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

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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.

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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

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    Authors and Affiliations

    1. College of Agronomy and Biotechnology, Southwest University, Chongqing, 400716, China

      Jing Zeng, Hecui Zhang, Song Chen & Liquan Zhu

    2. College of Horticulture and landscape Architecture, Southwest University, Chongqing, 400716, China

      Qiguo Gao, Songmei Shi, Xiaoping Lian, Xiaohong Yang & Xuesong Ren

    3. Department of Biological Sciences, University of Cincinnati, Cincinnati, OH, 45267-0524, USA

      Richard Converse

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    1. Jing Zeng
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    Zeng, J., Gao, Q., Shi, S. et al. Dissecting Pistil Responses to Incompatible and Compatible Pollen in Self-Incompatibility Brassica oleracea Using Comparative Proteomics. Protein J 36, 123–137 (2017). https://doi.org/10.1007/s10930-017-9697-y

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