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Screening proteins interacting with VpPR10.1 of Chinese wild grapevine using the yeast two-hybrid system

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Abstract

Among the 17 plant pathogenesis-related (PR) protein families, only PR10 family is intracellular and cytosolic. PR proteins are expressed in response to pathogen challenge and abiotic stresses in higher plants. However, the molecular mechanisms of their actions remain poorly understood. In a previous work, we isolated a PR10 gene from Erysiphe necator-resistant Chinese wild Vitis sp. (Baihe-35-1) and it was designated as VpPR10.1. In this study, yeast two-hybrid method was used to screen proteins interacting with VpPR10.1 proteins. Twenty-one ESTs were isolated and sequenced. All sequences were compared using BLASTx to identify presumptive orthologs. Several proteins associated with VpPR10.1 protein were screened, including CNR8, UFGT6, HSP, DEAD-box, Trx h2, Grx C9 and GLOX. These proteins are closely related to defensive action of plants against pathogens and abiotic stresses. Our results reveal that VpPR10.1 gene may be involved in hormone signaling, programmed cell death and defense responses of grapevine.

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Abbreviations

PR:

Pathogenesis-related

P-loop:

Phosphate-binding loop

VpPR10.1:

Vitis pseudoreticulata pathogenesis-related protein gene 10.1

Trp:

Tryptophan

His:

Histidine

Ade:

Adenine

Leu:

Leucine

SD:

Medium for yeast comprised of a nitrogen base, a carbon source and a DO supplement

DO:

Dropout (supplement or solution)

YPD:

A blend of yeast extract, peptone, and dextrose

YPDA:

YPD medium supplemented with adenine hemisulfate

X-α-Gal:

5-Bromo-4-chloro-3-indolyl-α-d-galactopyranoside

CNR8:

Cell number regulator 8-like

UFGT6:

UDP-glucose flavonoid 3-O-glucosyltransferase 6

HSP:

Heat shock protein

PDC2:

Pyruvate decarboxylase 2

Trx h2:

Thioredoxin H2

Grx C9:

Glutaredoxin-C9

GLOX:

Glyoxal oxidase

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 31272125), and also supported by program for New Century Excellent Talents in University (NCET-10-0692) and the program for Young Talents in Northwest A&F University (QN2011052) to Y. Xu.

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Teng-Fei Xu, Jiang Xiang, Feng-Ju Li, Tie-Mei Li, Yi-He Yu, Yue-Jin Wang & Yan Xu

  2. Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Teng-Fei Xu, Jiang Xiang, Feng-Ju Li, Tie-Mei Li, Yi-He Yu, Yue-Jin Wang & Yan Xu

  3. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Teng-Fei Xu, Jiang Xiang, Feng-Ju Li, Tie-Mei Li, Yi-He Yu, Yue-Jin Wang & Yan Xu

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  1. Teng-Fei Xu
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  2. Jiang Xiang
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Correspondence to Yan Xu.

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Communicated by M. Hajduch.

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Xu, TF., Xiang, J., Li, FJ. et al. Screening proteins interacting with VpPR10.1 of Chinese wild grapevine using the yeast two-hybrid system. Acta Physiol Plant 35, 2355–2364 (2013). https://doi.org/10.1007/s11738-013-1269-y

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  • DOI: https://doi.org/10.1007/s11738-013-1269-y

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