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Identification and functional characterization of APRR2 controlling green immature fruit color in cucumber (Cucumis sativus L.)

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Abstract

Our previous studies have found that gene APRR2 is the candidate gene of w controlling the green color of immature fruit in cucumber, and its allele aprr2, which controls the white color, has a 101-amino acid residues deletion at the C-terminus compared with APRR2. Here, we characterized the APRR2 and analyzed its function in cucumber. qPCR analysis and prokaryotic expression of APRR2 and aprr2 clarified the relationship between APRR2 and aprr2 and confirmed that APRR2/aprr2 is a gene controlling cucumber color. Sequence and phylogenetic relationship analysis revealed that gene APRR2 and aprr2 with REC domain and DNA-binding B-motif are placed into subclass I of PRR subfamily. Subcellular localization assay indicated that APRR2 is a nuclear protein, but aprr2 is not. These results suggest APRR2 gene is a transcription factor in cucumber. Additionally, the expression analysis of Class-I KNOTTED1-like homeobox genesTKN4 and TKN2 involved in the chloroplast development showed they were in line with APRR2 expression, therefore, APRR2 together withTKN4, TKN2 likely contribute to the mechanisms related to chloroplast development.

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Abbreviations

TCS:

Two-component regulatory systems

RR:

Response regulator

PRR:

Pseudo-response regulators

APRR2 :

Arabidopsis pseudo-response regulator 2 gene;

KNOX :

Class-I KNOTTED1-like homeobox

IPTG:

Isopropyl-β-d-thiogalactoside

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

YEB:

Agrobacterium rhizogenes culture medium

LB:

Lysogeny broth

PCR:

Polymerase chain reaction

Rt-PCR:

Real-time quantitative PCR

cDNA:

DNA complementary to RNA

GLK2 :

Golden2-like

REC:

Receiver

RLD:

Receiver-like domain

PRRs:

Pseudo-response regulators

ARRS:

Response regulators

CTK:

Cytokinin

CLM9:

Calmodulin9-like

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Acknowledgements

This research was supported by grants from the National Natural Science Foundation of China (Project#31372074), National Key Research and Development Plan (2016YFD0101705)of China, Xi’an Agricultural Science and Technology Innovation Project of China (NC1501(1)) and Shaanxi Province Agricultural Science and Technology Innovation and Research Project of China (2016NY-048). We acknowledge the assistance provided by Dr. Fang Cheng (Northwest A&F University of Shaanxi).

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    Authors and Affiliations

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

      Jianqing Jiao, Hanqiang Liu, Jia Liu, Mingming Cui, Jing Xu, Huanwen Meng, Yuhong Li, Shuxia Chen & Zhihui Cheng

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    1. Jianqing Jiao
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    2. Hanqiang Liu
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    Contributions

    JL and HL conceived and designed the study. JL and HL constructed the prokaryotic expression, qPCR analysis, sequence and phylogenetic relationship analysis, and wrote the paper. JJ performed the subcellular localization. MC and JX. performed some of the field work. HM, SC and YL provided valuable research ideas. ZC and HL led and supervised the study.

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    Correspondence to Huanwen Meng or Zhihui Cheng.

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    Jianqing Jiao, Hanqiang Liu and Jia Liu have contributed equally to this work.

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    Jiao, J., Liu, H., Liu, J. et al. Identification and functional characterization of APRR2 controlling green immature fruit color in cucumber (Cucumis sativus L.). Plant Growth Regul 83, 233–243 (2017). https://doi.org/10.1007/s10725-017-0304-1

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