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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (11): 2983-2995.doi: 10.3724/SP.J.1006.2025.55029

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Functional study of the phosphate transporter protein BnaPT48 in Brassica napus

ZHANG Wen(), LI Yu, WANG Chuang, SHI Lei, DING Guang-Da()   

  1. College of Resources and Environment, Huazhong Agricultural University / Microelement Research Center / Key Laboratory of Arable Land Conservation in the Middle and Lower Reaches of Yangtze River, Ministry of Agriculture and Rural Affairs, Wuhan 430070, Hubei, China
  • Received:2025-04-18 Accepted:2025-08-13 Online:2025-11-12 Published:2025-08-19
  • Contact: *E-mail: dgd@mail.hzau.edu.cn
  • Supported by:
    National Key R&D Program for Agricultural Biological Breeding(2023ZD04072)

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Abstract:

Phosphorus (P) is an essential macronutrient required for plant growth and development. Rapeseed (Brassica napus), a major oilseed crop in China, is characterized by a high demand for P and strong sensitivity to P deficiency. This study investigates the gene BnaPT48, which is highly expressed in the root system of Brassica napus and is strongly induced under P-deficient conditions. Through analyses of its expression pattern, protein subcellular localization, and phenotypes of transgenic materials, the role of BnaPT48 in P uptake was elucidated. BnaPT48 was found to localize to the plasma membrane and was significantly upregulated by P deficiency in various root tissues. The BnaPT48 protein complemented the P uptake defect of the Arabidopsis thaliana mutant atpt1/2, indicating its functional role in enhancing P acquisition. Overexpression of BnaPT48 in A. thaliana promoted plant growth under both normal and low P conditions. In rapeseed, overexpression of BnaPT48 significantly increased the shoot dry weight under low P treatment, reduced inorganic P concentration in cotyledons, accelerated chlorophyll degradation and cotyledon senescence, and increased inorganic P accumulation in roots under normal P conditions. These findings reveal the function of BnaPT48 in regulating phosphate uptake and redistribution, and provide a theoretical foundation and genetic resource for improving P use efficiency in B. napus through genetic approaches.

Key words: Brassica napus, BnaPT48, transport protein, low phosphorus stress, P concentration, P absorption

Table 1

Primers used in this study"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 引物作用
Function of primers
BnaPT48-GFP-F ATCGATTCTAGAGCC ATGGCTGATAATCAGCAGCTAGGAG 构建亚细胞定位载体
Construct a subcellular localization vector
BnaPT48-GFP-R TTCTCCTTTGCCCAT GTTTCTTCCGGAGGTAG
BnaPT48p-GUS-F CAGGTCGACGGATCCATATCAACTTTATGGAAAGATGACAA 构建组织定位载体
Construct a tissue-specific targeting vector
BnaPT48p-GUS-R TAAGGGACTGACCACTTGCCTTCTTCTAGATTATCTACA
BnPT48OX-F CCCCCGGG ATGGCTGATAATCAGCAGCTAGGAG 构建过表达载体
Construct an overexpression vector
BnPT48OX-R GCGTCGAC TCAGTTTCTTCCGGAGGTAG
BnaPT48-qPCR-F TATTATTGGCGAATGAAGATGC 定量基因表达水平
Quantify gene expression levels
BnaPT48-qPCR-R CCTTGGAGAACAAGCCGTAG

Fig. 1

Evolutionary and sequence similarity analysis of BnaPT48 and homologous genes A: phylogenetic relationship analysis; B: amino acid sequence alignment; C: protein tertiary structure analysis. Merge: the merge of AtPHT1;3 and BnaPT48."

Fig. 2

Spatiotemporal expression patterns of BnaPT48 A: spatiotemporal expression patterns of roots at seedling stage under normal phosphorus and low phosphorus treatments (with the 0 h expression level in roots under normal phosphorus treatment as baseline); B: spatiotemporal expression patterns of shoots at seedling stage under normal phosphorus and low phosphorus treatment (with the 0 h expression level in shoots under normal phosphorus treatment as baseline); C: gene expression pattern analysis at seedling stage (with the expression level in various tissue at seeding stage under control phosphorus as baseline); D: analysis of gene expression patterns in mature tissues (with the expression level in various tissue at mature stage under control phosphorus as baseline, CK: 90 kg hm-2; LP: 15 kg hm-2). FEL: fully expanded leaf; CL: cauline leaf; IS: inflorescence stem; B: bud; F: flower; Hu: husk; S: seed; Ca: carpopodium. Seeding stage, CK: 1 mmol L-1 P; -P: 0 mmol L-1 P. Data are shown as means ± SD (n = 3), t-test. *, **, *** mean significant difference at the 0.05, 0.01, and 0.001 probability levels, respectively."

Fig. 3

Subcellular and tissue localization of the BnaPT48 protein A: subcellular localization of BnaPT48 protein; B: GUS staining of transgenic BnaPT48p::GUS line. a-g: root under normal phosphorus conditions (a), root under low phosphorus conditions (b), and cotyledon (c), true leaf (d), hypocotyl (e), flower (f), and silique (g) under low phosphorus conditions. CK: normal P treatment (1 mmol L-1 P); LP: low P treatment (5 μmol L-1 P). The scale bar for cotyledon, true leaves, hypocotyl, flower, and siliques is 0.2 cm; the scale bar for root is 0.5 mm."

Fig. 4

Phenotypic analysis of BnaPT48 complementary phosphorus (P) absorbing mutant atpt1/2 under different P levels in Arabidopsis A: identification of A. thaliana supplementary materials; B: growth of overexpressing BnaPT48 materials and wild-type Col-0 in Arabidopsis; C: fresh weight under normal P conditions and low P conditions; D: inorganic P concentration under normal P conditions and low P conditions. t-test (Student’s t-test). *, **, *** mean significant difference at the 0.05, 0.01, and 0.001 probability level, respectively. CK: normal P treatment (625 μmol L-1 P); LP: low P treatment (15 μmol L-1 P). The scale bar in figure B is 0.5 cm. Using wild-type Col-0 under identical conditions as the control."

Fig. 5

Phenotypic analysisof Arabidopsis overexpressing BnaPT48 and wild-type materials under different phosphorus (P) levels A: identification of overexpressing BnaPT48 material in Arabidopsis; B: growth of overexpressing BnaPT48 material and wild-type Col-0 in Arabidopsis; C: root and shoot fresh weight; D: root and shoot dry weight; E: root and shoot inorganic P concentration; F: root and shoot total P concentration; G: total root length; H: root total surface area; I: root and shoot P accumulation. Data are shown as the means ± SD (n = 3). t-test (Student’s t-test). *, **, *** mean significant difference at the 0.05, 0.01, and 0.001 probability level, respectively. CK: normal P treatment (200 μmol L-1 P); LP: low P treatment (5 μmol L-1 P). The scale bar in figure B is 3 cm. Using wild-type Col-0 under identical conditions as the control."

Fig. 6

Phenotypic analysis of rapeseed overexpressing BnaPT48 and wild-type materials under different phosphorus (P) levels A: the growth of each material under normal P conditions; B: growth of each material under low P conditions; C: identification of overexpressing BnaPT48 material; D: dry weight under normal P conditions and low P conditions; E: inorganic P concentration under normal P conditions and low P conditions; F: shoot P distribution ratio; G: P concentration under normal P conditions and low P conditions; H: transport coefficient; I: P content under normal P conditions and low P conditions. Scale = 5 cm. a represents cotyledons, b represents the first true leaf (leaf 1). Data are shown as the means ± SD (n = 3). t-test is used here. *, **, *** mean significant difference at the 0.05, 0.01, and 0.001 probability level, respectively. CK: normal P treatment (1 mmol L-1 P); LP: low P treatment (10 μmol L-1 P). Using wild-type Col-0 under identical conditions as the control."

Fig. 7

Effect of overexpression of BnaPT48 on chlorophyll and carotenoid concentrations in rapeseed leaves A: chlorophyll a concentration under normal P conditions and low P conditions; B: chlorophyll b concentration under normal P conditions and low P conditions; C: carotenoid concentration under normal P conditions and low P conditions; D: total chlorophyll concentration under normal P conditions and low P conditions; E: SPAD value under normal P conditions and low P conditions. Data are shown as the means ± SD (n = 3), t-test is used here. *, **, *** mean significant difference at the 0.05, 0.01, and 0.001 probability level, respectively. CK: normal P treatment (1 mmol L-1 P); LP: low P treatment (10 μmol L-1 P). Using wild-type Col-0 under identical conditions as the control."

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