Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (1): 166-174.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0489

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Expression and Functional Analysis of OsPT1 Gene in Rice Under Cadmium Stress

JIANG Nan1(), SHI Yang1, ZHAO Zhi-hui1, LI Bin1, ZHAO Yi-hui1, YANG Jun-biao1, YAN Jia-ming1, JIN Yu-fan1, CHEN Ji2, HUANG Jin1()   

  1. 1. College of Ecology and Environment, Chengdu University of Technology, Chengdu 610059
    2. College of Agronomy, Sichuan Agricultural University, Chengdu 611130
  • Received:2022-04-21 Online:2023-01-26 Published:2023-02-02
  • Contact: HUANG Jin E-mail:2091077656@qq.com;huangjin18@cdut.edu.cn

Abstract:

OsPT1-encoded rice phosphate transporter protein plays important regulatory roles in growth and abiotic stress response in rice. Previous data showed that the expression of OsPT1 gene was induced by cadmium(Cd)stress, however, the function and mechanism of OsPT1 under Cd stress remains unclear. Therefore, the aim of this study is to clarify the function of OsPT1 under Cd stress and lay a foundation for the breeding of low-Cd rice cultivars. By using bioinformatics methods, the sequence feature, structure and function of OsPT1 gene were analyzed or predicted. The expression profiles of OsPT1 in different tissues and at different time points under Cd stress were examined by quantitative real-time PCR(RT-qPCR)as well. Simultaneously, PCR was used to clone the encoding sequence of OsPT1, and the recombinant plasmid pGADT7-OsPT1 was constructed and transferred into the the Cd-sensitive yeast strain ∆ycf BY4741 for determining the effect of OsPT1 on the tolerance of yeast to Cd. The results showed that OsPT1 is a protein of 527 amino acids with the molecular weight of 57.46 kD, which was encoded by a CDS of 1 584 bp. In rice genome, the cis-acting elements such as environmental factors like light and anaerobic, as well as hormones including methyl jasmonate were found in the promoter region of this gene. The results of phylogenetic analysis showed that OsPT1 was most closely related with sorghum SbPT1. The results of Cd-responsive gene expression analysis showed that the expression levels of OsPT1 in rice shoots increased to 1.31, 1.34 and 2.46 folds comparing to the untreated control under 100 μmol/L Cd treatment for 1, 6 and 12 h, respectively. The expressions in the roots increased to 1.28 and 1.14 folds comparing the untreated control under 100 μmol/L Cd treatment for 1 or 6 h, Cd treatment, however, it was down-regulated to 0.62 folds after 12 h Cd treatment. The result of Cd tolerance examination showed that the yeast transfered with OsPT1 demonstrated the decreased tolerance to Cd when treated with 25 μmol/L Cd when compared with the control(0 Cd). All together, OsPT1 may play a certain role in the respondses of rice to Cd stress.

Key words: rice, OsPT1, cadmium stress, yeast, gene expression