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Inhibition of isoflavone biosynthesis enhanced T-DNA delivery in soybean by improving plant–Agrobacterium tumefaciens interaction

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Abstract

Soybean is a difficult crop to manipulate through Agrobacterium tumefaciens-mediated genetic transformation. Plant–bacterium interaction plays an important role in the transformation process. Being rich with isoflavones, soybean may have an adverse effect on the A. tumefaciens-mediated genetic transformation. To investigate whether high content of endogenous isoflavones in soybean is a serious obstacle in achieving high efficient Agrobacterium-mediated transformation in soybean, a series of experiments on inhibition of isoflavone biosynthesis were conducted to improve upon soybean transformation efficiency. Results indicated that soybean isoflavones inhibited A. tumefaciens growth and respiration, the transformation efficiency [β-glucuronidase (GUS) transient expression] was negatively correlated with the phenylalanine ammonia-lyase activity and isoflavones content. The biosynthesis of soybean isoflavones was partially inhibited by sonication treatment and applying antagonists in co-culture medium and thereby decreased the adverse effects of isoflavones on Agrobacterium infection. A discernible improvement in transformation efficiency was achieved when sonication at 40 kHz for 3 min was applied along with Agro-infection and the explants were cultured on co-culture medium containing 20 μM α-aminooxyacetic acid (AOA), with the percentage of GUS transient expression as 41.4 %, being 3.6 times higher than that not sonicated and co-cultured on medium without AOA. Sonication was found not only to simply make micro-wounds for Agrobacterium to penetrate or releasing phenolic compounds for induced Agrobacterium vir gene expression; it disturbed the biosynthesis of isoflavones at the transcription level and decreased the adverse effects of isoflavones on soybean transformation, and thereby improving soybean transformation efficiency.

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Abbreviations

PAL:

Phenylalanine ammonia-lyase

AOA:

α-Aminooxyacetic acid

AS:

Acetosyringone

SAAT:

Sonication-assisted Agrobacterium-mediated transformation

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Acknowledgments

This work was supported by National Major Project for Transgenic Crops of Chinese Agriculture Ministry (Grant No. 2014ZX0800402B); Natural Science Foundation of Hebei Province, China (Grant No. C2013301033); Key project for fundamental research of Hebei Province, China (Grant No. 14962903D).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences, Plant Genetic Engineering Center of Hebei Province, Shijiazhuang, 050051, China

    Yan-Min Zhang, Hong-Mei Zhang, Zi-Hui Liu, Xiu-Lin Guo, Hui-Cong Li & Guo-Liang Li

  2. Institute of Food and Oil Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang, 050035, China

    Chun-Zhi Jiang & Meng-Chen Zhang

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  1. Yan-Min Zhang
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Zhang, YM., Zhang, HM., Liu, ZH. et al. Inhibition of isoflavone biosynthesis enhanced T-DNA delivery in soybean by improving plant–Agrobacterium tumefaciens interaction. Plant Cell Tiss Organ Cult 121, 183–193 (2015). https://doi.org/10.1007/s11240-014-0693-z

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