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Lethal albinic seedling, encoding a threonyl-tRNA synthetase, is involved in development of plastid protein synthesis system in rice

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Abstract

Key message

An albinic rice is caused by mutation of threonyl-tRNA synthetase, which is essential for plant development by stabilizing of NEP and PEP gene expressions and chloroplast protein synthesis.

Abstract

Chloroplast biogenesis and development depend on complex genetic mechanisms. Apart from their function in translation, aminoacyl-tRNA synthetases (aaRSs) play additional role in gene expression regulation, RNA splicing, and cytokine activity. However, their detailed functions in plant development are still poorly understood. We isolated a lethal albinic seedling (las) mutant in rice. Physiological and ultrastructural analysis of las mutant plants revealed weak chlorophyll fluorescence, negligible chlorophyll accumulation, and defective thylakoid membrane development. By map based cloning we determined that the LAS allele gene encodes threonyl-tRNA synthetase (ThrRS). LAS was constitutively expressed with relatively high level in leaves. NEP-dependent gene transcripts accumulated in the developing chloroplasts, while PEP-dependent transcripts were reduced in the las mutant. This result indicated that PEP activity was impaired. Chloroplast-encoded protein levels were sharply reduced in the las mutant. Biogenesis of chloroplast rRNAs (16S and 23S rRNA) was arrested, leading to impaired translation and protein synthesis. Together, our findings indicated that LAS is essential not only for chloroplast development by stabilizing the NEP and PEP gene expression, but also for protein synthesis and construction of the ribosome system in rice chloroplasts.

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Abbreviations

aaRSs:

Aminoacyl-tRNA synthetases

Chl:

Chlorophyll

GFP:

Green fluorescent protein

ORF:

Open reading frame

RNAi:

RNA interference

SSR:

Simple sequence repeat

TEM:

Transmission electron microscopy

PEP:

Plastid-encoded RNA polymerase

NEP:

Nuclear-encoded RNA polymerase

RBCL:

Rubisco large subunits

AtpB:

ATP synthase CF1βsubunit

RBCS:

Rubisco small subunit

rpl2:

Ribosome large subunit 2

psaA1:

PS I P700 apoprotein A1

psaA2:

PS I P700 apoprotein A2

psbC:

PS II CP43-protein

cytf:

Cytochrome b6f complex

Hsp90:

Heat-shock protein 90

CBB:

Coomassie blue

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Acknowledgements

This research was supported by the Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in the Mid-lower Yangtze River, Ministry of Agriculture, P.R. China; Jiangsu Collaborative Innovation Center for Modern Crop Production; the Yangtze River Valley Hybrid Rice Collaboration Innovation Center; grants from the National 863 Program (2014AA10A 600), National Transformation Science and Technology Program (2014ZX08001004-002), National Science and Technology Support Program of China (2013BAD01B02-16), And Jiangsu Science and Technology Development Program (BE2014394, BE2015363).

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  1. State Key Laboratory for Crop Genetics and Germplasm Enhancement, Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, Nanjing, 210095, China

    Yuan-Yan Zhang, Yuan-Yuan Hao, Yi-Hua Wang, Chun-Ming Wang, Yun-Long Wang, Wu-Hua Long, Di Wang, Xi Liu, Ling Jiang & Jian-Min Wan

  2. National Key Facility for Crop Resources and Genetic Improvement, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Jian-Min Wan

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  1. Yuan-Yan Zhang
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Communicated by Kang Chong.

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299_2017_2136_MOESM1_ESM.tif

Fig. S1 Phenotypes of RNAi and complementation of transgenic plants of LAS. A, wt, wild type; las, mutant plant; RNAi, RNA interference transgenic T0 plant, las/1390:LAS, complementation transgenic T0 plant. B, Complementation transgenic T0 lines of las (TIF 9100 KB)

299_2017_2136_MOESM2_ESM.tif

Fig. S2 Transcript analyses of chlorophyll synthesis genes in wild type and las. Genes upregulated by more than two times compared to the mutant include HEML, HEMC, HEME, URO-D, HEMF, CHLI, CHLM, POR, CHLG, PS and FLU. Genes down-regulated in the mutant include CAO, HO, GUN4, PIF4 (TIF 1006 KB)

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Zhang, YY., Hao, YY., Wang, YH. et al. Lethal albinic seedling, encoding a threonyl-tRNA synthetase, is involved in development of plastid protein synthesis system in rice. Plant Cell Rep 36, 1053–1064 (2017). https://doi.org/10.1007/s00299-017-2136-x

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