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Overexpression of Citrus junos mitochondrial citrate synthase gene in Nicotiana benthamiana confers aluminum tolerance

Planta volume 230, pages 355–365 (2009)Cite this article

Abstract

Aluminum (Al) toxicity is one of the major factors that limit plant growth in acid soils. Al-induced release of organic acids into rhizosphere from the root apex has been identified as a major Al-tolerance mechanism in many plant species. In this study, Al tolerance of Yuzu (Citrus Junos Sieb. ex Tanaka) was tested on the basis of root elongation and the results demonstrated that Yuzu was Al tolerant compared with other plant species. Exposure to Al triggered the exudation of citrate from the Yuzu root. Thus, the mechanism of Al tolerance in Yuzu involved an Al-inducible increase in citrate release. Aluminum also elicited an increase of citrate content and increased the expression level of mitochondrial citrate synthase (CjCS) gene and enzyme activity in Yuzu. The CjCS gene was cloned from Yuzu and overexpressed in Nicotiana benthamiana using Agrobacterium tumefaciens-mediated methods. Increased expression level of the CjCS gene and enhanced enzyme activity were observed in transgenic plants compared with the wild-type plants. Root growth experiments showed that transgenic plants have enhanced levels of Al tolerance. The transgenic Nicotiana plants showed increased levels of citrate in roots compared to wild-type plants. The exudation of citrate from roots of the transgenic plants significantly increased when exposed to Al. The results with transgenic plants suggest that overexpression of mitochondrial CS can be a useful tool to achieve Al tolerance.

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Abbreviations

35S:

Cauliflower mosaic virus 35S promoter

CS:

Citrate synthase

DEPC:

Diethyl pyrocarbonate

GUS:

β-Glucuronidase

NPTII:

Neomycin phosphotransferase

ORF:

Open reading frame

RACE:

Rapid amplification of cDNA ends

YADE:

Y-shaped adaptor-dependent extension

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Affiliations

  1. Key Laboratory of Functional Gene and Regulation Technologies under Chongqing Municipal Education Commission, Genetic Engineering Research Center, Bioengineering College, Chongqing University, 400044, Chongqing, People’s Republic of China

    Wei Deng, Zhengguo Li, Yingwu Yang, Nan Hu & Yu Wu

  2. Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of Agriculture, Biotechnology Research Center, Southwest University, 400710, Chongqing, People’s Republic of China

    Keming Luo

Authors
  1. Wei Deng
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  2. Keming Luo
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  3. Zhengguo Li
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  4. Yingwu Yang
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  5. Nan Hu
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  6. Yu Wu
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Correspondence to Wei Deng.

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Deng, W., Luo, K., Li, Z. et al. Overexpression of Citrus junos mitochondrial citrate synthase gene in Nicotiana benthamiana confers aluminum tolerance. Planta 230, 355–365 (2009). https://doi.org/10.1007/s00425-009-0945-z

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  • DOI: https://doi.org/10.1007/s00425-009-0945-z

Keywords

  • Aluminum tolerance
  • Citrate
  • Citrate synthase
  • Overexpression
  • Yuzu