Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (11): 2329-2339.DOI: 10.3969/j.issn.1004-1524.2022.11.02
• Crop Science • Previous Articles Next Articles
Functional analysis of plastid phosphate transporter OsPPT family members in rice
WANG Jichun1(
), LI Ruili2, WANG Jiaoling3, SHAO Junwen1, ZHAO Hongyu2,*()
- 1. College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China
2. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100098, China
3. College of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang 473061, Henan, China
-
Received:2022-04-20Online:2022-11-25Published:2022-11-29 -
Contact:ZHAO Hongyu
CLC Number:
Cite this article
WANG Jichun, LI Ruili, WANG Jiaoling, SHAO Junwen, ZHAO Hongyu. Functional analysis of plastid phosphate transporter OsPPT family members in rice[J]. Acta Agriculturae Zhejiangensis, 2022, 34(11): 2329-2339.
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Table 1 Primers for qRT-PCR
| 基因Gene | 上游引物Forward primer(5'→3') | 下游引物Reverse primer(5'→3') |
|---|---|---|
| OsPPT1 | TTCCAATTGTTGGTGGTGTG | AGGTGACATTTGAGGCCATT |
| OsPPT2 | GCATCTCTTACTGAGGCTTCCT | ATTTGAAGCCATTGCACTCC |
| OsPPT3 | TCTACTTCAACATCTACAACAAGCAG | GATGACGAAGGAGCCGAAC |
| OsPPT4 | TCAATCGCGAAATGTTTTCA | GGTTTATATCATCCAATGTTTCCTC |
Table 1 Primers for qRT-PCR
| 基因Gene | 上游引物Forward primer(5'→3') | 下游引物Reverse primer(5'→3') |
|---|---|---|
| OsPPT1 | TTCCAATTGTTGGTGGTGTG | AGGTGACATTTGAGGCCATT |
| OsPPT2 | GCATCTCTTACTGAGGCTTCCT | ATTTGAAGCCATTGCACTCC |
| OsPPT3 | TCTACTTCAACATCTACAACAAGCAG | GATGACGAAGGAGCCGAAC |
| OsPPT4 | TCAATCGCGAAATGTTTTCA | GGTTTATATCATCCAATGTTTCCTC |
Table 2 Primers used for fragment amplification
| 基因Gene | 上游引物Forward primer(5'→3') | 下游引物Reverse primer(5'→3') |
|---|---|---|
| OsPPT1 | CGCGGATCCATGCAGAGCGCGGCGGCCGT | ACGCGTCGACGGCAGTCTTGGGCTTGGGTTT |
| OsPPT2 | CGCGGATCCATGCAGAGCGCCGCGGCGGCGTT | ACGCGTCGACCGCAGCCTTGGGCTTGGGCTT |
| OsPPT3 | CGCGGATCCATGCAGCGCGCGGCGGCGGCCT | ACGCGTCGACGGCATTCTTTGGTTTTGTTCTCTT |
| OsPPT4 | CGCGGATCCATGCAGGCCGTGGCGGCGGCGA | ACGCGTCGACTGCAGTCTTAGCCTTTGGTTTAGC |
Table 2 Primers used for fragment amplification
| 基因Gene | 上游引物Forward primer(5'→3') | 下游引物Reverse primer(5'→3') |
|---|---|---|
| OsPPT1 | CGCGGATCCATGCAGAGCGCGGCGGCCGT | ACGCGTCGACGGCAGTCTTGGGCTTGGGTTT |
| OsPPT2 | CGCGGATCCATGCAGAGCGCCGCGGCGGCGTT | ACGCGTCGACCGCAGCCTTGGGCTTGGGCTT |
| OsPPT3 | CGCGGATCCATGCAGCGCGCGGCGGCGGCCT | ACGCGTCGACGGCATTCTTTGGTTTTGTTCTCTT |
| OsPPT4 | CGCGGATCCATGCAGGCCGTGGCGGCGGCGA | ACGCGTCGACTGCAGTCTTAGCCTTTGGTTTAGC |
Table 3 Primers for yeast vector construction
| 引物名称Primer name | 引物序列Forward primer(5'→3') |
|---|---|
| OsPPT1-BamHI-F | GCAGCCCGGGGGATCCATGCAGAGCGCGGCGGCC |
| OsPPT1-SacI-R | GGGAACAAAAGCTGGAGCTCTCAGGCAGTCTTGGGCTTGGGT |
| OsPPT2-BamHI-F | GCAGCCCGGGGGATCCATGCAGAGCGCCGCGGCG |
| OsPPT2-SacI-R | GGGAACAAAAGCTGGAGCTCTTACGCAGCCTTGGGCTTGG |
| OsPPT3-BamHI-F | GCAGCCCGGGGGATCCATGCAGCGCGCGGCGGCG |
| OsPPT3-SacI-R | GGGAACAAAAGCTGGAGCTCTCAGGCATTCTTTGGTTTTGTTCTC |
| OsPPT4-BamHI-F | GCAGCCCGGGGGATCCATGCAGGCCGTGGCGGCG |
| OsPPT4-SacI-R | GGGAACAAAAGCTGGAGCTCTCATGCAGTCTTAGCCTTTGG |
Table 3 Primers for yeast vector construction
| 引物名称Primer name | 引物序列Forward primer(5'→3') |
|---|---|
| OsPPT1-BamHI-F | GCAGCCCGGGGGATCCATGCAGAGCGCGGCGGCC |
| OsPPT1-SacI-R | GGGAACAAAAGCTGGAGCTCTCAGGCAGTCTTGGGCTTGGGT |
| OsPPT2-BamHI-F | GCAGCCCGGGGGATCCATGCAGAGCGCCGCGGCG |
| OsPPT2-SacI-R | GGGAACAAAAGCTGGAGCTCTTACGCAGCCTTGGGCTTGG |
| OsPPT3-BamHI-F | GCAGCCCGGGGGATCCATGCAGCGCGCGGCGGCG |
| OsPPT3-SacI-R | GGGAACAAAAGCTGGAGCTCTCAGGCATTCTTTGGTTTTGTTCTC |
| OsPPT4-BamHI-F | GCAGCCCGGGGGATCCATGCAGGCCGTGGCGGCG |
| OsPPT4-SacI-R | GGGAACAAAAGCTGGAGCTCTCATGCAGTCTTAGCCTTTGG |
Fig.1 Phylogenetic tree analysis of PPT family members The different colours indicated different plastidic phosphate translocators family: red, GPT and XPT; grey, TPT; green, PPT. Letters in the codes represented species names as follows: Os, Oryza sativa; AT, Arabidopsis thaliana; ATR, Amborella trichopoda; Bradi, Brachypodium distachyon; Cre, Chlamydomonas reinhardtii; Mapoly, Marchantia polymorpha; Potri, Populus trichocarpa; PP, Physcomitrella patens; SMO, Selaginella moellendorffii; Soly, Solanum lycopersicum; ZM, Zea mays.
Fig.1 Phylogenetic tree analysis of PPT family members The different colours indicated different plastidic phosphate translocators family: red, GPT and XPT; grey, TPT; green, PPT. Letters in the codes represented species names as follows: Os, Oryza sativa; AT, Arabidopsis thaliana; ATR, Amborella trichopoda; Bradi, Brachypodium distachyon; Cre, Chlamydomonas reinhardtii; Mapoly, Marchantia polymorpha; Potri, Populus trichocarpa; PP, Physcomitrella patens; SMO, Selaginella moellendorffii; Soly, Solanum lycopersicum; ZM, Zea mays.
Fig.2 Expression pattern of OsPPT family members in various tissues in rice VS, Vegetative stage, 21-day-old plants; FS, Flowering stage, 48-day-old plants; GFS, Grain filling stage, 60-day-old plants. Data are x -±s(n=3). Leaf1-Leaf7 represents the number of leaves from below, leaf1 refers to the number of the second leaf from below (the first leaf has fallen off), and leaf7 refers to the number of the eighth leaf from below (flag leaf). Different letters represent significant differences, P<0.05.
Fig.2 Expression pattern of OsPPT family members in various tissues in rice VS, Vegetative stage, 21-day-old plants; FS, Flowering stage, 48-day-old plants; GFS, Grain filling stage, 60-day-old plants. Data are x -±s(n=3). Leaf1-Leaf7 represents the number of leaves from below, leaf1 refers to the number of the second leaf from below (the first leaf has fallen off), and leaf7 refers to the number of the eighth leaf from below (flag leaf). Different letters represent significant differences, P<0.05.
Fig.3 Subcellular localization of OsPPT1-OsPPT4 proteins in rice protoplasts The green signals indicated green fluorescent protein (GFP), and the red signals indicated autofluorescence of chlorophyll. Bright, bright area; Scale bar=10 μm.
Fig.3 Subcellular localization of OsPPT1-OsPPT4 proteins in rice protoplasts The green signals indicated green fluorescent protein (GFP), and the red signals indicated autofluorescence of chlorophyll. Bright, bright area; Scale bar=10 μm.
Fig.4 OsPPT genes can confer phosphate transport in yeast PHO84 was a high-affinity phosphate transporter and served as a positive control, and empty vector was a negative control.
Fig.4 OsPPT genes can confer phosphate transport in yeast PHO84 was a high-affinity phosphate transporter and served as a positive control, and empty vector was a negative control.
Fig.5 Dynamic changes in transcript levels of OsPPT gene family members during phosphate deprivation in shoots of rice seeding
Fig.5 Dynamic changes in transcript levels of OsPPT gene family members during phosphate deprivation in shoots of rice seeding
Fig.6 Dynamic changes of transcript levels of OsPPT gene family members in response to NaCl, SA and ABA stresses of rice seedings Fourteen-day-old seedlings grown under normal conditions were exposed to different chemical treatments for 24 h. ABA, 100 μmol·L-1; NaCl, 100 mmol·L-1; SA, 500 μmol·L-1. Data were x -±s(n=3). *, P<0.05; **, P<0.01; ***, P<0.001.
Fig.6 Dynamic changes of transcript levels of OsPPT gene family members in response to NaCl, SA and ABA stresses of rice seedings Fourteen-day-old seedlings grown under normal conditions were exposed to different chemical treatments for 24 h. ABA, 100 μmol·L-1; NaCl, 100 mmol·L-1; SA, 500 μmol·L-1. Data were x -±s(n=3). *, P<0.05; **, P<0.01; ***, P<0.001.
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