Downregulation of the auxin transporter gene SlPIN8 results in pollen abortion in tomato

  • Zengyu Gan
  • Yi Feng
  • Ting Wu
  • Yi Wang
  • Xuefeng Xu
  • Xinzhong Zhang
  • Zhenhai HanEmail author


Key message

SlPIN8 is expressed specifically within tomato pollen, and that it is involved in tomato pollen development and intracellular auxin homeostasis.


The auxin (IAA) transport protein PIN-FORMED (PIN) plays key roles in various aspects of plant development. The biological role of the auxin transporter SlPIN8 in tomato development remains unclear. Here, we examined the expression pattern of the SlPIN8 gene in vegetative and reproductive organs of tomato. RNA interference (RNAi) transgenic lines specifically silenced for the SlPIN8 gene were generated to identify the role of SlPIN8 in pollen development. We found that SlPIN8 mRNA is expressed specifically within tomato pollen. In the anthers, the highest mRNA expression and β-glucuronidase (GUS) activity of promoter-SlPIN8-GUS was detected during late stages of anther development, when pollen maturation occurred. The downregulation of SlPIN8 did not drastically affect the vegetative growth of tomato. However, in SlPIN8-RNAi transgenic plants, approximately 80% of the pollen grains were identified to be abnormal and lack viability; they were shriveled and flattened. Furthermore, the downregulation of SlPIN8 affected the gene expression of some anther development-specific proteins. SlPIN8-RNAi transgenic plants induced seedless fruits because of defective pollen function rather than defective female gametophyte function. In addition, SlPIN8 was found to localize to the endoplasmic reticulum, consistent with the changes in the auxin levels of SlPIN8-RNAi lines, whereas the level of free IAA was increased in SlPIN8-overexpressing protoplasts, indicating that SlPIN8 is involved in intracellular auxin homeostasis.


Auxin SlPIN8 Pollen RNAi Endoplasmic reticulum 



This work was supported by the Modern Agricultural Industry Technology System (CARS-27); Special Fund for Agro-Scientific Research in the Public Interest (201203075); Key Laboratory of Biology and Genetic Improvement of Horticultural Crop (Nutrition and Physiology) in Ministry of Agriculture, and Key Laboratory of Stress Physiology and Molecular Biology for Fruit Trees in Beijing Municipality.

Author Contributions

ZG designed and conducted the experiments, analyzed the data, accomplished pictures and wrote the manuscript. YF, YW, TW, XX contributed in design of the experiments. XZ designed the experiments and finalized the manuscript. ZH conceived and designed the experiments, and finalized the manuscript. All authors read and approved the final manuscript.

Supplementary material

11103_2019_836_MOESM1_ESM.docx (6.5 mb)
Figure S1. Expression analysis of tomato PIN genes in pollen at anthesis by qPCR. Error bars represent the means ± SD from three independent biological replicates. Figure S2. SlPIN8 expression patterns by qPCR. R root, S stem, L leaf, Fl flower, Se sepal, Pe petal, An anther, Po pollen. Figure S3. SlPIN8 gene promoter cis-element analysis. Figure S4. Inflorescence stem with GUS histochemical staining in Arabidopsis under the control of the SlPIN8 promoter. Bars = 0.5 mm. Figure S5. The construction and analysis of RNA interference (RNAi) SlPIN8 tomato lines. A, The RNAi tomato lines were generated by Agrobacterium tumefaciens-mediated transformation with the SlPIN8-RNAi T-DNA construct. B, Relative SlPIN8 mRNA levels in anthers collected from WT and SlPIN8-RNAi transgenic flowers at anthesis. Error bars represent the means ± SD from three independent biological replicates. Figure S6. The expression of SlPINs in SlPIN8-RNAi line and WT. Error bars represent the means ± SD from three independent biological replicates. Figure S7. Morphological observation of normal pollen germination in the WT and SlPIN8-RNAi transgenic plants. Bar = 25 μm. Figure S8. Paraffin section comparison of pollen development between the WT plants and SlPIN8-RNAi lines. Table S1. Primer for qRT-PCR. (DOCX 6627 KB)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Zengyu Gan
    • 1
  • Yi Feng
    • 1
  • Ting Wu
    • 1
  • Yi Wang
    • 1
  • Xuefeng Xu
    • 1
  • Xinzhong Zhang
    • 1
  • Zhenhai Han
    • 1
    Email author
  1. 1.Institute of Horticultural Plants, College of HorticultureChina Agricultural UniversityBeijingPeople’s Republic of China

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