Structure and function of class III pistil-specific extensin-like protein in interspecific reproductive barriers

Camila M.L. Alves, Andrzej K. Noyszewski, Alan G. Smith

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Background: The transmitting tissue of the style is the pathway for pollen tube growth to the ovules and has components that function in recognizing and discriminating appropriate pollen genotypes. In Nicotiana tabacum, the class III pistil extensin-like (PELPIII) arabinogalactan protein is essential for the inhibition of N. obtusifolia pollen tube growth. The transmitting tissue-specific (TTS) arabinogalactan protein amino acid sequence and expression pattern is similar to PELPIII, but it facilitates self-pollinated N. tabacum. The TTS and PELPIII arabinogalactan protein can be divided into the less conserved N-terminal (NTD) and the more conserved C-terminal (CTD) domains. This research tested whether the NTD is the key domain in determining PELPIII function in the inhibition of interspecific pollen tube growth. Three variant PELPIII gene constructs were produced where the PELPIII NTD was exchanged with the TTS NTD and a single amino acid change (cysteine to alanine) was introduced into the PELPIII NTD. The PELPIII variants of N. tabacum were tested for activity by measuring the inhibition N. obtusifolia pollen tube growth by using them to complement a 3'UTR RNAi transgenic line with reduced PELPIII mRNA. Results: The RNAi N. tabacum line had reduced PELPIII mRNA accumulation and reduced inhibition of N. obtusifolia pollen tube growth, but had no effect on self-pollen tube growth or pollen tube growth of 12 other Nicotiana species. The NTD of PELPIII with either the PELPIII or TTS CTDs complemented the loss PELPIII activity in the RNAi transgenic line as measured by inhibition of N. obtusifolia pollen tube growth. The TTS NTD with the PELPIII CTD and a variant PELPIII with a cysteine to alanine mutation in its NTD failed to complement the loss of PELPIII activity and did not inhibit N. obtusifolia pollen tube growth. Conclusion: The NTD is a key determinant in PELPIII's function in regulating interspecific pollen tube growth and is a first step toward understanding the mechanism of how PELPIII NTD regulates pollen tube growth.

Original languageEnglish (US)
Article number118
JournalBMC plant biology
Volume19
Issue number1
DOIs
StatePublished - Mar 29 2019

Bibliographical note

Funding Information:
This work was supported by Coordenação de Aperfeicoamento de Pessoal de Nível Superior - CAPES (Science without Borders), Minnesota Agricultural Experiment Station, the University of Minnesota, Applied Plant Science Metric Fellowship and Bridging funds, and an Alexander and Lydia Anderson grant. The funding bodies had no role in the design of the study and no role in the collection, analysis, and interpretation of data or in writing the manuscript.

Publisher Copyright:
© 2019 The Author(s).

Keywords

  • 3'UTR RNAi
  • Arabinogalactan proteins
  • Gene complementation
  • Nicotiana obtusifolia
  • Nicotiana tabacum
  • PELPIII
  • Pollen tube growth
  • TTS

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