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Young Leaf Chlorosis 2 encodes the stroma-localized heme oxygenase 2 which is required for normal tetrapyrrole biosynthesis in rice

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Abstract

Main conclusion

Rice heme oxygenase 2 (OsHO2) mutants are chlorophyll deficient with distinct tetrapyrrole metabolite and transcript profiles, suggesting a potential regulatory role of the stromal-localized OsHO2 in tetrapyrrole biosynthesis.

In plants, heme oxygenases (HOs) are classified into the subfamilies HO1 and HO2. HO1 are highly conserved plastid enzymes required for synthesizing the chromophore in phytochromes which mediate a number of light-regulated responses. However, the physiological and biochemical functions of HO2, which are distantly related to HO1, are not well understood, especially in crop plants. From a population of 60Coγ-irradiated rice mutants, we identified the ylc2 (young leaf chlorosis 2) mutant which displays a chlorosis phenotype in seedlings with substantially reduced chlorophyll content. Normal leaf pigmentation is gradually restored in older plants while newly emerged leaves remain yellow. Transmission electron microscopy further revealed defective chloroplast structures in the ylc2 seedlings. Map-based cloning located the OsYLC2 gene on chromosome 3 and it encodes the OsHO2 protein. The gene identification was confirmed by complementation and T-DNA mutant analyses. Subcellular localization and chloroplast fractionation experiments indicated that OsHO2 resides in the stroma. However, recombinant enzyme assay demonstrated that OsHO2 is not a functional HO enzyme. Analysis of tetrapyrrole metabolites revealed the reduced levels of most chlorophyll and phytochromobilin precursors in the ylc2 mutant. On the other hand, elevated accumulation of 5-aminolevulinic acid and Mg-protoporphyrin IX was observed. These unique metabolite changes are accompanied by consistent changes in the expression levels of the corresponding tetrapyrrole biosynthesis genes. Taken together, our work suggests that OsHO2 has a potential regulatory role for tetrapyrrole biosynthesis in rice.

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Abbreviations

ALA:

5-Aminoleuvlinic acid

BAC:

Bacterial artificial chromosome

BV:

Biliverdin

Car:

Carotenoid

Chl:

Chlorophyll

Chlide:

Chlorophyllide

EYFP:

Enhanced yellow fluorescent protein

His:

Histidine

HM:

Homozygous

HO:

Heme oxygenase

MgCH:

Mg chelatase

PΦB:

Phytochromobilin

Pchlide:

Protochlorophyllide

PG:

Plastoglobules

Proto IX:

Protoporphyrin IX

RT:

Reverse transcription

STS:

Sequence-tagged site

TEM:

Transmission electron microscopy

Urogen III:

Uroporphyrinogen III

WT:

Wild type

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Acknowledgments

This work is supported by the National Key Laboratory of Plant Molecular Genetics, the National Natural Science Foundation of China (grant no. 31171530), Zhejiang Provincial Natural Science Foundation of China (grant no. Y3100591), and the HKU Seed Funding for Basic Research (grant no. 201111159147).

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Author notes

  1. Yi Sun

    Present address: Department of Biology, Concordia University, Montreal, QC, H4B 1R6, Canada

Authors and Affiliations

  1. State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Crops and Nuclear Technology Utilization, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China

    Qingzhu Li, Xiaoli Gao, Sujuan Li, Yuezhi Tao & Hongjia Liu

  2. School of Biological Sciences, The University of Hong Kong, Pokfulam, Hong Kong, China

    Qingzhu Li, Fu-Yuan Zhu, Yi Sun & Clive Lo

  3. College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, 311121, China

    Xiaoli Gao

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  1. Qingzhu Li
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Correspondence to Clive Lo or Hongjia Liu.

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Q. Li and F.-Y. Zhu have contributed equally to this work.

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Li, Q., Zhu, FY., Gao, X. et al. Young Leaf Chlorosis 2 encodes the stroma-localized heme oxygenase 2 which is required for normal tetrapyrrole biosynthesis in rice. Planta 240, 701–712 (2014). https://doi.org/10.1007/s00425-014-2116-0

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