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The Protein Journal

, Volume 36, Issue 2, pp 123–137 | Cite as

Dissecting Pistil Responses to Incompatible and Compatible Pollen in Self-Incompatibility Brassica oleracea Using Comparative Proteomics

  • Jing Zeng
  • Qiguo Gao
  • Songmei Shi
  • Xiaoping Lian
  • Richard Converse
  • Hecui Zhang
  • Xiaohong Yang
  • Xuesong Ren
  • Song Chen
  • Liquan Zhu
Article

Abstract

Angiosperms have developed self-incompatibility (SI) systems to reject self-pollen, thereby promoting outcrossing. The Brassicaceae belongs to typical sporophytic system, having a single S-locus controlled SI response, and was chosen as a model system to study SI-related intercellular signal transduction. In this regard, the downstream factor of EXO70A1 was unknown. Here, protein two-dimensional electrophoresis (2-DE) method and coupled with matrix-assisted laser desorption ionization/time of flight of flight mass spectrometry (MALDI-TOF -MS) and peptide mass fingerprinting (PMF) was used to further explore the mechanism of SI responses in Brassica oleracea L. var. capitata L. at protein level. To further confirm the time point of protein profile change, total proteins were collected from B. oleracea pistils at 0 min, 1 h, and 2 h after self-pollination. In total 902, 1088 and 1023 protein spots were separated in 0 min, 1 h and 2 h 2-DE maps, respectively. Our analyses of self-pollination profiles indicated that proteins mainly changed at 1 h post-pollination in B. oleracea. Moreover, 1077 protein spots were separated in cross-pollinated 1 h (CP) pistil 2-DE map. MALDI-TOF-MS and PMF successfully identified 34 differentially-expressed proteins (DEPs) in SP and CP 1 h 2-DE maps. Gene ontology and KEGG analysis revealed an array of proteins grouped in the following categories: stress and defense response (35%), protein metabolism (18%), carbohydrate and energy metabolism (12%), regulation of translation (9%), pollen tube development (12%), transport (9%) and cytoskeletal (6%). Sets of DEPs identified specifically in SP or only up-regulated expressed in CP pistils were chosen for funther investigating in floral organs and during the process of self- and cross-pollination. The function of these DEPs in terms of their potential involvement in SI in B. oleracea is discussed.

Keywords

Brasssic oleracea Incompatible and compatible pollination Pistil proteins Functional classification 

Abbreviations

SI

Self-incompatibility

2-DE

Two-dimension electrophoresis

MALDI-TOF-MS

Matrix assisted laser desorption/lionization time of flight mass spectrometry

PMF

Peptide mass fingerprinting

SP

Self-pollinated

CP

Cross-pollinated

DEPs

Differentially-expressed proteins

KEGG

Kyoto encyclopedia of genes and genomes

SCR

S-locus cysteine rich protein

SRK

S-locus receptor kinase protein

SLG

S-locus glycoprotein protein

ARC1

Armadillo repeat containing 1

EXO70A1

Exocyst 70 A1

MLPK

M-locus protein kinase protein

THL1/THL2

Thioredoxin-h-like proteins

TCA

Trichloroacetic acid

CHAPS

3-cholamidopropyl dimethylammonio propanesulfonate

DTT

Dithiothreitol

IEF

Isoelectric focusing

SDS-PAGEs

Sodium dodecyl sulfate polyacrylamide gels

SD

Standard deviation

Notes

Compliance with Ethical Standards

Conflict of interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled. And this article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10930_2017_9697_MOESM1_ESM.docx (848 kb)
Supplementary material 1 (DOCX 848 KB)

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Jing Zeng
    • 1
  • Qiguo Gao
    • 2
  • Songmei Shi
    • 2
  • Xiaoping Lian
    • 2
  • Richard Converse
    • 3
  • Hecui Zhang
    • 1
  • Xiaohong Yang
    • 2
  • Xuesong Ren
    • 2
  • Song Chen
    • 1
  • Liquan Zhu
    • 1
  1. 1.College of Agronomy and BiotechnologySouthwest UniversityChongqingChina
  2. 2.College of Horticulture and landscape ArchitectureSouthwest UniversityChongqingChina
  3. 3.Department of Biological SciencesUniversity of CincinnatiCincinnatiUSA

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