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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

In planta complementation assay of VuMATE1 in Fe nutrition in Arabidopsis mutant frd3-1 driven by 35S CaMV promoter. (A) Phenotype analysis of leaf chlorosis in WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). (B) Root ferric precipitation of WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). Bar, 100 μm.
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Figure 3: In planta complementation assay of VuMATE1 in Fe nutrition in Arabidopsis mutant frd3-1 driven by 35S CaMV promoter. (A) Phenotype analysis of leaf chlorosis in WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). (B) Root ferric precipitation of WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). Bar, 100 μm.

Mentions: We have previously demonstrated that VuMATE1 is a plasma membrane-localized citrate-permeable transporter protein (Yang et al., 2011). Thus, the inability of VuMATE1 to restore frd3-1 phenotype with respect to Fe nutrition suggests that the expression pattern but not gene function is responsible for the loss of its role in Fe nutrition. To test this hypothesis, we introduced VuMATE1 using 35S CaMV promoter into the frd3-1 mutant (35S::VuMATE1/frd3-1). The chlorosis was greatly, albeit not completely, restored in two independent transgenic lines, OX1 and OX2 (Figure 3A). In addition, Perls blue staining result also showed that the accumulation of Fe in root vasculature was decreased dramatically in comparison to frd3-1 mutant (Figure 3B).


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)

In planta complementation assay of VuMATE1 in Fe nutrition in Arabidopsis mutant frd3-1 driven by 35S CaMV promoter. (A) Phenotype analysis of leaf chlorosis in WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). (B) Root ferric precipitation of WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). Bar, 100 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835453&req=5

Figure 3: In planta complementation assay of VuMATE1 in Fe nutrition in Arabidopsis mutant frd3-1 driven by 35S CaMV promoter. (A) Phenotype analysis of leaf chlorosis in WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). (B) Root ferric precipitation of WT, frd3-1, and two transgenic lines overexpressing VuMATE1 (OX1 and OX2). Bar, 100 μm.
Mentions: We have previously demonstrated that VuMATE1 is a plasma membrane-localized citrate-permeable transporter protein (Yang et al., 2011). Thus, the inability of VuMATE1 to restore frd3-1 phenotype with respect to Fe nutrition suggests that the expression pattern but not gene function is responsible for the loss of its role in Fe nutrition. To test this hypothesis, we introduced VuMATE1 using 35S CaMV promoter into the frd3-1 mutant (35S::VuMATE1/frd3-1). The chlorosis was greatly, albeit not completely, restored in two independent transgenic lines, OX1 and OX2 (Figure 3A). In addition, Perls blue staining result also showed that the accumulation of Fe in root vasculature was decreased dramatically in comparison to frd3-1 mutant (Figure 3B).

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