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Silencing of glycerol-3-phosphate acyltransferase 6 (GPAT6) gene using a newly established virus induced gene silencing (VIGS) system in cucumber alleviates autotoxicity mimicked by cinnamic acid (CA)

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Abstract

Background and aims

Autotoxicity is a serious problem for continuous cropping of cucumber, it can limit its own growth and that of nearby cucumber plants and decrease yield and quality. Cinnamic acid (CA) derivatives are a class of autotoxins that can mimic autotoxic effects in cucumber.

Methods

We screened an Arabidopsis long hairpin RNA (lhRNA) library under CA-mimicked autotoxicity stress to obtain autotoxicity-resistant mutant lines. The glycerol-3-phosphate 2-O-acyltransferase 6 (GPAT6) gene was found to be silenced in one resistant mutant line, and GPAT6 homologous gene in cucumber was cloned and named CuGPAT6. To test gene function quickly, a new tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) system was established in cucumber using a special agroinfiltration solution and whole cotyledonary nodes.

Results

Cucumber phytoene desaturase gene (CuPDS) and CuGPAT6 gene were successfully silenced with a newly established tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) system in cucumber showing photobleaching and decreased CuPDS expression in leaves; and silencing of CuGPAT6 enhanced the tolerance to the autotoxin stress mimicked by CA in cucumber.

Conclusions

It indicated that the alleviation of the damage caused by CA treatment was probably through maintaining the integrity of cellular structure, supporting cell vitality and promoting growth in roots, and increasing the integrate chloroplast and thylakoid numbers in leaves, which restored the growth and net photosynthesis rate of CA treated CuGPAT6 silenced plants to the similar level to those of control plants. The results also demonstrated that the newly established TRV mediated VIGS system is functional, rapid, convenient and highly efficient in cucumber.

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Abbreviations

TRV-VIGS:

Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS)

PDS:

Phytoene desaturase

lhRNA:

Long hairpin RNA

RMHR:

Rolling-circle amplification (RCA)-mediated hpRNA

GPAT6:

Glycerol-3-phosphate 2-O-acyltransferase 6

CA:

Cinnamic acid

WT:

Wild-type

TEM:

Transmission electron microscopy

Pn:

Net photosynthetic rate

NCBI:

National Center for Biotechnology Information

BLAST:

Basic Local Alignment Search Tool

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Acknowledgments

We thank professor Lei Wang and the Key Laboratory of Plant Genetics and Molecular Breeding at Zhoukou Normal University for providing RNAi mutant library of Arabidopsis thaliana and pTRV2-SlPDS vector. The authors gratefully acknowledge Prof. Yule Liu (Tsinghua University, China) and Prof. David Baulcombe (University of Cambridge, UK) for providing VIGS vectors. This research was supported by the National Natural Science Foundation Funded of China (31872129 and 31071807), Henan Natural Science Fund (162300410107); University-Level Technology Leader (2016BZ01); China Postdoctoral Science Foundation Funded Project (2018 M630821); Henan Postdoctoral Science Foundation Funded Project (103020218003/002).

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Authors and Affiliations

  1. College of Life Science, Henan Agricultural University, Zhengzhou, 450000, China

    Ruifang Bu & Jianyu Wu

  2. Henan Engineering Research Center of Crop Genome Editing, College of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang, 453003, China

    Ruifang Bu, Runhao Wang, Qichao Wei, Haiyan Hu, Haili Sun, Puwen Song, Yongang Yu, Qili Liu, Zichao Zheng, Tong Li, Dongxiao Li, Shijun Chen, Liuliu Wu & Chengwei Li

  3. Postdoctoral Research Base, Henan Institute of Science and Technology, Xinxiang, 453003, China

    Ruifang Bu

  4. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Lei Wang

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  10. Tong Li
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  11. Dongxiao Li
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  12. Lei Wang
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  13. Shijun Chen
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  14. Liuliu Wu
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  15. Jianyu Wu
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  16. Chengwei Li
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Corresponding authors

Correspondence to Jianyu Wu or Chengwei Li.

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Bu, R., Wang, R., Wei, Q. et al. Silencing of glycerol-3-phosphate acyltransferase 6 (GPAT6) gene using a newly established virus induced gene silencing (VIGS) system in cucumber alleviates autotoxicity mimicked by cinnamic acid (CA). Plant Soil 438, 329–346 (2019). https://doi.org/10.1007/s11104-019-03996-0

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