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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (2): 394-402.doi: 10.3724/SP.J.1006.2023.34064

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

Phosphorus transporter StPHO1.2 improving heat tolerance in potato

LI Wan1,*(), LI Cheng2, CHENG Min3, WU Fang4   

  1. 1School of Biomedicine and Food Engineering, Shangluo University, Shangluo 726000, Shaanxi, China
    2Xi’an Runzhuangjia Agricultural Technology Co., Ltd, Xi’an 710003, Shaanxi, China
    3Science and Technology Division, Shangluo University, Shangluo 726000, Shaanxi, China
    4Domestic Cooperation and the Schoolfellow Bureau, Shangluo University, Shangluo 726000, Shaanxi, China
  • Received:2023-03-24 Accepted:2023-06-29 Online:2024-02-12 Published:2023-07-20
  • Contact: *E-mail: 599092122@qq.com
  • Supported by:
    Natural Science Basic Research Program of Shaanxi(2022JQ-223);Young Talent fund of University Association for Science and Technology in Shaanxi, China(20220213);Research Fund for the Doctoral Program of Shangluo University(21SKY112);Science and Technology Innovation Team Project of Shaanxi(2022TD-56)

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Abstract:

Plants absorb and transport phosphorus through phosphorus transporters, and sufficient phosphorus can improve crop yield, quality, and stress resistance. High temperature is an important environmental factor affecting the growth and development of potato. In this study, we overexpressed the phosphorus transporter StPHO1.2 in potato mediated by Agrobacterium tumefaciens, and then compared the growth of transgenic strains and wild-type strains at high temperature (35℃) and normal temperature (22℃±1℃). The results showed that overexpression of StPHO1.2 could improve the heat resistance and promote the growth of potato. And the higher the phosphorus concentration, the stronger the resistance, the better the growth. Subcellular localization in tobacco showed that StPHO1.2 was expressed on cell membrane. Therefore, the membrane system library plasmid was selected to screen the interacting proteins of StPHO1.2. This study demonstrated that the phosphorus transporter StPHO1.2 interacted with both the calcium ion transporter associated protein (StCAX1) and the photosynthetic system II protein subunit (StPsbR) by the yeast two-hybrid and BiFC. In conclusion, overexpression of StPHO1.2 may improve the heat tolerance and promote the growth of potato by affecting photosynthesis and signal transduction in potato. These results provide theoretical basis and reference for further understanding of the function of phosphorus transporters and promote the breeding of new potato varieties.

Key words: Solanum tuberosum L., phosphorus transporter, high temperature stress, yeast two-hybrid, BiFC

Table 1

Primers, endonuclease site, recombinant plasmid name, and vector resistance"

引物名称
Primer name		 引物序列
Primer sequence (5'-3')		 酶切位点
Endonuclease
site		 重组质粒名称
Recombinant
plasmid name		 载体抗性
Vector resistance
Ox2F GAACACGGGGGACTCTAGAGGATCCATGGTGAAGTTTTCTAAAGAACTTG BamH I
Sac I		 pBI121-StPHO1.2 Kan+
Ox2R AACGATCGGGGAAATTCGAGCTCCTAGCCATCAGAGTCTGTCTCG
RFP2F GTTCCAGATTACGCTGAGCTCATGGTGAAGTTTTCTAAAGAACTTG Sac I
Sac I		 p1300-RFP-StPHO1.2 Kan+
RFP2R CGATCGGGGAAATTCGAGCTCCTAGCCATCAGAGTCTGTCTCG
STE2F TTTTATGTAATGGCCATTACGGCCATGGTGAAGTTTTCTAAAGAACTT Sfi I
Sfi I		 pBT3-STE-StPHO1.2 Kan+
STE2R# TCCTGCAGATGGCCGAGGCGGCGCCATCAGAGTCTGTCTCGCG
SUC2F TATCTGCAATGGCCATTACGGCCATGGTGAAGTTTTCTAAAGAACTT Sfi I
Sfi I		 pBT3-SUC-StPHO1.2 Kan+
SUC2R# TCCTGCAGATGGCCGAGGCGGCGCCATCAGAGTCTGTCTCGCG
pBT3-N2F ATTCCTGCAGGGCCATTACGGCCATGGTGAAGTTTTCTAAAGAACTTGA Sfi I
Sfi I		 pBT3-N-StPHO1.2 Kan+
pBT3-N2R# ACTTACCATGGGGCCGAGGCGGCGCCATCAGAGTCTGTCTCGCG
pBT3-C2F TAATCTAGACGGCCATTACGGCCATGGTGAAGTTTTCTAAAGAACTTGA Sfi I
Sfi I		 pBT3-C-StPHO1.2 Kan+
pBT3-C2R# TGGAGGCCTTTGGCCGAGGCGGCGCCATCAGAGTCTGTCTCGCG
NY2F CGCCACAACATCGAGGGATCCATGGTGAAGTTTTCTAAAGAACTTG BamH I
Sma I		 NY-StPHO1.2 Kan+
NY2R GAATTCGAGCTCTATCCCGGGCTAGCCATCAGAGTCTGTCTCG

Table 2

The selection of vector"

序号
Serial number		 质粒1
Plasmid 1		 质粒2
Plasmid 2		 目的
Purpose
1 pNubG-Fe65 pTSU2-APP 阳性对照 Positive control
2 NMY51 — 阴性对照 Negative control
3 pPR3N pBT3-STE-StPHO1.2 自激活检测 Self-activation detection
4 pPR3N pBT3-SUC-StPHO1.2 自激活检测 Self-activation detection
5 pPR3N pBT3-N-StPHO1.2 自激活检测 Self-activation detection
6 pPR3N pBT3-C-StPHO1.2 自激活检测 Self-activation detection
7 pOST1-NubI pBT3-STE-StPHO1.2 功能检测 Functional detection
8 pOST1-NubI pBT3-SUC-StPHO1.2 功能检测 Functional detection
9 pOST1-NubI pBT3-N-StPHO1.2 功能检测 Functional detection
10 pOST1-NubI pBT3-C-StPHO1.2 功能检测 Functional detection

Fig. 1

Identification of positive transgenic lines of resistance gene by PCR M: DNA marker; 1: StPHO1.2-Desiree; 2: pBI121 empty vector; 3: Desiree."

Fig. 2

Identification of positive transgenic lines by qPCR"

Fig. 3

Phenotypic analysis of StPHO1.2-Desiree and Desiree A: no phosphorus (0 mmol L-1 Pi); B: phosphorus deficiency (0.1 mmol L-1 Pi); C: rich in phosphorus (1.0 mmol L-1 Pi); 1: Desiree; 2: StPHO1.2-Desiree. In “1” and “2”, the left plant grew at high temperature (35℃), and the right plant grew at normal temperature (22℃±1℃)."

Fig. 4

Subcellular localization of StPHO1.2"

Fig. 5

Screening of vector for yeast double-hybrid experiment"

Fig. 6

Screening of interacting proteins of StPHO1.2 by yeast double-hybrid Arrow indicates the representative color-developing colony."

Fig. 7

“One-to-one” verification of StPHO1.2 and interacting proteins 1-5 indicate different replicates."

Fig. 8

BiFC identification of StPHO1.2 and interacting proteins"

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