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WHITE STRIPE LEAF8, encoding a deoxyribonucleoside kinase, is involved in chloroplast development in rice

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Abstract

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WSL8 encoding a deoxyribonucleoside kinase (dNK) that catalyzes the first step in the salvage pathway of nucleotide synthesis plays an important role in early chloroplast development in rice.

Abstract

The chloroplast is an organelle that converts light energy into chemical energy; therefore, the normal differentiation and development of chloroplast are pivotal for plant survival. Deoxyribonucleoside kinases (dNKs) play an important role in the salvage pathway of nucleotides. However, the relationship between dNKs and chloroplast development remains elusive. Here, we identified a white stripe leaf 8 (wsl8) mutant that exhibited a white stripe leaf phenotype at seedling stage (before the four-leaf stage). The mutant showed a significantly lower chlorophyll content and defective chloroplast morphology, whereas higher reactive oxygen species than the wild type. As the leaf developed, the chlorotic mutant plants gradually turned green, accompanied by the restoration in chlorophyll accumulation and chloroplast ultrastructure. Map-based cloning revealed that WSL8 encodes a dNK on chromosome 5. Compared with the wild type, a C-to-G single base substitution occurred in the wsl8 mutant, which caused a missense mutation (Leu 349 Val) and significantly reduced dNK enzyme activity. A subcellular localization experiment showed the WSL8 protein was targeted in the chloroplast and its transcripts were expressed in various tissues, with more abundance in young leaves and nodes. Ribosome and RNA-sequencing analysis indicated that some components and genes related to ribosome biosynthesis were down-regulated in the mutant. An exogenous feeding experiment suggested that the WSL8 performed the enzymic activity of thymidine kinase, especially functioning in the salvage synthesis of thymidine monophosphate. Our results highlight that the salvage pathway mediated by the dNK is essential for early chloroplast development in rice.

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Abbreviations

dNK:

Deoxyribonucleoside kinase

dNMP:

Deoxyribonucleoside monophosphate

dNDP:

Deoxyribonucleoside diphosphate

dNTP:

Deoxyribonucleoside triphosphate

TK:

Thymidine kinase

dCK:

Deoxycytidine kinase

dGK:

Deoxyguanosine kinase

TEM:

Transmission electron microscope

ROS:

Reactive oxygen species

NBT:

Nitroblue tetrazolium

DAB:

Diaminobenzidine

ORF:

Open reading frame

dAMP:

Deoxyadenosine monophosphate

dGMP:

Deoxyguanosine monophosphate

dUMP:

Deoxyuridine monophosphate

dCMP:

Deoxycytidine monophosphate

dTMP:

Thymidine monophosphate

dTTP:

Thymidine triphosphate

dT:

Thymidine

dTDP:

Thymidine diphosphate

TMPK:

Thymidylate kinase

NDK:

Diphosphate kinase

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Acknowledgments

This research was supported by Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River, Ministry of Agriculture, P. R. China, Jiangsu Collaborative Innovation Center for Modern Crop Production, and the grants from The National Key Research and Development Program of China (2017YFD0100401, 2016YFD0100700), a project (2016ZX08001004) from the Ministry of Agriculture of China for transgenic research, Jiangsu Science and Technology Development Program (BE2017368), and the National Natural Science Foundation of China Grant (31571629).

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Liu, L.L., You, J., Zhu, Z. et al. WHITE STRIPE LEAF8, encoding a deoxyribonucleoside kinase, is involved in chloroplast development in rice. Plant Cell Rep 39, 19–33 (2020). https://doi.org/10.1007/s00299-019-02470-6

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