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CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis

Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (Glycine max). Breeding soybean varieties that are resistant to leaf-chewing insects can minimize the need for insecticide use and reduce yield loss. The marker gene for QTL-M, Glyma.07g110300 (LOC1007...

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Autores principales: Zhang, Yongxing, Guo, Wei, Chen, Limiao, Shen, Xinjie, Yang, Hongli, Fang, Yisheng, Ouyang, Wenqi, Mai, Sihua, Chen, Haifeng, Chen, Shuilian, Hao, Qingnan, Yuan, Songli, Zhang, Chanjuan, Huang, Yi, Shan, Zhihui, Yang, Zhonglu, Qiu, Dezhen, Zhou, Xinan, Cao, Dong, Li, Xia, Jiao, Yongqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8902248/
https://www.ncbi.nlm.nih.gov/pubmed/35273623
http://dx.doi.org/10.3389/fpls.2022.802716
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author Zhang, Yongxing
Guo, Wei
Chen, Limiao
Shen, Xinjie
Yang, Hongli
Fang, Yisheng
Ouyang, Wenqi
Mai, Sihua
Chen, Haifeng
Chen, Shuilian
Hao, Qingnan
Yuan, Songli
Zhang, Chanjuan
Huang, Yi
Shan, Zhihui
Yang, Zhonglu
Qiu, Dezhen
Zhou, Xinan
Cao, Dong
Li, Xia
Jiao, Yongqing
author_facet Zhang, Yongxing
Guo, Wei
Chen, Limiao
Shen, Xinjie
Yang, Hongli
Fang, Yisheng
Ouyang, Wenqi
Mai, Sihua
Chen, Haifeng
Chen, Shuilian
Hao, Qingnan
Yuan, Songli
Zhang, Chanjuan
Huang, Yi
Shan, Zhihui
Yang, Zhonglu
Qiu, Dezhen
Zhou, Xinan
Cao, Dong
Li, Xia
Jiao, Yongqing
author_sort Zhang, Yongxing
collection PubMed
description Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (Glycine max). Breeding soybean varieties that are resistant to leaf-chewing insects can minimize the need for insecticide use and reduce yield loss. The marker gene for QTL-M, Glyma.07g110300 (LOC100775351) that encodes a UDP-glycosyltransferase (UGT) is the major determinant of resistance against leaf-chewing insects in soybean; it exhibits a loss of function in insect-resistant soybean germplasms. In this study, Agrobacterium-mediated transformation introduced the CRISPR/Cas9 expression vector into the soybean cultivar Tianlong No. 1 to generate Glyma.07g110300-gene mutants. We obtained two novel types of mutations, a 33-bp deletion and a single-bp insertion in the GmUGT coding region, which resulted in an enhanced resistance to Helicoverpa armigera and Spodoptera litura. Additionally, overexpressing GmUGT produced soybean varieties that were more sensitive to H. armigera and S. litura. Both mutant and overexpressing lines exhibited no obvious phenotypic changes. The difference in metabolites and gene expression suggested that GmUGT is involved in imparting resistance to leaf-chewing insects by altering the flavonoid content and expression patterns of genes related to flavonoid biosynthesis and defense. Furthermore, ectopic expression of the GmUGT gene in the ugt72b1 mutant of Arabidopsis substantially rescued the phenotype of H. armigera resistance in the atugt72b1 mutant. Our study presents a strategy for increasing resistance against leaf-chewing insects in soybean through CRISPR/Cas9-mediated targeted mutagenesis of the UGT genes.
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spelling pubmed-89022482022-03-09 CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis Zhang, Yongxing Guo, Wei Chen, Limiao Shen, Xinjie Yang, Hongli Fang, Yisheng Ouyang, Wenqi Mai, Sihua Chen, Haifeng Chen, Shuilian Hao, Qingnan Yuan, Songli Zhang, Chanjuan Huang, Yi Shan, Zhihui Yang, Zhonglu Qiu, Dezhen Zhou, Xinan Cao, Dong Li, Xia Jiao, Yongqing Front Plant Sci Plant Science Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (Glycine max). Breeding soybean varieties that are resistant to leaf-chewing insects can minimize the need for insecticide use and reduce yield loss. The marker gene for QTL-M, Glyma.07g110300 (LOC100775351) that encodes a UDP-glycosyltransferase (UGT) is the major determinant of resistance against leaf-chewing insects in soybean; it exhibits a loss of function in insect-resistant soybean germplasms. In this study, Agrobacterium-mediated transformation introduced the CRISPR/Cas9 expression vector into the soybean cultivar Tianlong No. 1 to generate Glyma.07g110300-gene mutants. We obtained two novel types of mutations, a 33-bp deletion and a single-bp insertion in the GmUGT coding region, which resulted in an enhanced resistance to Helicoverpa armigera and Spodoptera litura. Additionally, overexpressing GmUGT produced soybean varieties that were more sensitive to H. armigera and S. litura. Both mutant and overexpressing lines exhibited no obvious phenotypic changes. The difference in metabolites and gene expression suggested that GmUGT is involved in imparting resistance to leaf-chewing insects by altering the flavonoid content and expression patterns of genes related to flavonoid biosynthesis and defense. Furthermore, ectopic expression of the GmUGT gene in the ugt72b1 mutant of Arabidopsis substantially rescued the phenotype of H. armigera resistance in the atugt72b1 mutant. Our study presents a strategy for increasing resistance against leaf-chewing insects in soybean through CRISPR/Cas9-mediated targeted mutagenesis of the UGT genes. Frontiers Media S.A. 2022-02-22 /pmc/articles/PMC8902248/ /pubmed/35273623 http://dx.doi.org/10.3389/fpls.2022.802716 Text en Copyright © 2022 Zhang, Guo, Chen, Shen, Yang, Fang, Ouyang, Mai, Chen, Chen, Hao, Yuan, Zhang, Huang, Shan, Yang, Qiu, Zhou, Cao, Li and Jiao. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Zhang, Yongxing
Guo, Wei
Chen, Limiao
Shen, Xinjie
Yang, Hongli
Fang, Yisheng
Ouyang, Wenqi
Mai, Sihua
Chen, Haifeng
Chen, Shuilian
Hao, Qingnan
Yuan, Songli
Zhang, Chanjuan
Huang, Yi
Shan, Zhihui
Yang, Zhonglu
Qiu, Dezhen
Zhou, Xinan
Cao, Dong
Li, Xia
Jiao, Yongqing
CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title_full CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title_fullStr CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title_full_unstemmed CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title_short CRISPR/Cas9-Mediated Targeted Mutagenesis of GmUGT Enhanced Soybean Resistance Against Leaf-Chewing Insects Through Flavonoids Biosynthesis
title_sort crispr/cas9-mediated targeted mutagenesis of gmugt enhanced soybean resistance against leaf-chewing insects through flavonoids biosynthesis
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8902248/
https://www.ncbi.nlm.nih.gov/pubmed/35273623
http://dx.doi.org/10.3389/fpls.2022.802716
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