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Cloning and elucidation of the functional role of apple MdLBD13 in anthocyanin biosynthesis and nitrate assimilation

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Abstract

The plant specific lateral organ boundaries domain (LBD) family of transcription factors (TFs) is involved in many aspects of plant growth and development. In this study, MdLBD13, a nitrate-induced LBD family gene from the apple (Malus × domestica), was isolated and characterized. Overexpression of MdLBD13 repressed anthocyanin biosynthesis by reducing expression of the structural genes associated with the flavonoid pathway. Overexpression ofMdLBD13 also repressed expression of the N–responsive genes that are required for nitrate uptake, transport and assimilation, resulting in reduced nitrate content and nitrate reductase activity in apple calli, as well as in Arabidopsis. Ectopic expression of MdLBD13 also promoted lateral root development in transgenic Arabidopsis. These results demonstrate that MdLBD13acts as a negative regulator in anthocyanin biosynthesis and nitrate utilization.

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Abbreviations

LBD:

Lateral organ boundaries domain

MBW:

R2R3 MYB TFS, basic helix–loop–helix TFS and WD40 proteins

RT-qPCR:

Real-time quantitative polymerase chain reaction

MYB1:

MYB domain protein 1

PAP1/2:

Production of anthocyanins pigment 1/2

CHS:

Chalcone synthase (CHS; EC 2.3.1.74)

CHI:

Chalcone flavanone isomerase (CHI; EC 5.5.1.6)

F3H:

Flavanone 3-hydroxylase (F3H, EC 1.14.11.9)

DFR:

Dihydroflavonol 4-reductase (DFR, EC 1.1.1.219)

ANR:

Anthocyanidin reductase (ANR, EC 1.3.1.77)

UFGT:

UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT; EC 2.4.1.9 1)

NRT:

Nitrate response transporter

NIA1/2 :

Nitrate reductase1/2

NR:

Nitrate reductase (NR, EC 1.7.1.1)

NRA:

Nitrate reductase activity

WT:

Wild type

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Acknowledgements

This work was supported by grants from NSFC (31325024, 31601742 and 31272142), Ministry of Education of China (IRT15R42) and Shandong Province (SDAIT-06-03).

Author contributions

Chun-Xiang You and Xiao-Fei Wang designed the experiments. Hao-Hao Li and Xin-Liu carried out the experiments and analyzed the data. Jian-Ping An carried out analysis of the content of nitrate and NRA. Hao-Hao Li, Xiao-Fei Wang and Yu-Jin Hao wrote the manuscript. Hao-Hao Li and Xin-Liu contributed to revised paper.

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    1. National Key Laboratory of Crop Biology, MOA Key Laboratory of Horticultural Crop Biology and Germplasm Innovation, College of Horticulture Science and Engineering, Shandong Agricultural University, Tai-An, 271018, Shandong, China

      Hao-Hao Li, Xin Liu, Jian-Ping An, Yu-Jin Hao, Xiao-Fei Wang & Chun-Xiang You

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    1. Hao-Hao Li
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    Correspondence to Xiao-Fei Wang or Chun-Xiang You.

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    Hao-Hao Li and Xin Liu have contributed equally to the article.

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    Li, HH., Liu, X., An, JP. et al. Cloning and elucidation of the functional role of apple MdLBD13 in anthocyanin biosynthesis and nitrate assimilation. Plant Cell Tiss Organ Cult 130, 47–59 (2017). https://doi.org/10.1007/s11240-017-1203-x

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