Abstract
Purpose
Purple acid phosphatases (PAPs) includ the largest classes of non-specific plant acid phosphatases. Most characterized PAPs were found to play physiological functions in phosphorus metabolism. In this study, we investigated the function of AtPAP17 gene encoding an important purple acid phosphatase in Arabidopsis thaliana.
Methods
The full-length cDNA sequence of AtPAP17 gene under the control of CaMV-35S promoter was transferred to the A. thaliana WT plant. The generated homozygote AtPAP17-overexpressed plants were compared by the types of analyses with corresponding homozygote atpap17-mutant plant and WT in both + P (1.2 mM) and − P (0 mM) conditions.
Results
In the + P condition, the highest and the lowest amount of Pi was observed in AtPAP17-overexpressed plants and atpap17-mutant plants by 111% increase and 38% decrease compared with the WT plants, respectively. Furthermore, under the same condition, APase activity of AtPAP17-overexpressed plants increased by 24% compared to the WT. Inversely, atpap17-mutant plant represented a 71% fall compared to WT plants. The comparison of fresh weight and dry weight in the studied plants showed that the highest and the lowest amount of absorbed water belonged to OE plants (with 38 and 12 mg plant−1) and Mu plants (with 22 and 7 mg plant−1) in + P and − P conditions, respectively.
Conclusion
The lack of AtPAP17 gene in the A. thaliana genome led to a remarkable reduction in the development of root biomass. Thus, AtPAP17 could have an important role in the root but not shoot developmental and structural programming. Consequently, this function enables them to absorb more water and eventually associated with more phosphate absorption.
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Data availability
All data generated or analyzed during this study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.
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Acknowledgements
This research was supported by a grant from the Tarbiat Modares University of I.R. Iran. We gratefully acknowledge the National Institute of Genetic Engineering and Biotechnology (NIGEB) for kindly supporting us in preparing of AtPAP17 construct. In the end, we should appreciate their valuable technical assistance.
Supplementary information
Table S1—Chi-square test for genotypes conformity from Mendelian 3:1 ratio (kanamycin resistant: sensitive).
Figure S1—Semi-quantitative RT-PCR test. The comparison of AtPAP17 transcript levels in (A) root or shoot of Arabidopsis WT against homozygous atpap17-mutant (Mu) line, or (B) some of distinct homozygous T3 overexpressed (OE) plants versus WT (samples prepared from whole of plant), or (C) root or shoot of WT and homozygous OE plant. RT-PCR reactions were done for 35 cycles for all experiments with the specific primers for AtPAP17 (Unique PAP17 F, R). The α-Tubulin transcript level was selected as an internal control (bottom panel). Samples were obtained by the whole of plant or shoot/root pool of WT plants, Mu line and OE plants.
Figure S2—Phenotypically comparison of 21-day-old seedlings. The AtPAP17-overexpressed (OE), atpap17-mutant (Mu) and WT plants have grown for 14 days on MS medium culture containing (A) 1.2 mM Pi (+ P) or (B) no Pi (− P). Unit scale is 1 cm.
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This study was funded by a grant for a Master’s thesis from the Tarbiat Modares University of I.R. Iran (Grant No. 9085326).
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All the experiments, data analysis, and manuscript writing were done by AJL. MSS supervised the project and provided editorial input into the writing. MJ-J, KZ, TL, and MAM advised the project. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Jamali Langeroudi, A., Sabet, M.S., Jalali-Javaran, M. et al. Functional assessment of AtPAP17; encoding a purple acid phosphatase involved in phosphate metabolism in Arabidopsis thaliana. Biotechnol Lett 45, 719–739 (2023). https://doi.org/10.1007/s10529-023-03375-x
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DOI: https://doi.org/10.1007/s10529-023-03375-x