Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate

doi:10.1016/S2095-3119(19)62747-4

Journal of Integrative Agriculture ›› 2020, Vol. 19 ›› Issue (5): 1186-1196.DOI: 10.1016/S2095-3119(19)62747-4

收稿日期:2019-01-04 出版日期:2020-04-01 发布日期:2020-03-25

Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate

GUO Bing-fu^{1, 2}, HONG Hui-long¹, HAN Jia-nan¹, ZHANG Li-juan¹, LIU Zhang-xiong¹, GUO Yong¹, QIU Li-juan¹

¹ National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI)/MOA Key Lab of Soybean Biology/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
² Jiangxi Province Key Laboratory of Oilcrops Biology/Crops Research Institute, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, P.R.China

Received:2019-01-04 Online:2020-04-01 Published:2020-03-25
Contact: Correspondence QIU Li-juan, Tel/Fax: +86-10-82105840, E-mail: qiulijuan@caas.cn; GUO Yong, Tel/Fax: +86-10-82105840, E-mail: guoyong@caas.cn
About author: GUO Bing-fu, E-mail: gbfhq@163.com;
Supported by:
This work was supported by the National Natural Science Foundation of China (31601326), the National Transgenic Major Program of China (2016ZX08004001 and 2016ZX08011003), and the China Postdoctoral Science Foundation (2016M591300).

摘要/Abstract

Abstract:

Glyphosate-tolerant soybean is the most widely planted genetically modified crop worldwide. However, soybean remains recalcitrant to routine transformation because of the low infection efficiency of Agrobacterium to soybean and lack of useful selectable markers. In this study, several Agrobacterium strains and cell densities were compared by transient expression of the GUS gene. The results showed that Agrobacterium strain Ag10 at cell densities of OD₆₀₀of 0.6–0.9 yielded the highest infection efficiency in Agrobacterium-mediated soybean cotyledonary node transformation system. Meanwhile, a simple and rapid method was developed for identification of glyphosate tolerance in putative T₀ transgenic plants, consisting of spotting plantlets with 1 µL Roundup®. The whole cycle of genetic transformation could be shortened to about 3 mon by highly efficient selection with glyphosate during the transformation process and application of the spot assay in putative T₀ transgenic plantlets. The transformation frequency ranged from 2.9 to 5.6%. This study provides an improved protocol for development and identification of glyphosate-tolerant transgenic soybeans.

Key words: soybean , transformation , spot assay , glyphosate selection , transgenic plants

. [J]. Journal of Integrative Agriculture, 2020, 19(5): 1186-1196.

GUO Bing-fu, HONG Hui-long, HAN Jia-nan, ZHANG Li-juan, LIU Zhang-xiong, GUO Yong, QIU Li-juan. Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate[J]. Journal of Integrative Agriculture, 2020, 19(5): 1186-1196.

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Development and identification of glyphosate-tolerant transgenic soybean via direct selection with glyphosate

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