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Journal of Integrative Agriculture  2020, Vol. 19 Issue (3): 612-623    DOI: 10.1016/S2095-3119(19)62710-3
Special Issue: 水稻遗传育种合辑Rice Genetics · Breeding · Germplasm Resources
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OsHemA gene, encoding glutamyl-tRNA reductase (GluTR) is essential for chlorophyll biosynthesis in rice (Oryza sativa)
ZENG Zhao-qiong1, 2*, LIN Tian-zi1*, ZHAO Jie-yu1, ZHENG Tian-hui1, XU Le-feng1, WANG Yi-hua1, LIU Ling-long1, JIANG Ling1, CHEN Sai-hua3, WAN Jian-min 
1 State Key Laboratory for Crop Genetics & Germplasm Enhancement/Jiangsu Provincial Center of Plant Gene Engineering/College of Agriculture, Nanjing Agricultural University, Nanjing 210095, P.R.China
2 Nanchong Academy of Agricultural Sciences, Nanchong 637000, P.R.China
3 Key Laboratory of Plant Functional Genomics, Ministry of Education/Co?Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, P.R.China
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Chlorophyll (Chl) biosynthesis is essential for photosynthesis and plant growth.  Glutamyl-tRNA reductase (GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde (GSA) and initiates the chlorophyll biosynthesis.  Even though the main role of GluTR has been established, the effects caused by natural variations in its corresponding gene remain largely unknown.  Here, we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency, designated as cbd1.  With intact thylakoid lamellar structure, the cbd1 plant showed light green leaves and reduced Chl and carotenoids (Cars) content significantly compared to the wild type.  By map-based gene cloning, the mutation was restricted within a 57-kb region on chromosome 10, in which an mPingA miniature inverted-repeat transposable element (MITE) inserted in the promoter region of OsHemA gene.  Both leaf color and the pigment contents in cbd1 were recovered in a complementation test, confirming OsHemA was responsible for the mutant phenotype.  OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1.  Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus.  A majority of Chl biosynthesis genes, except for POR and CHLG, were down-regulated synchronously by the repression of OsHemA, suggesting that an attenuation occurred in the Chl biosynthesis pathway.  Interestingly, we found major agronomic traits involved in rice yield were statistically unaffected, except for the number of full grains per panicle was increased in cbd1.  Collectively, OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.
Keywords:  OsHemA        GluTR        chlorophyll biosynthesis  
Received: 29 November 2018   Accepted:
Fund: This research was supported by the National Key Research and Development Program of China (2016YFD0101801), the National Excellent Doctoral Dissertation of China (201262), the Key Laboratory of Biology, Genetics and Breeding of Japonica Rice in Mid-lower Yangtze River, Ministry of Agriculture and Rural Affairs, China, the Collaborative Innovation Center for Hybrid Rice in Yangtze River, China, and the Jiangsu Collaborative Innovation Center for Modern Crop Production, China, the National High-Tech R&D Program of China (2014AA10A603-15), the National Key Technologies R&D Program of China during the 12th Five-Year Plan period (2013BAD01B02-16), and the Jiangsu Science and Technology Development Program, China (BE2014394 and BE2015363).
Corresponding Authors:  Correspondence CHEN Sai-hua, Tel/Fax: +86-514-87972178, E-mail:; WAN Jian-min, E-mail:   
About author:  * These authors contributed equally to this study.

Cite this article: 

ZENG Zhao-qiong, LIN Tian-zi, ZHAO Jie-yu, ZHENG Tian-hui, XU Le-feng, WANG Yi-hua, LIU Ling-long, JIANG Ling, CHEN Sai-hua, WAN Jian-min . 2020. OsHemA gene, encoding glutamyl-tRNA reductase (GluTR) is essential for chlorophyll biosynthesis in rice (Oryza sativa). Journal of Integrative Agriculture, 19(3): 612-623.

Arnon D I. 1949. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiology, 24, 1–15.
Bougri O, Grimm B. 1996. Members of a low-copy number gene family encoding glutamyl-tRNA reductase are differentially expressed in barley. The Plant Journal, 9, 867–878.
Camp R O D, Przybyla D, Ochsenbein C, Laloi C, Kim C, Danon A, Wagner D, Hideg E, Gobel C, Feussner I. 2003. Rapid induction of distinct stress responses after the release of singlet oxygen in Arabidopsis. The Plant Cell, 15, 2320–2332.
Carugo O, Argos P. 1997. NADP-dependent enzymes. I: Conserved stereochemistry of cofactor binding. Proteins, 28, 10–28.
Czarnecki O, Hedtke B, Melzer M, Rothbart M, Richter A, Schroter Y, Pfannschmidt T, Grimm B. 2012. An Arabidopsis GluTR binding protein mediates spatial separation of 5-aminolevulinic acid synthesis in chloroplasts. The Plant Cell, 23, 4476–4491.
Goslings D, Meskauskiene R, Kim C, Lee K P, Nater M, Apel K. 2004. Concurrent interactions of heme and FLU with Glu tRNA reductase (HEMA1), the target of metabolic feedback inhibition of tetrapyrrole biosynthesis, in dark- and light-grown Arabidopsis plants. The Plant Journal, 40, 957–967.
Hiei Y, Ohta S, Komari T, Kumashiro T. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. The Plant Journal, 6, 271–282.
Howitt C A, Pogson B J. 2006. Carotenoid accumulation and function in seeds and non-green tissues. Plant Cell and Environment, 29, 435–445.
Ilag L L, Kumar A M, Soll D. 1994. Light regulation of chlorophyll biosynthesis at the level of 5-aminolevulinate formation in Arabidopsis. The Plant Cell, 6, 265–275.
Kumar A M, Csankovszki G, Soll D. 1996. A second and differentially expressed glutamyl-tRNA reductase gene from Arabidopsis thaliana. Plant Molecular Biology, 30, 419–426.
Lichtenthaler H K. 1987. Chlorophylls and carotenoids - pigments of phytosynthetic biomembranes. Methods in Enzymology, 148, 350–382.
Liu W, Fu Y, Hu G, Si H, Zhu L, Wu C, Sun Z. 2007. Identification and fine mapping of a thermo-sensitive chlorophyll deficient mutant in rice (Oryza sativa L.). Planta, 226, 785–795.
Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods, 25, 402–408.
Macqueen D J, Johnston I A. 2009. Evolution of the multifaceted eukaryotic akirin gene family. BMC Evolutionary Biology, 9, 34.
Matsumoto F. 2004. Gene expression profiling of the tetrapyrrole metabolic pathway in Arabidopsis with a mini-array system. Plant Physiology, 135, 2379–2391.
Meier S, Tzfadia O, Vallabhaneni R, Gehring C, Wurtzel E T. 2011. A transcriptional analysis of carotenoid, chlorophyll and plastidial isoprenoid biosynthesis genes during development and osmotic stress responses in Arabidopsis thaliana. BMC Systems Biology, 5, 77.
Meskauskiene R, Nater M, Goslings D, Kessler F, Camp R O D, Apel K. 2001. FLU: A negative regulator of chlorophyll biosynthesis in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the United States of America, 98, 12826–12831.
Mochizuki N, Tanaka R, Grimm B, Masuda T, Moulin M, Smith A G, Tanaka A, Terry M J. 2010. The cell biology of tetrapyrroles: A life and death struggle. Trends in Plant Science, 15, 488–498.
Moser J, Schubert W D, Beier V, Bringemeier I, Jahn D, Heinz D W. 2001. V-shaped structure of glutamyl-tRNA reductase, the first enzyme of tRNA-dependent tetrapyrrole biosynthesis. The EMBO Journal, 20, 6583–6590.
Rebeiz C A, Montazerzouhoor A, Mayasich J M, Tripathy B C, Wu S M, Rebeiz C C. 1988. Phytodynamic herbicides-recent developments and molecular-basis of selectivity. Critical Reviews in Plant Sciences, 6, 385–436.
Sangwan I, O’Brian M R. 1999. Expression of a soybean gene encoding the tetrapyrrole-synthesis enzyme glutamyl-tRNA reductase in symbiotic root nodules. Plant Physiology, 119, 593–598.
Sasarman A, Surdeanu M, Szegli G, Horodniceanu T, Greceanu V, Dumitrescu A. 1968. Hemin-deficient mutants of Escherichia coli K-12. Journal of Bacteriology, 96, 570–572.
Su N, Hu M, Wu D, Wu F, Fei G, Lan Y, Chen X, Shu X, Zhang X, Guo X. 2012. Disruption of a rice pentatricopeptide repeat protein causes a seedling-specific albino phenotype and its utilization to enhance seed purity in hybrid rice production. Plant Physiology, 159, 227–238.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731–2739.
Tanaka R, Tanaka A. 2007. Tetrapyrrole biosynthesis in higher plants. Annual Review of Plant Biology, 58, 321–346.
Tanaka R, Yoshida K, Nakayashiki T, Masuda T, Tsuji H, Inokuchi H, Tanaka A. 1996. Differential expression of two hemA mRNAs encoding glutamyl-tRNA reductase proteins in greening cucumber seedlings. Plant Physiology, 110, 1223–1230.
Tanaka R, Yoshida K, Nakayashiki T, Tsuji H, Inokuchi H, Okada K, Tanaka A. 1997. The third member of the hemA gene family encoding glutamyl-tRNA reductase is primarily expressed in roots in Hordeum vulgare. Photosynthesis Research, 53, 161–171.
Terry M J, Smith A G. 2013. A model for tetrapyrrole synthesis as the primary mechanism for plastid-to-nucleus signaling during chloroplast biogenesis. Frontiers in Plant Science, 4, 14.
Ujwal M L, McCormac A C, Goulding A, Kumar A M, Soll D, Terry M J. 2002. Divergent regulation of the HEMA gene family encoding glutamyl-tRNA reductase in Arabidopsis thaliana: Expression of HEMA2 is regulated by sugars, but is independent of light and plastid signalling. Plant Molecular Biology, 50, 83–91.
Verkamp E, Jahn M, Jahn D, Kumar A M, Söll D. 1992. Glutamyl-tRNA reductase from Escherichia coli and Synechocystis 6803. Gene structure and expression. Journal of Biological Chemistry, 267, 8275–8280.
Vothknecht U C, Kannangara C G. 1998. Barley glutamyl tRNAglu reductase: Mutations affecting haem inhibition and enzyme activity*. Phytochemistry, 4, 513–519.
Vothknecht U C, Kannangara C G, von Wettstein D. 1996. Expression of catalytically active barley glutamyl tRNAGlu reductase in Escherichia coli as a fusion protein with glutathione S-transferase. Proceedings of the National Academy of Sciences of the United States of America, 93, 9287–9291.
Zhang H, Liu L, Cai M, Zhu S, Zhao J, Zheng T, Xu X, Zeng Z, Niu J, Jiang L. 2015. A point mutation of magnesium chelatase OsCHLI gene dampens the interaction between CHLI and CHLD subunits in rice. Plant Molecular Biology Reporter, 33, 1975–1987.
Zhang M, Zhang F, Fang Y, Chen X, Chen Y, Zhang W, Dai H, Lin R, Liu L. 2015. The non-canonical tetratricopeptide repeat (TPR) domain of fluorescent (FLU) mediates complex formation with glutamyl-tRNA reductase. Journal of Biological Chemistry, 290, 17559–17565.
Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D. 2011. A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods, 7, 30.
Zhao A, Fang Y, Chen X, Zhao S, Dong W, Lin Y, Gong W, Liu L. 2014. Crystal structure of Arabidopsis glutamyl-tRNA reductase in complex with its stimulator protein. Proceedings of the National Academy of Sciences of the United States of America, 111, 6630–6635.
Zhou K, Ren Y, Lv J, Wang Y, Liu F, Zhou F, Zhao S, Chen S, Peng C, Zhang X. 2013. Young Leaf Chlorosis 1, a chloroplast-localized gene required for chlorophyll and lutein accumulation during early leaf development in rice. Planta, 237, 279–292.
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