Cloning ThPCS1 gene of Tamarix hispida to improve cadmium tolerance

WANG Peilong, YANG Ni, ZHANG Aoran, Tangnver•Sailike , LI Shuang, GAO Caiqiu

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (3) : 71-78.

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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 ›› 2021, Vol. 45 ›› Issue (3) : 71-78. DOI: 10.12302/j.issn.1000-2006.202011014

Cloning ThPCS1 gene of Tamarix hispida to improve cadmium tolerance

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Abstract

【Objective】 This study aimed to improve the cadmium tolerance of Tamarix hispida. 【Method】The cadmium stress response of the phytochelatin synthase gene (ThPCS1) in T. hispida was explored. We cloned the ThPCS1 gene of T. hispida by using reverse transcription-polymerase chain reaction (RT-PCR). The bioinformatics information of the ThPCS1 protein was analyzed using BioEdit, MEGA 5.0 and other software. The expression of ThPCS1 in the roots and leaves of T. hispida under cadmium stress was analyzed by real-time quantitative (RT-q) PCR. We constructed a pROKII-ThPCS1 overexpression vector and used it to obtain ThPCS1 transgenic T. hispida by transient infection. Physiological indicators and staining of transgenic and control T. hispida under the Cd stress response were compared. 【Result】We sequenced the full length of the ThPCS1 gene from T. hispida transcriptome data. The open reading frame (ORF) of ThPCS1 gene was 1 581 bp, encoding 526 amino acids with a relative molecular mass of 130.75 ku, and the theoretical isoelectric point pI was 5.0. Conserved domains and multiple sequence alignment analyses showed that the ThPCS1 protein has a phytochelatin synthase (PCS) domain at the N-terminus. The RT-qPCR results revealed that ThPCS1 expression was induced in the roots of T. hispida under cadmium stress, and the expression was obviously tissue-specific. Furthermore, ThPCS1 was overexpressed in T. hispida and under Cd stress. The results of relevant physiological determination showed less reactive oxygen species and cadmium ions in plants overexpressing ThPCS1 than in the control. Accordingly, the cell membrane was less damaged in transgenic as compared to the control plants. 【Conclusion】The response of the T. hispida ThPCS1 gene to cadmium stress was significant, confirming its role in the response of T. hispida to cadmium stress. The ThPCS1 gene might improve the cadmium tolerance of T. hispida and serve as a candidate gene for the molecular breeding of T. hispida with excellent cadmium tolerance.

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Tamarix hispida / phytochelatin synthase / cadmium stress / gene expression / RT-qPCR

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WANG Peilong , YANG Ni , ZHANG Aoran , et al . Cloning ThPCS1 gene of Tamarix hispida to improve cadmium tolerance[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2021, 45(3): 71-78 https://doi.org/10.12302/j.issn.1000-2006.202011014

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