Sains Malaysiana 52(2)(2023): 513-531

http://doi.org/10.17576/jsm-2023-5202-15

UHPLC-Q-Orbitrap HRMS-Based Metabolomic Show Biological Pathways Involved in Rice (Oryza sativa L.) under Fe Toxicity Stress

(Metabolomik UHPLC-Q-Orbitrap Berasaskan HRMS menunjukkan Laluan Biologi Terlibat untuk Beras (Oryza sativa L.) di bawah Tekanan Ketoksikan Fe)

TURHADI TURHADI^1,2, HAMIM HAMIM³, MUNIF GHULAMAHDI⁴ & MIFTAHUDIN MIFTAHUDIN^3,*

¹Plant Biology Graduate Program, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Kampus IPB Dramaga 16680 Bogor, West Java, Indonesia

²Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Jl. Veteran, Malang 65415, East Java, Indonesia

³Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Kampus IPB Dramaga 16680 Bogor, West Java, Indonesia

⁴Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Kampus IPB Dramaga 16680 Bogor, West Java, Indonesia

Received: 19 July 2022/Accepted: 30 November 2022

Abstract

The iron (Fe) toxicity stress is still a serious problem in rice cultivation, especially on land with high Fe content. The Fe toxicity stress affects various complex physiological aspects of plants. The metabolomic analysis using LC-MS is expected to provide information about rice's metabolism regulation under Fe toxicity stress. The objective of this study was to show the biological pathway signature in rice after exposure to Fe toxicity stress using UHPLC-Q-Orbitrap HRMS-based metabolomic analysis. The two rice varieties, i.e., IR64 (Fe-sensitive) and Pokkali (Fe-tolerant) were analyzed their metabolites using UHPLC-Q-Orbitrap HRMS. The metabolite profiles of both varieties were analyzed using MetaboAnalyst 5.0 software. The results showed that Fe toxicity stress affected the metabolite profile in both root and shoot tissues of two rice varieties. A number of 102 metabolites were detected in root and shoot tissues of rice. The comprehensive univariate and multivariate analyses showed that 1-aminocyclopropane-1-carboxylate (ACC) in shoot tissues and galactose in root tissues was suggested as metabolite markers for Fe tolerance character of rice var. Pokkali.  The genes encoded the enzymes involved in biosynthetic pathway of both metabolite markers could be a target to be explored for Fe toxicity tolerance in rice.

Keywords: Galactose; metabolism; metabolite markers; 1-aminocyclopropane-1-carboxylate

Abstrak

Tekanan ketoksikan besi (Fe) masih menjadi masalah serius dalam penanaman padi, terutamanya pada tanah yang mempunyai kandungan Fe yang tinggi. Tekanan ketoksikan Fe mempengaruhi pelbagai aspek fisiologi tumbuhan yang kompleks. Analisis metabolomik menggunakan LC-MS dijangka memberikan maklumat tentang peraturan metabolisme beras di bawah tekanan ketoksikan Fe. Objektif kajian ini adalah untuk menunjukkan pengenalan laluan biologi dalam beras selepas terdedah kepada tekanan ketoksikan Fe menggunakan analisis metabolomik berasaskan UHPLC-Q-Orbitrap HRMS. Kedua-dua varieti beras, iaitu IR64 (Fe-sensitif) dan Pokkali (Fe-toleransi) telah dianalisis metabolitnya menggunakan UHPLC-Q-Orbitrap HRMS. Profil metabolit kedua-dua jenis varieti dianalisis menggunakan perisian MetaboAnalyst 5.0. Keputusan menunjukkan bahawa tekanan ketoksikan Fe mempengaruhi profil metabolit dalam kedua-dua tisu akar dan pucuk kedua-dua varieti padi. Sejumlah 102 metabolit telah dikesan dalam tisu akar dan pucuk padi. Analisis komprehensif univariat dan multivariat menunjukkan bahawa 1-aminosiklopropana-1-karboksilat (ACC) dalam tisu pucuk dan galaktosa dalam tisu akar telah dicadangkan sebagai penanda metabolit untuk sifat toleransi Fe bagi beras var. Pokkali. Gen yang mengekod enzim yang terlibat dalam laluan biosintetik kedua-dua penanda metabolit boleh menjadi sasaran untuk diterokai untuk toleransi ketoksikan Fe dalam beras.

Kata kunci: Galaktosa; metabolisme; penanda metabolit; 1-aminosiklopropana-1-karboksilat

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^*Corresponding author; email: miftahudin@apps.ipb.ac.id