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Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response
BACKGROUND: The Agrobacterium-mediated transient transformation is a versatile and indispensable way of rapid analyzing gene function in plants. Despite this transient expression system has been successfully applied in a number of plant species, it is poorly developed in Caragana intermedia. RESULTS...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6554893/ https://www.ncbi.nlm.nih.gov/pubmed/31170915 http://dx.doi.org/10.1186/s12870-019-1800-4 |
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author | Liu, Kun Yang, Qi Yang, Tianrui Wu, Yang Wang, Guangxia Yang, Feiyun Wang, Ruigang Lin, Xiaofei Li, Guojing |
author_facet | Liu, Kun Yang, Qi Yang, Tianrui Wu, Yang Wang, Guangxia Yang, Feiyun Wang, Ruigang Lin, Xiaofei Li, Guojing |
author_sort | Liu, Kun |
collection | PubMed |
description | BACKGROUND: The Agrobacterium-mediated transient transformation is a versatile and indispensable way of rapid analyzing gene function in plants. Despite this transient expression system has been successfully applied in a number of plant species, it is poorly developed in Caragana intermedia. RESULTS: In this study, we established an Agrobacterium-mediated transient expression system in C. intermedia leaves and optimized the effect of different Agrobacterial strains, several surfactants and the concentration of Silwet L-77, which would affect transient expression efficiency. Among the 5 Agrobacterial strains examined, GV3101 produced the highest GUS expression level. Besides, higher level of transient expression was observed in plants infiltrated with Silwet L-77 than with Triton X-100 or Tween-20. Silwet L-77 at a concentration of 0.001% greatly improved the level of GUS transient expression. Real-time PCR showed that expression of CiDREB1C was highly up-regulated in transiently expressed plants and reached the highest level at the 2nd day after infiltration. Based on this optimized transient transformation method, we characterized CiDREB1C function in response to drought, salt and ABA treatment. The results showed that transiently expressed CiDREB1C in C. intermedia leaves could enhance the survival rate and chlorophyll content, and reduce the lodging rate compared with the control seedlings under drought, salt and ABA treatments. Furthermore, the rate of leaf shedding of CiDREB1C transient expression seedlings was lower than that of the control under ABA treatment. CONCLUSIONS: The optimized transient expression condition in C. intermedia leaves were infiltrated with Agrobacterial strains GV3101 plus Silwet L-77 at a concentration of 0.001% added into the infiltration medium. Transiently expressed CiDREB1C enhanced drought, salt and ABA stress tolerance, indicated that it was a suitable and effective tool to determine gene function involved in abiotic stress response in C. intermedia. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12870-019-1800-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6554893 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-65548932019-06-10 Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response Liu, Kun Yang, Qi Yang, Tianrui Wu, Yang Wang, Guangxia Yang, Feiyun Wang, Ruigang Lin, Xiaofei Li, Guojing BMC Plant Biol Methodology Article BACKGROUND: The Agrobacterium-mediated transient transformation is a versatile and indispensable way of rapid analyzing gene function in plants. Despite this transient expression system has been successfully applied in a number of plant species, it is poorly developed in Caragana intermedia. RESULTS: In this study, we established an Agrobacterium-mediated transient expression system in C. intermedia leaves and optimized the effect of different Agrobacterial strains, several surfactants and the concentration of Silwet L-77, which would affect transient expression efficiency. Among the 5 Agrobacterial strains examined, GV3101 produced the highest GUS expression level. Besides, higher level of transient expression was observed in plants infiltrated with Silwet L-77 than with Triton X-100 or Tween-20. Silwet L-77 at a concentration of 0.001% greatly improved the level of GUS transient expression. Real-time PCR showed that expression of CiDREB1C was highly up-regulated in transiently expressed plants and reached the highest level at the 2nd day after infiltration. Based on this optimized transient transformation method, we characterized CiDREB1C function in response to drought, salt and ABA treatment. The results showed that transiently expressed CiDREB1C in C. intermedia leaves could enhance the survival rate and chlorophyll content, and reduce the lodging rate compared with the control seedlings under drought, salt and ABA treatments. Furthermore, the rate of leaf shedding of CiDREB1C transient expression seedlings was lower than that of the control under ABA treatment. CONCLUSIONS: The optimized transient expression condition in C. intermedia leaves were infiltrated with Agrobacterial strains GV3101 plus Silwet L-77 at a concentration of 0.001% added into the infiltration medium. Transiently expressed CiDREB1C enhanced drought, salt and ABA stress tolerance, indicated that it was a suitable and effective tool to determine gene function involved in abiotic stress response in C. intermedia. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12870-019-1800-4) contains supplementary material, which is available to authorized users. BioMed Central 2019-06-06 /pmc/articles/PMC6554893/ /pubmed/31170915 http://dx.doi.org/10.1186/s12870-019-1800-4 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Liu, Kun Yang, Qi Yang, Tianrui Wu, Yang Wang, Guangxia Yang, Feiyun Wang, Ruigang Lin, Xiaofei Li, Guojing Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title | Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title_full | Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title_fullStr | Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title_full_unstemmed | Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title_short | Development of Agrobacterium-mediated transient expression system in Caragana intermedia and characterization of CiDREB1C in stress response |
title_sort | development of agrobacterium-mediated transient expression system in caragana intermedia and characterization of cidreb1c in stress response |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6554893/ https://www.ncbi.nlm.nih.gov/pubmed/31170915 http://dx.doi.org/10.1186/s12870-019-1800-4 |
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