Protein replenishment in pitcher fluids of Nepenthes × ventrata revealed by quantitative proteomics (SWATH-MS) informed by transcriptomics

Wan Nor Adibah Wan Zakaria, Wan Mohd Aizat Wan Kamaruddin, Hoe Han Goh, Normah Mohd. Noor

Research output: Contribution to journalArticle

Abstract

Carnivorous plants capture and digest insects for nutrients, allowing them to survive in soil deprived of nitrogenous nutrients. Plants from the genus Nepenthes produce unique pitchers containing secretory glands, which secrete enzymes into the digestive fluid. We performed RNA-seq analysis on the pitcher tissues and liquid chromatography-tandem mass spectrometry (LC–MS/MS) analysis on the pitcher fluids of Nepenthes × ventrata to study protein expression in this carnivory organ during early days of pitcher opening. This transcriptome provides a sequence database for pitcher fluid protein identification. A total of 32 proteins of diverse functions were successfully identified in which 19 proteins can be quantified based on label-free quantitative proteomics (SWATH-MS) analysis while 16 proteins were not reported previously. Our findings show that certain proteins in the pitcher fluid were continuously secreted or replenished after pitcher opening, even without any prey or chitin induction. We also discovered a new aspartic proteinase, Nep6, secreted into pitcher fluid. This is the first SWATH-MS analysis of protein expression in Nepenthes pitcher fluid using a species-specific reference transcriptome. Taken together, our study using a gel-free shotgun proteomics informed by transcriptomics (PIT) approach showed the dynamics of endogenous protein secretion in the digestive organ of N. × ventrata and provides insights on protein regulation during early pitcher opening prior to prey capture.

Original languageEnglish
Pages (from-to)681-694
Number of pages14
JournalJournal of Plant Research
Volume132
Issue number5
DOIs
Publication statusPublished - 2 Sep 2019

Fingerprint

Nepenthes
transcriptomics
proteomics
proteins
transcriptome
protein synthesis
carnivorous plants
aspartic proteinases
protein secretion
digestive enzymes
nutrients
chitin
carnivores
liquid chromatography
fluids
gels
RNA
insects

Keywords

  • Aspartic proteinase
  • Carnivorous plant
  • Nepenthesin
  • Proteomics informed by transcriptomics
  • RNA-seq
  • Shotgun proteomics
  • SWATH

ASJC Scopus subject areas

  • Plant Science

Cite this

Protein replenishment in pitcher fluids of Nepenthes × ventrata revealed by quantitative proteomics (SWATH-MS) informed by transcriptomics. / Wan Zakaria, Wan Nor Adibah; Wan Kamaruddin, Wan Mohd Aizat; Goh, Hoe Han; Mohd. Noor, Normah.

In: Journal of Plant Research, Vol. 132, No. 5, 02.09.2019, p. 681-694.

Research output: Contribution to journalArticle

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