Cloning of EoNLA gene in Eremochloa ophiuroides and the transgenic Arabidopsis phenotypic characterization under various phosphorus levels

HE Qingqing, LIU Chuanqiang, LI Jianjian, WANG Jingjing, YAO Xiang, ZHOU Shenghao, CHEN Ying, WANG Haoran

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (3) : 134-142.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2022, Vol. 46 ›› Issue (3) : 134-142. DOI: 10.12302/j.issn.1000-2006.2021006013

Cloning of EoNLA gene in Eremochloa ophiuroides and the transgenic Arabidopsis phenotypic characterization under various phosphorus levels

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Abstract

【Obiectives】 Phosphorus is a macronutrient of plants which is essential for growth and development. Nitrogen limitation adaptation (NLA), a RING-type E3 ubiquitin ligase involved in the ubiquitin regulation of phosphorus transporters, plays an important role in the regulation of phosphorus balance in plants. 【Method】In this study, we used Eremochloa ophiuroides, which naturally adapts to low-phosphorus soil conditions, to clone the EoNLA gene using RACE. 【Result】The EoNLA gene is 1 353 bp long and encodes a protein of 331 amino acids. Amino acid sequence analysis showed that the NLA protein had a typical RING domain and an SPX domain. Cell membrane localization of the EoNLA protein was observed using a protoplast transient expression system. Agrobacterium-mediated transformation of Arabidopsis thaliana was carried out to verify gene function 【Conclusion】The qRT-PCR analysis showed that expression levels of EoNLA were significantly higher in roots than in stems and leaves, suggesting that EoNLA expression is root-specific. Furthermore, functional identification based on Arabidopsis thaliana transgenic plants suggested that EoNLA is associated with phosphorus regulation in plants.

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Eremochloa ophiuroides / phosphorus regulation / EoNLA / functional analysis

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HE Qingqing , LIU Chuanqiang , LI Jianjian , et al . Cloning of EoNLA gene in Eremochloa ophiuroides and the transgenic Arabidopsis phenotypic characterization under various phosphorus levels[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2022, 46(3): 134-142 https://doi.org/10.12302/j.issn.1000-2006.2021006013

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