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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

Histochemical analysis of GUS expression in transgenic Arabidopsis lines driven by different 5′ deletion sequence of VuMATE1 promoter. (A) Induction of VuMATE1 by Al in transgenic Arabidopsis carrying VuMATE1p::GUS constructs. The representative images show GUS staining in roots after 9 h of exposure to 0 (-Al) or 1 μM Al (+Al) activity in nutrient solution. (B) GUS expression in root apex and maturation root zone of transgenic Arabidopsis carrying the shortest promoter of VuMATE1 (-192 bp::GUS). At least two independent transgenic lines were used to analyze GUS expression. Bar, 100 μm.
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Figure 6: Histochemical analysis of GUS expression in transgenic Arabidopsis lines driven by different 5′ deletion sequence of VuMATE1 promoter. (A) Induction of VuMATE1 by Al in transgenic Arabidopsis carrying VuMATE1p::GUS constructs. The representative images show GUS staining in roots after 9 h of exposure to 0 (-Al) or 1 μM Al (+Al) activity in nutrient solution. (B) GUS expression in root apex and maturation root zone of transgenic Arabidopsis carrying the shortest promoter of VuMATE1 (-192 bp::GUS). At least two independent transgenic lines were used to analyze GUS expression. Bar, 100 μm.

Mentions: To characterize in detail the regulation of VuMATE1 transcription by Al stress and Fe status, we analyzed GUS expression in VuMATE1p::GUS transgenic lines carrying different lengths of the 5′ region of the VuMATE1 promoter. In the absence of Al stress, GUS staining was not observed in the root apex of all promoter-GUS reporter transgenic lines carrying different 5′ deletion promoters (Figure 6A). However, after 9-h exposure to Al stress, promoter-GUS reporter lines, -1720 and -1228 bp::GUS, exhibited Al-inducible and root apex staining of GUS activity, whereas GUS staining was not observed in -574 and -192 bp::GUS transgenic lines (Figure 6A). This result indicated that cis-acting elements responsible for root-tip specific and Al-inducible expression resided in VuMATE1 promoter sequence between -1228 and -574 bp. On the other hand, all promoter-GUS reporter transgenic lines examined showed constitutive expression in maturation root zone, even in the shortest -192 bp::GUS transgenic line (Figure 6B).


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)

Histochemical analysis of GUS expression in transgenic Arabidopsis lines driven by different 5′ deletion sequence of VuMATE1 promoter. (A) Induction of VuMATE1 by Al in transgenic Arabidopsis carrying VuMATE1p::GUS constructs. The representative images show GUS staining in roots after 9 h of exposure to 0 (-Al) or 1 μM Al (+Al) activity in nutrient solution. (B) GUS expression in root apex and maturation root zone of transgenic Arabidopsis carrying the shortest promoter of VuMATE1 (-192 bp::GUS). At least two independent transgenic lines were used to analyze GUS expression. Bar, 100 μm.
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Figure 6: Histochemical analysis of GUS expression in transgenic Arabidopsis lines driven by different 5′ deletion sequence of VuMATE1 promoter. (A) Induction of VuMATE1 by Al in transgenic Arabidopsis carrying VuMATE1p::GUS constructs. The representative images show GUS staining in roots after 9 h of exposure to 0 (-Al) or 1 μM Al (+Al) activity in nutrient solution. (B) GUS expression in root apex and maturation root zone of transgenic Arabidopsis carrying the shortest promoter of VuMATE1 (-192 bp::GUS). At least two independent transgenic lines were used to analyze GUS expression. Bar, 100 μm.
Mentions: To characterize in detail the regulation of VuMATE1 transcription by Al stress and Fe status, we analyzed GUS expression in VuMATE1p::GUS transgenic lines carrying different lengths of the 5′ region of the VuMATE1 promoter. In the absence of Al stress, GUS staining was not observed in the root apex of all promoter-GUS reporter transgenic lines carrying different 5′ deletion promoters (Figure 6A). However, after 9-h exposure to Al stress, promoter-GUS reporter lines, -1720 and -1228 bp::GUS, exhibited Al-inducible and root apex staining of GUS activity, whereas GUS staining was not observed in -574 and -192 bp::GUS transgenic lines (Figure 6A). This result indicated that cis-acting elements responsible for root-tip specific and Al-inducible expression resided in VuMATE1 promoter sequence between -1228 and -574 bp. On the other hand, all promoter-GUS reporter transgenic lines examined showed constitutive expression in maturation root zone, even in the shortest -192 bp::GUS transgenic line (Figure 6B).

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