title: GmPAP4, a novel purple acid phosphatase gene isolated from soybean (Glycine max), enhanced extracellular phytate utilization in Arabidopsis thaliana.
abstract: GmPAP4 , a novel plant PAP gene in soybean, has phytase activity. Over-expressing GmPAP4 can enhance Arabidopsis growth when phytate is the sole P source in culture. Phosphorus (P) is an important macronutrient for plant growth and development. However, most of the total P in soils is fixed into organic phosphate (Po). Purple acid phosphatase (PAP) can hydrolyze Po in the soil to liberate inorganic phosphate and enhance plant P utilization. We isolated a novel PAP gene, GmPAP4, from soybean (Glycine max). It had an open reading frame of 1,329 bp, encoding 442 amino acid residues. Sequence alignment and phylogenetics analysis indicated that GmPAP4 was similar to other plant PAPs with large molecular masses. Quantitative real-time PCR analysis showed that the induced expression of GmPAP4 was greater in P-efficient genotype Zhonghuang15 (ZH15) than in P-inefficient genotype Niumaohuang (NMH) during the periods of flowering (28-35 days post phytate stress; DPP) and pod formation (49-63 DPP). Moreover, peak expression, at 63 DPP, was about 3-fold higher in 'ZH15' than in 'NMH'. Sub-cellular localization showed that GmPAP4 might be on plasma membrane or in cytoplasm. Over-expressing GmPAP4 in Arabidopsis resulted in significant rises in P acquisition and utilization compared with the wild-type (WT). Under phytate condition, transgenic Arabidopsis plants showed increases of approximately 132.7 % in dry weight and 162.6 % in shoot P content compared with the WT. Furthermore, when phytate was added as the sole P source in cultures, the activity of acid phosphatase was significantly higher in transgenic plants. Therefore, GmPAP4 is a novel PAP gene that functions in plant's utilization of organic phosphate especially under phytate condition.
authors.affiliation: North China Key Laboratory for Germplasm Resources of Education Ministry, Department of Plant Genetics and Breeding, Hebei Agricultural University, Baoding, 071001, People's Republic of China.
authors.forename: Youbin
authors.initials: Y
authors.lastname: Kong
authors.forename: Xihuan
authors.initials: X
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.forename: Wenlong
authors.initials: W
authors.lastname: Li
authors.forename: Guijun
authors.initials: G
authors.lastname: Yan
authors.forename: Caiying
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000135:Acid Phosphatase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D019295:Computational Biology; D057075:Enzyme Assays; D004926:Escherichia coli / Q000378:metabolism; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D010758:Phosphorus / Q000494:pharmacology; D010802:Phylogeny; D010833:Phytic Acid / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics; D030821:Plants, Genetically Modified; D021381:Protein Transport / Q000187:drug effects; D016415:Sequence Alignment; D013025:Glycine max / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology
chemical_list: D006023:Glycoproteins; D010940:Plant Proteins; D010758:Phosphorus; D010833:Phytic Acid; C091137:purple acid phosphatase; D000135:Acid Phosphatase