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Effects of the wheat UDP-glucosyltransferase gene TaUGT-B2 on Agrobacterium-mediated plant transformation

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Abstract

UDP-glucosyltransferase enzymes (UGTs) are known to function in plant disease defense responses. In our previous RNA-seq analysis, TaUGT expression was found to be strongly up-regulated in Agrobacterium-infected wheat immature embryos, as compared to untreated controls. In this study, we isolated the TaUGT-B2, located on wheat 2BS chromosome, and characterized its function in tobacco, Arabidopsis, and wheat. Transient overexpression of TaUGT-B2 reduced the number of Agrobacterium cells attached to the surface of wheat cells, and hindered Agrobacterium-mediated GUS transient expression in wheat cells. Transiently silenced TaUGT-B2 expression resulted in an increased rate of shoot regeneration from Agrobacterium-infected wheat tissues. Therefore, we concluded that the overexpression of TaUGT-B2 impeded T-DNA transformation by reducing number of Agrobacterium cells attached to wheat tissues and lowering the regeneration efficiency of transgenic plantlets. In order to further investigate the function of TaUGT-B2, it was introduced into tobacco and Arabidopsis, and transgenic plants were then re-transformed with another vector carrying the GUS gene. The results of these experiments indicated that the expression of TaUGT-B2 in tobacco and Arabidopsis led to a reduction in the efficiency of Agrobacterium-mediated transformation. GUS transient expression was also impeded following overexpression of TaUGT-B2 in tobacco and Arabidopsis. Therefore, we conclude that TaUGT-B2 plays a negative role in Agrobacterium-mediated transformation of plants, especially in wheat.

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Abbreviations

PCR:

Polymerase chain reaction

SAR:

Systemic acquired resistance

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEM:

Scanning electron microscopy

TOGT:

UDP-glucose:phenylpropanoid glucosyltransferase

UGGT:

UDP-glucose:glycoprotein glycosyltransferase

UGTs:

UDP-glucosyltransferases

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Acknowledgements

This research was financially supported in part by the Ministry of Agriculture of China (2014ZX08010-004) and the National Natural Science Foundation of China (31401380, 31371621), and the Agricultural Science and Technology Innovation Program (ASTIP) of Chinese Academy of Agricultural Sciences.

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    Authors and Affiliations

    1. Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

      Xiaohong Zhou, Ke Wang, Lipu Du, Zhishan Lin & Xingguo Ye

    2. Institute of Genetics and Physiology/Plant Genetic Engineering Center of Hebei Province, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050051, People’s Republic of China

      Yongwei Liu

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    1. Xiaohong Zhou
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    2. Ke Wang
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    Correspondence to Xingguo Ye.

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    Communicated by J-H Liu.

    Xiaohong Zhou and Ke Wang contributed equally to this work.

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    Zhou, X., Wang, K., Du, L. et al. Effects of the wheat UDP-glucosyltransferase gene TaUGT-B2 on Agrobacterium-mediated plant transformation. Acta Physiol Plant 39, 15 (2017). https://doi.org/10.1007/s11738-016-2317-1

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