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Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)

Eleusine indica (goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report...

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Autores principales: Luo, Qiyu, Chen, Shu, Nian, Hai, Ma, Qibing, Ding, Yuyao, Hao, Qinwen, Wei, Jiping, Patel, Jinesh D., McElroy, Joseph Scott, Liu, Yaoguang, Chen, Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10095498/
https://www.ncbi.nlm.nih.gov/pubmed/37047599
http://dx.doi.org/10.3390/ijms24076629
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author Luo, Qiyu
Chen, Shu
Nian, Hai
Ma, Qibing
Ding, Yuyao
Hao, Qinwen
Wei, Jiping
Patel, Jinesh D.
McElroy, Joseph Scott
Liu, Yaoguang
Chen, Yong
author_facet Luo, Qiyu
Chen, Shu
Nian, Hai
Ma, Qibing
Ding, Yuyao
Hao, Qinwen
Wei, Jiping
Patel, Jinesh D.
McElroy, Joseph Scott
Liu, Yaoguang
Chen, Yong
author_sort Luo, Qiyu
collection PubMed
description Eleusine indica (goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report a successful Agrobacterium-mediated transformation method for goosegrass. Firstly, we optimized conditions for breaking seed dormancy and increasing seed germination rate. A higher callus induction rate from germinated seeds was obtained in N6 than in MS or B5 medium. Then the optimal transformation efficiency of the gus reporter gene was obtained by infection with Agrobacterium tumefaciens culture of OD(600) = 0.5 for 30 min, followed by 3 days of co-cultivation with 300 μmol/L acetosyringone. Concentrations of 20 mg L(−1) kanamycin and 100 mg L(−1) timentin were used to select the transformed calli. The optimal rate of regeneration of the calli was generated by using 0.50 mg L(−1) 6-BA and 0.50 mg L(−1) KT in the culture medium. Then, using this transformation method, we overexpressed the paraquat-resistant EiKCS gene into a paraquat-susceptible goosegrass biotype MZ04 and confirmed the stable inheritance of paraquat-resistance in the transgenic goosegrass lines. This approach may provide a potential mechanism for the evolution of paraquat-resistant goosegrass and a promising gene for the manipulation of paraquat-resistance plants. This study is novel and valuable in future research using similar methods for herbicide resistance.
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spelling pubmed-100954982023-04-13 Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.) Luo, Qiyu Chen, Shu Nian, Hai Ma, Qibing Ding, Yuyao Hao, Qinwen Wei, Jiping Patel, Jinesh D. McElroy, Joseph Scott Liu, Yaoguang Chen, Yong Int J Mol Sci Article Eleusine indica (goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report a successful Agrobacterium-mediated transformation method for goosegrass. Firstly, we optimized conditions for breaking seed dormancy and increasing seed germination rate. A higher callus induction rate from germinated seeds was obtained in N6 than in MS or B5 medium. Then the optimal transformation efficiency of the gus reporter gene was obtained by infection with Agrobacterium tumefaciens culture of OD(600) = 0.5 for 30 min, followed by 3 days of co-cultivation with 300 μmol/L acetosyringone. Concentrations of 20 mg L(−1) kanamycin and 100 mg L(−1) timentin were used to select the transformed calli. The optimal rate of regeneration of the calli was generated by using 0.50 mg L(−1) 6-BA and 0.50 mg L(−1) KT in the culture medium. Then, using this transformation method, we overexpressed the paraquat-resistant EiKCS gene into a paraquat-susceptible goosegrass biotype MZ04 and confirmed the stable inheritance of paraquat-resistance in the transgenic goosegrass lines. This approach may provide a potential mechanism for the evolution of paraquat-resistant goosegrass and a promising gene for the manipulation of paraquat-resistance plants. This study is novel and valuable in future research using similar methods for herbicide resistance. MDPI 2023-04-01 /pmc/articles/PMC10095498/ /pubmed/37047599 http://dx.doi.org/10.3390/ijms24076629 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Luo, Qiyu
Chen, Shu
Nian, Hai
Ma, Qibing
Ding, Yuyao
Hao, Qinwen
Wei, Jiping
Patel, Jinesh D.
McElroy, Joseph Scott
Liu, Yaoguang
Chen, Yong
Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title_full Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title_fullStr Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title_full_unstemmed Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title_short Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)
title_sort establishment of an efficient agrobacterium-mediated genetic transformation system to enhance the tolerance of the paraquat stress in engineering goosegrass (eleusine indica l.)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10095498/
https://www.ncbi.nlm.nih.gov/pubmed/37047599
http://dx.doi.org/10.3390/ijms24076629
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