Abstract
Background
In response to various abiotic stressors such as drought, many plants engage different protein phosphatases linked to several physiological and developmental processes. However, comprehensive analysis of this gene family is lacking for soybean.
Objective
This study was performed to identify the TOPP-type protein phosphatase family in soybean and investigate the gene’s role under drought stress.
Methods
Soybean genome sequences and transcriptome data were downloaded from the Phytozome v.12, and the microarray data were downloaded from NCBI GEO datasets GSE49537. Expression profiles of GmTOPP13 were obtained based on qRT-PCR results. GmTOPP13 gene was transformed into tobacco plants via Agrobacterium mediated method, and the drought tolerance was analyzed by water deficit assay.
Results
15 GmTOPP genes were identified in the soybean genome database (GmTOPP1–15). GmTOPP genes were distributed on 9 of 20 chromosomes, with similar exon-intron structure and motifs arrangement. All GmTOPPs contained Metallophos and STPPase_N domains as well as the core catalytic sites. Cis-regulatory element analysis predicted that GmTOPPs were widely involved in plant development, stress and hormone response in soybean. Expression profiles showed that GmTOPPs expressed in different tissues and exhibited divergent expression patterns in leaf and root in response to drought stimulus. Moreover, GmTOPP13 gene was isolated and expression pattern analysis indicated that this gene was highly expressed in seed, root, leaf and other tissues detected, and intensively induced upon PEG6000 treatment. In addition, overexpression of GmTOPP13 gene enhanced the drought tolerance in tobacco plants. The transgenic tobacco plants showed regulation of stress-responsive genes including CAT, SOD, ERD10B and TIP during drought stress.
Conclusions
This study provides valuable information for the study of GmTOPP gene family in soybean, and lays a foundation for further functional studies of GmTOPP13 gene under drought and other abiotic stresses.
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (31971832), Natural Science Foundation of Heilongjiang Province of China (YQ2019C006) and the Academic Backbone Project of Northeast Agricultural University in China (19XG16).
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Supplementary Fig. S1
Construction of plant expression vector pCAMBIA2300-GmTOPP13. (A) Schematic diagram of expression vector construction (B) PCR identification of pCAMBIA2300-GmTOPP13 M: DL2000 Marker; 1: Positive control; 2: Negative control; 3–5: Recombinant molecule candidates. (PNG 116 KB)
Fig. S2
Positions of 15 GmTOPP genes on the soybean chromosomes. Genes were mapped to the soybean chromosomes by TBtools. (PNG 36 KB)
Supplementary Fig. S3
PCR identification of transgenic tobacco plants. M: DL2000 Marker; 1: Positive control; 2: Wild type control; 3–21: GmTOPP13 transgenic tobacco seedlings. (TIFF 2039 KB)
Fig. S4
Motif analysis of GmTOPP genes. The different colored boxes represented different motifs and their positions in each TOPP protein sequence. (PNG 1067 KB)
Supplementary Fig. S5
Expression profiles of GmTOPP genes under drought stress in soybean based on dataset GSE153951. (JPEG 2721 KB)
Supplementary Fig. S6
Expression profiles of GmTOPP genes under repeated drought stress in soybean based on dataset GSE153660. (JPEG 3288 KB)
Supplementary Fig. S7
Genetic transformation and culture of transgenic tobacco plants. (A) Pre-culture of leaf discs (B) Co-culture of leaf discs and Agrobacterium (C) Differentiation of resistance buds (D) Rooting of resistant seedlings (E) Flowering of transgenic tobacco plants (F) Seeds of transgenic tobacco. (PNG 857 KB)
Fig. S8
Phylogenetic relationships of TOPP proteins from soybean and Arabidopsis using the Neighbor-Joining method with p-distance correction. Bootstrap values were derived from 1000 replicate runs. (PNG 139 KB)
Supplementary Fig. S9
RT-PCR identification of transgenic tobacco plants. M: DL2000 Marker; 1: Positive control; 2: Negative control; 3: Wild type control; 4–22: T1 generation of GmTOPP13 transgenic tobacco lines. (PNG 145 KB)
Supplementary Table S1
The primer sequences. (DOCX 15 KB)
Supplementary Table S2
FPKM (Fragments/kilobase/million) of GmTOPP genes in different soybean tissues. (XLSX 11 KB)
Supplementary Table S3
Microarray data of GmTOPP genes under drought treatment. (XLSX 23 KB)
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Wang, S., Guo, J., Zhang, Y. et al. Genome‐wide characterization and expression analysis of TOPP-type protein phosphatases in soybean (Glycine max L.) reveal the role of GmTOPP13 in drought tolerance. Genes Genom 43, 783–796 (2021). https://doi.org/10.1007/s13258-021-01075-2
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DOI: https://doi.org/10.1007/s13258-021-01075-2