Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

doi:10.1016/S2095-3119(14)60907-2

Journal of Integrative Agriculture ›› 2015, Vol. 14 ›› Issue (7): 1242-1250.DOI: 10.1016/S2095-3119(14)60907-2

Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

GUO Bing-fu, GUO Yong, WANG Jun, ZHANG Li-juan, JIN Long-guo, HONG Hui-long, CHANG, Ru-zheng, QIU Li-juan

1、College of Agriculture, Northeast Agricultural University, Harbin 150030, P.R.China
2、The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
3、College of Agriculture, Yangtze University, Jingzhou 434025, P.R.China

收稿日期:2014-09-05 出版日期:2015-07-03 发布日期:2015-07-06
通讯作者: QIU Li-juan, Tel: +86-10-82105843, E-mail:qiulijuan@caas.cn
作者简介:GUO Bing-fu, Tel: +86-10-82105836, E-mail: gbfhg@163.com,These authors contributed equally to this work.
基金资助:
This work was supported by the National Transgenic Major Program of China (2014ZX08004001).

Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

GUO Bing-fu, GUO Yong, WANG Jun, ZHANG Li-juan, JIN Long-guo, HONG Hui-long, CHANG, Ru-zheng, QIU Li-juan

1、College of Agriculture, Northeast Agricultural University, Harbin 150030, P.R.China
2、The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI), Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
3、College of Agriculture, Yangtze University, Jingzhou 434025, P.R.China

Received:2014-09-05 Online:2015-07-03 Published:2015-07-06
Contact: QIU Li-juan, Tel: +86-10-82105843, E-mail:qiulijuan@caas.cn
About author:GUO Bing-fu, Tel: +86-10-82105836, E-mail: gbfhg@163.com,These authors contributed equally to this work.
Supported by:
This work was supported by the National Transgenic Major Program of China (2014ZX08004001).

摘要/Abstract

摘要： Soybean is a widely planted genetically modified crop around the world. However, it is still one of the most recalcitrant crops for genetic transformation due to the difficulty of regeneration via organogenesis and some factors that affect the transformation efficiency. The percentages of resistant bud formation and transient expression efficiency are important indexes reflecting the regeneration and transformation efficiency of soybean. In this study, the percentages of resistant bud formation and transient expression of β-glucuronidase (GUS) were compared after treatment with sonication or surfactant and co-treatment with both. The results showed that treatment with either sonication or surfactant increased the percentage of resistant bud formation and transient expression efficiency. The highest percentages were acquired and significantly improved when cotyledon node explants were co-treated with sonication for 2 s and surfactant at 0.02% (v:v) using two different soybean genotypes, Jack and Zhonghuang 10. The improved transformation efficiency of this combination was also evaluated by development of herbicide-tolerant soybeans with transformation efficiency at 2.5–5.7% for different genotypes, which was significantly higher than traditional cotyledonary node method in this study. These results suggested that co-treatment with surfactant and sonication significantly improved the percentages of resistance bud formation, transient expression efficiency and stable transformation efficiency in soybean.

关键词: soybean , genetic transformation , sonication , surfactant , co-treatment

Abstract: Soybean is a widely planted genetically modified crop around the world. However, it is still one of the most recalcitrant crops for genetic transformation due to the difficulty of regeneration via organogenesis and some factors that affect the transformation efficiency. The percentages of resistant bud formation and transient expression efficiency are important indexes reflecting the regeneration and transformation efficiency of soybean. In this study, the percentages of resistant bud formation and transient expression of β-glucuronidase (GUS) were compared after treatment with sonication or surfactant and co-treatment with both. The results showed that treatment with either sonication or surfactant increased the percentage of resistant bud formation and transient expression efficiency. The highest percentages were acquired and significantly improved when cotyledon node explants were co-treated with sonication for 2 s and surfactant at 0.02% (v:v) using two different soybean genotypes, Jack and Zhonghuang 10. The improved transformation efficiency of this combination was also evaluated by development of herbicide-tolerant soybeans with transformation efficiency at 2.5–5.7% for different genotypes, which was significantly higher than traditional cotyledonary node method in this study. These results suggested that co-treatment with surfactant and sonication significantly improved the percentages of resistance bud formation, transient expression efficiency and stable transformation efficiency in soybean.

Key words: soybean , genetic transformation , sonication , surfactant , co-treatment

GUO Bing-fu, GUO Yong, WANG Jun, ZHANG Li-juan, JIN Long-guo, HONG Hui-long, CHANG, Ru-zheng, QIU Li-juan. Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean[J]. Journal of Integrative Agriculture, 2015, 14(7): 1242-1250.

GUO Bing-fu, GUO Yong, WANG Jun, ZHANG Li-juan, JIN Long-guo, HONG Hui-long, CHANG , Ru-zheng , QIU Li-juan. Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean[J]. Journal of Integrative Agriculture, 2015, 14(7): 1242-1250.

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Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

Co-treatment with surfactant and sonication significantly improves Agrobacterium-mediated resistant bud formation and transient expression efficiency in soybean

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