Cargando…
Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants
Agrobacterium-mediated sorghum transformation frequency has been enhanced significantly via medium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the rest...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4544465/ https://www.ncbi.nlm.nih.gov/pubmed/26316679 http://dx.doi.org/10.1007/s11627-013-9583-z |
_version_ | 1782386670816985088 |
---|---|
author | Wu, Emily Lenderts, Brian Glassman, Kimberly Berezowska-Kaniewska, Maya Christensen, Heather Asmus, Tracy Zhen, Shifu Chu, Uyen Cho, Myeong-Je Zhao, Zuo-Yu |
author_facet | Wu, Emily Lenderts, Brian Glassman, Kimberly Berezowska-Kaniewska, Maya Christensen, Heather Asmus, Tracy Zhen, Shifu Chu, Uyen Cho, Myeong-Je Zhao, Zuo-Yu |
author_sort | Wu, Emily |
collection | PubMed |
description | Agrobacterium-mediated sorghum transformation frequency has been enhanced significantly via medium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the resting and selection media. Using Agrobacterium strain LBA4404, the transformation frequency reached over 10% using either of two different selection marker genes, moPAT or PMI, and any of three different vectors in large-scale transformation experiments. With Agrobacterium strain AGL1, the transformation frequencies were as high as 33%. Using quantitative PCR analyses of 1,182 T(0) transgenic plants representing 675 independent transgenic events, data was collected for T-DNA copy number, intact or truncated T-DNA integration, and vector backbone integration into the sorghum genome. A comparison of the transformation frequencies and molecular data characterizing T-DNA integration patterns in the transgenic plants derived from LBA4404 versus AGL1 transformation revealed that twice as many transgenic high-quality events were generated when AGL1 was used compared to LBA4404. This is the first report providing molecular data for T-DNA integration patterns in a large number of independent transgenic plants in sorghum. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11627-013-9583-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4544465 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-45444652015-08-25 Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants Wu, Emily Lenderts, Brian Glassman, Kimberly Berezowska-Kaniewska, Maya Christensen, Heather Asmus, Tracy Zhen, Shifu Chu, Uyen Cho, Myeong-Je Zhao, Zuo-Yu In Vitro Cell Dev Biol Plant Biotechnology/Genetic Transformation/Functional Genomics Agrobacterium-mediated sorghum transformation frequency has been enhanced significantly via medium optimization using immature embryos from sorghum variety TX430 as the target tissue. The new transformation protocol includes the addition of elevated copper sulfate and 6-benzylaminopurine in the resting and selection media. Using Agrobacterium strain LBA4404, the transformation frequency reached over 10% using either of two different selection marker genes, moPAT or PMI, and any of three different vectors in large-scale transformation experiments. With Agrobacterium strain AGL1, the transformation frequencies were as high as 33%. Using quantitative PCR analyses of 1,182 T(0) transgenic plants representing 675 independent transgenic events, data was collected for T-DNA copy number, intact or truncated T-DNA integration, and vector backbone integration into the sorghum genome. A comparison of the transformation frequencies and molecular data characterizing T-DNA integration patterns in the transgenic plants derived from LBA4404 versus AGL1 transformation revealed that twice as many transgenic high-quality events were generated when AGL1 was used compared to LBA4404. This is the first report providing molecular data for T-DNA integration patterns in a large number of independent transgenic plants in sorghum. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11627-013-9583-z) contains supplementary material, which is available to authorized users. Springer US 2013-12-13 2014 /pmc/articles/PMC4544465/ /pubmed/26316679 http://dx.doi.org/10.1007/s11627-013-9583-z Text en © The Author(s) 2013 |
spellingShingle | Biotechnology/Genetic Transformation/Functional Genomics Wu, Emily Lenderts, Brian Glassman, Kimberly Berezowska-Kaniewska, Maya Christensen, Heather Asmus, Tracy Zhen, Shifu Chu, Uyen Cho, Myeong-Je Zhao, Zuo-Yu Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title | Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title_full | Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title_fullStr | Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title_full_unstemmed | Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title_short | Optimized Agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
title_sort | optimized agrobacterium-mediated sorghum transformation protocol and molecular data of transgenic sorghum plants |
topic | Biotechnology/Genetic Transformation/Functional Genomics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4544465/ https://www.ncbi.nlm.nih.gov/pubmed/26316679 http://dx.doi.org/10.1007/s11627-013-9583-z |
work_keys_str_mv | AT wuemily optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT lendertsbrian optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT glassmankimberly optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT berezowskakaniewskamaya optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT christensenheather optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT asmustracy optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT zhenshifu optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT chuuyen optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT chomyeongje optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants AT zhaozuoyu optimizedagrobacteriummediatedsorghumtransformationprotocolandmoleculardataoftransgenicsorghumplants |