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Plant Cell Reports

, Volume 37, Issue 12, pp 1667–1679 | Cite as

OsNDUFA9 encoding a mitochondrial complex I subunit is essential for embryo development and starch synthesis in rice

  • Tingting Hu
  • Yunlu Tian
  • Jianping Zhu
  • Yunlong Wang
  • Ruonan Jing
  • Jie Lei
  • Yinglun Sun
  • Yanfang Yu
  • Jingfang Li
  • Xiaoli Chen
  • Xiaopin Zhu
  • Yuanyuan Hao
  • Linglong Liu
  • Yihua Wang
  • Jianmin Wan
Original Article

Abstract

Key message

Loss of function of a mitochondrial complex I subunit (OsNDUFA9) causes abnormal embryo development and affects starch synthesis by altering the expression of starch synthesis-related genes and proteins.

Abstract

Proton-pumping NADH: ubiquinone oxidoreductase (also called complex I) is thought to be the largest and most complicated enzyme of the mitochondrial respiratory chain. Mutations of complex I subunits have been revealed to link with a number of growth inhibitions in plants. However, the function of complex I subunits in rice remains unclear. Here, we isolated a rice floury endosperm mutant (named flo13) that was embryonic lethal and failed to germinate. Semi-thin sectioning analysis showed that compound starch grain development in the mutant was greatly impaired, leading to significantly compromised starch biosynthesis and decreased 1000-grain weight relative to the wild type. Map-based cloning revealed that FLO13 encodes an accessory subunit of complex I protein (designated as OsNDUFA9). A single nucleotide substitution (G18A) occurred in the first exon of OsNDUFA9, introducing a premature stop codon in the flo13 mutant gene. OsNDUFA9 was ubiquitously expressed in various tissues and the OsNDUFA9 protein was localized to the mitochondria. Quantitative RT-PCR and protein blotting indicated loss of function of OsNDUFA9 altered gene expression and protein accumulation associated with respiratory electron chain complex in the mitochondria. Moreover, transmission electron microscopic analysis showed that the mutant lacked obvious mitochondrial cristae structure in the mitochondria of endosperm cell. Our results demonstrate that the OsNDUFA9 subunit of complex I is essential for embryo development and starch synthesis in rice endosperm.

Keywords

OsNDUFA9 Mitochondrial complex I subunit Embryo development Starch synthesis Rice (Oryza sativa L.) 

Abbreviations

AOX

Alternative oxidase

DAF

Days after flowering

dCAPS

Derived cleaved amplified polymorphic sequence

EMS

Ethyl methane sulfonate

ETC

Electron transfer chain

GFP

Green fluorescent protein

InDel

Insertion–deletion

nad

NADH dehydrogenase subunit

NDUFA9

Mitochondrial NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9

ORF

Open reading frame

OXPHOS

Oxidative phosphorylation

SEM

Scanning electron microscope

TTC

2,3,5-Triphenyltetrazolium chloride

TEM

Transmission electron microscope

RT-PCR

Real-time PCR

Notes

Acknowledgements

This research was supported by the Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River, Ministry of Agriculture of China, The Yangtze River Valley Hybrid Rice Collaboration Innovation Center, Jiangsu Collaborative Innovation Center for Modern Crop Production, and the grants from The National Key Research and Development Program of China (2017YFD0100400), a project (2016ZX08001006) from the Ministry of Agriculture of China for transgenic research, Jiangsu Science and Technology Development Program (BE2015355-1), and Jiangsu Province Self-Innovation Program [CX(16)1029]. This work was also supported by the Fundamental Research Funds for the Central Universities (KYTZ201601).

Compliance with ethical standards

Conflict of interest

These authors declare that they have no conflict of interest.

Supplementary material

299_2018_2338_MOESM1_ESM.docx (1.1 mb)
Supplementary material 1 (DOCX 1151 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research CenterNanjing Agricultural UniversityNanjingChina
  2. 2.National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijingChina
  3. 3.Xuzhou Institute of Agricultural Sciences in Jiangsu Xuhuai AreaXuzhouChina

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