Abstract

Both zinc (Zn) and iron (Fe) are essential micro-nutrients for plant growth and development, yet their levels in plants are tightly regulated to prevent either deficiency or phytotoxicity. In agronomic reality, such an imbalance of metal bioavailability to crops occurs frequently. Thus, mining genetic resources to improve crop traits relevant to metal homeostasis is a great challenge to ensure crop yield and food quality. This study functionally identified an uncharacterized metallochaperone family HIPP protein gene Heavy Metal Associated Isoprenylated Plant Proteins 33 (OsHIPP33) in rice (Oryza sativa). OsHIPP33 resides in the nucleus and plasma membrane and constitutively expresses throughout the lifespan. Transcription of OsHIPP33 is not induced by deprivation of Zn and Fe but upregulated under excessive Zn and Fe stress. In a short-term (one month) hydroponic study with the normal Zn and Fe supply, there were no significant changes in the growth and metal accumulation between the knockout (OsHIPP33) or knockdown (RNA interference) mutant lines and wild-type, while the long-term field trials (for two successive years) demonstrated that the mutation of OsHIPP33 significantly compromised the rice growth and development (such as rice leave tissues, panicle length, spikelet fertility, seed weight per plant, 1000-grain weight, etc.), with the mature grain yield of OsHIPP33 and RNAi lines reduced by 52% and 12–15% respectively, compared with wild-type. Furthermore, the accumulation of Zn and Fe in rice straw, husk and brown rice was also reduced. These results suggest that the disruption of OsHIPP33 can dampen rice agronomic traits, signifying that OsHIPP33 expression is required for Zn and Fe homeostasis and subsequent production of rice grains.

Highlights

  • Zinc is an essential micro-nutritional element for diverse biological functions in plants.One central role is as key cofactor for many metalloenzymes and other proteins actively participating in biological processes including conformation of structured proteins, activation of transcription factors or mediation of protein–protein interactions [1]

  • We identified a new Heavy metal-associated isoprenylated plant proteins (HIPPs) family gene, OsHIPP33, which was isolated from our previous metal-stress responsive transcriptome [16]

  • Phylogenetic analysis of rice OsHIPP33 homologous genes revealed that its close homolog is OsHIPP32 (Figure S1D)

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Summary

Introduction

Zinc is an essential micro-nutritional element for diverse biological functions in plants.One central role is as key cofactor for many metalloenzymes and other proteins actively participating in biological processes including conformation of structured proteins, activation of transcription factors or mediation of protein–protein interactions [1]. Zinc is an essential micro-nutritional element for diverse biological functions in plants. Zinc starvation symptoms appear with stunted plant growth, manifested by shorter internodes, smaller size of leaves, necrosis of root tips and young leaf yellowing [4]. Iron is another Zn-equivalent essential metal and serves as a cofactor of diverse enzymes, playing an indispensable role in photosynthesis, respiration and many other metabolic processes [5]. Most soils on Earth have sufficient iron elements, their real availability to plants is limited. Iron deficiency causes the arrest of plant growth, leaf chlorosis, reduced reproductively, and low quality of seed [6]

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