Disruption of an amino acid transporter LHT1 leads to growth inhibition and low yields in rice

Xiaohu Wang,Dongli Hao,Zhigang Wang,Mingxing Shi,Qiuxing Wei,Shan Fu,Yanhua Su,Jixing Xia,Guangzhe Yang

doi:10.1186/s12870-019-1885-9

Abstract

BackgroundResearch on plant amino acid transporters was mainly performed in Arabidopsis, while our understanding of them is generally scant in rice. OsLHT1 (Lysine/Histidine transporter) has been previously reported as a histidine transporter in yeast, but its substrate profile and function in planta are unclear. The aims of this study are to analyze the substrate selectivity of OsLHT1 and influence of its disruption on rice growth and fecundity.ResultsSubstrate selectivity of OsLHT1 was analyzed in Xenopus oocytes using the two-electrode voltage clamp technique. The results showed that OsLHT1 could transport a broad spectrum of amino acids, including basic, neutral and acidic amino acids, and exhibited a preference for neutral and acidic amino acids. Two oslht1 mutants were generated using CRISPR/Cas9 genome-editing technology, and the loss-of-function of OsLHT1 inhibited rice root and shoot growth, thereby markedly reducing grain yields. QRT-PCR analysis indicated that OsLHT1 was expressed in various rice organs, including root, stem, flag leaf, flag leaf sheath and young panicle. Transient expression in rice protoplast suggested OsLHT1 was localized to the plasma membrane, which is consistent with its function as an amino acid transporter.ConclusionsOur results indicated that OsLHT1 is an amino acid transporter with wide substrate specificity and with preference for neutral and acidic amino acids, and disruption of OsLHT1 function markedly inhibited rice growth and fecundity.

Highlights

Research on plant amino acid transporters was mainly performed in Arabidopsis, while our understanding of them is generally scant in rice
This study will further our understanding of the substrate specificity of OsLHT1 and its roles in rice growth promotion and yield formation
Broad substrate specificity of OsLHT1 To analyze the substrate selectivity of OsLHT1, OsLHT1 cRNA was injected into Xenopus oocytes, and twoelectrode voltage clamp (TEVC) was employed to record the currents induced by different amino acids [23, 24]

Summary

Introduction

Research on plant amino acid transporters was mainly performed in Arabidopsis, while our understanding of them is generally scant in rice. Plants absorb N mainly in the inorganic form (nitrate and ammonium) from soils, while the organic N such as amino acids could be absorbed [1, 2]. The reduced N is transported mainly in the form of amino acids to developing vegetative or reproductive sink organs, where it is used for growth and development. The nitrate and Physiological studies using isolated membrane vesicles and plant tissues have suggested the presence of multiple amino acid transporters with wide substrate specificity in plants [5,6,7]. In recent years, sequencing the full genomes of different plants gave us a chance to grasp the abundance of amino acid transporters in plant genomes

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