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Characterization of VuMATE1 Expression in Response to Iron Nutrition and Aluminum Stress Reveals Adaptation of Rice Bean (Vigna umbellata) to Acid Soils through Cis Regulation.

Liu M, Xu J, Lou H, Fan W, Yang J, Zheng S - Front Plant Sci (2016)

Bottom Line: The results showed that the expression of VuMATE1 was regulated by Al stress but not Fe status.We demonstrated that cis regulation of VuMATE1 expression is involved in Al tolerance mechanism, while not involved in Fe nutrition.These results reveal the evolution of VuMATE1 expression for better adaptation of rice bean to acid soils where Al stress imposed but Fe deficiency pressure released.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang UniversityHangzhou, China; Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural SciencesHangzhou, China.

ABSTRACT
Rice bean (Vigna umbellata) VuMATE1 appears to be constitutively expressed at vascular system but root apex, and Al stress extends its expression to root apex. Whether VuMATE1 participates in both Al tolerance and Fe nutrition, and how VuMATE1 expression is regulated is of great interest. In this study, the role of VuMATE1 in Fe nutrition was characterized through in planta complementation assays. The transcriptional regulation of VuMATE1 was investigated through promoter analysis and promoter-GUS reporter assays. The results showed that the expression of VuMATE1 was regulated by Al stress but not Fe status. Complementation of frd3-1 with VuMATE1 under VuMATE1 promoter could not restore phenotype, but restored with 35SCaMV promoter. Immunostaining of VuMATE1 revealed abnormal localization of VuMATE1 in vasculature. In planta GUS reporter assay identified Al-responsive cis-acting elements resided between -1228 and -574 bp. Promoter analysis revealed several cis-acting elements, but transcription is not simply regulated by one of these elements. We demonstrated that cis regulation of VuMATE1 expression is involved in Al tolerance mechanism, while not involved in Fe nutrition. These results reveal the evolution of VuMATE1 expression for better adaptation of rice bean to acid soils where Al stress imposed but Fe deficiency pressure released.

No MeSH data available.


Related in: MedlinePlus

Physiological and molecular responses of rice bean to Fe deficiency. Seedlings of rice bean were subject to nutrient solution with 20 μM Fe (+Fe) or without (-Fe) for 12 days. (A) SPAD value of newly expanded leaves. Data are means ± SD (n = 8). (B) Ferric chelate reductase (FCR) activity of roots. Data are means ± SD (n = 4). (C) RT-PCR analysis of the expression of VuMATE1 and VuIRT1 in both roots and shoots. (D) Quantitative real-time PCR analysis of the expression of VuIRT1 in roots. Data are means ± SD (n = 3). (E) Quantitative real-time PCR analysis of the expression of VuMATE1 in roots. Data are means ± SD (n = 3). Asterisks represent significant differences between +Fe and -Fe treatment at P < 0.05.
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Figure 1: Physiological and molecular responses of rice bean to Fe deficiency. Seedlings of rice bean were subject to nutrient solution with 20 μM Fe (+Fe) or without (-Fe) for 12 days. (A) SPAD value of newly expanded leaves. Data are means ± SD (n = 8). (B) Ferric chelate reductase (FCR) activity of roots. Data are means ± SD (n = 4). (C) RT-PCR analysis of the expression of VuMATE1 and VuIRT1 in both roots and shoots. (D) Quantitative real-time PCR analysis of the expression of VuIRT1 in roots. Data are means ± SD (n = 3). (E) Quantitative real-time PCR analysis of the expression of VuMATE1 in roots. Data are means ± SD (n = 3). Asterisks represent significant differences between +Fe and -Fe treatment at P < 0.05.

Mentions: To examine whether the expression of VuMATE1 is regulated by Fe status, we analyzed the response of VuMATE1 expression to Fe nutrition by RT- and qRT-PCR. After 12 days culture of rice bean seedlings in nutrient solution with (+Fe) or without Fe (-Fe), newly expanded leaves of -Fe plants displayed obvious chlorosis symptom (Supplementary Figure S3). SPAD values (indications of chlorophyll content) of newly expanded leaves decreased by ∼60% following this treatment compared to +Fe conditions (Figure 1A) and the ferric chelate reductase (FCR) activity in roots increased by almost threefold (Figure 1B), demonstrating that the plants were Fe-deficient after 12 days treatment. This is supported by RT-PCR and qRT-PCR analysis indicating that expression of VuIRT1 (Iron Regulated Transporter 1), the ferrous Fe transporter, was fifteen times greater in the roots of -Fe plants compared to +Fe controls (Figures 1C,D). By contrast, VuMATE1 expression in roots was unaffected by Fe status since expression levels were similar in the -Fe and +Fe plants (Figure 1E).


Characterization of VuMATE1 Expression in Response to Iron Nutrition and Aluminum Stress Reveals Adaptation of Rice Bean (Vigna umbellata) to Acid Soils through Cis Regulation.

Liu M, Xu J, Lou H, Fan W, Yang J, Zheng S - Front Plant Sci (2016)

Physiological and molecular responses of rice bean to Fe deficiency. Seedlings of rice bean were subject to nutrient solution with 20 μM Fe (+Fe) or without (-Fe) for 12 days. (A) SPAD value of newly expanded leaves. Data are means ± SD (n = 8). (B) Ferric chelate reductase (FCR) activity of roots. Data are means ± SD (n = 4). (C) RT-PCR analysis of the expression of VuMATE1 and VuIRT1 in both roots and shoots. (D) Quantitative real-time PCR analysis of the expression of VuIRT1 in roots. Data are means ± SD (n = 3). (E) Quantitative real-time PCR analysis of the expression of VuMATE1 in roots. Data are means ± SD (n = 3). Asterisks represent significant differences between +Fe and -Fe treatment at P < 0.05.
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Figure 1: Physiological and molecular responses of rice bean to Fe deficiency. Seedlings of rice bean were subject to nutrient solution with 20 μM Fe (+Fe) or without (-Fe) for 12 days. (A) SPAD value of newly expanded leaves. Data are means ± SD (n = 8). (B) Ferric chelate reductase (FCR) activity of roots. Data are means ± SD (n = 4). (C) RT-PCR analysis of the expression of VuMATE1 and VuIRT1 in both roots and shoots. (D) Quantitative real-time PCR analysis of the expression of VuIRT1 in roots. Data are means ± SD (n = 3). (E) Quantitative real-time PCR analysis of the expression of VuMATE1 in roots. Data are means ± SD (n = 3). Asterisks represent significant differences between +Fe and -Fe treatment at P < 0.05.
Mentions: To examine whether the expression of VuMATE1 is regulated by Fe status, we analyzed the response of VuMATE1 expression to Fe nutrition by RT- and qRT-PCR. After 12 days culture of rice bean seedlings in nutrient solution with (+Fe) or without Fe (-Fe), newly expanded leaves of -Fe plants displayed obvious chlorosis symptom (Supplementary Figure S3). SPAD values (indications of chlorophyll content) of newly expanded leaves decreased by ∼60% following this treatment compared to +Fe conditions (Figure 1A) and the ferric chelate reductase (FCR) activity in roots increased by almost threefold (Figure 1B), demonstrating that the plants were Fe-deficient after 12 days treatment. This is supported by RT-PCR and qRT-PCR analysis indicating that expression of VuIRT1 (Iron Regulated Transporter 1), the ferrous Fe transporter, was fifteen times greater in the roots of -Fe plants compared to +Fe controls (Figures 1C,D). By contrast, VuMATE1 expression in roots was unaffected by Fe status since expression levels were similar in the -Fe and +Fe plants (Figure 1E).

Bottom Line: The results showed that the expression of VuMATE1 was regulated by Al stress but not Fe status.We demonstrated that cis regulation of VuMATE1 expression is involved in Al tolerance mechanism, while not involved in Fe nutrition.These results reveal the evolution of VuMATE1 expression for better adaptation of rice bean to acid soils where Al stress imposed but Fe deficiency pressure released.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang UniversityHangzhou, China; Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural SciencesHangzhou, China.

ABSTRACT
Rice bean (Vigna umbellata) VuMATE1 appears to be constitutively expressed at vascular system but root apex, and Al stress extends its expression to root apex. Whether VuMATE1 participates in both Al tolerance and Fe nutrition, and how VuMATE1 expression is regulated is of great interest. In this study, the role of VuMATE1 in Fe nutrition was characterized through in planta complementation assays. The transcriptional regulation of VuMATE1 was investigated through promoter analysis and promoter-GUS reporter assays. The results showed that the expression of VuMATE1 was regulated by Al stress but not Fe status. Complementation of frd3-1 with VuMATE1 under VuMATE1 promoter could not restore phenotype, but restored with 35SCaMV promoter. Immunostaining of VuMATE1 revealed abnormal localization of VuMATE1 in vasculature. In planta GUS reporter assay identified Al-responsive cis-acting elements resided between -1228 and -574 bp. Promoter analysis revealed several cis-acting elements, but transcription is not simply regulated by one of these elements. We demonstrated that cis regulation of VuMATE1 expression is involved in Al tolerance mechanism, while not involved in Fe nutrition. These results reveal the evolution of VuMATE1 expression for better adaptation of rice bean to acid soils where Al stress imposed but Fe deficiency pressure released.

No MeSH data available.


Related in: MedlinePlus