blotting analysis showed that the T-DNA was inserted into the genome at precise insertion sites and was stably inherited by the progeny. Expression analysis using qRT-PCR and Western blotting indicated that <i>GmWRI1a</i> and <i>bar</i> driven by the CaMV <i>35S</i> promoter were significantly upregulated in the transgenic plants at different developmental stages. Transcriptome sequencing results showed there were obvious differences in gene expression between transgenic line and transgenic receptor during seed developmental stages. KEGG analysis found that the differentially expressed genes mainly annotated to metabolic pathways, such as carbohydrated metabolism and lipid metabolism. A 2-year single-location field trial revealed that three transgenic lines overexpressing <i>GmWRI1a</i> (GmWRI1a-OE) showed a stable increase in seed oil content of 4.97–10.35%. Importantly, no significant effect on protein content and yield was observed. Overexpression of <i>GmWRI1a</i> changed the fatty acid composition by increasing the linoleic acid (C18:2) content and decreasing the palmitic acid (C16:0) content in the seed. The three GmWRI1a-OE lines showed no significant changes in agronomic traits. The results demonstrated that the three <i>GmWRI1a</i> overexpression lines exhibited consistent increases in seed oil content compared with that of the wild type and did not significantly affect the seed yield and agronomic traits. The genetic engineering of <i>GmWRI1a</i> will be an effective strategy for the improvement of seed oil content and value in soybean.
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spelling doaj.art-f9b6d87948364fcf89cd77daf0cd01ee2023-11-23T08:26:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-01239508410.3390/ijms23095084Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in SoybeanZhikun Wang0Yuanzhuo Wang1Ping Shang2Chao Yang3Mingming Yang4Jinxiu Huang5Baizheng Ren6Zhaohui Zuo7Qingyan Zhang8Wenbin Li9Bo Song10Key Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaKey Laboratory of Soybean Biology of Ministry of Education China, Key Laboratory of Soybean Biology and Breeding (Genetics) of Ministry of Agriculture and Rural Affairs, Northeast Agricultural University, Harbin 150030, ChinaWRINKLED1 (WRI1), an APETALA2/ethylene-responsive-element-binding protein (AP2/EREBP) subfamily transcription factor, plays a crucial role in the transcriptional regulation of plant fatty acid biosynthesis. In this study, <i>GmWRI1a</i> was overexpressed in the soybean cultivar ‘Dongnong 50’ using <i>Agrobacterium</i>-mediated transformation to generate three transgenic lines with high seed oil contents. PCR and Southern blotting analysis showed that the T-DNA was inserted into the genome at precise insertion sites and was stably inherited by the progeny. Expression analysis using qRT-PCR and Western blotting indicated that <i>GmWRI1a</i> and <i>bar</i> driven by the CaMV <i>35S</i> promoter were significantly upregulated in the transgenic plants at different developmental stages. Transcriptome sequencing results showed there were obvious differences in gene expression between transgenic line and transgenic receptor during seed developmental stages. KEGG analysis found that the differentially expressed genes mainly annotated to metabolic pathways, such as carbohydrated metabolism and lipid metabolism. A 2-year single-location field trial revealed that three transgenic lines overexpressing <i>GmWRI1a</i> (GmWRI1a-OE) showed a stable increase in seed oil content of 4.97–10.35%. Importantly, no significant effect on protein content and yield was observed. Overexpression of <i>GmWRI1a</i> changed the fatty acid composition by increasing the linoleic acid (C18:2) content and decreasing the palmitic acid (C16:0) content in the seed. The three GmWRI1a-OE lines showed no significant changes in agronomic traits. The results demonstrated that the three <i>GmWRI1a</i> overexpression lines exhibited consistent increases in seed oil content compared with that of the wild type and did not significantly affect the seed yield and agronomic traits. The genetic engineering of <i>GmWRI1a</i> will be an effective strategy for the improvement of seed oil content and value in soybean.https://www.mdpi.com/1422-0067/23/9/5084soybean<i>GmWRI1a</i>overexpressionoil contentfatty acid composition
spellingShingle Zhikun Wang
Yuanzhuo Wang
Ping Shang
Chao Yang
Mingming Yang
Jinxiu Huang
Baizheng Ren
Zhaohui Zuo
Qingyan Zhang
Wenbin Li
Bo Song
Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
International Journal of Molecular Sciences
soybean
<i>GmWRI1a</i>
overexpression
oil content
fatty acid composition
title Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
title_full Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
title_fullStr Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
title_full_unstemmed Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
title_short Overexpression of Soybean <i>GmWRI1a</i> Stably Increases the Seed Oil Content in Soybean
title_sort overexpression of soybean i gmwri1a i stably increases the seed oil content in soybean
topic soybean
<i>GmWRI1a</i>
overexpression
oil content
fatty acid composition
url https://www.mdpi.com/1422-0067/23/9/5084
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