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. 2014 Feb;65(2):683-96.
doi: 10.1093/jxb/ert442.

Overexpression of VP, a vacuolar H+-pyrophosphatase gene in wheat (Triticum aestivum L.), improves tobacco plant growth under Pi and N deprivation, high salinity, and drought

Xiaojuan Li  1 , Chengjin Guo, Juntao Gu, Weiwei Duan, Miao Zhao, Chunying Ma, Xiaoming Du, Wenjing Lu, Kai Xiao
Affiliations

Overexpression of VP, a vacuolar H+-pyrophosphatase gene in wheat (Triticum aestivum L.), improves tobacco plant growth under Pi and N deprivation, high salinity, and drought

Xiaojuan Li et al. J Exp Bot. 2014 Feb.

Retraction in

Abstract

Establishing crop cultivars with strong tolerance to P and N deprivation, high salinity, and drought is an effective way to improve crop yield and promote sustainable agriculture worldwide. A vacuolar H+-pyrophosphatase (V-H+-PPase) gene in wheat (TaVP) was functionally characterized in this study. TaVP cDNA is 2586-bp long and encodes a 775-amino-acid polypeptide that contains 10 conserved membrane-spanning domains. Transcription of TaVP was upregulated by inorganic phosphate (Pi) and N deprivation, high salinity, and drought. Transgene analysis revealed that TaVP overexpression improved plant growth under normal conditions and specifically under Pi and N deprivation stresses, high salinity, and drought. The improvement of growth of the transgenic plants was found to be closely related to elevated V-H+-PPase activities in their tonoplasts and enlarged root systems, which possibly resulted from elevated expression of auxin transport-associated genes. TaVP-overexpressing plants showed high dry mass, photosynthetic efficiencies, antioxidant enzyme activities, and P, N, and soluble carbohydrate concentrations under various growth conditions, particularly under the stress conditions. The transcription of phosphate and nitrate transporter genes was not altered in TaVP-overexpressing plants compared with the wild type, suggesting that high P and N concentrations regulated by TaVP were caused by increased root absorption area instead of alteration of Pi and NO3- acquisition kinetics. TaVP is important in the tolerance of multiple stresses and can serve as a useful genetic resource to improve plant P- and N-use efficiencies and to increase tolerance to high salinity and drought.

Keywords: Abiotic stresses; gene expression; physiological and biochemical property; plant growth; transgene analysis; vacuolar H+-pyrophosphatase..

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Figures

Fig. 1.
Fig. 1.
Diagram of the membrane-spanning domains of TaVP. I to X, conserved transmembrane domains in TaVP.
Fig. 2.
Fig. 2.
Expression patterns of TaVP under normal growth and under Pi and N deprivation, high salinity, and drought. (A and B) Reverse-transcription PCR in roots (A) and leaves (B). (C) Quantitative PCR in roots and leaves; data are mean ± SE of four independent assays; different letters indicate significant difference in each stressor setup (P < 0.01).
Fig. 3.
Fig. 3.
TaVP expression and vacuolar H+-PPase activities in the wild-type and transgenic plants. (A) Northern blot analysis of TaVP expression. (B) Vacuolar H+-PPase activity; data are mean ± SE of four independent assays; different letters indicate significant difference (P < 0.01). Lines 1 to 8, eight T3 transgenic lines transformed with TaVP.
Fig. 4.
Fig. 4.
Plant phenotypic features of the wild-type, control, and the transgenic lines. (A) normal growth. (B) Pi deprivation. (C) N deprivation. (D) High salinity. (E) Drought. WT, wild-type; CTR, control that transformed an empty vector; line 5 and line 7, two TaVP-overexpressing transgenic lines (this figure is available in colour at JXB online).
Fig. 5.
Fig. 5.
Plant dry mass, root parameters, nutrient and soluble carbohydrate concentrations, and nutrient accumulative amounts in TaVP-overexpressing tobacco plants. (A) Dry masses of aerial part and root. (B) Root volume. (C) Root total absorption surface area (TASA) and effective absorption surface area (EASA). (D) P and N concentrations. (E) Soluble carbohydrate content. (F) Accumulated amounts of P and N. Line 5 and line 7, two TaVP-overexpressing transgenic lines. Asterisks indicate significant difference between tissues or stressor setup (P < 0.01).
Fig. 6.
Fig. 6.
Expression of auxin transport-associated genes (A), phosphate transporter genes (B), and nitrate transporter genes (C) in TaVP-overexpressing tobacco plants. Line 5 and line 7, two TaVP-overexpressing transgenic lines. Data are mean ± SE of four independent assays; different letters indicate significant difference in each stressor setup (P < 0.01).
Fig. 7.
Fig. 7.
Photosynthetic parameters in TaVP-overexpressing tobacco plants. (A) Photosynthetic rate (P n). (B) PSII efficiency (Φ PSII). (C) Nonphotochemical quenching (NPQ). Line 5 and line 7, two TaVP-overexpressing transgenic lines. Data are mean ± SE of four independent assays; different letters indicate significant difference in each stressor setup (P < 0.01).
Fig. 8.
Fig. 8.
Antioxidant enzyme activities and malondialdehyde contents in TaVP-overexpressing tobacco plants. (A) SOD activity. (B) CAT activity. (C) POD activity. (D) Malondialdehyde content. Line 5 and line 7, two TaVP-overexpressing transgenic lines. Data are mean ± SE of four independent assays; different letters indicate significant difference in each stressor setup (P < 0.01).
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